黄连温胆汤对IGT大鼠炎症反应与肝细胞焦亡的影响  

Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats

作　　者：李寒 黄云蕾 董婉茹 马伯艳[4] LI Han;HUANG Yun-lei;DONG Wan-ru;MA Bo-yan(Shifang People's Hospital,Shifang 618400,China;Aviation University of Air Force,Changchun 130000,China;Drug Evaluation Center of Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]什邡市人民医院,四川什邡618400 [2]空军航空大学,长春130000 [3]黑龙江中医药大学药物安全性评价中心,哈尔滨150040 [4]黑龙江中医药大学,哈尔滨150040

出　　处：《中国实验方剂学杂志》2021年第8期25-32,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金应急管理项目(81641145);国家自然科学基金面上项目(81873231)。

摘　　要：目的:探讨高脂饮食诱导糖耐量低减(IGT)大鼠是否存在炎症反应与细胞焦亡及黄连温胆汤对其干预作用。方法:健康雄性SD大鼠予45%脂肪含量饲料喂养20周复制IGT模型。将符合模型标准的大鼠随机分为3组,每组10只,另取10只空白鼠作为正常组。黄连温胆汤组予7.8 g·kg^(-1)·d^(-1)复方水煎液,阳性药组予盐酸二甲双胍水溶液0.05 g·kg^(-1)·d^(-1),体积为10 mL·kg^(-1)·d^(-1),正常组和模型组予以同体积蒸馏水。连续灌胃4周后,测体质量、体长、腹围,计算Lee’s指数;测空腹血浆葡萄糖(FPG)及餐后2 h血糖(2 h PG)。酶联免疫吸附测定(ELISA)检测血清白介素-6(IL-6)含量;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)测肝组织核转录因子-κB(NF-κB),天冬氨酸特异性半胱氨酸蛋白酶-1(Caspase-1)及NOD样受体蛋白3(NLRP3)mRNA及蛋白表达;免疫荧光法测肝组织Caspase-1及NLRP3蛋白表达;苏木素-伊红(HE)染色观察肝组织形态。结果:与正常组比较,模型组大鼠体质量、腹围、体长及Lee’s指数明显增长(P<0.05,P<0.01),FPG及2 h PG水平均明显升高(P<0.05,P<0.01),血清中IL-6含量和肝组织NF-κB,Caspase-1及NLRP3基因与蛋白表达均显著增加(P<0.01),免疫荧光显示IGT大鼠肝组织Caspase-1和NLRP3表达明显上调,HE染色见IGT大鼠肝组织存在炎症细胞浸润;与模型组比较,黄连温胆汤和盐酸二甲双胍组均可有效减轻IGT大鼠体质量(P<0.01),腹型肥胖,纠正FPG和2 h PG水平(P<0.01),同时能有效降低IGT大鼠血清IL-6含量和肝组织中NF-κB,Caspase-1及NLRP3基因与蛋白表达,减轻肝组织炎症细胞浸润程度。结论:高脂饮食诱导IGT大鼠存在明显炎症反应及细胞焦亡,黄连温胆汤可明显减轻其炎症反应,缓解肝细胞焦亡。Objective: To investigate whether there is inflammatory reaction and cell pyroptosis in impaired glucose tolerance(IGT) rats induced by high-fat diet and the intervention effect of Huanglian Wendantang. Method: Healthy male SD rats were fed with 45% fat content diet for 20 weeks to replicate the IGT model. The rats in line with the model establish criteria were randomly divided into 3 groups,with 10 rats in each group,another 10 rats were selected as the blank control group. Huanglian Wendantang group was given7.8 g·kg^(-1)·d^(-1) compound decoction of Huanglian Wendantang, and the positive control group was given0.05 g·kg^(-1)·d^(-1) aqueous solution of metformin hydrochloride,with the dose volume of 10 mL·kg^(-1)·d^(-1). The blank group and the model group were given the same volume of distilled water. After continuous intragastric administration for 4 weeks,the body weight,body length and abdominal circumferences were measured,the Lee’s obesity index was calculated,and the levels of fasting plasma glucose(FPG)and 2-hour plasma glucose(2 h PG)were detected in each group. Serum interleukin-6(IL-6)content was detected by enzyme-linked immunosorbent assay(ELISA). The mRNA and protein expressions of nuclear factor kappa B(NF-κB),cysteinyl aspartate specific proteinase-1(Caspase-1)and NOD-like receptor protein 3(NLRP3)in liver tissues were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot. Immunofluorescence was used to detect the expression of Caspase-1 and NLRP3 in rat liver. Hematoxylin and eosin(HE)staining was used to observe the morphology of liver tissue. Result: Compared with blank group, the body weight, abdominal circumference, body length and Lee’s index in model group were significantly increased(P<0.05,P<0.01),the levels of FPG and 2 h PG were significantly increased(P<0.05,P<0.01),serum IL-6 content and NF-κB,Caspase-1 and NLRP3 gene and protein expressions in liver tissue were significantly increased(P<0.01),immunofluorescence techniqu

关 键 词：黄连温胆汤 肝细胞焦亡 炎症反应 糖耐量低减(IGT) 

分 类 号：R2-0[医药卫生—中医学] R22