温肾健脾方通过p70S6K信号通路调控自噬减轻糖尿病肾病大鼠足细胞损伤的机制  

Study on mechanism of Wenshen Jianpi recipe regulatingautophagy by p70S6K signaling pathway on alleviatingpodocyte injury in diabetic nephropathy rats

作　　者：史波[1] 李如瑶 金汀龙 王金 曹晓丹[1] SHI Bo;LI Ru-yao;JIN Ting-long;WANG Jin;CAO Xiao-dan(Ningbo Hospital of Traditional Chinese Medicine Affiliated to Zhejiang Chinese Medical University,Ningbo Zhejiang 315010,China)

机构地区：[1]浙江中医药大学附属宁波市中医院,浙江宁波315010

出　　处：《中国药理学通报》2025年第3期567-573,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82205063);宁波市医疗卫生高端团队重大攻坚项目(No 2022030309)。

摘　　要：目的探究基于70 ku核糖体S6激酶(phosphoprotein 70 S6 kinase,p70S6K)信号通路的温肾健脾方调控自噬减轻糖尿病肾病(diabetic nephropathy,DN)大鼠足细胞损伤的作用机制。方法将链脲佐菌素诱导的DN模型大鼠分为5组,每组6只,即DN对照组、温肾健脾方低剂量组(7.5 g·kg^(-1)·d^(-1))、温肾健脾方中剂量组(15 g·kg^(-1)·d^(-1))、温肾健脾方高剂量组(30 g·kg^(-1)·d^(-1))、缬沙坦阳性对照组(25 mg·kg^(-1)·d^(-1)),另以6只正常大鼠作为阴性对照组(等渗NaCl溶液10 mL·kg^(-1)·d^(-1)),各组大鼠连续灌胃8周。测定各组大鼠空腹血糖、尿液白蛋白/肌酐比值(urine albumin/creatinine ratio,UACR)和血液黏滞性,透射电镜观察各组大鼠足细胞结构和自噬小体变化,Western blot检测各组大鼠肾脏组织信号通路因子p70S6K和自噬因子p62的表达水平,免疫组化检测各组大鼠肾脏组织p62的表达水平。结果温肾健脾方可降低DN大鼠空腹血糖和UACR比值,改善血液黏滞性。透射电镜提示,温肾健脾方能够明显改善DN模型大鼠足细胞结构,提高足细胞自噬水平。Western blot结果提示,DN模型大鼠肾脏细胞信号通路因子p70S6K和自噬因子p62表达水平比空白对照组大鼠明显升高,差异有统计学意义( P <0.01)。与模型对照组相比,不同浓度温肾健脾方干预后p70S6K和p62表达水平均有所下降( P <0.05),其中温肾健脾方中剂量组变化最为显著。免疫组化结果显示,与对照组大鼠相比,DN大鼠肾脏组织自噬因子p62水平升高,温肾健脾方对DN大鼠p62的表达有一定的抑制作用。 结论 温肾健脾方可能通过抑制DN大鼠肾脏组织p70S6K水平,降低自噬因子p62表达,诱导肾脏细胞自噬增强进而恢复细胞稳态。Aim To detect the mechanism of Wenshen Jianpi recipe(WSJPR)regulating the autophagy by p70S6K signaling pathway on alleviating podocyte injury in diabetic nephropathy(DN)rats.Methods DN model rats induced by streptozotocin were divided into five groups with six rats in each group:model control group,low dose group(7.5 g·kg^(-1)·d^(-1)),medium dose group(15 g·kg^(-1)·d^(-1)),high dose group(30 g·kg^(-1)·d^(-1)),and positive control group(25 mg·kg^(-1)·d^(-1)).In addition,six normal rats were used as negative control group(isotonic NaCl solution 10 mL·kg^(-1)·d^(-1)).All the rats were given continuous gavage for eight weeks.Fasting blood glucose,urine albumin/creatinine ratio(UACR)and blood viscosity were determined.The changes of podocyte ultrastructure and autophagosome in each group were observed by transmission electron microscopy(TEM).The protein levels of signaling pathway factor p70S6K and autophagy factor p62 in renal tissues of rats in each group were detected by Western blot.Besides,p62 expression was observed by immunohistochemistry.Results WSJPR could decrease fasting blood glucose and UACR,and improve the indexes of blood viscosity in rats.TEM indicated that WSJPR could significantly improve the podocyte ultrastructure and autophagy level in DN rats.Western blot showed that the expression level of signaling pathway factor p70S6K and autophagy factor p62 in the kidney of DN rats increased significantly compared with blank control group(P<0.01).The expression level of p70S6K and p62 in WSJPR groups decreased compared with model control group(P<0.05).Among them,the medium-dose group of WSJPR had the most significant change.Immunohistochemical results showed that the level of autophagy factor p62 in kidney tissue of DN rats increased compared with the control group.WSJPR had a certain inhibitory effect on p62 expression in DN rats.Conclusion WSJPR might restore cell homeostasis by inhibiting p70S6K level,reducing the expression of autophagy factor p62 and enhancing autophagy level in renal tissue

关 键 词：温肾健脾方 糖尿病肾病 自噬 足细胞 P70S6K P62 

分 类 号：R-332[医药卫生] R289.5R322.61R329.24R345.57R392R587.2