温肾健脾方由mTOR信号通路调控自噬-内质网应激平衡减轻DN大鼠肾脏损伤的机制研究  

Study on the Mechanism of Wenshen Jianpi Recipe Regulating the Balance of Autophagy-endoplasmic Reticulum Stress by mTOR Signaling Pathway on Alleviating Renal Injury in DN Rats

作　　者：周俊 陈霞波 魏任雄 曹晓丹 ZHOU Jun;CHEN Xiabo;WEI Renxiong(Department of Clinical Laboratory,Ningbo Hospital of Traditional Chinese Medicine,Affiliated to Zhejiang Chinese Medical University,Ningbo,315010)

机构地区：[1]浙江中医药大学附属宁波中医院检验科,宁波315010 [2]浙江中医药大学附属宁波中医院内分泌科,宁波315010

出　　处：《中国中西医结合肾病杂志》2023年第12期1049-1052,共4页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金资助项目(No.82205063)。

摘　　要：目的:探究温肾健脾方由mTOR信号通路调控自噬-内质网应激平衡减轻DN大鼠肾脏损伤的作用机制。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周后,分别采用Western blot、RT-PCR等方法检测各组大鼠肾脏组织内Ca^(2+)浓度、mTOR蛋白磷酸化水平、自噬因子LC3、Beclin-1蛋白表达水平及ERS分子伴侣GRP78 mRNA水平。结果:与正常对照组大鼠相比,DN模型组大鼠肾脏细胞内Ca2+浓度和自噬因子LC3、Beclin-1蛋白水平降低,磷酸化mTOR(p-mTOR)/mTOR比值及ERS分子伴侣GRP78 mRNA水平升高,温肾健脾方干预组大鼠肾脏细胞内Ca^(2+)浓度和LC3、Beclin-1蛋白表达水平升高,p-mTOR/mTOR比值及GRP78基因转录水平显著下降。结论:温肾健脾方可能通过提高肾脏细胞内Ca2+浓度,抑制DN大鼠肾脏细胞mTOR活性,诱导肾脏细胞自噬增强,改善ERS进而恢复细胞稳态。Objective:To detect the mechanism of Wenshen Jianpi recipe regulating the balance of autophagy-endoplasmic reticulum stress by mTOR signaling pathway on alleviating renal injury in DN rats.Methods:DN model rats induced by streptozotocin were divided into 5 groups with 6 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)),positive control group(valsartan,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)).After continuous intragastric administration for 8 weeks,Western blot and RT-PCR were used to detect Ca^(2+) concentration,mTOR phosphorylation level,protein levels of autophagy factor LC3,Beclin-1 and mRNA level of ERS molecular chaperone GRP78 in renal tissue of rats in each group.Results:Compared with the normal control group,the intracellular Ca^(2+) concentration and the protein levels of autophagy factors LC3 and Beclin-1 in the renal tissues of DN model group were decreased,the p-mTOR/mTOR ratio and the mRNA level of ERS molecular chaperone GRP78 were increased.Compared with DN group,intracellular Ca^(2+) concentration and protein expression levels of LC3 and Beclin-1 were increased,and the p-mTOR/mTOR ratio and GRP78 transcription level were significantly decreased in the group of Wenshenjianpi recipe intervention.Conclusion:Wenshenjianpi recipe might increase intracellular Ca^(2+) concentration,inhibite the activity of mTOR and improve the balance of autophagy and ERS to restore cell homeostasis in DN rats.

关 键 词：温肾健脾方 糖尿病肾病 自噬 内质网应激 MTOR 

分 类 号：R285.5[医药卫生—中药学]