摘要
目的探究参芪地黄汤(Shenqi Dihuang decoction,SQDH)通过调控AMPK/Sirt1/Nrf2介导的铁死亡对过敏性紫癜性肾炎(henoch-schonlein purpura nephritis,HSPN)大鼠的影响及可能的作用机制。方法30只SD大鼠随机分为对照组(Control)、HSPN组、SQDH低剂量组(SQDH-L,5.4 g·kg^(-1))、SQDH高剂量组(SQDH-H,21.6 g·kg^(-1))、SQDH高剂量+AMPK抑制剂组(SQDH-H+CC,20 mg·kg^(-1)),每组各6只。除Control组外,其余各组大鼠均采取腹腔注射卵白蛋白和完全弗氏佐剂构建HSPN大鼠模型。待模型构建成功后,各组大鼠按照对应给药剂量或生理盐水灌胃,每日1次,持续5周。HE染色观察肾脏组织病理学变化;免疫组化检测肾脏组织IgA、IgG沉积情况;检测肾功能生化指标及尿蛋白水平;试剂盒测定肾组织氧化应激指标、Fe^(2+)水平;Western blot检测肾脏组织铁死亡及AMPK/Sirt1/Nrf2信号通路蛋白表达。结果与Control组相比,HSPN组大鼠肾脏组织出现明显的病理性损伤,且评分明显升高,肾组织IgA、IgG沉积增加,血清总蛋白(total protein,TP)、白蛋白(albumin,ALB)降低,血清肌酐(creatinine,Cre)、尿素氮(urea nitrogen,BUN)、尿蛋白水平均升高(P<0.05),肾组织MDA、ROS、Fe^(2+)水平以及ACSL4蛋白表达均明显升高,SOD活性、SLC7A11、GPX4、p-AMPK/AMPK、Sirt1、Nrf2蛋白表达均明显降低(P<0.05)。与HSPN组相比,SQDH各剂量组大鼠肾脏组织出病理性损伤减轻,肾组织IgA、IgG沉积减少,TP、ALB降低,Cre、BUN、尿蛋白水平均降低(P<0.05),肾组织MDA、ROS、Fe^(2+)水平以及ACSL4蛋白表达均明显降低,SOD活性、SLC7A11、GPX4、p-AMPK/AMPK、Sirt1、Nrf2蛋白表达均明显升高(P<0.05)。与SQDH-H组相比,AMPK抑制剂CC能够抑制AMPK/Sirt1/Nrf2信号通路诱导铁死亡,逆转SQDH对HSPN大鼠肾损伤的保护作用。结论SQDH能够改善HSPN大鼠肾脏损伤,其机制可能与激活AMPK/Sirt1/Nrf2信号通路抑制细胞铁死亡有关。
Aim To investigate the effect of Shenqi Dihuang decoction(SQDH)on henoch-schonlein purpura nephritis(HSPN)rats by regulating AMPK/Sirt1/Nrf2 mediated ferroptosis and the possible mechanism.Methods Thirty SD rats were randomly divided into a control group(Control),HSPN group,SQDH low-dose group(SQDH-L,5.4 g·kg^(-1)),SQDH high-dose group(SQDH-H,21.6 g·kg^(-1)),and SQDH high-dose+AMPK inhibitor group(SQDH-H+CC,20 mg·kg^(-1)),with six rats in each group.Except for the control group,all other groups of rats were injected intraperitoneally with ovalbumin and complete Freund’s adjuvant to establish HSPN rat models.After the successful construction of the model,each group of rats was orally administered with the corresponding dosage or physiological saline once a day for five weeks.HE staining was used to observe the histopathological changes in renal tissues;immunohistochemistry(IHC)was employed to detect the deposition of IgA and IgG in renal tissues;serum biochemical parameters of renal function and urinary protein levels were measured;commercial assay kits were utilized to determine oxidative stress markers and Fe^(2+)levels in renal tissues;Western blot analysis was performed to assess ferroptosis-related proteins and the protein expression of the AMPK/Sirt1/Nrf2 signaling pathway in renal tissues.Results Compared with the control group,the HSPN group showed significant pathological damage in kidney tissue,with significantly increased scores,increased deposition of IgA and IgG in kidney tissue,decreased serum total protein(TP)and albumin(ALB),and average increases in serum creatinine(Cre),urea nitrogen(BUN),and urinary protein levels(P<0.05).The levels of MDA,ROS,Fe^(2+),and ACSL4 protein expression in the kidney tissue significantly increased,while SOD activity,SLC7A11,GPX4,p-AMPK/AMPK,Sirt1,and Nrf2 protein expression significantly decreased(P<0.05).Compared with the HSPN group,the pathological damage to renal tissue was reduced,IgA and IgG deposition in renal tissue decreased,TP and ALB decreased,Cre,BUN,and urinary protein levels were all reduced(P<0.05),and the levels of MDA,ROS,Fe^(2+),and ACSL4 protein expression in renal tissue were significantly reduced in each dose group of SQDH rats.SOD activity,SLC7A11,GPX4,p-AMPK/AMPK,Sirt1,and Nrf2 protein expression significantly increased(P<0.05).Compared with the SQDH-H group,the AMPK inhibitor CC could inhibit the AMPK/Sirt1/Nrf2 signaling pathway induced ferroptosis and reverse the protective effect of SQDH on renal injury in HSPN rats.Conclusions SQDH can improve renal injury in HSPN rats,and its mechanism may be related to the activation of AMPK/Sirt1/Nrf2 signaling pathway to inhibit ferroptosis.
作者
徐芳丽
徐可
郭利芹
布海霞
王焕
XU Fang-li;XU Ke;Guo Li-qin;BU Hai-xia;WANG Huan(Dept of Nephrology,Xinxiang Central Hospital,The Fourth Clinical College of Xinxiang Medical University,Xinxiang Henan 453000,China)
出处
《中国药理学通报》
北大核心
2025年第12期2379-2385,共7页
Chinese Pharmacological Bulletin
基金
河南省医学科技攻关计划联合共建项目(No LHGJ20220988)。
