Simultaneous determination of geniposidic acid. chlorogenic acid and geniposide in eucommia by HPLC

A high performance liquid chromatography (HPLC) method was established for simultaneous determina-tion of geniposidic acid, chlorogenic acid and geniposide in eucommia. Detection at 240 nm with a reversed-phasecolumn, CH3OH volume fraction, acidic additive and pH value of mobile phase were studied for their effects on theseparability of the compounds. The most suitable separation was obtained with isocratic gradient elution systemusing CH3OH-H2O-H3 PO4 (12.00: 87.96: 0.04, volume ratio) at a flow-rate of 1.0 mL/min. Under the experi-mental conditions, the capacity factors of three compounds are in 3-13. The sample is separated rightly. Theanalysis time is 30 min and the retention time of genfposidic acid, chlorogenic acid and geniposide are 6. 7 min,10.5 min and 21 min, respectively.

Baicalin and geniposide inhibit the development of atherosclerosis by increasing Wntl and inhibiting dickkopf-related protein-1 expression

Background Our previous study showed that the combined Chinese herbs containing scutellaria baicalensis georgi and gardenia jasminoids ellis inhibited atherosclerosis. In this study, we sought to determine if baicalin and geniposide could inhibit atherosclerosis through Wntl and dickkopf-related protein-1 （DKK1）. Methods The wild-type and ApoE-/- mice were treated with baicalin, geniposide, and baicalin plus geniposide daily by gavage for 12 weeks. Blood lipid levels were measured with an automatic biochemistry analyzer. Aortic atherosclerotic lesion areas were analyzed with Image-ProPlus software. The mRNA and protein expression of DKK1, Wntt and nuclear factor-r,B （NF-κB） were measured with RT-PCR and Westem Blot. Serum levels of interleukin-12 （IL-12） were quantified with ELISA. Results The baicalin or geniposide monotherapy as well as combination therapy inhibited the development of atherosclerotic lesions, increased Wntl and decreased DKKI expression and elevated the ratio of Wntl/DKK1 compared with high-lipid diet group. However, only baicalin or geniposide monotherapy decreased NF-κB expression. Moreover, baicalin and geniposide monoor combination therapy lowered IL-12 levels. Geniposide reduced both serum total cholesterol and low density lipoprotein levels, while baicalin either alone or in combination with geniposide did not affect serum lipid levels. In human, umbilical vein endothelial ceils stimulated by oxidized low density lipoprotein, baicalin and geniposide also increased Wntl and decreased DKK1 expression and elevated the ratio of Wntl/DKK1. Condusions Baicalin and geniposide exert inflammation-regulatory effects and may prevent atherosclerotic lesions through enhancing Wntl and inhibit- ing DKK1 expression.

Geniposide protects human neuroblastoma SH-SY5Y cells against corticosterone-induced injury

In vitro cultured human neuroblastoma SH-SY5Y cells were pretreated with 50 or 5 ug/mL geniposide for 12 hours and exposed to 400 umol/L corticosterone. Corticosterone exposure in cultures not pretreated with geniposide resulted in inhibited cell growth, reduced cell survival, and increased P53 and P21 protein expression. However, in geniposide pretreated SH-SY5Y cells, cell viability and the number of cells in the G2 phase of the cell cycle were significantly increased, P21 and P53 protein expression was reduced, and cell apoptosis was inhibited following corticosterone exposure. These results indicate that geniposide can protect SH-SY5Y cells against high-dose corticosterone-induced injury.

Geniposide, the component of the Chinese herbal formula Tongluojiunao, protects amyloid-β peptide(1–42)-mediated death of hippocampal neurons via the non-classical estrogen signaling pathway

Tongluojiunao （TLJN） is an herbal medicine consisting of two main components, geniposide and ginsenoside Rg1. TLJN has been shown to protect primary cultured hippocampal neurons. How-ever, its mechanism of action remains unclear. In the present study, primary cultured hippocampal neurons treated with Aβ1-42 （10 μmol/L） signiifcantly increased the release of lactate dehydroge-nase, which was markedly reduced by TLJN （2 μL/mL）, speciifcally by the component geniposide （26 μmol/L）, but not ginsenoside Rg1 （2.5 μmol/L）. hTe estrogen receptor inhibitor, ICI182780 （1 μmol/L）, did not block TLJN-or geniposide-mediated decrease of lactate dehydrogenase under Aβ1-42-exposed conditions. However, the phosphatidyl inositol 3-kinase or mitogen-activated protein kinase pathway inhibitor, LY294002 （50 μmol/L） or U0126 （10 μmol/L）, respectively blo cked the decrease of lactate dehydrogenase mediated by TLJN or geniposide. hTerefore, these results suggest that the non-classical estrogen pathway （i.e., phosphatidyl inositol 3-kinase or mitogen-activated protein kinase） is involved in the neuroprotective effect of TLJN, speciifcally its component, geniposide, against Aβ1-42-mediated cell death in primary cultured hippocampal neurons.

Idiopathic mesenteric phlebosclerosis associated with long-term oral intake of geniposide

BACKGROUND Idiopathic mesenteric phlebosclerosis(IMP)is a rare disease,and its etiology and risk factors remain uncertain.AIM To investigate the possible influence of Chinese herbal liquid containing geniposide on IMP.METHODS The detailed formula of herbal liquid prescriptions of all patients was studied,and the herbal ingredients were compared to identify the toxic agent as a possible etiological factor.Abdominal computed tomography(CT)and colonoscopy images were reviewed to determine the extent and severity of mesenteric phlebosclerosis and the presence of findings regarding colitis.The disease CT score was determined by the distribution of mesenteric vein calcification and colon wall thickening on CT images.The drinking index of medicinal liquor was calculated from the daily quantity and drinking years of Chinese medicinal liquor.Subsequently,Spearman’s correlation analysis was conducted to evaluate the correlation between the drinking index and the CT disease score.RESULTS The mean age of the 8 enrolled patients was 75.7 years and male predominance was found(all 8 patients were men).The patients had histories of 5-40 years of oral Chinese herbal liquids containing geniposide and exhibited typical imaging characteristics(e.g.,threadlike calcifications along the colonic and mesenteric vessels or associated with a thickened colonic wall in CT images).Calcifications were confined to the right-side mesenteric vein in 6 of the 8 patients(75%)and involved the left-side mesenteric vein of 2 cases(25%)and the calcifications extended to the mesorectum in 1 of them.The thickening of colon wall mainly occurred in the right colon and the transverse colon.The median disease CT score was 4.88(n=7)and the median drinking index was 5680(n=7).After Spearman’s correlation analysis,the median CT score of the disease showed a significant positive correlation with the median drinking index(r=0.842,P<0.05).CONCLUSION Long-term oral intake of Chinese herbal liquid containing geniposide may play a role in the pathogenesis of IMP.

Effects of Pulse Ultrasound on Adsorption of Geniposide on Resin 1300 in a Fixed Bed

The effects of pulse ultrasound with different pulse parameters on the breakthrough curves of Geniposide on Resin 1300 were studied. The mass transfer model describing the adsorption process was constructed. Adsorption capability and the overall mass-transfer coefficient were obtained by fitting the constructed mass-transfer model and the experimental data. The effects of pulse ultrasound on adsorption of Geniposide on Resin1300 in a fixed bed were studied and compared. Amount of Geniposide adsorbed on Resin 1300 in the presence of ultrasound is lower than that in the absence of ultrasound, but the mass-transfer rate with ultrasonic irradiation is higher than that without ultrasound. Furthermore, mass transfer rate is enhanced by pulse modulation. In the conditions studied, the adsorption equilibrium constant decreases with increasing ultrasonic power, while the overall mass-transfer co-efficient increases. With increasing pulse duty ratio, adsorption equilibrium constant decreases initially, reaches a minimum when pulse duty ratio is 0.5, and then increases. On the contrary, the overall mass-transfer coefficient in-creases initially and reaches a maximum when pulse ratio is 0.5, and then decreases. Effects of pulse period on ad-sorption equilibrium and mass transfer rate reached the peak at pulse period of 28.6 ms.

Geniposide prevents rotenone-induced apoptosis in primary cultured neurons

Geniposide, a monomer extracted from gardenia and widely used in Chinese medicine, is a novel agonist at the glucagon-like peptide-1 receptor. This receptor is involved in neuroprotection. In the present study, we sought to identify an anti-apoptotic mechanism for the treatment of neurodegenerative diseases. Primary cultured neurons were treated with different concentrations of rotenone for 48 hours. Morphological observation, cell counting kit-8 assay, lactate dehydrogenase detection and western blot assay demonstrated that 0.5 n M rotenone increased lactate dehydrogenase release, decreased the expression of procaspase-3 and Bcl-2, and increased cleaved caspase-3 expression in normal neurons. All these effects were prevented by geniposide. Our results indicate that geniposide diminished rotenone-induced injury in primary neurons by suppressing apoptosis. This may be one of the molecular mechanisms underlying the efficacy of geniposide in the treatment of neurodegenerative diseases.

Synergistic use of geniposide and baicalin on BV2 cell activation damage caused by LPS

OBJECTIVE To explore the synergistic effect of baicalin and geniposide(BG)on BV2 cell activation damage caused by lipopolysaccharide(LPS).METHODS BV2 murine microglial cell line was cultured in vitro,LPS(final concentration 500 ng·m L-1)and various concentrationof Baicalin and Geniposide(BG)(final concentration12.5,25 and 50μg·m L-1)were added tointerven,the negative control was establised.MTT method was used to value the effect of LPS on the viability of BV2 cell line.The accumulated nitrite was assayed utilizing the Griess reaction method.RESULTS(1)Morphological observation:The common marphological of quesient microglia is circle,cell bodies smaller and synaptic slender.The enlargement of microglial cell bodies and an amoeboid morphology with retraction of extensions are generally induced by LPS.BG markedly suppressed the LPS-activated BV2 microglia morphological variations,meanwhile the dose-dependent was dramaticaly performed.(2)MTT test showed that LPS-stimulated BV2 cells viability was significantly decreased compared to the control group;compared to LPS treated cells,drug group(LPS+BG)effectively improves the LPS-stimulated BV2 cells viability.(3)The Griess reaction method indicated that LPS could obviously promoted the BV2 cells′NO generation contrasted to control group;while the drug group(LPS+BG)can effectively inhibited the generation of NO which activated by LPS.CONCLUSION The treatment group could significantly enhance survival rate of LPSstimulated BV2 cells,while,the level of NO was markedly decreased in BV2 induced by LPS.These findings suggest that combination of BG could attenuate BV2 microglial cells activation and injury which induced by LPS,possessed the capacity of neuroprotective.

Investigation of paeonol-geniposide on acute alcoholic liver injury based on uniform design method

Objective:To explore the optimal ratio and compatibility effect of paeonol-geniposide combination on acute alcoholic liver injury by uniform design.Methods:Lieber-DeCarli alcoholic liquid feed was used to induce acute alcoholic liver injury in mice.Uniform design was used to select the best dosage combination of paeonol and geniposide,and the related indexes of liver injury and oxidative stress were detected by kit.Serum inflammatory factors were detected by ELISA,and the expressions of p38 MAPK,JNK and NF-κB P65 related proteins in liver were detected by Western-blot.Results:The regression equation suggested that paeonol:geniposide=220:20 was the best ratio of paeonol and geniposide to resist alcoholic liver injury.Compared with the model group,the liver injury indexes and oxidation products of the paeonol+geniposide group decreased significantly,the antioxidant activity of liver tissue increased significantly,and the expression levels of p-p38 MAPK,p-JNK and NF-κB P65 protein decreased significantly.Conclusion:The optimal dosage of paeonolgeniposide was effectively optimized by uniform design and pharmacodynamic analysis.The combination of the two drugs could reduce the alcoholic liver injury by reducing the oxidative stress injury and inflammatory response in the liver tissue of mice,and its effect might be related to the targeting of p38 MAPK/JNK/NF-κB channel.

ERβ modulation and non-modulation of ERα by administration of geniposide and panax notoginseng saponins in SH-SY5Y cells

Objective:To illustrate the effect of geniposide (GP) and panax notoginseng saponins (PNS) on estrogen receptors (ER) including ERα and ERβ within the cytoplasm and nucleus of SH-SY5Y cells.Methods:Immunofluorescence was used to observe the distribution of ERα and ERβ in cytoplasm and nucleus,but Western blot was only for ERβ detection.q-PCR was applied to detect NR3C1,S100A6 and LGALS1downstream mRNA gene expression levels of ER.Results:Through analyzing fluorescence intensity under the administration of GP and PNS in SHSY5Y cells,we found that the distribution of ERα has not been affected.We also discovered that GP and/or PNS significantly stimulated the transportation of ERβ into the nucleus in a timedependent manner (all P <.001).When SH-SY5Y cells were treated with supplements of GP,PNS,GP + PNS at 15 minutes,30 minutes and 45 minutes,the distribution of ERβ in the nucleus significantly increased compared with that in control group (all P <.001).Evidently,treatment with GP,PNS,GP + PNS was able to significantly increase the levels of ERβ protein within the nucleus compared with control group at both 30 minutes and 45 minutes intervals (all P <.001).Furthermore,GP and PNS showed signs of activating to NR3C1 and LGALS1,two genes downstream of ER.It is possible that the 5100A6 gene mainly encoded the downstream gene in ERα's signaling pathway,which was not affected after the treatment of GP and/or PNS.Conclusion:The distribution and expression of ERβ has been modulated under the administration of GP + PNS within the SH-SY5Y cells,whereas ERα has not.GP and PNS in combination may play an estrogenic-like effect with selectivity on ERβ modulation.

Effect and mechanism of baicalin and geniposide on excitotoxicity of acute cerebral ischemia

OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech.nology,the mechanism of Baicalin and Geniposide(BC/GP) against excitatory amino acid toxicity in ce.rebral ischemia was studied.This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS(1) Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia.(1)18 SD rats with body weight of(280±20) g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg^(-1),45 mg·kg^(-1) and 60 mg·kg^(-1) respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model.Then,microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia.(2) First,we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi.alysis samples and observed their changes in animals.(2) The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique.(1) 16 SD rats with body weight of 240±20 g were randomly divided into sham group,model group,treatment group of BC(60 mg·kg^(-1)),treatment group of GP(60 mg·kg^(-1)) and treatment group of BC/GP(7:3)(60 mg·kg^(-1)).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model.Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃ for further detection.(2) After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of ΔΔCt.RESULTS(1) Only three compounds includ.ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmax and t1/2 z in high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2)89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol.ogy.There were 22 genes with |Fold Regulation| >1.5 in the model group,compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2 c(2.9026),Chrna7(-1.5877),and Tacr2(-1.7695).Htr3 a(-1.8172) and Grm6(-2.3527).There were 5 genes with |Fold Regulation|>1.5 in the BC group,compared with the model group,Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2 c(-1.7979).There were 14 genes with |Fold Reg.ulation| >1.5 in the GP group,compared with the model group.Three of them displayed statistically significant differences,including Hcrtr2(-1.6584),Sctr(-3.8524) and Grin2 c(-4.8408).Compared with model group,the genes of |Fold Regulation| >1.5 in BC/GP(7:3) group are 5,and only one of them showed a significant differences.CONCLUSION(1) After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus.(2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2 c gene.(3) BC/GP may exert their brain protection effect by reducing the release of excit.atory amino acids and inhibiting the expression of excitatory amino acid receptors.

芩栀鱼合剂质量标准的建立

目的:建立芩栀鱼合剂的质量标准。方法:运用薄层鉴别(TLC)的方法对芩栀鱼合剂中鱼腥草、甘草、黄芩三味中药进行鉴别;运用高效液相(HPLC)的方法对芩栀鱼合剂中特征性成分黄芩苷、栀子苷的含量进行测定,色谱柱C18(250 mm×4.6 mm,5μm,Wondasil);流动相为乙腈-0.1%甲酸水溶液(梯度洗脱);柱温为30℃;流速为1.0 mL·min^(-1);进样量为10μL;检测波长分别为283 nm(黄芩苷)、238 nm(栀子苷)。结果:鱼腥草、甘草、黄芩薄层色谱(TLC)图专属性强、特征明显且阴性对照无干扰;黄芩苷、栀子苷分别在浓度26.14~261.40μg·mL^(-1)(r=0.9999,n=6)、20.40~204.00μg·mL^(-1)(r=0.9997,n=6)范围内与对应浓度的峰面积线性关系良好;黄芩苷和栀子苷平均加样回收率分别为99.50%(RSD=1.39%,n=6)和98.97%(RSD=1.10%,n=6);方法学考察验证,两种特征性成分含量测定方法均符合要求。结论:所建方法操作简单、可行性强且结果准确,适用于芩栀鱼合剂的质量控制。

Development of a method for the quantitative determination of geniposide in rat plasma by liquid chromatography-tandem mass spectrometry with positive/negative ion-switching electrospray ionization and its application in a pharmacokinetic study in rats

Geniposide is a major bioactive constituent isolated from Gardeniajasminoides Ellis. To evaluate the pharmacokinetics of geniposide in pre-clinical studies, a rapid and specific liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was developed and validated. After simple protein precipitation, geniposide was analyzed on a DiamonsilR C18 column with a mobile phase of 10 mM ammonium acetate and methanol （20：80, v/v） at a flow rate of 0.6 mL/min. Detection was performed in ＂Truncated＂ multiple-reaction monitoring （MRM） mode with positive electrospray ionization （ESI） at m/z 411→411 for geniposide, and MRM mode with negative ESI ionization at m/z 415→295 for puerarin （internal standard, IS）. Linearity was established in the concentration range from 10.0 to 5000 ng/mL. The extraction recoveries ranged from 84.8% to 90.5% at concentrations of 10.0, 500 and 4.5x 103 ng/mL. The lower limit of quantification （LLOQ） was 10.0 ng/mL with 50 ~tL plasma. The validated method was successfully applied to the pharmacokinetic study of geniposide in rats at a dose of 200 mg/kg by oral administration.

栀子苷调节PI3K/AKT/mTOR信号通路在动脉粥样硬化形成过程中对Th17/Treg功能的影响

目的:观察栀子苷对载脂蛋白E缺乏(ApoE^(-/-))小鼠Th17/调节性T(Treg)细胞失衡的影响及其作用机制。方法:将50只纯合子ApoE^(-/-)雌性小鼠随机分为对照组、模型组和栀子苷低剂量组、栀子苷中剂量组、栀子苷高剂量组。对照组小鼠喂养普通饲料,模型组和栀子苷组小鼠喂养高脂饲料。从第8周开始,栀子苷各剂量组每日灌胃栀子苷(25、50、100 mg/kg),连续8周。试验结束时,采用油红O染色评估主动脉及其根部动脉粥样硬化(AS)病变面积比。采用定量逆转录聚合酶链式反应(RT-PCR)分析主动脉组织肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-17A和IL-10 mRNA表达;采用流式细胞仪分析脾脏中Th17和Treg细胞百分比;蛋白免疫印迹法(Western Blot)检测主动脉组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路相关蛋白表达。结果:油红O染色病变显示,栀子苷中剂量组、栀子苷高剂量组病变百分比低于模型组(P<0.05)。与对照组比较,模型组主动脉TNF-α、IL-6和IL-17A mRNA表达水平升高(P<0.05);栀子苷各剂量组主动脉TNF-α、IL-6和IL-17A mRNA表达水平降低(P<0.05)。与对照组比较,模型组主动脉抗炎细胞因子IL-10 mRNA表达水平降低(P<0.05);栀子苷各剂量组主动脉抗炎细胞因子IL-10 mRNA表达水平升高(P<0.05)。与对照组比较,模型组小鼠脾脏中Th17细胞百分比升高,Treg细胞百分比降低(P<0.05)。栀子苷处理恢复了AS小鼠Th17和Treg细胞的平衡。栀子苷抑制PI3K的表达及AKT和mTOR的磷酸化,MHY1485(mTOR活化剂)减弱了栀子苷对T细胞分化的影响。结论:栀子苷抗AS作用机制可能与抑制PI3K/AKT/mTOR信号引起的Treg细胞增多和Th17细胞减少有关。

基于5-LOX/LTB4信号通路探讨黄芩苷/栀子苷对PM_(2.5)暴露致血管内皮功能障碍的干预作用及其机制

目的基于5-LOX/LTB4信号通路探究黄芩苷/栀子苷(BC/GD)对PM_(2.5)诱导的血管内皮功能障碍的改善作用及其机制。方法利用离体肌张力描记技术测定BC/GD对不同状态下血管环张力的影响。将32只大鼠随机分为对照组、PM_(2.5)组、30 mg·kg^(-1)BC/GD组和60 mg·kg^(-1)BC/GD组,利用气管滴注法构建PM_(2.5)暴露模型,持续2个月,造模1个月后灌胃给予对应药物,持续1个月,末次给药后,HE染色观察肠系膜动脉血管内皮状态;化学发光法检测肠系膜动脉活性氧(ROS)水平;硝酸还原酶法检测一氧化氮(NO)水平;ELISA法检测血清炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、转化生长因子β1(TGF-β1)、白三烯B4(LTB4)水平;Western blot法检测5-脂氧酶(5-LOX)、内皮细胞一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)蛋白表达。结果BC/GD对基础状态的肠系膜动脉血管环张力无明显影响,但能明显舒张由5-HT预收缩的血管环;一氧化氮合酶抑制剂L-NAME、环氧合酶抑制剂INDO或L-NAME+INDO均能明显抑制BC/GD的血管舒张作用,但L-NAME的抑制作用更明显;50、100 mg·mL^(-1)PM_(2.5)能降低血管环对乙酰胆碱(ACh)的舒张血管效应,而BC/GD能明显改善PM_(2.5)诱导的舒张效应减弱。与对照组相比,PM_(2.5)组大鼠肠系膜动脉内皮完整性严重受损、内皮皱缩,大鼠血清中TNF-α、IL-1β、IL-6、LTB4水平显著升高、TGF-β1水平显著降低,肠系膜动脉组织中5-LOX、iNOS蛋白表达明显增加,eNOS蛋白表达降低,ROS水平升高,NO水平降低。与PM_(2.5)组相比,BC/GD能改善内皮损伤程度和完整性,可显著降低血清中TNF-α、IL-1β、IL-6、LTB4水平,升高TGF-β1水平;明显降低肠系膜动脉5-LOX、iNOS表达,升高eNOS蛋白表达;降低ROS水平,升高NO水平。结论BC/GD可通过抑制5-LOX/LTB4信号通路表达,从而降低ROS水平,抑制炎性损伤,上调eNOS表达,下调iNOS表达,升高血管中NO水平,改善PM_(2.5)诱导的血管内皮功能障碍。

HPLC-MS/MS法同时测定清开灵口服液中7种成分的含量

目的 建立高效液相色谱-质谱联用(HPLC-MS/MS)法同时测定清开灵口服液中7种成分含量。方法 采用Waters ACQUITY UPLC BEH C_(18)色谱柱(2.1 mm×10 mm,1.7μm);以含10 mmol·L^(-1)乙酸铵和0.1%甲酸的水溶液(A)及甲醇(B)为流动相,梯度洗脱;电喷雾离子源,正、负离子多反应监测(MRM)模式进行定量分析。结果 腺苷、绿原酸、咖啡酸、栀子苷、黄芩苷、猪去氧胆酸和胆酸质量浓度分别在0.100 4~3.213、0.784 5~8.982、0.998~3.194、0.622 5~19.92、25.05~300.6、2.513~30.15和7.775~93.30μg·m L^(-1)范围内与峰面积呈良好线性关系(r≥0.999 0);平均回收率(n=6)分别为100.9%、98.74%、101.2%、100.2%、100.8%、99.97%和98.94%,RSD分别为1.58%、0.59%、1.78%、1.25%、0.65%、1.69%和1.07%。15批样品中腺苷、绿原酸、咖啡酸、栀子苷、黄芩苷、猪去氧胆酸和胆酸的含量范围分别为0.12~0.18、0.19~0.24、0.06~0.09、0.34~0.37、4.54~4.85、0.49~0.67和1.82~2.19 mg·m L^(-1)。15批样品测定结果较为接近。结论 该方法具有良好的灵敏度、准确性和重复性,可为清开灵口服液的质量控制和后续研究提供参考及数据支持。

栀子苷调节AMPK/SIRT1/NF-κB信号通路对急性呼吸窘迫综合征大鼠肺损伤的影响

目的:探讨栀子苷(GE)通过调节AMP活化蛋白激酶(AMPK)/沉默信息调节因子1(SIRT1)/核因子κB(NF-κB)信号通路对急性呼吸窘迫综合征(ARDS)大鼠肺损伤的影响。方法:通过气管滴注脂多糖(LPS)方法建立ARDS大鼠模型。将建模后的50只大鼠随机分为ARDS组、GE低剂量(GE-L,12.5 mg/kg GE)组、GE中剂量(GE-M,25 mg/kg GE)组、GE高剂量(GE-H,50 mg/kg GE)组、GE-H+Compound C(AMPK抑制剂,50 mg/kg GE+250μg/kg Compound C)组,另取10只正常大鼠为对照组。干预后,分别取出各组大鼠支气管肺泡灌洗液(BALF)及肺组织,检测肺湿干重比(W/D);ELISA法检测BALF中炎症因子IL-6、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)水平;免疫组化法检测肺组织中血管细胞黏附因子(VCAM-1)、血管内皮细胞生长因子(VEGF)阳性表达;HE染色观察肺组织病理变化;Western blot检测肺组织中AMPK/SIRT1/NF-κB通路蛋白表达水平。结果:ARDS组W/D、IFN-γ、IL-6、TNF-α水平、p-NF-κB p65/NF-κB p65、VCAM-1表达较对照组显著升高,p-AMPK/AMPK、SIRT1、VEGF表达显著降低(P<0.05);经不同剂量GE治疗后,W/D、IFN-γ、IL-6、TNF-α水平、p-NF-κB p65/NF-κB p65、VCAM-1表达较ARDS组逐渐降低;p-AMPK/AMPK、SIRT1、VEGF表达逐渐升高(P<0.05);Compound C逆转了GE-H组对ARDS大鼠的保护作用(P<0.05)。结论:GE可以改善ARDS大鼠肺损伤状况,降低炎症因子水平,可能与激活AMPK/SIRT1/NF-κB信号通路有关。

栀子治疗肝脏疾病作用机制研究进展

肝脏疾病是严重威胁我国人民生命健康的普遍疾病,目前其治疗仍面临缺乏特效药物和药物不良反应的困境。中医药在防治肝脏疾病方面表现出独特的优势,其中栀子及其有效成分在不同类型肝脏疾病治疗中疗效显著。现代药理学研究表明,栀子具有抗炎、抗血栓、抗病毒、抗氧化、调节血脂等作用,可减轻非酒精性脂肪性肝病、肝硬化、肝损伤、肝癌等多种疾病的症状。对栀子在多种肝脏疾病的作用机制相关的新近研究进行总结,以期为栀子在防治肝脏疾病中更深入的研究及临床应用提供参考依据。

京尼平苷对代谢综合征大鼠的干预作用及对脂肪因子的影响

目的探讨京尼平苷对高脂肪/高果糖饮食诱导代谢综合征(HFFD-MS)大鼠的干预作用及对脂肪因子的影响。方法高脂肪/高果糖饮食持续16周,构建大鼠HFFD-MS模型,随机分为模型组、京尼平苷三个剂量组(分别按30、60、120 mg/kg)、阳性对照组(二甲双胍50 mg/kg);另取同期给予普通饲料和纯水饲养的大鼠为正常组,每组10只。灌胃给药8周,计算脂体比及胰岛素抵抗模型评估指数(HOMA-IR),检测空腹血糖(FBG)及血清总胆固醇(TC)、三酰甘油(TG)、游离脂肪酸(FFA)及胰岛素(INS)水平,观察肾周脂肪细胞病理学改变,放射免疫法检测血清脂肪因子瘦素(LP)、内脂素(VIS)、脂联素(APN)、抵抗素(RES)、肿瘤坏死因子α(TNF-α)、白介素6(IL-6)的表达水平。结果与模型组相比,京尼平苷各剂量组肾周脂肪细胞体积更小,排列更紧密;且可明显降低HFFD-MS大鼠的体质量、内脏脂肪质量及脂体比,差异有统计学意义(P<0.05)。另外,京尼平苷各剂量组可显著下调TC、TG、FFA、FBG、INS水平及HOMA-IR指数(P<0.01),可使血清LP、RES、TNF-α、IL-6水平明显降低(P<0.05),且京尼平苷高剂量组可显著上调APN的表达,差异有统计学意义(P<0.01),对VIS无显著性影响。结论京尼平苷具有调控脂肪因子的异常分泌、减少内脏脂肪堆积、纠正糖脂代谢紊乱等作用,可明显改善HFFD-MS病理进程。