Caspase1、8、9在高脂饮食小鼠肝组织中的表达研究  

Expression of Caspase1,8,9 in liver tissue of high-fat diet mice

作　　者：李娜 王雅光 LI Na;WANG Yaguang(Department of Histology and Embryology,Jinzhou Medical University,Jinzhou 121000,China)

机构地区：[1]锦州医科大学组织学与胚胎学教研室,锦州121000

出　　处：《中国体视学与图像分析》2024年第4期352-359,共8页Chinese Journal of Stereology and Image Analysis

基　　金：大学生创新创业训练项目(国家级)(No.2022G012X)。

摘　　要：目的利用小鼠模型观察高脂饮食(high fat diet,HFD)对肝的损伤以及对半胱氨酸蛋白酶(Caspases)表达的影响,并探讨其可能的机制。方法60只C57BL/6J雄性小鼠在适应环境1周后随机分为2组,分别给予普通饲料(正常组)和高脂饲料(高脂组),饲养4周、8周和12周,生化法检测谷草转氨酶(AST)、谷丙转氨酶(ALT)、甘油三酯(TG)和胆固醇(TC)等指标;ELISA法检测各组小鼠血清及肝脏组织中IL-1、IL-6和TNF-α炎性因子表达水平;另取部分肝脏组织做石蜡包埋,通过苏木精-伊红染色(HE)检测各组小鼠肝细胞形态;免疫组织化学染色和Western Blotting方法观察各组小鼠肝脏Caspase-1、Caspase-8和Caspase-9蛋白表达情况。结果随着喂养周数的增加,与正常组相较,高脂组小鼠肝组织形态学病理改变受损严重,且AST、ALT、TG、TC、IL-1、IL-6和TNF-α水平均升高;在HE染色实验中,正常组小鼠肝细胞形态结构正常,而高脂组小鼠肝脏组织可见泡状脂滴;在免疫组织化学实验中,肝细胞的胞质是Caspase-1、Caspase-8和Caspase-9的表达定位点,阳性表达为棕黄色或棕褐色,高脂组与正常组比较,棕色加深;在Western Blot实验中,高脂组Caspase-1、Caspase-8和Caspase-9的蛋白水平明显高于正常组(P<0.01)。结论高脂饮食引起肝损伤可能与Caspase-1、Caspase-8和Caspase-9的表达上调相关。Objective The effects of high fat diet(HFD)on liver injury and Caspases expression were observed in mouse models,and the possible mechanisms were discussed.Methods C57BL/6J male mice were randomly divided into two groups after one week of environmental adaptation,and were given high-fat diet(high-fat group)or ordinary diet(normal group),respectively.The mice were fed for 4,8 and 12 weeks,and the levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),triglyceride(TG)and cholesterol(TC)were determined using biochemical methods.The expression levels of IL-1,IL-6 and TNF-α in serum and liver tissues were detected by ELISA.Another portion of liver tissue was embedded in paraffin,and the morphology of liver cells in each group was detected by hematoxylin-eosin staining(HE).The expressions of caspase-1,caspase-8 and caspase-9 proteins in liver of each group were observed using immunohistochemical staining and Western blotting.Results As the feeding weeks increased,the liver histopathological and pathological changes in the high-fat group were more severe compared with the normal group.The levels of AST,ALT,TG,TC,IL-1,IL-6,and TNF-α were increased.In the HE staining experiment,the morphology and structure of liver cells in the normal group were normal,while bulbous lipid droplets were seen in the liver tissue of mice in the high-fat group.In the immunohistochemical experiments,the cytoplasm of hepatocytes served as the anchor point for the expression of Caspase-1,Caspase-8 and Caspase-9,with positive expression appearing brownish yellow or brown.The staining was more pronounced in the high-fat group than in the normal group.In Western blot assay,the protein levels of caspase-1,caspase-8 and caspase-9 in the high-fat group were significantly higher than those in the normal group(P<0.01).Conclusions Liver injury induced by high-fat diet may be related to the up-regulation of Caspase-1,Caspase-8 and Caspase-9 expression.

关 键 词：半胱酸蛋白酶 高脂饮食 肝损伤 炎症因子 

分 类 号：R589.2[医药卫生—内分泌]