摘要
目的探讨儿茶素(catechin,CA)是否通过影响SCD1表达抗四氯化碳(carbon tetrachloride,CCl_(4))诱导的小鼠肝纤维化。方法48只C57bl/6雄性小鼠被随机分为6组:正常组、CCl_(4)组、CA 100、200、400 mg·kg^(-1)组(CA-L/M/H)、水飞蓟宾组(200 mg·kg^(-1))。腹腔注射CCl_(4)造模,共6周,前3周给小鼠灌胃CA和水飞蓟宾。处死后,观察肝大体形态、测量肝指数,通过HE染色、AST、ALT检测肝损,Masson染色检测胶原沉积及免疫组织化学检测α-SMA、collagenⅠ的表达观察肝纤维化程度,ELISA检测甘油三酯、油红O染色观察脂质累积,进而QPCR、Western blot检测CA对脂肪酸限速酶SCD1表达的影响并用分子对接技术检测CA与SCD1相互作用模式。结果较正常组,CA明显降低ALT、AST、α-SMA、collagenⅠ、甘油三酯的含量,抑制了CCl_(4)引起的肝纤维化,减少脂质累积(P<0.05),量效依赖地降低SCD1mRNA和蛋白表达(P<0.05),与SCD1通过4个氢键(Arg74,Asn148,Trp184和Val187)和6个氨基酸形成的范德华力相互作用,结合自由能为-5.4 kcal·mol^(-1)。结论CA可量效依赖地对抗CCl_(4)引起的小鼠肝纤维化,其机制可能与抑制SCD1表达从而减少脂质合成有关。
Aim To investigate whether catechin can play against CCl_(4)induced-hepatic fibrosis in mice by affecting the expression of SCD1.Methods Forty-eight MALE C57bl/6 mice were randomly divided into six groups:normal group,CCl_(4)group,catechin 100,200,400 mg·kg^(-1) groups(CA-L/M/H),silybin group(200 mg·kg^(-1)).CCl_(4)was i.p.for six weeks,and mice were given catechin and silybin for the first three weeks orally.After death gross liver morphology was observed and liver index was measured.Liver damage was detected by HE staining,AST and ALT tests.Collagen deposition was detected by Masson staining,and expression ofα-SMA and collagen I was detected by immunohistochemistry to observe the degree of liver fibrosis,triglyceride was detected by Elisa,and lipid accumulation was observed by oil red O staining.Then the effect of catechin on the expression of SCD1 was detected by QPCR and Western blot,and the interaction mode between catechin and SCD1 was assessed by molecular docking technology.Results Compared with normal group,catechin significantly decreased the contents of ALT,AST,α-SMA collagen I and triglyceride,inhibited the hepatic fibrosis induced by CCl_(4),decreased lipid accumulation(P<0.05),and decreased the mRNA and protein expression of SCD1 in a dose-effect dependent manner(P<0.05).It interacted with SCD1 through four hydrogen bonds(Arg74,Asn148,Trp184 and Val187)and van der Waals forces formed by six amino acids,resulting in a binding free energy of-5.4 kcal·mol^(-1).Conclusions Catechin can inhibit CCl_(4)-induced hepatic fibrosis in mice dose-dependently,and the mechanism may be related to the inhibition of SCD1 expression and lipid synthesis.
作者
万星
陶柏楠
邵文翠
金铮钰
黄德斌
袁林
WAN Xing;TAO Bo-nan;SHAO Wen-cui;JIN Zheng-yu;HUANG De-bin;YUAN Lin(Dept of Medicine,Hubei Minzu University,Enshi,Hubei 445000,China;Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi,Hubei 445000,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2023年第2期305-310,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金地区科学基金(No 82060745)。
