基于ITGB4/FAK/p38信号通路蒙药蓝盆花总黄酮提取物体内外抗肝纤维化的作用研究  

Study on the Efects of Total Flavonoids Extract of Scabiosa Comosao on Hepatic Fibrosis in vitro and in vivo Based on ITGB4/FAK/p38 Signaling Pathway

作　　者：赵晓璐 孟根斯立木 张春艳 颜羽昕 高晓阳 马月宏 ZHAO Xiaolu;Menggenslimu;ZHANG Chunyan(Department of Basic Medical,The Inner Mongolia Medical University,Inner Mongolia 010000,China)

机构地区：[1]内蒙古医科大学基础医学院,呼和浩特010000

出　　处：《医学研究杂志》2023年第8期55-62,共8页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81960759,81560706);内蒙古自治区自然科学基金资助项目(2019MS08010,2014MS0841);内蒙古自治区草原英才培养计划项目;内蒙古医科大学致远人才项目(ZY0201012);内蒙古人才开发基金资助项目(2022056);内蒙古医科大学重点项目(YKD2022ZD019);内蒙古医科大学蒙药抗肝纤维化作用研究科技创新团队项目。

摘　　要：目的探讨蒙药蓝盆花总黄酮提取物(total flavonoid extract of Scabiosa comosa,TFS)对肝纤维化体内外的影响,以及通过对ITGB4/FAK/p38信号通路的调控对肝纤维化的作用。方法体内实验:选取50只Wistar大鼠随机分为空白组,模型组,TFS高(200mg/kg)、中(100mg/kg)、低(50mg/kg)剂量组,除对照组外均给予2ml/kg CCl_(4)灌胃。期间对TFS各剂量组给药,而对照组和模型组不做处理,灌胃10周。检测血清丙氨酸氨基转移酶(glutamic-pyruvic transaminase,ALT)、天冬氨酸氨基转移酶(glutamic-oxaloacetic transaminase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)含量及肝组织羟脯氨酸(hydroxyproline,HYP)含量;苏木精-伊红(hematoxylin eosin,HE)染色、Masson染色观察肝组织病理学改变;肝组织经转录组测序筛选与肝纤维化相关的通路;实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)与Western blot法分别检测肝组织中α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、collagenⅠ及ITGB4/FAK/p38信号通路相关mRNA及蛋白表达情况。体外实验:MTT法检测TFS对HSC-T6细胞的抑制率;RT-qPCR与Western blot法分别检测HSC-T6细胞中α-SMA、collagenⅠ及ITGB4/FAK/p38信号通路相关mRNA及蛋白表达;流式细胞术检测不同浓度TFS对HSC-T6细胞凋亡的影响。结果与空白组比较,TFS显著增加了模型组中ALT、AST、ALP及HYP的含量(P<0.01);与模型组比较,TFS显著降低了TFS剂量组中ALT、AST、ALP、HYP的含量(P<0.01);肝组织病理变化显示,肝纤维化增生明显改善;GO、KEGG富集分析表明,ITGB4/FAK/p38信号通路在模型组相关性显著。不同剂量的TFS对HSC-T6细胞均有抑制作用;体内外RT-qPCR与Western blot法检测结果显示,TFS各剂量组α-SMA、collagenⅠ、ITGB4/FAK/p38信号通路相关的mRNA及蛋白表达显著下降(P<0.05);凋亡结果亦显示,TFS各剂量组细胞凋亡数明显增加(P<0.01)。结论蒙药蓝盆花总黄酮提取物可显著抑�Objective To investigate the ffects of Total flavonoids extract of Scabiosa comosao(TFS)from on liver fibrosis in vitro and in vivo,and the efeets of TFS on liver fibrosis through the regulation of ITGB4/FAK/p38signaling pathway.Methods In vivo ex-periment:50W istar rats were randomly divided into 5 groups:control group,model group,high TFS(200mg/kg)group,medium TFS(100mg/kg)group and low TFS(50mg/kg)group.AlI rats were given 2ml/kg CCl_(4),by gavage except the control group.During this pe-rid,the TFS dose groups were administered,while the control group and the model group were not treated,and were gavaged for 10 weeks.The contents of glutamic-pyruvie transaminase(ALT),glutamie-oxaloacetie transaminase(AST),alkaline phosphatase(ALP)in serum and hydroxyproline(HYP)in liver were detected;hematoxylin eosin(HE)staining and Masson staining were used to observe the pathological changes of liver tissue;transcriptome sequencing was used to screen the pathways related to liver fibrosis;the mRNA and protein expression ofα-SMA,collagenⅠand ITCB4/FAK/p38signaling pathway in the liver tissue were delected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot.In vitro experiments:MITT assay was used to deteet the in-hibilory rale of TFS on HSC-T6 cell;the mRNA and prolein epression of a-SMA,cllagen 1 and ITGB4/FAK/p38 signaling pathway in HSC-T6 cells were detected by RT-qPCR and Western blot;the effects of different concentrations of TFS on apoptosis of HSC-T6 cells was detected by flow cytometry.Results Compared with the control group,TFS significantly increased the contents of ALT,AST,ALP and HYP in the model group(P<O.O1).Compared with model group,TFS significantly decreased the contents of ALT,AST,ALP and HYP in the TFS concentrations groups(P<O.O1).The pathological changes of liver tissue showed that hepatic fibrosis and hyperplasia were significantly improved.The enrichment of GO and KECG analysis showed that ITGB4/FAK/p38signaling pathway was significantly correlated with each other in the mo

关 键 词：蓝盆花 总黄酮 ITGB4/FAK/p38信号通路 肝纤维化 

分 类 号：R3[医药卫生—基础医学]