蒙药蓝盆花抑制肝星状细胞增殖的作用及机制初探  

Role and mechanism of action of the Mongolian medicine Scabiosa atropurea in inhibiting the proliferation of hepatic stellate cells

作　　者：颜羽昕 高晓阳 张春艳 金蓉 袁宏伟 马月宏 YAN Yuxin;GAO Xiaoyang;ZHANG Chunyan;JIN Rong;YUAN Hongwei;MA Yuehong(School of Basic Medical Sciences,Inner Mongolia Medical University,Hohhot 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：《临床肝胆病杂志》2023年第6期1351-1357,共7页Journal of Clinical Hepatology

基　　金：国家自然科学基金(81960759,81560706);内蒙古自治区自然科学基金(2019MS08010,2014MS0841);内蒙古自治区草原英才培养计划(内人社办发[2016]348号);内蒙古医科大学致远人才项目(2020-11);内蒙古医科大学蒙药抗肝纤维化作用研究科技创新团队(2022-01);内蒙古医科大学重点项目(YKD2022ZD019);内蒙古人才开发基金(2022-22056)。

摘　　要：目的利用细胞实验探究蓝盆花对肝星状细胞增殖的作用及机制。方法取20只Wistar大鼠随机分为对照组和给药组,每组10只,对照组以生理盐水灌胃,给药组予蓝盆花灌胃制备含药血清。分别加入对照组血清(10%)、蓝盆花含药血清低剂量(10%)、蓝盆花含药血清中剂量(15%)及蓝盆花含药血清高剂量(20%),用以孵育HSC-T6细胞。MTT法检测不同药物浓度在不同时间段对细胞的影响;流式细胞术检测细胞凋亡情况;qRT-PCR及Western blot检测HSC细胞中纤维化标志物(α-SMA、CollagenⅠ)及PI3K/Akt信号通路相关因子mRNA、蛋白表达情况。多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与对照组比较,蓝盆花含药血清低、中、高剂量组细胞OD值均显著降低(P值均<0.05),细胞总凋亡率均显著增高(P值均<0.05)。qRT-PCR结果显示,与对照组比较,蓝盆花含药血清低、中、高剂量组α-SMA、CollagenⅠ、PI3K、Akt mRNA表达显著下调,PTEN mRNA表达显著增高(P值均<0.05);Western blot结果显示,与对照组比较,蓝盆花含药血清低、中、高剂量组α-SMA、CollagenⅠ、PI3K、Akt、p-Akt蛋白表达显著下调,PTEN蛋白表达显著增高(P值均<0.05)。结论蒙药蓝盆花可抑制HSC-T6细胞增殖且促进其凋亡,其机制可能是通过调控纤维化标志物和PI3K/Akt信号通路来发挥抗肝纤维化作用。Objective To investigate the role and mechanism of action of Scabiosa atropurea in inhibiting the proliferation of hepatic stellate cells using cell experiment.Methods A total of 20 Wistar rats were randomly divided into control group and administration group,with 10 rats in each group.The rats in the control group were given normal saline by gavage,and those in the administration group were given Scabiosa atropurea by gavage to prepare drug-containing serum.HSC-T6 cells were incubated with the serum from the control group(10%)or the low-,middle-,and high-dose serum containing Scabiosa atropurea(10%,15%,and 20%,respectively).MTT assay was used to observe the effect of different drug concentrations on cells in different periods of time;flow cytometry was used to measure cell apoptosis;qRT-PCR and Western blot were used to measure the mRNA and protein expression levels of fibrosis markers(α-SMA,collagenⅠ)and PI3K/Akt signaling pathway-related factors in hepatic stellate cells(HSCs).A one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the control group,the low-,middle-,and high-dose serum containing Scabiosa atropurea groups had a significant reduction in the OD value of cells(all P<0.05)and a significant increase in the overall apoptosis rate of cells(all P<005).The results of qRT-PCR showed that compared with the control group,the low-,middle-,and high-dose serum containing Scabiosa atropurea groups had significant reductions in the mRNA expression levels ofα-SMA,collagenⅠ,PI3K,and Akt and a significant increase in the mRNA expression level of PTEN(all P<0.05);Western blot showed that compared with the control group,the low-,middle-,and high-dose serum containing Scabiosa atropurea groups had significant reductions in the protein expression levels ofα-SMA,collagenⅠ,PI3K,Akt,and p-Akt and a significant increase in the protein expression level of PTEN(all P<0.05).Con

关 键 词：肝纤维化 肝星状细胞 蓝盆花 信号传导 

分 类 号：R29[医药卫生—民族医学]