Objective:To explore the mechanism of action of Dahuang Lingxian Formula in alleviating the inflammatory response of bile duct cells in LPS-induced intrahepatic bile duct inflammation model rats based on IL-6/JAK/STAT...Objective:To explore the mechanism of action of Dahuang Lingxian Formula in alleviating the inflammatory response of bile duct cells in LPS-induced intrahepatic bile duct inflammation model rats based on IL-6/JAK/STAT3 signaling pathway.Methods:Fifty SD rats were randomly divided into five groups,blank group,model group,choling tablets(0.5 g/kg),and low and high concentration groups(2.4 g/kg and 4.8 g/kg)of Dahuang Lingxian Formula,ten rats in each group.Except for the blank group,the rats in each group were injected with 1.25 mg/kg LPS at the common bile duct at one time to construct an animal model of intrahepatic bile duct infection.After gavage on day 8,liver tissues were taken from rats at the hepatic hilum,and the histopathological changes of the hepatic hilum and biliary tree were observed by HE staining.The expression levels of serum glutamic alanine transaminase(ALT),glutamic oxalacetic transaminase(AST),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by biochemical method.The expression levels of interleukin 6(IL-6),Janus protein tyrosine kinase 2(JAK2),signal transducer and activator of transcription 3(STAT3)in rat serum were measured by enzyme-linked immunosorbent assay(ELISA).Protein immunoblotting(WB)and real-time fluorescence quantitative PCR(RT-qPCR)were used to detect the expression levels of IL-6,JAK2,STAT3 protein and mRNA in biliary tree tissues.Results:①Compared with the blank group,the structures such as interlobular bile ducts in the hepatic sinusoids and portal duct area of the model rats were destroyed,and inflammatory cells infiltrated around them.The expression of ALT,AST,MDA,IL-6,JAK2 and STAT3 in the serum increased significantly,the expression level of SOD decreased,and the expression levels of IL-6,JAK2 and STAT3 proteins and mRNA increased.②Compared with the model group,the degree of liver pathological damage in rats in the Chiling Ning tablet group and the low and high concentration groups of Dahuang Lingxian Formula were improved,which could significantly reduce the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and up-regulate SOD in serum,and down-regulate the expression of IL-6,JAK2,STAT3 protein and mRNA,with the best effect in the high concentration group of Dahuang Lingxian Formula.③Compared with the choling tablet group,the rats in the low and high concentration groups of Dahuang Lingxian Formula tended to normalize the degree of liver pathological damage,without obvious inflammatory cell infiltration,and the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and the expression levels of IL-6,JAK2,STAT3 protein and mRNA in serum were reduced,and the expression levels of SOD were increased,with the best effect of Dahuang Lingxian Formula The treatment effect was best in the high concentration group.Conclusion:The mechanism may be related to the down-regulation of IL-6/JAK/STAT3 signaling pathway activation,and the best therapeutic effect was achieved by the high concentration group of Dahuang Lingxian Formula.展开更多
Objective: To investigate the mechanism of inhibiting the secretion of “lithogenic bile” and treating gallbladder distention with the TCM prescription Dahuang Lingxian. Methods: Forty SPF-grade SD male mice were ran...Objective: To investigate the mechanism of inhibiting the secretion of “lithogenic bile” and treating gallbladder distention with the TCM prescription Dahuang Lingxian. Methods: Forty SPF-grade SD male mice were randomly divided into blank control group, model group, ursodeoxycholic acid group and TCM group, 10 mice in each group. The blank control group was kept normally, and the model group, ursodeoxycholic acid group and TCM group were established as gallbladder distention models. The blank control group and the model group were given 1.5 ml of saline by gavage daily;the ursodeoxycholic acid group was given 100 mg/kg/d of a mixture of saline and ursodeoxycholic acid capsules by gavage daily;the TCM group was given 39 g/kg/d of aqueous decoction of Dahuang Lingxian Prescription by gavage daily. After 6 weeks of continuous intervention, all mice were executed and the lithogenesis rate, ultrastructure and mRNA expression were observed. Results: The lithogenesis rate of mice in the TCM group was significantly reduced (P β1/Smads pathway was significantly improved (P < 0.05), which could achieve the same therapeutic effect as ursodeoxycholic acid capsules. Conclusion: Dahuang Lingxian Prescription can reduce the secretion of “lithogenic bile” by inhibiting the inflammatory reaction and fibrosis of biliary system. Dahuang Lingxian Prescription has certain advantages in the treatment of gallbladder distention.展开更多
Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photoli...Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photolipin fluorescent staining,and the arrangement of cytoskeleton was observed under laser confocal microscope.Western blotting was used to detect the expression of f-actin.Results:After LPS intervention,the biliary epithelial cells showed nuclear shrinkage or damage,and the skeleton was broken or lumped.The cytoskeleton was partially repaired after the intervention of pathway blocking preparation combined with RHUbarb Lingxianfang.All the other groups had different degree of cytoskeleton fracture.Compared with normal group,the expression of F-actin protein in LPS group was decreased(P<0.05);Compared with LPS group,the expression of F-actin in LPS+TCM group,LPS+PDTC+TCM group,LPS+SB203580+TCM group and LPS+PDTC+SB203580+TCM group was significantly increased(P<0.05);Compared with traditional Chinese medicine group,the expression of F-actin in LPS+PDTC+traditional Chinese medicine group,LPS+SB203580+traditional Chinese medicine group and LPS+PDTC+SB203580+traditional Chinese medicine group had no significant difference(P>0.05).Conclusion:RHUbarb Lingxianfang can restore the sequence of biliary epithelial cytoskeleton and protect its microfilament structure under inflammation,and its mechanism may be related to the regulation of NF-KB/MAPK signaling pathway.展开更多
目的:通过网络药理学方法分析大黄灵仙胶囊治疗胆石症的作用机制。方法:使用中药系统药理分析平台提取大黄灵仙胶囊的有效活性成分及对应作用靶点,结合UniProt数据库对靶点进行标准化。通过GeneCards数据库检索得到胆石症相关靶点,借助...目的:通过网络药理学方法分析大黄灵仙胶囊治疗胆石症的作用机制。方法:使用中药系统药理分析平台提取大黄灵仙胶囊的有效活性成分及对应作用靶点,结合UniProt数据库对靶点进行标准化。通过GeneCards数据库检索得到胆石症相关靶点,借助R语言筛选大黄灵仙胶囊与胆石症的交集靶点,借助Perl语言映射出交集靶点对应的有效活性成分,构建药物治疗疾病的“有效活性成分-靶点”数据库,利用Cytoscape软件对数据库绘制可视化图并筛选出核心有效成分;借助STRING数据库对交集靶点进行蛋白相互作用分析,DAVID数据库对交集靶点进行基因本体论(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。结果:得到160个大黄灵仙胶囊有效活性成分,116个对应作用靶点,其中与胆石症的共同靶点28个。大黄灵仙胶囊治疗胆石症的核心成分有槲皮素、β-谷甾醇、山柰酚、异鼠李素等,治疗的主要核心靶点有白细胞介素6、血管内皮生长因子A、表皮生长因子受体等,主要通过细胞增殖调控、一氧化氮生物合成过程调控等生物作用在细胞核浆、内吞体膜、胞外间隙等细胞位置,发挥类固醇结合、酶结合、转录因子结合等分子功能参与调控癌变通路、缺氧诱导因子1(hypoxia inducible factor-1,HIF-1)信号通路等,发挥治疗胆石症的效用。结论:网络药理学构建了大黄灵仙胶囊治疗胆石症多活性成分、多基因靶点、多治疗通路相互作用的特点,筛选出大黄灵仙胶囊作用于胆石症的基因靶点和信号通路,为进一步探索大黄灵仙胶囊治疗胆石症的作用机制提供参考依据。展开更多
基金Guangxi Natural Science Foundation(No.2020GXNSFAA238012)Research on Traditional Chinese Medicine Prevention and Treatment of Liver and Bile Related Diseases in the 2021"Qihuang Project"High Level Talent Team Cultivation Project(No.2021006)+1 种基金2020 Guangxi University of Traditional Chinese Medicine First Affiliated Hospital Hospital Hospital Level Doctoral Initiation Fund Project(No.2020BS004)2020 Guangxi University of Traditional Chinese Medicine Introduction Doctoral Research Initiation Fund Project(No.2020BS030)。
文摘Objective:To explore the mechanism of action of Dahuang Lingxian Formula in alleviating the inflammatory response of bile duct cells in LPS-induced intrahepatic bile duct inflammation model rats based on IL-6/JAK/STAT3 signaling pathway.Methods:Fifty SD rats were randomly divided into five groups,blank group,model group,choling tablets(0.5 g/kg),and low and high concentration groups(2.4 g/kg and 4.8 g/kg)of Dahuang Lingxian Formula,ten rats in each group.Except for the blank group,the rats in each group were injected with 1.25 mg/kg LPS at the common bile duct at one time to construct an animal model of intrahepatic bile duct infection.After gavage on day 8,liver tissues were taken from rats at the hepatic hilum,and the histopathological changes of the hepatic hilum and biliary tree were observed by HE staining.The expression levels of serum glutamic alanine transaminase(ALT),glutamic oxalacetic transaminase(AST),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by biochemical method.The expression levels of interleukin 6(IL-6),Janus protein tyrosine kinase 2(JAK2),signal transducer and activator of transcription 3(STAT3)in rat serum were measured by enzyme-linked immunosorbent assay(ELISA).Protein immunoblotting(WB)and real-time fluorescence quantitative PCR(RT-qPCR)were used to detect the expression levels of IL-6,JAK2,STAT3 protein and mRNA in biliary tree tissues.Results:①Compared with the blank group,the structures such as interlobular bile ducts in the hepatic sinusoids and portal duct area of the model rats were destroyed,and inflammatory cells infiltrated around them.The expression of ALT,AST,MDA,IL-6,JAK2 and STAT3 in the serum increased significantly,the expression level of SOD decreased,and the expression levels of IL-6,JAK2 and STAT3 proteins and mRNA increased.②Compared with the model group,the degree of liver pathological damage in rats in the Chiling Ning tablet group and the low and high concentration groups of Dahuang Lingxian Formula were improved,which could significantly reduce the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and up-regulate SOD in serum,and down-regulate the expression of IL-6,JAK2,STAT3 protein and mRNA,with the best effect in the high concentration group of Dahuang Lingxian Formula.③Compared with the choling tablet group,the rats in the low and high concentration groups of Dahuang Lingxian Formula tended to normalize the degree of liver pathological damage,without obvious inflammatory cell infiltration,and the expression levels of ALT,AST,MDA,IL-6,JAK2,STAT3 and the expression levels of IL-6,JAK2,STAT3 protein and mRNA in serum were reduced,and the expression levels of SOD were increased,with the best effect of Dahuang Lingxian Formula The treatment effect was best in the high concentration group.Conclusion:The mechanism may be related to the down-regulation of IL-6/JAK/STAT3 signaling pathway activation,and the best therapeutic effect was achieved by the high concentration group of Dahuang Lingxian Formula.
文摘Objective: To investigate the mechanism of inhibiting the secretion of “lithogenic bile” and treating gallbladder distention with the TCM prescription Dahuang Lingxian. Methods: Forty SPF-grade SD male mice were randomly divided into blank control group, model group, ursodeoxycholic acid group and TCM group, 10 mice in each group. The blank control group was kept normally, and the model group, ursodeoxycholic acid group and TCM group were established as gallbladder distention models. The blank control group and the model group were given 1.5 ml of saline by gavage daily;the ursodeoxycholic acid group was given 100 mg/kg/d of a mixture of saline and ursodeoxycholic acid capsules by gavage daily;the TCM group was given 39 g/kg/d of aqueous decoction of Dahuang Lingxian Prescription by gavage daily. After 6 weeks of continuous intervention, all mice were executed and the lithogenesis rate, ultrastructure and mRNA expression were observed. Results: The lithogenesis rate of mice in the TCM group was significantly reduced (P β1/Smads pathway was significantly improved (P < 0.05), which could achieve the same therapeutic effect as ursodeoxycholic acid capsules. Conclusion: Dahuang Lingxian Prescription can reduce the secretion of “lithogenic bile” by inhibiting the inflammatory reaction and fibrosis of biliary system. Dahuang Lingxian Prescription has certain advantages in the treatment of gallbladder distention.
基金Guangxi Natural Science Foundation Project(No.2020GXNSFAA238012)2021"Qihuang Project"High-level Talent Team Cultivation Project(2021006)2021 Graduate Education Innovation Project(No.YCXJ2021047)。
文摘Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photolipin fluorescent staining,and the arrangement of cytoskeleton was observed under laser confocal microscope.Western blotting was used to detect the expression of f-actin.Results:After LPS intervention,the biliary epithelial cells showed nuclear shrinkage or damage,and the skeleton was broken or lumped.The cytoskeleton was partially repaired after the intervention of pathway blocking preparation combined with RHUbarb Lingxianfang.All the other groups had different degree of cytoskeleton fracture.Compared with normal group,the expression of F-actin protein in LPS group was decreased(P<0.05);Compared with LPS group,the expression of F-actin in LPS+TCM group,LPS+PDTC+TCM group,LPS+SB203580+TCM group and LPS+PDTC+SB203580+TCM group was significantly increased(P<0.05);Compared with traditional Chinese medicine group,the expression of F-actin in LPS+PDTC+traditional Chinese medicine group,LPS+SB203580+traditional Chinese medicine group and LPS+PDTC+SB203580+traditional Chinese medicine group had no significant difference(P>0.05).Conclusion:RHUbarb Lingxianfang can restore the sequence of biliary epithelial cytoskeleton and protect its microfilament structure under inflammation,and its mechanism may be related to the regulation of NF-KB/MAPK signaling pathway.
文摘目的:通过网络药理学方法分析大黄灵仙胶囊治疗胆石症的作用机制。方法:使用中药系统药理分析平台提取大黄灵仙胶囊的有效活性成分及对应作用靶点,结合UniProt数据库对靶点进行标准化。通过GeneCards数据库检索得到胆石症相关靶点,借助R语言筛选大黄灵仙胶囊与胆石症的交集靶点,借助Perl语言映射出交集靶点对应的有效活性成分,构建药物治疗疾病的“有效活性成分-靶点”数据库,利用Cytoscape软件对数据库绘制可视化图并筛选出核心有效成分;借助STRING数据库对交集靶点进行蛋白相互作用分析,DAVID数据库对交集靶点进行基因本体论(gene ontology,GO)功能富集分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。结果:得到160个大黄灵仙胶囊有效活性成分,116个对应作用靶点,其中与胆石症的共同靶点28个。大黄灵仙胶囊治疗胆石症的核心成分有槲皮素、β-谷甾醇、山柰酚、异鼠李素等,治疗的主要核心靶点有白细胞介素6、血管内皮生长因子A、表皮生长因子受体等,主要通过细胞增殖调控、一氧化氮生物合成过程调控等生物作用在细胞核浆、内吞体膜、胞外间隙等细胞位置,发挥类固醇结合、酶结合、转录因子结合等分子功能参与调控癌变通路、缺氧诱导因子1(hypoxia inducible factor-1,HIF-1)信号通路等,发挥治疗胆石症的效用。结论:网络药理学构建了大黄灵仙胶囊治疗胆石症多活性成分、多基因靶点、多治疗通路相互作用的特点,筛选出大黄灵仙胶囊作用于胆石症的基因靶点和信号通路,为进一步探索大黄灵仙胶囊治疗胆石症的作用机制提供参考依据。
