基于NLRP3/Caspase-1/GSDMD通路探讨葛连调糖丸对2型糖尿病db/db小鼠肾脏纤维化的干预作用  

Gelian Tiaotang Pills Ameliorate Renal Fibrosis in db/db Mice via NLRP3/Caspase-1/GSDMD Pathway

作　　者：樊俐慧 王志刚 杨霞[2] 梅小龙 张坤 FAN Lihui;WANG Zhigang;YANG Xia;MEI Xiaolong;ZHANG Kun(Clinical College of Traditional Chinese Medicine(TCM),Gansu University of TCM,Lanzhou 730000,China;Tianshui Hospital of TCM,Tianshui 741000,China)

机构地区：[1]甘肃中医药大学中医临床学院,兰州730000 [2]天水市中医医院,甘肃天水741000

出　　处：《中国实验方剂学杂志》2025年第6期136-145,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：甘肃省中医药科研课题项目(GZKP-2021-46);甘肃省中医药综合防治重大疑难疾病科技攻关项目(GZKZD-2018-01);天水市科技支撑计划项目(2020-SHFZKJK-4207)。

摘　　要：目的:基于NOD样受体蛋白3(NLRP3)/胱天蛋白酶-1(Caspase-1)/消皮素D(GSDMD)信号通路探讨葛连调糖丸对2型糖尿病db/db小鼠肾脏纤维化的干预作用。方法:将成模的40只db/db小鼠随机分为模型组、达格列净组和葛连调糖丸高、中、低剂量组,每组8只。另取8只db/m小鼠作为正常组。正常组和模型组给予等体积纯水,中药高、中、低剂量组分别给予葛连调糖丸混悬液灌胃(3.12、1.56、0.78 g·kg·d^(-1)),达格列净组给予0.0013 g·kg·d^(-1)达格列净混悬液,各组小鼠每日灌胃1次,连续给药12周。每日观察小鼠一般情况;每4周检测小鼠空腹血糖及体质量,取材后称肾重,计算肾脏指数;全自动生化仪检测血清中甘油三酯(TG)、血清总胆固醇(TC)、血肌酐(SCr)、血尿素氮(BUN)的表达情况;采用苏木素-伊红(HE)、过碘酸雪夫(PAS)和马松(Masson)染色法观察肾组织病理形态、细胞外基质沉积情况及纤维化程度;酶联免疫吸附测定法(ELISA)检测肾组织白细胞介素(IL)-1β、IL-18的含量;免疫组化法(IHC)检测肾组织纤维连接蛋白(Fibronectin)、骨胶原Ⅰ(CollagenⅠ)的蛋白定位表达;蛋白免疫印迹法(Western blot)检测肾组织NLRP3、Caspase-1、剪切的(cleaved)Caspase-1、GSDMD、GSDMD-N的蛋白表达情况。结果:与正常组比较,模型组小鼠一般状态较差,体质量、肾重、肾脏指数、空腹血糖(FBG)、TG、TC、SCr、BUN表达水平显著升高(P<0.01);肾小球固缩,基质增多,肾小管上皮细胞空泡样变性,间质伴有炎性细胞浸润(P<0.01);肾组织IL-1β、IL-18水平均显著升高(P<0.01);肾组织NLRP3、Caspase-1、cleaved Caspase-1、GSDMD、GSDMD-N、Fibronectin、CollagenⅠ蛋白表达水平均显著升高(P<0.01)。经药物干预12周后,与模型组比较,各药物组体质量、肾重、肾脏指数、FBG、TG、TC、SCr、BUN表达水平及肾组织IL-1β、IL-18水平均明显降低(P<0.05,P<0.01),肾组织病变程度得到改善,且肾组织NLRPObjective:To investigate the effect of Gelian Tiaotang pills on renal fibrosis in db/db mice based on the nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3)/cysteinyl aspartate-specific proteinase(Caspase)-1/gasdermin D(GSDMD)signaling pathway.Methods:Forty db/db mice were randomly assigned into model,positive control(0.0013 g·kg·d^(-1)dapagliflozin suspension),and high-,medium-,and low-dose Gelian Tiaotang pills(3.12,1.56,and 0.78 g·kg·d^(-1)suspension of Gelian Tiaotang pills,respectively)groups,with 8 mice in each group.Eight db/m mice were selected as the normal group.The normal group and model group were given equal volumes of pure water,while the drug interventions groups were administrated with corresponding agents by gavage once a day for 12 consecutive weeks.The general conditions of mice were observed daily.The fasting blood glucose(FBG)and body mass were measured every 4 weeks.Kidneys were weighed after sampling,and the kidney index was calculated.An automatic biochemical analyzer was used to measure the serum levels of triglyceride(TG),total cholesterol(TC),serum creatinine(SCr),and blood urea nitrogen(BUN).The pathological changes,extracellular matrix deposition,and renal fibrosis degree were examined by hematoxylin-eosin,periodic acid-schiff(PAS),and Masson staining,respectively.Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin(IL)-1β and IL-18 in the renal tissue.Immunohistochemistry(IHC)was used to detect the localization and expression of fibronectin and collagen Ⅰ in the renal tissue.Western blot was employed to determine the protein levels of NLRP3,Caspase-1,cleaved Caspase-1,GSDMD,and GSDMD-N in the renal tissue.Results:Compared with the normal group,the model group generally had poor general states and increases in the body mass,kidney weight,kidney index,and levels of FBG,TG,TC,SCr,and BUN(P<0.01).In addition,glomerular pyknosis,increased matrix,vacuolar degeneration of renal tubular epithelial cells,and inters

关 键 词：葛连调糖丸 糖尿病肾脏病 细胞焦亡 炎症反应 肾脏纤维化 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R587.1