摘要
目的探讨蒙古山萝卜酸(SCA)对肝星状细胞(HSC)的基质金属蛋白酶(MMP1)和组织金属蛋白酶抑制酶(TIMP1)的活性,细胞外基质Ⅲ型前胶原(PC-Ⅲ)和Ⅳ型胶原(C-Ⅳ)的合成以及纤维化肝组织细胞外基质(ECM)与HSC体密度、HSC的凋亡及抗氧化作用。方法采用HSC初次传代后,将HSC随机分为4组:空白对照组、SCA高剂量组(15.6μg·mL^-1)、SCA中剂量组(7.81μg·mL^-1)和SCA低剂量组(3.9μg·mL^-1)。加药孵育48h后,采用Elisa法测定HSC中PC-Ⅲ、MMP1与TIMP1活性,WesternBlot法检测C-Ⅳ表达,透射电镜扫描SCA对纤维化肝组织ECM与HSC体密度、HSC的凋亡形态及抗氧化测定。结果SCA可抑制HSC中TIMP1活性与细胞外基质PC-Ⅲ和C-Ⅳ合成,对MMP1活性作用差异无统计学意义,呈抑制趋势。SCA质量浓度在3.9~15.6μg·mL^-1范围内呈量效关系;SCA可明显减少大鼠肝脏汇管区纤维化肝组织ECM与HSC体密度、诱导HSC结构转变呈凋亡形态及抑制肝组织氧化应激反应。结论SCA对体外培养的HSC分泌合成TIMP1活性及PC-Ⅲ、C-Ⅳ合成有抑制作用,可减少纤维化肝组织ECM与HSC体密度、促进HSC的凋亡,抑制丙二醛(MDA)合成、提高超氧化物歧化酶(SOD)和谷胱甘肽氧化酶(GSH)活性,其作用机制与SCA抑制肝组织氧化应激作用有关。
Objective To study the effect of Scabiosa comosa acid(SCA)on matrix metalloproteinases(MMP1)and tissue inhibitor of metalloproteinases(TIMP1),extracellular matrix precollagenⅢ(PC-Ⅲ),Ⅳtype collagen(C-Ⅳ)synthesis in hepatic stellate cells(HSC)and the effect of SCA on extracellular matrix(ECM)and HSC density,HSC apoptosis and antioxidant ability in fibrotic liver tissue.Methods After wedding for the first time,HSC were randomly divided into 4 groups:blank control group,SCA high dose group(15.6μg·mL^-1),SCA middle dose group(7.81μg·mL^-1)and SCA low dose group(3.9μg·mL^-1),dosed 48 h after incubation.Using Elisa method was used to determine PC-Ⅲ,MMP1 and TIMP1 active in HSC.The C-Ⅳcollagen expression was determined by Western Blot test.The ECM and HSC density were observed by transmission electron microscope.The HSC apoptosis and antioxidant ability were assayed.Results The SCA inhibited the TIMP1 enzyme activity in HSC and PC-Ⅲ,C-Ⅳsynthesis in extracellular matrix but showed no significant effect on the MMP1 enzyme activity.Within the designed concentration range(3.9-15.6μg·mL^-1),SCA showed the concentration-response relationship.SCA can significantly reduce the ECM and HSC density,induce the apoptosis of HSC structure change and inhibit the hepatic tissue oxidative stress in fibrosis hepatic tissue.Conclusion SCA has the inhibitory effect on TIMP1 enzyme activity and PC-Ⅲ,C-Ⅳsynthesis in vitro culture of HSC,can reduce the ECM and HSC density,promote the HSC apoptosis,inhibit the malondialdehyde(MDA)synthesis and increase the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH)in fibrosis liver tissue.The action mechanism is related to the inhibition of oxidative stress.
作者
常亮
白音夫
杨宏昕
陈广荣
董珉翔
米裕青
刘静
CHANG Liang;BAI Yinfu;YANG Hongxin;CHEN Guangrong;DONG Minxiang;MI Yuqing;LIU Jing(The Inner Mongolia Autonomous Region Hospital of International Mongolian Medicine,Hohhot 010065,China;The Inner Mongolia Autonomous Region Institute of Traditional Chinese Medicine,Hohhot 010020,China;The Inner Mongolia Autonomous Region People′s Hospital,Hohhot 010017,China;People′s Hospital Affiliated to Medical University of Inner Mongolia Autonomous Region,Hohhot 010010,China;The First Hospital of Inner Mongolia Autonomous Region Bureau of Prisons,Hohhot 010050,China)
出处
《西北药学杂志》
CAS
2019年第4期513-518,共6页
Northwest Pharmaceutical Journal
基金
国家自然科学基金项目(编号:81260659)
内蒙古自然科学基金项目(编号:2014MS0860)
关键词
蒙古山萝卜酸
肝星状细胞
细胞外基质
基质金属蛋白酶
组织金属蛋白酶抑制酶
Scabiosa comosa acid
hepatic stellate cells
extracellular matrix
matrix metalloproteinases
tissue inhibitor of metalloproteinases
