清热消癥方对db/db糖尿病肾脏病小鼠NLRP3/Caspase-1/GSDMD焦亡通路的影响  

Effects of Qingre Xiaozheng Formula on NLRP3/Caspase-1/GSDMD pyroptosis pathway in db/db mice with diabetic kidney disease

作　　者：高亚斌 王峥[1] 曹博宁 李丹婷 王耀献[3] 王珍[2] GAO Yabin;WANG Zheng;CAO Boning;LI Danting;WANG Yaoxian;WANG Zhen(The First Affiliated Hospital of Henan University of CM,Zhengzhou 450000,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]北京中医药大学东直门医院,北京100700 [3]河南中医药大学,郑州450000

出　　处：《中华中医药杂志》2025年第1期159-163,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82274460);河南省中医药科学研究专项(No.2019JDZX2116);河南省中医药拔尖人才培养项目(No.2022ZYBJ08);河南省自然科学基金项目(No.242300421523)。

摘　　要：目的:研究清热消癥方对db/db糖尿病肾脏病小鼠NOD样受体热蛋白结构域3(NLRP3)/半胱氨酸天冬氨酸蛋白酶-1(Caspase-1)/消皮素D(GSDMD)焦亡通路的影响。方法:7周龄雄性db/db小鼠20只按体质量随机分为模型组及清热消癥方组,每组10只;另取雄性db/m小鼠8只为正常组。连续灌胃8周,监测小鼠体质量、血糖,ELISA法检测尿液尿微量白蛋白及肾损伤分子1(KIM-1)、血总胆固醇(TC)、血甘油三酯(TG)、血游离脂肪酸(NEFA)、血肌酐(Scr)、血尿素氮(BUN)、血谷丙转氨酶(ALT)及血谷草转氨酶(AST);肾脏切片行HE及PAS染色;Western Blot及免疫组化检测肾组织NLRP3、GSDMD-N、Cleaved-Caspase1、IL-18及IL-1β蛋白表达。结果:与正常组比较,模型组血糖、体质量、8 h尿微量白蛋白定量、尿KIM-1、TC、TG、NEFA、Scr、ALT及AST均显著升高(P<0.01,P<0.05),肾脏病理表现为肾小球体积增大,系膜细胞增加及系膜基质增多,肾小管出现空泡变性及扩张,肾脏NLRP3、Cleaved-Caspase1及GSDMD-N蛋白表达均显著升高(P<0.01),免疫组化提示肾脏NLRP3、IL-18及IL-1β蛋白表达显著增加(P<0.01)。与模型组比较,清热消癥方组8 h尿微量白蛋白定量、KIM-1、TC、TG及NEFA均显著降低(P<0.05),肾脏病理损伤改善,NLRP3、Cleaved-Caspase1及GSDMD-N蛋白表达均显著下降(P<0.05),免疫组化提示NLRP3、IL-18及IL-1β蛋白表达均显著减少(P<0.05)。结论:清热消癥方可减轻db/db糖尿病肾脏病小鼠肾损伤,其机制可能与抑制NLRP3/Caspase-1/GSDMD焦亡通路相关。Objective:To explore the Qingre Xiaozheng Formula(QRXZF) in inhibiting pyroptosis in db/db mice with diabetic kidney disease via NLRP3/Caspase-1/GSDMD pathway.Methods:Twenty 7-week-old male db/db mice were randomly divided into model group and QRXZF group(n=10) according to the body weight.And 8 db/m mice were used as a control group.Mice in each group were treated by gavage once a day for 8 consecutive weeks.The body weight,random blood glucose,the urine microalbumin,kidney injury molecule 1(KIM-1),serum triglyceride(TG),total cholesterol(TC),free fatty acids(NEFA),blood urea nitrogen(BUN),serum creatinine(Scr),serum enzyme activities of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) were measured.HE and PAS staining were used to observe kidney pathological tissue morphology;Western Blot and immunohistochemical were used to assess protein expression of NLRP3,Cleaved-Caspase1 and GSDMD-N,IL-18 and IL-1β in kidney tissue.Results:Compared with normal group,the levels of body weight,random blood glucose,8-hour urine microalbumin,KIM-1,TC,TG,NEFA,Scr,ALT and AST in model group were increased(P<0.01,P<0.05),histological features of kidneys from mice in the model group include glomerular hypertrophy,increasing glomerular mesangial cells,mesangial matrix expansion and vacuolar degeneration and expansion of tubular epithelial cells;higher protein levels of NLRP3,Cleaved-Caspase1 and GSDMD-N in model group renal tissue(P<0.01),also the expression of NLRP3,IL-18 and IL-1β were increased in renal tissues by immunohistochemistry in model group(P<0.01).Compared with the model group,8-hour urine microalbumin,KIM-1,TC,TG and NEFA levels of the QRXZF group were decreased(P<0.05).And glomerular hypertrophy,mesangial matrix expansion and tubulointerstitial injury were partially ameliorated.Also,lower protein levels of NLRP3,CleavedCaspase1 and GSDMD-N in QRXZF group renal tissue(P<0.05),and the expression of NLRP3,IL-18 and IL-1β were decreased in renal tissues by immunohistochemistry in QRXZF group(P<0.05).C

关 键 词：清热消癥方 糖尿病肾脏病 NOD样受体热蛋白结构域3 半胱氨酸天冬氨酸蛋白酶-1 消皮素D 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]