二乙基二硫代氨基甲酸对肝星状细胞脂滴蛋白Perilipin 5表达及活化的影响  

The effect of diethyldithiocarbamate on the expression of Perilipin 5 and the activation of hepatic stellate cells

作　　者：孙祥云 康碧莲 于庆红 刘琳[1] 丛敏[1] 王萍[1] 刘天会[1] SUN Xiang-yun;KANG Bi-lian;YU Qing-hong(Liver Research Center,Beijing Friendship Hospital,Capital Medical University,Beijing Key Laboratory of Translational Medicine in Liver Cirrhosis,National Clinical Research Center of Digestive Diseases,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院肝病中心肝硬化转化医学北京市重点实验室暨国家消化系统疾病临床医学研究中心,北京100050

出　　处：《临床和实验医学杂志》2022年第8期785-788,共4页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金(编号:82070618,81770596)。

摘　　要：目的 探讨二乙基二硫代氨基甲酸(DDC)对肝星状细胞(HSCs)中脂滴蛋白Perilipin 5(Plin5)表达的调控作用及对HSCs活化的影响。方法 将雄性C57BL/6小鼠按随机数字表法分为3组:对照组给予正常饮食喂养;模型组给予胆碱缺乏、L-氨基酸替代蛋白饲料(CDAA)喂养,建立非酒精性脂肪性肝炎(NASH)纤维化模型;治疗组给予DDC灌胃治疗。GFAP和Plin5免疫荧光双染观察小鼠肝组织HSCs中Plin5表达,明确DDC对HSCs中Plin5表达的调控作用。应用DDC处理人肝星状细胞系LX-2,Real-time PCR方法检测Plin5表达,脂滴染色观察脂滴变化,明确DDC对LX-2细胞Plin5表达及脂滴形成的影响;Real-time PCR方法检测α-平滑肌肌动蛋白(α-SMA)及Ⅲ型胶原等的表达,明确DDC对LX-2细胞活化的影响。结果 在CDAA饮食诱导的NASH肝纤维化小鼠肝组织中,Plin5在HSCs中的表达较对照组显著降低;与模型组相比,DDC治疗组小鼠肝组织中Plin5在HSCs中表达显著升高。与对照组相比,DDC组处理12 h及24 h时,Plin5的mRNA相对表达量均显著增高(1.00±0.15 vs.1.93±0.70;1.00±0.33 vs.1.86±0.09),差异有统计学意义(P <0.05)。同时,LX-2脂滴染色结果显示,与对照相比,DDC组处理后LX-2细胞中脂滴数量显著增多[(0.71±0.12)%vs.(1.32±0.15)%],差异有统计学意义(P <0.05),且脂滴多位于细胞核周围,提示DDC可促进LX-2细胞恢复静止状态。与对照组相比,DDC组处理12 h及24 h时可显著抑制LX-2细胞中α-SMA、Ⅲ型胶原表达(1.00±0.07 vs.0.36±0.21;1.00±0.12 vs.0.66±0.02;1.00±0.04 vs.0.63±0.26;1.00±0.15 vs.0.76±0.07),差异均有统计学意义(P <0.05),提示DDC可抑制LX-2活化。结论 DDC可以上调HSCs中Plin5表达,促进脂滴形成并抑制HSCs活化。Objective To investigate the effect of diethyldithiocarbamate(DDC)on the expression of Perilipin 5(Plin5)in hepatic stellate cells(HSCs),and its effect on the activation of HSCs.Methods Male C57BL/6 mice were assigned into three independent groups according to the random number table method:the control group received chow diet,the CDAA group received choline-deficient,L-amino acid-defined(CDAA)diet to establish NASH related liver fibrosis model,the CDAA+DDC group were administrated DDC by gavage.Double immunofluorescence of Plin5 and glial fibrillary acidic protein(GFAP)was used to observe the expression of Plin5 in HSCs.DDC was used to treat human hepatic stellate cell line LX-2.To investigate the effect of DDC on the expression of Plin5 and the formation of lipid droplets in LX-2 cells,the expression of Plin5 was detected by Real-time PCR,and lipid droplets were observed by lipid droplet staining.Real-time PCR was used to detect the expression ofα-SMA and typeⅢcollagen to investigate the effect of DDC on the activation of LX-2 cells.Results Compared with the control group,the expression of Plin5 was significantly decreased in HSCs of CDAA diet-induced NASH related liver fibrosis mice.Compared with CDAA group,DDC treatment significantly increased the expression of Plin5 in HSCs.Compared with the control group,the relative expressions of Plin5 mRNA in the DDC group were significantly increased at 12 h and 24 h(1.00±0.15 vs.1.93±0.70;1.00±0.33 vs.1.86±0.09),and the differences were statistically significant(P<0.05).At the same time,the results of LX-2 lipid droplet staining showed that compared with the control group,the number of lipid droplets in LX-2 cells in the DDC group was significantly increased[(0.71±0.12)%vs.(1.32±0.15)%],the difference was statistically significant(P<0.05),and most of the lipid droplets were located around the nucleus,suggesting that DDC promoted the quiescence of LX-2 cells.Compared with the control group,the DDC group significantly inhibited the expressions ofα-SMA and type�

关 键 词：二乙基二硫代氨基甲酸 肝星状细胞 Perilipin 5 脂滴 非酒精脂肪性肝炎 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统]