小柴胡汤对体外培养LX-2细胞增殖的影响及机制初探  

Observation of the effects and mechanism of a Xiaochaihu decoction on proliferation of LX-2 cells in vitro

作　　者：卢金 朱恬宇 张英[1,2,3] 许永良[1,2,3] 吴奇俊 刘新建[5] 黄玉政 LU Jin;ZHU Tian-yu;ZHANG Ying;XU Yong-liang;WU Qi-jun;LIU Xin-jian;HUANG Yu-zheng(National Health Commission Key Laboratory of Parasitic Disease Control and Prevention,Jiangsu Institute of Parasitic Diseases,Wuxi,Jiangsu 214064,China;Tropical Diseases Research Center,Nanjing Medical University;Public Health Research Center of Jiangnan University;Wuxi Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine;Department of Pathogenic Biology,Nanjing Medical University)

机构地区：[1]江苏省血吸虫病防治研究所国家卫生健康委员会寄生虫病预防与控制技术重点实验室,江苏无锡214064 [2]南京医科大学热带病研究中心 [3]江南大学公共卫生研究中心 [4]南京中医药大学附属无锡中医医院 [5]南京医科大学病原生物学系

出　　处：《中国病原生物学杂志》2021年第1期49-52,57,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金面上项目(No.81673673);江苏省卫生健康委医学科研重点项目(No.ZDB2020037);江南大学公共卫生研究中心课题(No.JUPH201811);江苏省"科教强卫工程"专项经费资助(No.ZDXKA2015016);江苏省省属公益类科研院所自主科研经费项目资助(No.BM2018020)。

摘　　要：目的观察小柴胡汤对人肝星状细胞(LX-2细胞)增殖的影响。方法体外培养LX-2细胞,用不同浓度的小柴胡汤(2%,1%,0.5%,0.2%,0.1%)处理24 h,对照组不做处理。显微镜下观察药物作用后细胞形态改变,采用四甲基偶氮唑盐(MTT)法和5-乙炔基-2’脱氧尿嘧啶核苷(Edu)法检测细胞增殖,实时荧光定量PCR(RT-PCR)检测I型胶原(COL-Ⅰ)mRNA的表达情况。结果对照组细胞,贴壁生长良好,形态未见明显异常。经不同浓度小柴胡汤(2%、1%、0.5%、0.2%、0.1%)处理24 h后,LX-2细胞均出现皱缩,变圆,界限不清,细胞核固缩等不同程度形态改变,MTT检测显示其增殖抑制率分别为100%、54.9%、41.2%、14.8%、12.4%。经1%、0.5%、0.1%小柴胡汤作用24 h后,Edu法检测LX-2细胞增殖率分别为0.1%、5.3%、10.1%;RT-PCR检测其COL-ⅠmRNA的表达量,与对照组1.04±0.35相比,均显著下降,分别为0.45±0.15(P<0.05)、0.27±0.16(P<0.01)和0.38±0.24(P<0.01)。MTT和Edu结果表明小柴胡汤能够抑制体外培养LX-2细胞的增殖,并呈剂量依赖效应。结论小柴胡汤通过抑制LX-2细胞增殖,并抑制COL-ⅠmRNA的表达,从而发挥抗纤维化活性。Objectives To observe the effects of a Xiaochaihu decoction(XCH)on the proliferation of human hepatic stellate cells(LX-2).Methods LX-2 cells were cultured and treated with different concentrations of XCH(2%,1%,0.5%,0.2%,and 0.1%)in vitro for 24 h.The control group was not treated.Changes in cell morphology were observed under a microscope.Methyl thiazolyl tetrazolium(MTT)and 5-ethynyl-2’deoxyuridine(Edu)were used to evaluate LX-2 cell proliferation.RT-PCR was used to evaluate expression of type I collagen(COL-I)mRNA in cultured cells.Results In the control group,the cells grew well and had no obvious abnormalities.After treatment with different concentrations of XCH(2%,1%,0.5%,0.2%,and 0.1%),LX-2 cells displayed various morphological changes,such as shrinkage,roundness,indistinct boundaries,and nuclear pyknosis.An MTT assay indicated that the rate of inhibited proliferation was 100%,54.9%,41.2%,14.8%,and 12.4%,respectively.After treatment with 1%,0.5%,or 0.1%XCH for 24 h,the rate of LX-2 cell proliferation was respectively 0.1%,5.3%,and 10.1%according to Edu.The level of expression of COL-I mRNA in LX-2 cells was 0.45±0.15(P<0.05),0.27±0.16(P<0.01),and 0.38±0.24(P<0.01),reflecting a significant decrease compared to 1.04±0.35 in the control group.MTT and Edu results indicated that XCH inhibited the proliferation of LX-2 cells in a dose-dependent manner.Conclusion XCH plays an anti-fibrotic role by inhibiting the proliferation of LX-2 cells and the expression of COL-I mRNA in LX-2 cells.

关 键 词：小柴胡汤 血吸虫病 肝纤维化 LX-2细胞 细胞增殖 

分 类 号：R392[医药卫生—免疫学]