Non-alcoholic fatty liver disease(NAFLD)is the main cause of chronic liver disease worldwide.Bupleurum is widely used in the treatment of non-alcoholic fatty liver,and saikosaponin D(SSD)is one of the main active comp...Non-alcoholic fatty liver disease(NAFLD)is the main cause of chronic liver disease worldwide.Bupleurum is widely used in the treatment of non-alcoholic fatty liver,and saikosaponin D(SSD)is one of the main active components of Bupleurum.The purpose of this study was to investigate the efficacy of SSD in the treatment of NAFLD and to explore the mechanism of SSD in the improvement of NAFLD based on“gut-liver axis”.Our results showed that SSD dose-dependently alleviated high fat diet-induced weight gain in mice,improved insulin sensitivity,and also reduced liver lipid accumulation and injury-related biomarkers aspartate aminotransferase(AST)and alanine aminotransferase(ALT).Further exploration found that SSD inhibited the mRNA expression levels of farnesoid X receptor(Fxr),small heterodimer partner(Shp),recombinant fibroblast growth factor 15(Fgf15)and apical sodium dependent bile acid transporter(Asbt)in the intestine,suggesting that SSD improved liver lipid metabolism by inhibiting intestinal FXR signaling.SSD can significantly reduce the gut microbiota associated with bile salt hydrolase(BSH)expression,such as Clostridium.Decreased BSH expression reduced the ratio of unconjugated to conjugated bile acids,thereby inhibiting the intestinal FXR.These data demonstrated that SSD ameliorated NAFLD potentially through the gut microbiota-bile acidintestinal FXR pathway and suggested that SSD is a promising therapeutic agent for the treatment of NAFLD.展开更多
A new saikosaponin was isolated from Bupleurum chinense DC., and its structure was identified as 3β,16α,23,28,30_pentahydroxy_olean_11,13(18)_dien_3_O_β_D_glucopyranosyl(1→6)_[α_L_rhamnopyranosyl (1→4)]_β_D...A new saikosaponin was isolated from Bupleurum chinense DC., and its structure was identified as 3β,16α,23,28,30_pentahydroxy_olean_11,13(18)_dien_3_O_β_D_glucopyranosyl(1→6)_[α_L_rhamnopyranosyl (1→4)]_β_D_glucopyranoside on the basis of chemical and spectral evidence, named as saikosaponin q_1. In addition, two known saikosaponins, 3″_O_acetyl_saikosaponin d and 3″_O_acetyl_saikosaponin b 2, were also isolated and identified from this plant for the first time.展开更多
Objective To investigate the anticancer effects and detailed mechanisms of Saikosaponin D(SSD)in human hepatoma HepG2 cells.Methods Cell proliferation and apoptosis were tested by MTT assay and Annexin-V/PI assay resp...Objective To investigate the anticancer effects and detailed mechanisms of Saikosaponin D(SSD)in human hepatoma HepG2 cells.Methods Cell proliferation and apoptosis were tested by MTT assay and Annexin-V/PI assay respectively.The expressions of CCAAT enhancer binding protein β(C/EBPβ)and p53 were detected by RT-PCR and Western blotting.Results SSD inhibited cell proliferation in a dose-dependent manner and induced apoptosis at the concentration of 5.0 mg/L.SSD significantly increased the mRNA and protein levels of C/EBPβ and p53 in a dose-dependent manner.Conclusion SSD exerts its anticancer effect by inhibiting cell proliferation and inducing apoptosis partly through C/EBPβ-p53 signal pathway in HepG2 cells.展开更多
Nerve scarring after peripheral nerve injury can severely hamper nerve regeneration and functional recovery.Further,the anti-inflammatory cytokine,interleukin-10,can inhibit nerve scar formation.Saikosaponin a(SSa) ...Nerve scarring after peripheral nerve injury can severely hamper nerve regeneration and functional recovery.Further,the anti-inflammatory cytokine,interleukin-10,can inhibit nerve scar formation.Saikosaponin a(SSa) is a monomer molecule extracted from the Chinese medicine,Bupleurum.SSa can exert anti-inflammatory effects in spinal cord injury and traumatic brain injury.However,it has not been shown whether SSa can play a role in peripheral nerve injury.In this study,rats were randomly assigned to three groups.In the sham group,the left sciatic nerve was directly sutured after exposure.In the sciatic nerve injury(SNI) + SSa and SNI groups,the left sciatic nerve was sutured and continuously injected daily with SSa(10 mg/kg) or an equivalent volume of saline for 7 days.Enzyme linked immunosorbent assay results demonstrated that at 7 days after injury,interleukin-10 level was considerably higher in the SNI + SSa group than in the SNI group.Masson staining and western blot assay demonstrated that at 8 weeks after injury,type I and III collagen content was lower and nerve scar formation was visibly less in the SNI + SSa group compared with the SNI group.Simultaneously,sciatic functional index and nerve conduction velocity were improved in the SNI + SSa group compared with the SNI group.These results confirm that SSa can increase the expression of the anti-inflammatory factor,interleukin-10,and reduce nerve scar formation to promote functional recovery of injured sciatic nerve.展开更多
Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after longterm use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an...Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after longterm use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saikosaponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-kB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RBinduced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to saikogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NFkB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosaponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.展开更多
Saikosaponins(SSs)are the main active components extracted from Bupleuri Radix(BR)which has been used as an important herbal drug in Asian countries for thousands of years.It has been reported that the intestinal bact...Saikosaponins(SSs)are the main active components extracted from Bupleuri Radix(BR)which has been used as an important herbal drug in Asian countries for thousands of years.It has been reported that the intestinal bacteria plays an important role in the in vivo disposal of oral SSs.Although the deglycosylated derivatives(saikogenins,SGs)of SSs metabolized by the intestinal bacteria are speculated to be the main components absorbed into the blood after oral administration of SSs,no studies have been reported on the characteristics of SGs for their intestinal absorption,and those for SSs are also limited.Therefore,a rapid UHPLC-MS/MS method was developed to investigate and compare the apparent permeability of three common SSs(SSa,SSd,SSb2)and their corresponding SGs(SGF,SGG,SGD)through a bidirectional transport experiment on Caco-2 cell monolayer model.The method was validated according to the latest FDA guidelines and applied to quantify the six analytes in transport medium samples extracted via liquid-liquid extraction(LLE).The apparent permeability coefficient(Papp)determined in this study indicated that the permeability of SGs improved to the moderate class compared to the corresponding parent compounds,predicting a higher in vivo absorption.Moreover,the efflux ratio(ER)value demonstrated an active uptake of SSd and the three SGs,while a passive diffusion of SSa and SSb2.展开更多
OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosap...OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.展开更多
Three new saikosaponin-like compounds (named prostratoside F-H) were isolated from the whole plants of Polycarpon prostratum (Forssk.) Aschers. et Schwein. ex Aschers. By detailed spectroscopic analysis, their structu...Three new saikosaponin-like compounds (named prostratoside F-H) were isolated from the whole plants of Polycarpon prostratum (Forssk.) Aschers. et Schwein. ex Aschers. By detailed spectroscopic analysis, their structures were determined as 13 beta, 28-epoxy-16-keto-22 alpha, 23-dihydroxyolean-11-en-3 beta -yl-alpha -L-arabinopyranoside, 13 beta, 28-epoxy-16-keto-23-hydroxy-olean-11-en-3 beta -yl-alpha -L-arabinopyranoside and 13 beta, 28-epoxy-16-keto-22 alpha -hydroxyolean-11-en-3 beta -yl-alpha -L-arabinopyranoside, respectively.展开更多
Saikosaponin v-2(1). was isolated li om the roots of the title plant and thc structure was identified on rhs basis of spectral anal? sis. Saikosaponin v-2 is a new compound. which was identified as 3 beta .16 alpha .2...Saikosaponin v-2(1). was isolated li om the roots of the title plant and thc structure was identified on rhs basis of spectral anal? sis. Saikosaponin v-2 is a new compound. which was identified as 3 beta .16 alpha .23.28-tetrahydroxy-olean-11.13(18)-dien-30-oic acid-3-O-beta -D-glucopyranosyl- (1 -->2)glucopyranosyl-(1 -->3)-beta -D-fucopyranosol-30-O-xylitol ester.展开更多
Objective:To investigate the Effect of saikosaponin A on Treg and Th17 immune balance in depressive rats.Methods:The rat depression model was established with reference to the Katz method,and the rats were randomly di...Objective:To investigate the Effect of saikosaponin A on Treg and Th17 immune balance in depressive rats.Methods:The rat depression model was established with reference to the Katz method,and the rats were randomly divided into control group,model group,western medicine group,and saikosaponin A group.The western medicine group was given 1.2 mg/kg/d of fluoxetine,and the saikosaponin A group was given 25 mg/kg/d of saikosaponin A,while the control group and model group were given the same volume of normal saline.The evaluation of depression in Rats was analyzed by Openfield-test and sugar water preference test.Flow cytometry was used to detect the expression of Th17 and Treg cells.And the expression of IL-17,IL-23,TNF-α,IL-10,TGF-βwere detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the control group,the horizontal exercise score,vertical exercise score,and sugar preference of the model group decreased significantly(P<0.05).Compared with the model group,the above indicators were significantly increased in the western medicine group and saikosaponin A group(P<0.05).Flow cytometry showed that compared with the control group,the Th17 cells,Th17/Treg cell ratio in model group increased significantly,whereas the Treg cells decreased significantly(P<0.05).Compared with the model group,The Th17 cells and Th17/Treg ratio in western medicine group and saikosaponin A group decreased,while the Treg cells increased significantly(P<0.05).ELISA showed that compared with control group,the serum levels of IL-17,IL-23 and TNF-αin model group increased,while the levels of IL-10 and TGF-βdecreased(P<0.05).Compared with model group,the levels of IL-17,IL-23 and TNF-αdecreased,while the levels of IL-10 and TGF-βincreased in western medicine group and saikosaponin A group(P<0.05).Conclusion:Saikosaponin A can reduce the degree of depression by regulating the imbalance of Th17/Treg cells and the secretion of inflammatory cytokines in depressed rats.展开更多
OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin...OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin-b(SS-b)on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway.METHODS HepG2 cells were treated with SS-b(10-800 g·L^(-1))for 48 h in vitro.The CCK-8 assay was used to assess cell proliferation,and cell apoptosis was determined by Hoechst33258 staining,AnnexinⅤ/PI staining and caspase 3 assay.RT-PCR was used to examine the expression of MACC1,c-MET and hepatocyte growth factor(HGF)mR NA.MACC1 protein was detected by Western blot and immunohistochemistry.The protein expressions of p-cMET,c-MET,p-AKT,AKT,p-BAD,BAD were measured by Western blot.RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly.HepG2 cells showed karyopyknosis,fragmentation and fluorescence highlight in SS-b treatment group.FCM results showed that apoptosis rate of HepG2 cells increased with SS-b concentration.The immunofluorescence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b.The expression levels of MACC1,c-MET and HGF mR NA in HepG2 cells were significantly inhibited by SS-b.SS-b also significantly decreased the protein expressions of MACC1,p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05).CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.展开更多
Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and...Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and apoptosis of Huh7 cells were detected by MTT and flow cytometry, and then constructed recombinant plasmid pEGFP-N1-LC3B and transfected into Huh7 cells. After intervened by SSA culture medium, the autophagy level was observed under confocal microscope. The expression of apoptosis proteins Bax, Bcl-2, PCNA and autophagy-related proteins LC3B, Beclin1, and Apg12-Apg5 were detected by Western Blot. Results: SSA can significantly inhibit the proliferation of Huh7 cells, promote apoptosis, increase the number of autophagy bodies in the cytoplasm, up-regulate the expression of Bax, LC3B-II, Beclin1, Apg12-Apg5 and down-regulate the expression of Bcl-2, PCNA. Conclusion:SSA induced apoptosis of Huh7 cells in vitro and upregulated the autophagy level.展开更多
Anatomical, histochemical and phytochemical methods were used to investigate the structure, the localization and content changes of total saikosaponin and saikosaponin-a of the roots of Bupleurum chinense DC. at diffe...Anatomical, histochemical and phytochemical methods were used to investigate the structure, the localization and content changes of total saikosaponin and saikosaponin-a of the roots of Bupleurum chinense DC. at different developmental stages. Results showed that saikosaponin was mainly distributed in pericycle and primary phloem in the young root; but in the mature root, it was mainly distributed in vascular cambium and secondary phloem. During the whole growth period from the pre-blossom, blossom, fruit, and fruit mature periods until the pre-withering period, it was in the fruit mature period that both the total saikosaponin content and the saikosaponin-a content reached the highest level. So the last 20 d of October was considered as the right collecting season for the drug of B. chinense. In addition, the quality of 1-year-old drug was better than that of 2-year-old drug due to its higher saikosaponin content. On the other hand, judging from the external characteristics of the drug, the one with an acerose taproot and more lateral roots was of better quality. The results offered theoretical bases for selecting medicinal material of high quality and determining the most appropriate harvesting stage and part of B. chinense.展开更多
Nonalcoholic fatty liver disease(NAFLD)has become one of the most prominent causes of chronic liver diseases and malignancies.However,few therapy has been approved.Radix Bupleuri(RB)is the most frequently used herbal ...Nonalcoholic fatty liver disease(NAFLD)has become one of the most prominent causes of chronic liver diseases and malignancies.However,few therapy has been approved.Radix Bupleuri(RB)is the most frequently used herbal medicine for the treatment of liver diseases.In the current study,we aim to systemically evaluate the therapeutic effects of saikosaponin A(SSa)and saikosaponin D(SSd),the major bioactive monomers in RB,against NAFLD and to investigate the underlying mechanisms.Our results demonstrated that both SSa and SSd improved diet-induced NAFLD.Integrative lipidomic and transcriptomic analysis revealed that SSa and SSd modulated glycerolipid metabolism by regulating related genes,like Lipe and Lipg.SSd profoundly suppressed the fatty acid biosynthesis by downregulating Fasn and Acaca expression and promoted fatty acid degradation by inducing Acox1 and Cpt1 a expression.Bioinformatic analysis further predicted the implication of master transcription factors,including peroxisome proliferator-activated receptor alpha(PPARα),in the protective effects of SSa and SSd.These results were further confirmed in vitro in mouse primary hepatocytes.In summary,our study uncoded the complicated mechanisms underlying the promising anti-steatosis activities of saikosaponins(SSs),and provided critical evidence inspiring the discovery of innovative therapies based on SSa and SSd for the treatment of NAFLD and related complications.展开更多
OBJECTIVE:To investigate effects of Saikosaponin D(SSd) on syndecan-2,matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases-2(TIMP-2) in livers of rat with hepatocellular carcinoma(HCC).METHODS:Ma...OBJECTIVE:To investigate effects of Saikosaponin D(SSd) on syndecan-2,matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases-2(TIMP-2) in livers of rat with hepatocellular carcinoma(HCC).METHODS:Male SD rats were divided into control(n=10),model(n=20) and SSd(n=20) groups,and model and SSd groups given intragastric 0.2%(w/v) N-diethylnitrosamine to induce HCC.SSd group received 0.03%(w/v) SSd in saline.Liver samples were analysed immunohistochemically for syndecan-2,MMP-2,MMP-13 and TIMP-2 at 16 weeks.RESULTS:The model group had more malignant nodules than the SSd group;all model-group HCC cells were grade III;SSd-group HCC cells were grades I-II.Controls showed normal hepatic cell phenotypes and no syndecan-2 + staining.Syndecan-2 + staining was greater in the model group(35.2%,P≤0.001) than in controls or the SSd group(16.5%,P ≤ 0.001).The model group had more intense MMP-2 + staining than controls(0.37 vs 0.27,P≤0.01) or the SSd group(0.31 vs 0.37,P≤0.05);and higher MMP-13 + staining(72.55%) than in controls(12.55%,P≤0.001) and SSd group(20.18%,P≤0.01).The model group also had more TIMP-2 + staining(57.2%) than controls(20.9%,P≤0.001) and SSd group(22.7%,P≤0.001).Controls and SSd group showed no difference in TIMP-2 + rates.CONCLUSION:SSd inhibited HCC development,and downregulated expression of syndecan-2,MMP-2,MMP-13 and TIMP-2 in rat HCC liver tissue.展开更多
As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri(Chai-Hu) is commonly used for the treatment of influenza,which is derived from the dried root...As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri(Chai-Hu) is commonly used for the treatment of influenza,which is derived from the dried roots of Bupleurum chinense DC.and Bupleurum scorzonerifolium Willd.Among of diverse chemical components,saikosaponins are the key active components of the herb medicine.In the present study,we established a method of high performance liquid chromatography(HPLC) coupled with evaporative light scattering detection(ELSD) for simultaneous determination of saikosaponin a,c and d in root,stem,leaf and flower of Bupleurum chinense(B chinense) collected from different areas of Shanxi Province,China.The results from 16 samples of root,stem,leaf and flower of B chinense demonstrated that the total contents of the three saikosaponins in the root of B chinense collected from Dongshan Taiyuan,Xishan,Tianlongshan and Pangquangou were 4.26 mg/g,3.22 mg/g,4.23 mg/g and 3.05 mg/g,respectively.However,there was scarcely any saikosaponins in the stem,leaf and flower of B chinense collected from above-mentioned areas.The method of HPLC coupled with ELSD was suitable for quality control of Radix Bupleuri.The result also confirmed that the root of B chinense was the best medicinal part.展开更多
5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was...5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was revealed to have agonistic activity on 5-HT2 C receptor, and the subsequent bioassay-guided isolation led to identification of several saikosaponins as the active constituents with 5-HT2 C receptor agonistic activity in vitro and anti-obesity activity in vivo. The new compound, 22-oxosaikosaponin d(1), was determined by extensive spectroscopic analyses(HR-ESI-MS, IR, and 1D and 2D NMR). The primary structure-activity relationship study suggested that the intramolecular ether bond between C-13 and C-28 and the number of sugars at C-3 position were closely related to the 5-HT2 C receptor agonistic activity. Saikosaponin a(3), the main saponin in B. chinense, showed obviously agonistic activity on 5-HT2 C receptor with an EC50 value of 21.08 ± 0.33 μmol×L^(–1) in vitro and could reduce food intake by 39.1% and 69.2%, and weight gain by 13.6% and 16.4%, respectively, at 3.0 and 6.0 mg×kg^(–1) in vivo. This investigation provided valuable information for the potential use of B. chinense as anti-obesity agent.展开更多
Objective To identify the genes of WRKY transcription factors(TFs) from roots of Bupleurum chinense and genes that potentially regulate saikosaponin(SS) biosynthesis.Methods Firstly,the subfamily cluster analysis ...Objective To identify the genes of WRKY transcription factors(TFs) from roots of Bupleurum chinense and genes that potentially regulate saikosaponin(SS) biosynthesis.Methods Firstly,the subfamily cluster analysis was mainly based on Arabidopsis thaliana WRKYs for 27 putative WRKY TFs selected from previous transcriptome sequencing data.Secondly,qPCR was used to screen such genes of WRKY TFs that could be induced by NaCI and PEG6000 in adventitious roots of B.chinense.Meanwhile,saikosaponins(SSs) in treated adventitious roots were determined by HPLC.The roots were collected at 0,2,4,8,12,24,48,and 72 h after treatments,and 120 h only for PEG.Finally,the tissue-specific expression was analyzed on screened genes by qPCR.Results The 27 genes were grouped into three categories:There were nine in Group Ⅰ,15 in Group Ⅱ,and two in Group Ⅲ.Four genes of WRKYTFs,BCWRKY6,BCWRKY16,BCWRKY32,and BCWRKY35 were obviously induced by NaCI in adventitious roots of B.chinense,while only BCWRKY32 was induced by PEG.The content of SSs increased at different levels in NaCI and PEG6000 treatment.Three genes including BCWRKY6,BCWRKY32,and BCWRKY35,expressed most in roots,were similar to the accumulation pattern of SS.Conclusion The three WRKY genes,BCWRKY6,BCWRKY32,and BCWRKY35,may be involved in the biosynthesis of SS.展开更多
基金supported by National Natural Science Foundation of China (82222071, 82273990, 82104253)the opening project of State Key Laboratory of Natural Medicines (SKLNMKF202208)
文摘Non-alcoholic fatty liver disease(NAFLD)is the main cause of chronic liver disease worldwide.Bupleurum is widely used in the treatment of non-alcoholic fatty liver,and saikosaponin D(SSD)is one of the main active components of Bupleurum.The purpose of this study was to investigate the efficacy of SSD in the treatment of NAFLD and to explore the mechanism of SSD in the improvement of NAFLD based on“gut-liver axis”.Our results showed that SSD dose-dependently alleviated high fat diet-induced weight gain in mice,improved insulin sensitivity,and also reduced liver lipid accumulation and injury-related biomarkers aspartate aminotransferase(AST)and alanine aminotransferase(ALT).Further exploration found that SSD inhibited the mRNA expression levels of farnesoid X receptor(Fxr),small heterodimer partner(Shp),recombinant fibroblast growth factor 15(Fgf15)and apical sodium dependent bile acid transporter(Asbt)in the intestine,suggesting that SSD improved liver lipid metabolism by inhibiting intestinal FXR signaling.SSD can significantly reduce the gut microbiota associated with bile salt hydrolase(BSH)expression,such as Clostridium.Decreased BSH expression reduced the ratio of unconjugated to conjugated bile acids,thereby inhibiting the intestinal FXR.These data demonstrated that SSD ameliorated NAFLD potentially through the gut microbiota-bile acidintestinal FXR pathway and suggested that SSD is a promising therapeutic agent for the treatment of NAFLD.
文摘A new saikosaponin was isolated from Bupleurum chinense DC., and its structure was identified as 3β,16α,23,28,30_pentahydroxy_olean_11,13(18)_dien_3_O_β_D_glucopyranosyl(1→6)_[α_L_rhamnopyranosyl (1→4)]_β_D_glucopyranoside on the basis of chemical and spectral evidence, named as saikosaponin q_1. In addition, two known saikosaponins, 3″_O_acetyl_saikosaponin d and 3″_O_acetyl_saikosaponin b 2, were also isolated and identified from this plant for the first time.
基金supported by the National Natural Science Foundation of China(No.30771895)the Sci-tch Project of Shaanxi Province(No.2007K16-07)
文摘Objective To investigate the anticancer effects and detailed mechanisms of Saikosaponin D(SSD)in human hepatoma HepG2 cells.Methods Cell proliferation and apoptosis were tested by MTT assay and Annexin-V/PI assay respectively.The expressions of CCAAT enhancer binding protein β(C/EBPβ)and p53 were detected by RT-PCR and Western blotting.Results SSD inhibited cell proliferation in a dose-dependent manner and induced apoptosis at the concentration of 5.0 mg/L.SSD significantly increased the mRNA and protein levels of C/EBPβ and p53 in a dose-dependent manner.Conclusion SSD exerts its anticancer effect by inhibiting cell proliferation and inducing apoptosis partly through C/EBPβ-p53 signal pathway in HepG2 cells.
基金financially supported by the National Natural Science Foundation of China,No.11672332,11102235,8167050417the National Key Research and Development Plan of China,No.2016YFC1101500+1 种基金the Key Science and Technology Support Foundation of Tianjin City of China,No.17YFZCSY00620the Natural Science Foundation of Tianjin City of China,No.15JCYBJC28600,17JCZDJC35400
文摘Nerve scarring after peripheral nerve injury can severely hamper nerve regeneration and functional recovery.Further,the anti-inflammatory cytokine,interleukin-10,can inhibit nerve scar formation.Saikosaponin a(SSa) is a monomer molecule extracted from the Chinese medicine,Bupleurum.SSa can exert anti-inflammatory effects in spinal cord injury and traumatic brain injury.However,it has not been shown whether SSa can play a role in peripheral nerve injury.In this study,rats were randomly assigned to three groups.In the sham group,the left sciatic nerve was directly sutured after exposure.In the sciatic nerve injury(SNI) + SSa and SNI groups,the left sciatic nerve was sutured and continuously injected daily with SSa(10 mg/kg) or an equivalent volume of saline for 7 days.Enzyme linked immunosorbent assay results demonstrated that at 7 days after injury,interleukin-10 level was considerably higher in the SNI + SSa group than in the SNI group.Masson staining and western blot assay demonstrated that at 8 weeks after injury,type I and III collagen content was lower and nerve scar formation was visibly less in the SNI + SSa group compared with the SNI group.Simultaneously,sciatic functional index and nerve conduction velocity were improved in the SNI + SSa group compared with the SNI group.These results confirm that SSa can increase the expression of the anti-inflammatory factor,interleukin-10,and reduce nerve scar formation to promote functional recovery of injured sciatic nerve.
基金This study is funded by the National Nature Science Foundation of China(Grant Nos.:82074323,and 81673572)Key Research and Development Program of Shanxi Province(Program No.:202102130501010)+2 种基金The major science and technology project for“Significant New Drugs Creation”(Project No.:2017ZX09301047)Research Project Supported by Shanxi Scholarship Council of China(Project No.:2020019)The special fund for Science and Technology Innovation Teams of Shanxi Province(Grant No.:202204051002011).
文摘Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after longterm use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saikosaponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-kB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RBinduced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to saikogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NFkB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosaponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.
基金project was financially supported by the National Natural Science Foundation of China(No.81573626)the Open Project Program of Guangxi Key Laboratory of Traditional Chinese Medicine Quality Standards,China(No.201503)+1 种基金the Natural Research Foundation of Jiangsu Province,China(No.BK20161456)the Qing Lan Project of Jiangsu Province,China(2017)。
文摘Saikosaponins(SSs)are the main active components extracted from Bupleuri Radix(BR)which has been used as an important herbal drug in Asian countries for thousands of years.It has been reported that the intestinal bacteria plays an important role in the in vivo disposal of oral SSs.Although the deglycosylated derivatives(saikogenins,SGs)of SSs metabolized by the intestinal bacteria are speculated to be the main components absorbed into the blood after oral administration of SSs,no studies have been reported on the characteristics of SGs for their intestinal absorption,and those for SSs are also limited.Therefore,a rapid UHPLC-MS/MS method was developed to investigate and compare the apparent permeability of three common SSs(SSa,SSd,SSb2)and their corresponding SGs(SGF,SGG,SGD)through a bidirectional transport experiment on Caco-2 cell monolayer model.The method was validated according to the latest FDA guidelines and applied to quantify the six analytes in transport medium samples extracted via liquid-liquid extraction(LLE).The apparent permeability coefficient(Papp)determined in this study indicated that the permeability of SGs improved to the moderate class compared to the corresponding parent compounds,predicting a higher in vivo absorption.Moreover,the efflux ratio(ER)value demonstrated an active uptake of SSd and the three SGs,while a passive diffusion of SSa and SSb2.
基金supported by Scientific and Technology Projects of Henan Province(142102310137)Science and Technology Development Project of Luoyang City(1603001A-3)
文摘OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma(HCC).In this study,we investigated the anti-proliferative activities and antiangiogenesis effects of saikosaponins(SS)-b on hepatocellular carcinoma(HCC)and its regulation on VEGF/ERK/HIF-1 αsignal pathway.METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro.Pathological change of tumor tissue was observed by HE staining,the microvascular changes were detected by immunohistochemical method.The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane(CAM)model.The effects of SS-b on proliferation,migration and invasion were investigated by MTT assay,scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell(HUVEC)and HepG2 cells in vitro.Vascular endothelial growth factor(VEGF),matrix metalloproteinase-2/9(MMP-2/9),hypoxia-inducible factor-1α(HIF-1α)expression and the phosphorylation of extracellular regulated kinase(ERK)were analyzed using RT-PCR and Westernblot.RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo.The inhibitory rate of tumor was 49.1%,50.7%,66.1%in SS-b 5,10 and 20 mg·kg-1group respectively.HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice.Moreover,SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF.SS-b had an obvious inhibitory effect on cell proliferation,migration and invasion of HUVEC cells and HepG-2 cells.These effects were associated with downregulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1αsignaling in H22 mice and Hep-G2 cells.CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibiting tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.
文摘Three new saikosaponin-like compounds (named prostratoside F-H) were isolated from the whole plants of Polycarpon prostratum (Forssk.) Aschers. et Schwein. ex Aschers. By detailed spectroscopic analysis, their structures were determined as 13 beta, 28-epoxy-16-keto-22 alpha, 23-dihydroxyolean-11-en-3 beta -yl-alpha -L-arabinopyranoside, 13 beta, 28-epoxy-16-keto-23-hydroxy-olean-11-en-3 beta -yl-alpha -L-arabinopyranoside and 13 beta, 28-epoxy-16-keto-22 alpha -hydroxyolean-11-en-3 beta -yl-alpha -L-arabinopyranoside, respectively.
基金This study was financially supported by the National Natural Science Foundation of China (29632050).
文摘Saikosaponin v-2(1). was isolated li om the roots of the title plant and thc structure was identified on rhs basis of spectral anal? sis. Saikosaponin v-2 is a new compound. which was identified as 3 beta .16 alpha .23.28-tetrahydroxy-olean-11.13(18)-dien-30-oic acid-3-O-beta -D-glucopyranosyl- (1 -->2)glucopyranosyl-(1 -->3)-beta -D-fucopyranosol-30-O-xylitol ester.
基金Hebei Science and technology program 2017 Hebei key R&D plan big health service and biomedical special project(No.17277782D)
文摘Objective:To investigate the Effect of saikosaponin A on Treg and Th17 immune balance in depressive rats.Methods:The rat depression model was established with reference to the Katz method,and the rats were randomly divided into control group,model group,western medicine group,and saikosaponin A group.The western medicine group was given 1.2 mg/kg/d of fluoxetine,and the saikosaponin A group was given 25 mg/kg/d of saikosaponin A,while the control group and model group were given the same volume of normal saline.The evaluation of depression in Rats was analyzed by Openfield-test and sugar water preference test.Flow cytometry was used to detect the expression of Th17 and Treg cells.And the expression of IL-17,IL-23,TNF-α,IL-10,TGF-βwere detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the control group,the horizontal exercise score,vertical exercise score,and sugar preference of the model group decreased significantly(P<0.05).Compared with the model group,the above indicators were significantly increased in the western medicine group and saikosaponin A group(P<0.05).Flow cytometry showed that compared with the control group,the Th17 cells,Th17/Treg cell ratio in model group increased significantly,whereas the Treg cells decreased significantly(P<0.05).Compared with the model group,The Th17 cells and Th17/Treg ratio in western medicine group and saikosaponin A group decreased,while the Treg cells increased significantly(P<0.05).ELISA showed that compared with control group,the serum levels of IL-17,IL-23 and TNF-αin model group increased,while the levels of IL-10 and TGF-βdecreased(P<0.05).Compared with model group,the levels of IL-17,IL-23 and TNF-αdecreased,while the levels of IL-10 and TGF-βincreased in western medicine group and saikosaponin A group(P<0.05).Conclusion:Saikosaponin A can reduce the degree of depression by regulating the imbalance of Th17/Treg cells and the secretion of inflammatory cytokines in depressed rats.
基金supported by Scientific and Technology Projects of Henan Province(142102310137)Science and Technology Development Project of Luoyang City(1603001A-3)
文摘OBJECTIVE Metastasis-associated in colon cancer-1(MACC1)is an oncogene that has been newly identified.It promotes tumor proliferation and invasion via the MET pathway.Our study investigated the effects of Saikosaponin-b(SS-b)on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway.METHODS HepG2 cells were treated with SS-b(10-800 g·L^(-1))for 48 h in vitro.The CCK-8 assay was used to assess cell proliferation,and cell apoptosis was determined by Hoechst33258 staining,AnnexinⅤ/PI staining and caspase 3 assay.RT-PCR was used to examine the expression of MACC1,c-MET and hepatocyte growth factor(HGF)mR NA.MACC1 protein was detected by Western blot and immunohistochemistry.The protein expressions of p-cMET,c-MET,p-AKT,AKT,p-BAD,BAD were measured by Western blot.RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly.HepG2 cells showed karyopyknosis,fragmentation and fluorescence highlight in SS-b treatment group.FCM results showed that apoptosis rate of HepG2 cells increased with SS-b concentration.The immunofluorescence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b.The expression levels of MACC1,c-MET and HGF mR NA in HepG2 cells were significantly inhibited by SS-b.SS-b also significantly decreased the protein expressions of MACC1,p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05).CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.
基金This study was supported by National Natural Science Foundation(81273745).
文摘Objective: It is discussed whether saikosaponin A induces apoptosis of human hepatoma Huh7 cells is related to the change of autophagy level.Methods: The effects of different concentrations of SSA on proliferation and apoptosis of Huh7 cells were detected by MTT and flow cytometry, and then constructed recombinant plasmid pEGFP-N1-LC3B and transfected into Huh7 cells. After intervened by SSA culture medium, the autophagy level was observed under confocal microscope. The expression of apoptosis proteins Bax, Bcl-2, PCNA and autophagy-related proteins LC3B, Beclin1, and Apg12-Apg5 were detected by Western Blot. Results: SSA can significantly inhibit the proliferation of Huh7 cells, promote apoptosis, increase the number of autophagy bodies in the cytoplasm, up-regulate the expression of Bax, LC3B-II, Beclin1, Apg12-Apg5 and down-regulate the expression of Bcl-2, PCNA. Conclusion:SSA induced apoptosis of Huh7 cells in vitro and upregulated the autophagy level.
基金the National Natural Science Foundation of China (30770122)the Shaanxi Provincial Natural Science Foundation (2004C102)
文摘Anatomical, histochemical and phytochemical methods were used to investigate the structure, the localization and content changes of total saikosaponin and saikosaponin-a of the roots of Bupleurum chinense DC. at different developmental stages. Results showed that saikosaponin was mainly distributed in pericycle and primary phloem in the young root; but in the mature root, it was mainly distributed in vascular cambium and secondary phloem. During the whole growth period from the pre-blossom, blossom, fruit, and fruit mature periods until the pre-withering period, it was in the fruit mature period that both the total saikosaponin content and the saikosaponin-a content reached the highest level. So the last 20 d of October was considered as the right collecting season for the drug of B. chinense. In addition, the quality of 1-year-old drug was better than that of 2-year-old drug due to its higher saikosaponin content. On the other hand, judging from the external characteristics of the drug, the one with an acerose taproot and more lateral roots was of better quality. The results offered theoretical bases for selecting medicinal material of high quality and determining the most appropriate harvesting stage and part of B. chinense.
基金supported by grants from the National Natural Science Foundation of China(Nos.81773997 and 81073148 to Rong SunNo.82004029 to Runping Liu+3 种基金No.82004045 to Xiaojiaoyang Li)Beijing University of Chinese Medicine(No.2020-JYB-ZDGG-038 to Runping Liu,China)supported by grants from Beijing Nova Program of Science&Technology(Nos.Z201100006820025 and Z191100001119088,China)supported by research fund‘Traditional Chinese medicine pharmacology and toxicology expert(No.ts201511107)’from the Taishan Scholar Project of Shandong Province(China)。
文摘Nonalcoholic fatty liver disease(NAFLD)has become one of the most prominent causes of chronic liver diseases and malignancies.However,few therapy has been approved.Radix Bupleuri(RB)is the most frequently used herbal medicine for the treatment of liver diseases.In the current study,we aim to systemically evaluate the therapeutic effects of saikosaponin A(SSa)and saikosaponin D(SSd),the major bioactive monomers in RB,against NAFLD and to investigate the underlying mechanisms.Our results demonstrated that both SSa and SSd improved diet-induced NAFLD.Integrative lipidomic and transcriptomic analysis revealed that SSa and SSd modulated glycerolipid metabolism by regulating related genes,like Lipe and Lipg.SSd profoundly suppressed the fatty acid biosynthesis by downregulating Fasn and Acaca expression and promoted fatty acid degradation by inducing Acox1 and Cpt1 a expression.Bioinformatic analysis further predicted the implication of master transcription factors,including peroxisome proliferator-activated receptor alpha(PPARα),in the protective effects of SSa and SSd.These results were further confirmed in vitro in mouse primary hepatocytes.In summary,our study uncoded the complicated mechanisms underlying the promising anti-steatosis activities of saikosaponins(SSs),and provided critical evidence inspiring the discovery of innovative therapies based on SSa and SSd for the treatment of NAFLD and related complications.
基金Supported by the National Natural Science Foundation of China(No.30471982)
文摘OBJECTIVE:To investigate effects of Saikosaponin D(SSd) on syndecan-2,matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases-2(TIMP-2) in livers of rat with hepatocellular carcinoma(HCC).METHODS:Male SD rats were divided into control(n=10),model(n=20) and SSd(n=20) groups,and model and SSd groups given intragastric 0.2%(w/v) N-diethylnitrosamine to induce HCC.SSd group received 0.03%(w/v) SSd in saline.Liver samples were analysed immunohistochemically for syndecan-2,MMP-2,MMP-13 and TIMP-2 at 16 weeks.RESULTS:The model group had more malignant nodules than the SSd group;all model-group HCC cells were grade III;SSd-group HCC cells were grades I-II.Controls showed normal hepatic cell phenotypes and no syndecan-2 + staining.Syndecan-2 + staining was greater in the model group(35.2%,P≤0.001) than in controls or the SSd group(16.5%,P ≤ 0.001).The model group had more intense MMP-2 + staining than controls(0.37 vs 0.27,P≤0.01) or the SSd group(0.31 vs 0.37,P≤0.05);and higher MMP-13 + staining(72.55%) than in controls(12.55%,P≤0.001) and SSd group(20.18%,P≤0.01).The model group also had more TIMP-2 + staining(57.2%) than controls(20.9%,P≤0.001) and SSd group(22.7%,P≤0.001).Controls and SSd group showed no difference in TIMP-2 + rates.CONCLUSION:SSd inhibited HCC development,and downregulated expression of syndecan-2,MMP-2,MMP-13 and TIMP-2 in rat HCC liver tissue.
基金Shanxi Educational Committee(Grant No.20111113)Shanxi Science and Technology Department(Grant No.2016ZD0201)
文摘As a traditional Chinese herbal medicine exhibiting analgesic,fever-reducing and anti-inflammatory effects,Radix Bupleuri(Chai-Hu) is commonly used for the treatment of influenza,which is derived from the dried roots of Bupleurum chinense DC.and Bupleurum scorzonerifolium Willd.Among of diverse chemical components,saikosaponins are the key active components of the herb medicine.In the present study,we established a method of high performance liquid chromatography(HPLC) coupled with evaporative light scattering detection(ELSD) for simultaneous determination of saikosaponin a,c and d in root,stem,leaf and flower of Bupleurum chinense(B chinense) collected from different areas of Shanxi Province,China.The results from 16 samples of root,stem,leaf and flower of B chinense demonstrated that the total contents of the three saikosaponins in the root of B chinense collected from Dongshan Taiyuan,Xishan,Tianlongshan and Pangquangou were 4.26 mg/g,3.22 mg/g,4.23 mg/g and 3.05 mg/g,respectively.However,there was scarcely any saikosaponins in the stem,leaf and flower of B chinense collected from above-mentioned areas.The method of HPLC coupled with ELSD was suitable for quality control of Radix Bupleuri.The result also confirmed that the root of B chinense was the best medicinal part.
基金supported by the National Science Foundation of China(No.81573322)the Hundred-Talent Program of CAS+1 种基金the CAS"Light of West China"Programthe Youth Innovation Promotion Association of CAS
文摘5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was revealed to have agonistic activity on 5-HT2 C receptor, and the subsequent bioassay-guided isolation led to identification of several saikosaponins as the active constituents with 5-HT2 C receptor agonistic activity in vitro and anti-obesity activity in vivo. The new compound, 22-oxosaikosaponin d(1), was determined by extensive spectroscopic analyses(HR-ESI-MS, IR, and 1D and 2D NMR). The primary structure-activity relationship study suggested that the intramolecular ether bond between C-13 and C-28 and the number of sugars at C-3 position were closely related to the 5-HT2 C receptor agonistic activity. Saikosaponin a(3), the main saponin in B. chinense, showed obviously agonistic activity on 5-HT2 C receptor with an EC50 value of 21.08 ± 0.33 μmol×L^(–1) in vitro and could reduce food intake by 39.1% and 69.2%, and weight gain by 13.6% and 16.4%, respectively, at 3.0 and 6.0 mg×kg^(–1) in vivo. This investigation provided valuable information for the potential use of B. chinense as anti-obesity agent.
基金CAMS Innovation Fund for Medical Sciences(CIFMS)(2016-I2M-2-003)
文摘Objective To identify the genes of WRKY transcription factors(TFs) from roots of Bupleurum chinense and genes that potentially regulate saikosaponin(SS) biosynthesis.Methods Firstly,the subfamily cluster analysis was mainly based on Arabidopsis thaliana WRKYs for 27 putative WRKY TFs selected from previous transcriptome sequencing data.Secondly,qPCR was used to screen such genes of WRKY TFs that could be induced by NaCI and PEG6000 in adventitious roots of B.chinense.Meanwhile,saikosaponins(SSs) in treated adventitious roots were determined by HPLC.The roots were collected at 0,2,4,8,12,24,48,and 72 h after treatments,and 120 h only for PEG.Finally,the tissue-specific expression was analyzed on screened genes by qPCR.Results The 27 genes were grouped into three categories:There were nine in Group Ⅰ,15 in Group Ⅱ,and two in Group Ⅲ.Four genes of WRKYTFs,BCWRKY6,BCWRKY16,BCWRKY32,and BCWRKY35 were obviously induced by NaCI in adventitious roots of B.chinense,while only BCWRKY32 was induced by PEG.The content of SSs increased at different levels in NaCI and PEG6000 treatment.Three genes including BCWRKY6,BCWRKY32,and BCWRKY35,expressed most in roots,were similar to the accumulation pattern of SS.Conclusion The three WRKY genes,BCWRKY6,BCWRKY32,and BCWRKY35,may be involved in the biosynthesis of SS.