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)细胞焦亡通路,探讨益气养阴活血方防治糖尿病肾病(DKD)肾脏损伤的可能作用机制。方法:随机将50只雄性SD大鼠分为正常组8只、造模组42只。造模组高糖高脂饮食6周后予一次性腹腔注射链脲佐菌素(STZ)诱导建立DKD大鼠模型。造模成功后随机分为模型组、缬沙坦组(8.33 mg·kg^(-1))、益气养阴活血方低、高剂量组(11、22 g·kg^(-1))。连续灌胃6周后检测各组大鼠空腹血糖(FBG)、总胆固醇(CHO)、甘油三酯(TG)、血尿素氮(BUN)、血肌酐(SCr)及24 h尿蛋白定量(24 h-UTP);苏木素-伊红(HE)染色观察肾组织病理形态学变化;酶联免疫吸附测定法(ELISA)检测血清白细胞介素^(-1)β(IL-1β)、白细胞介素^(-1)8(IL^(-1)8)水平;蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠肾组织NLRP3/Caspase^(-1)/GSDMD蛋白及其mRNA表达水平。结果:与正常组比较,模型组大鼠FBG、CHO、TG、BUN、SCr、24 h-UTP及血清IL-1β、IL^(-1)8水平均显著升高(P<0.01),肾组织病变程度严重,且肾组织NLRP3/Caspase^(-1)/GSDMD蛋白及其mRNA表达水平显著升高(P<0.01);与模型组比较,各药物组FBG、CHO、TG、BUN、SCr、24 h-UTP及血清IL-1β、IL^(-1)8水平均明显下降(P<0.05,P<0.01),肾组织病变程度得到改善,且肾组织NLRP3/Caspase^(-1)/GSDMD蛋白及其mRNA表达水平明显降低(P<0.05,P<0.01),以益气养阴活血方高剂量组效果最佳。结论:益气养阴活血方可通过调控NLRP3/Caspase^(-1)/GSDMD通路抑制细胞焦亡,缓解DKD大鼠炎症反应,减轻肾脏病理损伤。
Objective:To explore the possible mechanism of Yiqi Yangyin Huoxue prescription in the prevention and treatment of kidney injury of diabetic kidney disease(DKD)rats based on NOD-like receptor protein 3(NLRP3)/cysteine protease^(-1)(Caspase^(-1))/gasdermin D(GSDMD)pyroptosis pathway.Method:Fifty male SD rats were randomly divided into normal group(n=8)and modeling group(n=42).The modeling group was given a one-time intraperitoneal injection of streptozotocin(STZ)after high-sugar and high-fat diet for 6weeks to induce the establishment of a DKD rat model.After successful modeling,the rats were randomly divided into model group,valsartan group(8.33 mg·kg^(-1)),and Yiqi Yangyin Huoxue prescription low-dose and high-dose group(11,22 g·kg^(-1)).After continuous gavage for 6 weeks,the fasting blood glucose(FBG),total cholesterol(CHO),triglyceride(TG),blood urea nitrogen(BUN),serum creatinine(SCr)and 24-hour urine protein quantification(24 h-UTP)were detected in each group of rats.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of kidney tissue.Enzyme-linked immunosorbent assay(ELISA)was used to detect serum interleukin^(-1)β(IL-1β)and interleukin^(-1)8(IL^(-1)8)levels.The protein and mRNA expression levels of NLRP3/Caspase^(-1)/GSDMD in kidney tissue of rats in each group were determined by Western blot and real-time quantitative polymerase chain reaction(Real-time PCR).Result:Compared with the conditions in normal group,the levels of FBG,CHO,TG,BUN,SCr,24 h-UTP and serum IL-1βand IL^(-1)8 as well as the protein and mRNA expression levels of NLRP3/Caspase^(-1)/GSDMD in kidney tissue in model group were increased(P<0.01),and the kidney tissue lesions were severe.Compared with the conditions in model group,the levels of FBG,CHO,TG,BUN,SCr,24 h-UTP and serum IL-1βand IL^(-1)8 as well as the protein and mRNA expression levels of NLRP3/Caspase^(-1)/GSDMD in kidney tissue in each intervention group were decreased(P<0.05,P<0.01),and the degree of kidney tissue lesions was improved,with Yiqi Yangyin Huoxue prescription high-dose group showing the optimal effect.Conclusion:Yiqi Yangyin Huoxue prescription could inhibit pyroptosis by regulating the NLRP3/Caspase^(-1)/GSDMD pathway,and thus relieve the inflammatory response of DKD rats and alleviate the pathological damage of the kidneys.
作者
贾评评
宋纯东
段凤阳
王宁丽
丁樱
王耀献
任献青
翟文生
JIA Pingping;SONG Chundong;DUAN Fengyang;WANG Ningli;DING Ying;WANG Yaoxian;REN Xianqing;ZHAI Wensheng(Department of Pediatrics,The First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,China;Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第1期75-81,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82074493)
河南省自然科学基金项目(202300410266)
河南省中医药学科领军人才项目(豫卫中医函(2021)8号)
河南省卫生健康委员会中医药青苗人才项目(豫卫中医函[2021]16号)。