The changes in the expression of cardiac bradykinin B2 receptors (BKB2Rs) and endogenous nitrix oxide synthase (eNOs) mRNA were studied in rats with remnant kidneys. Thirty-two rats were divided into sham-operated and...The changes in the expression of cardiac bradykinin B2 receptors (BKB2Rs) and endogenous nitrix oxide synthase (eNOs) mRNA were studied in rats with remnant kidneys. Thirty-two rats were divided into sham-operated and experimental groups randomly (n=16 in each group). The remnant kidney model was established by 2-stage 5/6 nephrectomy. Blood pressure and serum Cr were measured before operation and 15, 30, 60, 120 days after 5/6 nephrectomy. Eight animals in each group were killed at the first month and 4th month after the operation. The expression of BKB2Rs and eNOs mRNAs was detected by using RT-real time PCR from isolated left ventricle, and their correlation was also analyzed. The results showed that blood pressure and serum Cr were increased significantly 15 days after 5/6 nephrectomy (both P<0.01), and the hypertension and azomia existed constantly till 120 days but had no significant fluctuation. Cardiac BKB2Rs and eNOs mRNA was declined time-dependently (both P<0.05). And there was a close positive correlation between cardiac BKB2Rs and eNOs mRNA (r=0.82, P<0.01). It was suggested that a significant chronic renal failure can be produced at least 15 days after 5/6 nephrotomy and can sustain more than 4 months. The expression of BKB2Rs and eNOs was down-regulated time-dependently in this model, and there was a significant correlation between them.展开更多
Chronic loss of sleep damages health and disturbs the quality of life.Long-lasting sleep deprivation(SD)as well as sleep abnormalities are substantial risk factors for major depressive disorder,although the underlying...Chronic loss of sleep damages health and disturbs the quality of life.Long-lasting sleep deprivation(SD)as well as sleep abnormalities are substantial risk factors for major depressive disorder,although the underlying mechanisms are not clear.Here,we showed that chronic SD in mice promotes a gradual elevation of extracellular ATP,which activates astroglial P2X7 receptors(P2X7Rs).Activated P2X7Rs,in turn,selectively down-regulated the expression of 5-HT2B receptors(5-HT2BRs)in astrocytes.Stimulation of P2X7Rs induced by SD selectively suppressed the phosphorylation of AKT and FoxO3 a in astrocytes,but not in neurons.The overexpression of FoxO3a in astrocytes inhibited the expression of 5-HT2BRs.Down-regulation of 5-HT2BsRs instigated by SD suppressed the activation of STAT3 and relieved the inhibition of Ca2+-dependent phospholipase A2.This latter cascade promoted the release of arachidonic acid and prostaglandin E2.The depression-like behaviors induced by SD were alleviated in P2X7R-KO mice.Our study reveals the mechanism underlying chronic SD-induced depression-like behaviors and suggests 5-HT2BRs as a key target for exploring therapeutic strategies aimed at the depression evoked by sleep disorders.展开更多
Colorectal cancer(CRC)is a type of malignant tumor that seriously threatens human health and life,and its treatment has always been a difficulty and hotspot in research.Herein,this study for the first time reports tha...Colorectal cancer(CRC)is a type of malignant tumor that seriously threatens human health and life,and its treatment has always been a difficulty and hotspot in research.Herein,this study for the first time reports that antipsychotic aripiprazole(Ari)against the proliferation of CRC cells both in vitro and in vivo,but with less damage in normal colon cells.Mechanistically,the results showed that5-hydroxytryptamine 2B receptor(HTR2B)and its coupling protein G protein subunit alpha q(Gaq)were highly distributed in CRC cells.Ari had a strong affinity with HTR2B and inhibited HTR2B downstream signaling.Blockade of HTR2B signaling suppressed the growth of CRC cells,but HTR2B was not found to have independent anticancer activity.Interestingly,the binding of Gaq to HTR2B was decreased after Ari treatment.Knockdown of Gaq not only restricted CRC cell growth,but also directly affected the antiCRC efficacy of Ari.Moreover,an interaction between Ari and Gaq was found in that the mutation at amino acid 190 of Gaq reduced the efficacy of Ari.Thus,these results confirm that Gaq coupled to HTR2B was a potential target of Ari in mediating CRC proliferation.Collectively,this study provides a novel effective strategy for CRC therapy and favorable evidence for promoting Ari as an anticancer agent.展开更多
Objective:To investigate the effects of Clean-DM1(C-DM1),a polyherbal formulation of Radix Scrophulariae,Radix Astragali,Rhizoma Atractylodis,and Radix Salviae Miltiorrhizae,on high-fat diet(HFD)-induced diabetes mice...Objective:To investigate the effects of Clean-DM1(C-DM1),a polyherbal formulation of Radix Scrophulariae,Radix Astragali,Rhizoma Atractylodis,and Radix Salviae Miltiorrhizae,on high-fat diet(HFD)-induced diabetes mice.Methods:The information about active components of C-DM1 extract and molecular mechanism was obtained from network pharmacology analysis.Main compounds of C-DM1 extract by high performance liquid chromatography-mass spectrometry(HPLC-MS)analysis were conducted for quality control.For in vivo study,mice were induced diabetes by HFD for 12 weeks.The mice in the normal group(Nor)were maintained with a regular diet and treated with saline by gavage.The HFD model mice were randomly divided into 3 groups,including a HFD diabetic model group,a C-DM1 extract-administered group(C-DM1,500 mg/kg),and metformin-administered groups(Met,500 mg/kg),8 mice in each group.Food intake,body weight(BW),and fasting blood glucose(FBG)levels were recorded weekly for 4 weeks.After 4 weeks of treatment,alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood glucose,low-density lipoprotein cholesterol(LDL-C)were determined using an automated clinical chemistry analyzer,and homeostatic model for assessing insulin resistance(HOMA-IR)levels and oral glucose tolerance test(OGTT)were detected.The histopathological changes of liver and pancreatic tissues were observed by hematoxylin-eosin staining.Insulin receptor substrate(IRS)/phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)and adenosine 5'-monophosphate-activated protein kinase(AMPK)expressions in liver and pancreas tissues were detected by Western blot analysis.Results:HPLC-MS identified dihydroisotanshinone,dihydroisotanshinone I,cryptotanshinone,harpagoside,and atractyloside A in C-DM1 extract.The administration of C-DM1 extract significantly decreased body weight,calorie intake,and the levels of blood glucose and insulin in the diabetic mice(P<0.05 or P<0.01).The C-DM1 extract administration improved the impaired glucose tolerance and insulin resistance in the diabetic mice and significantly decreased the levels of LDL-C,ALT and AST(P<0.01).The C-DM1 extract inhibited the histopathological changes of fatty liver and hyperplasia of pancreatic islets in the diabetic mice.The C-DM1 extract significantly increased the phosphorylation of IRS,AKT,and AMPK and the expression of PI3K in pancreas and liver tissues(P<0.05 or P<0.01),which was consistent with the analysis results of network pharmacology.Conclusion:C-DM1 extract improved diabetes symptoms in longterm HFD-induced mice by regulation of IRS/PI3K/AKT and AMPK expressions in pancreas and liver tissues,suggesting that C-DM1 formulation may help prevent the progression of T2DM.展开更多
Benign prostatic hyperplasia(BPH)is a chronic male disease characterized by the enlarged prostate.Celtis choseniana Nakai(C.choseniana)is medicinally used to alleviate pain,gastric disease,and lung abscess.In this stu...Benign prostatic hyperplasia(BPH)is a chronic male disease characterized by the enlarged prostate.Celtis choseniana Nakai(C.choseniana)is medicinally used to alleviate pain,gastric disease,and lung abscess.In this study,the effect of C.choseniana extract on BPH was investigated using testosterone-induced rats.Sprague Dawley rats were divided into five groups:control,BPH(testosterone 5 mg·kg^(−1)),Fina(finasteride 2 mg·kg^(−1)),and C.choseniana(50 and 100 mg·kg^(−1)).After four weeks of TP treatment with finasteride or C.choseniana,prostate weights and DHT levels were measured.In addition,the prostates were histopathologically examined and measured for protein kinase B(Akt)/nuclear factor-κB(NF-κB)/AR signaling,proliferation,apoptosis,and autophagy.Pro-state weight and epithelial thickness were reduced in the C.choseniana groups compared with that in the BPH group.The extract of C.choseniana acted as a 5αreductase inhibitor,reducing DHT levels in the prostate.Furthermore,the extract of C.choseniana blocked the activation of p-Akt,nuclear NF-κB activation and reduced the expression of AR and PSA compared with BPH.Moreover,the ex-pression of Bax,PARP-1,and p53 increased,while the expression of bcl-2 decreased.The present study demonstrated that C.choseni-ana extract alleviated testosterone-induced BPH by suppressing 5αreductase and Akt/NF-κB activation,reducing AR signaling and in-ducing apoptosis and autophagy in the prostate.These results suggested that C.choseniana probably contain potential herbal agents to alleviate BPH.展开更多
Background:N6-methyladenosine(m6A)RNA modification has been demonstrated to be a significant regulatory process in the progression of various tumors,including breast cancer.Fat mass and obesity-associated(FTO)enzyme,i...Background:N6-methyladenosine(m6A)RNA modification has been demonstrated to be a significant regulatory process in the progression of various tumors,including breast cancer.Fat mass and obesity-associated(FTO)enzyme,initially known as the obesity-related protein,is the first identified m6A demethylase.However,the relationship between FTO and breast cancer remains controversial.In this study,we aimed to elucidate the role and clinical significance of FTO in breast cancer and to explore the underlying mechanism.Methods:We first investigated the expression of FTO in breast cancer cell lines and tissues by quantitative reverse transcription-PCR(qRT-PCR),Western blotting,and immunohistochemistry.Wound healing assay and Transwell assay were performed to determine the migration and invasion abilities of SKBR3 and MDAMB453 cells with either knockdown or overexpression of FTO.RNA sequencing(RNA-seq)was conducted to decipher the downstream targets of FTO.qRT-PCR,luciferase reporter assay,and Western blotting were employed to confirm the existence of the FTO/miR-181b-3p/ARL5B axis.The biological function of ADP ribosylation factor like GTPase 5B(ARL5B)in breast cancer cells was evaluated by wound healing assay and Transwell invasion assay.Results:High FTO expression was observed in human epidermal growth factor receptor 2(HER2)-positive breast cancer,predicting advanced progression(tumor size[P<0.001],nuclear grade[P=0.001],peritumoral lymphovascular invasion[P<0.001),lymph node metastasis[P=0.002],and TNM stage[P=0.001])and poor prognosis.Moreover,FTO promoted cell invasion and migration in vitro.Mechanistically,RNA-seq and further confirmation studies suggested that FTO up-regulated ARL5B by inhibiting miR-181b-3p.We further verified that ARL5B also displayed carcinogenic activity in breast cancer cells.Conclusion:Our work demonstrated the carcinogenic activity of FTO in promoting the invasion and migration of breast cancer cells via the FTO/miR-181b-3p/ARL5B signaling pathway.展开更多
文摘The changes in the expression of cardiac bradykinin B2 receptors (BKB2Rs) and endogenous nitrix oxide synthase (eNOs) mRNA were studied in rats with remnant kidneys. Thirty-two rats were divided into sham-operated and experimental groups randomly (n=16 in each group). The remnant kidney model was established by 2-stage 5/6 nephrectomy. Blood pressure and serum Cr were measured before operation and 15, 30, 60, 120 days after 5/6 nephrectomy. Eight animals in each group were killed at the first month and 4th month after the operation. The expression of BKB2Rs and eNOs mRNAs was detected by using RT-real time PCR from isolated left ventricle, and their correlation was also analyzed. The results showed that blood pressure and serum Cr were increased significantly 15 days after 5/6 nephrectomy (both P<0.01), and the hypertension and azomia existed constantly till 120 days but had no significant fluctuation. Cardiac BKB2Rs and eNOs mRNA was declined time-dependently (both P<0.05). And there was a close positive correlation between cardiac BKB2Rs and eNOs mRNA (r=0.82, P<0.01). It was suggested that a significant chronic renal failure can be produced at least 15 days after 5/6 nephrotomy and can sustain more than 4 months. The expression of BKB2Rs and eNOs was down-regulated time-dependently in this model, and there was a significant correlation between them.
基金the National Natural Science Foundation of China(81871852,81200935,81671862,and 81871529)Liaoning Revitalization Talents Program(XLYC1807137)+1 种基金the Scientific Research Foundation for Overseas Scholars of the Education Ministry of China(20151098)the Natural Science Foundation of Liaoning Province,China(20170541030)。
文摘Chronic loss of sleep damages health and disturbs the quality of life.Long-lasting sleep deprivation(SD)as well as sleep abnormalities are substantial risk factors for major depressive disorder,although the underlying mechanisms are not clear.Here,we showed that chronic SD in mice promotes a gradual elevation of extracellular ATP,which activates astroglial P2X7 receptors(P2X7Rs).Activated P2X7Rs,in turn,selectively down-regulated the expression of 5-HT2B receptors(5-HT2BRs)in astrocytes.Stimulation of P2X7Rs induced by SD selectively suppressed the phosphorylation of AKT and FoxO3 a in astrocytes,but not in neurons.The overexpression of FoxO3a in astrocytes inhibited the expression of 5-HT2BRs.Down-regulation of 5-HT2BsRs instigated by SD suppressed the activation of STAT3 and relieved the inhibition of Ca2+-dependent phospholipase A2.This latter cascade promoted the release of arachidonic acid and prostaglandin E2.The depression-like behaviors induced by SD were alleviated in P2X7R-KO mice.Our study reveals the mechanism underlying chronic SD-induced depression-like behaviors and suggests 5-HT2BRs as a key target for exploring therapeutic strategies aimed at the depression evoked by sleep disorders.
基金supported by Chongqing basic research and frontier exploration project(cstc2022ycjh-bgzxm0119,China)。
文摘Colorectal cancer(CRC)is a type of malignant tumor that seriously threatens human health and life,and its treatment has always been a difficulty and hotspot in research.Herein,this study for the first time reports that antipsychotic aripiprazole(Ari)against the proliferation of CRC cells both in vitro and in vivo,but with less damage in normal colon cells.Mechanistically,the results showed that5-hydroxytryptamine 2B receptor(HTR2B)and its coupling protein G protein subunit alpha q(Gaq)were highly distributed in CRC cells.Ari had a strong affinity with HTR2B and inhibited HTR2B downstream signaling.Blockade of HTR2B signaling suppressed the growth of CRC cells,but HTR2B was not found to have independent anticancer activity.Interestingly,the binding of Gaq to HTR2B was decreased after Ari treatment.Knockdown of Gaq not only restricted CRC cell growth,but also directly affected the antiCRC efficacy of Ari.Moreover,an interaction between Ari and Gaq was found in that the mutation at amino acid 190 of Gaq reduced the efficacy of Ari.Thus,these results confirm that Gaq coupled to HTR2B was a potential target of Ari in mediating CRC proliferation.Collectively,this study provides a novel effective strategy for CRC therapy and favorable evidence for promoting Ari as an anticancer agent.
基金Supported by Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI),the Ministry of Health&Welfare,Republic of Korea (No. HF20C0121)Shanxi Key Laboratory of Tradition Herbal Medicines Processing (No. 20210901)the Innovation Team of Shanxi University of Chinese Medicine (No. 2022TD1014)
文摘Objective:To investigate the effects of Clean-DM1(C-DM1),a polyherbal formulation of Radix Scrophulariae,Radix Astragali,Rhizoma Atractylodis,and Radix Salviae Miltiorrhizae,on high-fat diet(HFD)-induced diabetes mice.Methods:The information about active components of C-DM1 extract and molecular mechanism was obtained from network pharmacology analysis.Main compounds of C-DM1 extract by high performance liquid chromatography-mass spectrometry(HPLC-MS)analysis were conducted for quality control.For in vivo study,mice were induced diabetes by HFD for 12 weeks.The mice in the normal group(Nor)were maintained with a regular diet and treated with saline by gavage.The HFD model mice were randomly divided into 3 groups,including a HFD diabetic model group,a C-DM1 extract-administered group(C-DM1,500 mg/kg),and metformin-administered groups(Met,500 mg/kg),8 mice in each group.Food intake,body weight(BW),and fasting blood glucose(FBG)levels were recorded weekly for 4 weeks.After 4 weeks of treatment,alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood glucose,low-density lipoprotein cholesterol(LDL-C)were determined using an automated clinical chemistry analyzer,and homeostatic model for assessing insulin resistance(HOMA-IR)levels and oral glucose tolerance test(OGTT)were detected.The histopathological changes of liver and pancreatic tissues were observed by hematoxylin-eosin staining.Insulin receptor substrate(IRS)/phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)and adenosine 5'-monophosphate-activated protein kinase(AMPK)expressions in liver and pancreas tissues were detected by Western blot analysis.Results:HPLC-MS identified dihydroisotanshinone,dihydroisotanshinone I,cryptotanshinone,harpagoside,and atractyloside A in C-DM1 extract.The administration of C-DM1 extract significantly decreased body weight,calorie intake,and the levels of blood glucose and insulin in the diabetic mice(P<0.05 or P<0.01).The C-DM1 extract administration improved the impaired glucose tolerance and insulin resistance in the diabetic mice and significantly decreased the levels of LDL-C,ALT and AST(P<0.01).The C-DM1 extract inhibited the histopathological changes of fatty liver and hyperplasia of pancreatic islets in the diabetic mice.The C-DM1 extract significantly increased the phosphorylation of IRS,AKT,and AMPK and the expression of PI3K in pancreas and liver tissues(P<0.05 or P<0.01),which was consistent with the analysis results of network pharmacology.Conclusion:C-DM1 extract improved diabetes symptoms in longterm HFD-induced mice by regulation of IRS/PI3K/AKT and AMPK expressions in pancreas and liver tissues,suggesting that C-DM1 formulation may help prevent the progression of T2DM.
文摘Benign prostatic hyperplasia(BPH)is a chronic male disease characterized by the enlarged prostate.Celtis choseniana Nakai(C.choseniana)is medicinally used to alleviate pain,gastric disease,and lung abscess.In this study,the effect of C.choseniana extract on BPH was investigated using testosterone-induced rats.Sprague Dawley rats were divided into five groups:control,BPH(testosterone 5 mg·kg^(−1)),Fina(finasteride 2 mg·kg^(−1)),and C.choseniana(50 and 100 mg·kg^(−1)).After four weeks of TP treatment with finasteride or C.choseniana,prostate weights and DHT levels were measured.In addition,the prostates were histopathologically examined and measured for protein kinase B(Akt)/nuclear factor-κB(NF-κB)/AR signaling,proliferation,apoptosis,and autophagy.Pro-state weight and epithelial thickness were reduced in the C.choseniana groups compared with that in the BPH group.The extract of C.choseniana acted as a 5αreductase inhibitor,reducing DHT levels in the prostate.Furthermore,the extract of C.choseniana blocked the activation of p-Akt,nuclear NF-κB activation and reduced the expression of AR and PSA compared with BPH.Moreover,the ex-pression of Bax,PARP-1,and p53 increased,while the expression of bcl-2 decreased.The present study demonstrated that C.choseni-ana extract alleviated testosterone-induced BPH by suppressing 5αreductase and Akt/NF-κB activation,reducing AR signaling and in-ducing apoptosis and autophagy in the prostate.These results suggested that C.choseniana probably contain potential herbal agents to alleviate BPH.
文摘Background:N6-methyladenosine(m6A)RNA modification has been demonstrated to be a significant regulatory process in the progression of various tumors,including breast cancer.Fat mass and obesity-associated(FTO)enzyme,initially known as the obesity-related protein,is the first identified m6A demethylase.However,the relationship between FTO and breast cancer remains controversial.In this study,we aimed to elucidate the role and clinical significance of FTO in breast cancer and to explore the underlying mechanism.Methods:We first investigated the expression of FTO in breast cancer cell lines and tissues by quantitative reverse transcription-PCR(qRT-PCR),Western blotting,and immunohistochemistry.Wound healing assay and Transwell assay were performed to determine the migration and invasion abilities of SKBR3 and MDAMB453 cells with either knockdown or overexpression of FTO.RNA sequencing(RNA-seq)was conducted to decipher the downstream targets of FTO.qRT-PCR,luciferase reporter assay,and Western blotting were employed to confirm the existence of the FTO/miR-181b-3p/ARL5B axis.The biological function of ADP ribosylation factor like GTPase 5B(ARL5B)in breast cancer cells was evaluated by wound healing assay and Transwell invasion assay.Results:High FTO expression was observed in human epidermal growth factor receptor 2(HER2)-positive breast cancer,predicting advanced progression(tumor size[P<0.001],nuclear grade[P=0.001],peritumoral lymphovascular invasion[P<0.001),lymph node metastasis[P=0.002],and TNM stage[P=0.001])and poor prognosis.Moreover,FTO promoted cell invasion and migration in vitro.Mechanistically,RNA-seq and further confirmation studies suggested that FTO up-regulated ARL5B by inhibiting miR-181b-3p.We further verified that ARL5B also displayed carcinogenic activity in breast cancer cells.Conclusion:Our work demonstrated the carcinogenic activity of FTO in promoting the invasion and migration of breast cancer cells via the FTO/miR-181b-3p/ARL5B signaling pathway.
