基于肝细胞焦亡的黄连温胆汤干预IGT IR的机制  

Mechanism of Huanglian Wendan Decoction in intervening IGT IR based on liver cell pyroptosis

作　　者：李寒 王宁[1] 马伯艳[1] LI Han;WANG Ning;MA Bo-yan(Department of Epidemic Febrile Diseases,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学温病教研室,黑龙江哈尔滨150040

出　　处：《中国中药杂志》2021年第13期3394-3401,共8页China Journal of Chinese Materia Medica

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：基于肝细胞焦亡途径,探讨黄连温胆汤干预糖耐量低减(impaired glucose tolerance,IGT)大鼠胰岛素抵抗(insulin resistance,IR)机制。高脂饮食长程(20周)喂养联合高温高湿环境及少动的不良生活方式建立IGT模型。实验分为空白组、模型组、盐酸二甲双胍组(阳性药组)(0.05 g·kg^(-1)·d^(-1))及黄连温胆汤组(7.8 g·kg^(-1)·d^(-1)),连续灌胃给药4周。测量大鼠体质量、体长和腹围,计算Lee’s肥胖指数;血糖仪检测空腹血浆葡萄糖(fasting plasma glucose,FPG)及餐后2 h血糖(2-hour plasma glucose,2hPG)水平;ELISA法检测血清胰岛素水平和血清中肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)水平,并计算胰岛素敏感指数(insulin sensitivity index,ISI)和胰岛素抵抗指数(insulin resistance index,IRI);Real time-PCR技术和Western blot技术检测肝组织炎性半胱氨酸天冬氨酸蛋白酶-1(cysteinyl aspartate specific proteinase-1,caspase-1)、gasdermin D(GSDMD)、白细胞介素-1β(interleukin-1β,IL-1β)及白细胞介素-18(interleukin-18,IL-18)mR NA和蛋白的表达;免疫荧光技术检测肝组织caspase-1、GSDMD蛋白表达。黄连温胆汤可有效调节IGT大鼠体质量和腹型肥胖,纠正2hPG水平,改善血清胰岛素水平和IRI,提高ISI。与空白组比较,模型组血清中TNF-α水平和肝组织caspase-1、GSDMD、IL-1β、IL-18 mR NA与蛋白表达显著升高;黄连温胆汤可有效降低IGT大鼠血清TNF-α水平和肝组织中caspase-1、GSDMD、IL-1β、IL-18 mR NA与蛋白表达。以上结果表明“肥胖-IR-糖代谢异常-2型糖尿病”的发生发展与炎症反应及caspase-1/GSDMD介导的经典细胞焦亡途径密切相关;黄连温胆汤改善IR的机制可能是通过降低炎症因子水平,缓解细胞焦亡状态,从而逆转IGT病程。To investigate the mechanism of Huanglian Wendan Decoction in intervening impaired glucose tolerance(IGT)rat insulin resistance(IR)based on the pyroptosis pathway of liver cells.The IGT rat model was established by high-fat diet(20 weeks)combined with high temperature,humidity environment and inactive lifestyle.The experiment was divided into blank group,model group,metformin hydrochloride group(positive group)(0.05 g·kg^(-1)·d^(-1))and Huanglian Wendan Decoction group(7.8 g·kg^(-1)·d^(-1)).After continuous intragastric administration for 4 weeks,the body weight,body length and abdominal circumferences of all the rats were measured,the Lee’s obesity indexes was calculated,and the levels of fasting plasma glucose(FPG)and 2-hour plasma glucose(2 hPG)in each group were measured.Serum insulin levels(FINS)and tumour necrosis factor-α(TNF-α)levels were detected by ELISA,and insulin sensitivity index(ISI)and insulin resistance index(IRI)values were calculated.The expressions of cysteinyl aspartate specific proteinase-1(caspase-1),gasdermin D(GSDMD),interleukin-1β(IL-1β)and interleukin-18(IL-18)m RNA and proteins in liver tissues were detected by Real-time PCR and Western blot.Immunofluorescence was used to detect the expressions of caspase-1 and GSDMD protein in liver tissues.Huanglian Wendan Decoction could effectively reduce the weight of model rats,improve abdominal obesity,FINS,IRI and ISI indexes and correct 2 hPG levels.Compared with blank group,TNF-α levels in serum and caspase-1,GSDMD,IL-1β and IL-18 expressions in liver tissue of model control group were increased significantly.Huanglian Wendan Decoction could effectively reduce TNF-αlevel in serum,regulate the expressions of caspase-1,GSDMD,IL-1β and IL-18 genes and proteins in liver tissues of IGT rats.The above results showed that the occurrence and development of"obesity-IR-abnormal glucose metabolism-type 2 diabetes mellitus"was closely related to inflammatory response and the classical pyroptosis pathway mediated by caspase-1/GSDMD.The mecha

关 键 词：肝细胞焦亡 黄连温胆汤 糖耐量低减(IGT) 胰岛素抵抗(IR) 

分 类 号：R285[医药卫生—中药学]