黄连多糖通过抑制肾小管上皮细胞EMT及焦亡减轻糖尿病肾病小鼠肾脏损伤的作用研究  

Effect of Coptis chinensis polysaccharide on alleviating renal injury by inhibiting EMT and pyroptosis of renal tubular epithelial cell in diabetic nephropathy mice

作　　者：葛斌 于澄 郭寰宇 兰辛键 姜爽 GE Bin;YU Cheng;GUO Huanyu;LAN Xinjian;JIANG Shuang(School of Clinical Medicine,Changchun University of Chinese Medicine,Jilin Changchun 130117,China;School of Basic Medicine,Changchun University of Chinese Medicine,Jilin Changchun 130117,China)

机构地区：[1]长春中医药大学临床医学院,吉林长春130117 [2]长春中医药大学基础医学院,吉林长春130117

出　　处：《中国医院药学杂志》2025年第4期396-401,共6页Chinese Journal of Hospital Pharmacy

基　　金：吉林省科技发展计划项目(编号:YDZJ202401104ZYTS);吉林省教育厅科学研究项目(编号:JJKH20241083KJ);吉林省卫生健康科技能力提升项目(编号:2023JC040);吉林省中医药科技项目(编号:202471)。

摘　　要：目的:探讨黄连多糖通过抑制肾小管上皮细胞上皮间充质转化(epithelial mesenchymal transformation,EMT)及焦亡减轻糖尿病肾病(diabetic nephropathy,DN)小鼠肾脏损伤的作用。方法:将db/db小鼠分为模型组,黄连多糖低、高剂量组(200、400 mg·kg^(-1))及恩格列净组(10 mg·kg^(-1)),db/m小鼠为正常组,每组均为10只。持续干预12周,检测血尿素氮(BUN)、血肌酐(SCr)、24 h尿蛋白定量(24 h-UTP)、胱抑素C(CysC)、N-酰基-β-D-葡萄糖(NAG)等生化指标,苏木素伊红及Masson染色观察肾组织病理形态,用以评价黄连多糖对肾脏损伤的减轻作用;实时qPCR法检测EMT标志物如肾组织纤维粘连蛋白(FN)、α-平滑肌肌动蛋白(α-SMA)及E-钙黏素(E-Cad)mRNA表达;酶联免疫吸附法检测血清及肾组织白细胞介素-1β(IL-1β)、IL-6及IL-18等焦亡相关促炎因子含量;蛋白免疫印迹法检测NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)及半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)等焦亡相关蛋白表达。结果:与模型组比较,黄连多糖低、高剂量组小鼠BUN、SCr、24 h-UTP、CysC及NAG下降(P<0.01),肾组织病理形态减轻,肾组织FN及α-SMA mRNA表达降低(P<0.05,P<0.01),E-Cad mRNA表达增加(P<0.01);除黄连多糖低剂量组小鼠肾组织IL-6含量无明显变化外(P>0.05),其余各组血清及肾组织IL-1β、IL-6及IL-18含量均降低(P<0.05,P<0.01);黄连多糖低、高剂量组小鼠肾组织NLRP3、ASC及Caspase-1蛋白表达均降低(P<0.05,P<0.01)。结论:黄连多糖能够抑制DN小鼠肾小管上皮细胞EMT及焦亡,该作用可能是其减轻肾脏损伤的潜在机制。OBJECTIVE To explore the effect of Coptis chinensis polysaccharide(CCPs)on alleviating renal injury by inhibiting epithelial mesenchymal transformation(EMT)and pyroptosis of renal tubular epithelial cell in diabetic nephropathy(DN)mice.METHODS Db/db mice were divided into the model group,low-dose and high-dose CCPs groups(200 mg·kg^(-1)and 400 mg kg^(-1))and englipzin group(10 mg kg^(-1)),and db/m mice were included in the normal group,with 10 mice in each group.The intervention lasted for 12 weeks.Biochemical indicators,such as blood urea nitrogen(BUN),serum creatinine(SCr),24 h urinary total protein quantification(24 h-UTP),cystatin C(CysC)and N-acetyl-β-D-glucose(NAG)were detected.Pathological morphology of renal tissue was observed by hematoxylin-eosin(H&E)and Masson's trichrome staining,so as to evaluate the effect of CCPs on alleviating renal injury.The mRNA expressions of EMT markers,such as fibronectin(FN),α-smooth muscle actin(α-SMA)and E-cadherin(E-Cad)in renal tissue were detected by real-time qPCR.Proinflammatory factors related to pyroptosis,such as interleukin-1β(IL-1β),IL-6 and IL-18 in serum and renal tissue were detected by enzyme-linked immunosorbent assay(ELISA).Proteins expressions related to pyroptosis,such as NOD-like receptor protein 3(NLRP3),apoptosisassociated spot-like protein(ASC)and Caspase-1 in renal tissue were detected by Western blot.RESULTS Compared with model groups,mice in the low-dose and high-dose CCPs groups had significantly lower BUN,SCr,24 h-UTP,CysC and NAG(P<0.01),alleviated pathological morphology of renal tissue,downregulated mRNA levels of FN andα-SMA(P<0.05,P<0.01),and upregulated mRNA level of E-Cad(P<0.01).In addition to the IL-6 level in mosque renal tissue of low-dose CCPs group(P>0.05),contents of IL-1β,IL-6 and IL-18 in serum and renal tissue were significantly reduced in other groups(P<0.05,P<0.01).Protein levels of NLRP3,ASC and Caspase-1 in mice renal tissue of low-dose and high-dose CCPs groups were significantly downregulated(P<0.05,P<0.01).CONCLUS

关 键 词：黄连多糖 上皮间充质转化 焦亡 糖尿病肾病 

分 类 号：R285[医药卫生—中药学] R966[医药卫生—中医学]