大黄糖络丸通过AMPK/mTOR/ULK1通路调控糖尿病肾病小鼠足细胞自噬的作用机制研究OA北大核心CSTPCD

AIM:To explore the intervention ef-fect of Dahuangtang pellets(DHT)on diabetic ne-phropathy(DN)based on the AMP-activated pro-tein kinase/mammalian target of rapamycin/unc-51-like kinase 1(AMPK/mTOR/ULK1)signaling path-way.METHODS:Eight mice were randomly as-signed to the model group,the dapagliflozin group,and the DHT(high,medium,and low dosage)group out of a total of 40 C57BL/KSJ-db/db(hereaf-ter referred to as db/db)mice;another 10 C57BL/KSJ-db/dm mice were used as the normal group,sa-line was provided to the normal and model groups,and the mice in the treatment group received the appropriate medications.The medications were giv-en for 10 consecutive weeks,once per day,to the mice in the treatment group.At weeks 0,4,8,and 10 of administration,fasting blood glucose(FBG)was assessed by drawing blood at a predetermined time from the tail vein;Urine samples were taken at 0,5,and 10 weeks after treatment to evaluate the levels of albumin and creatinine,and the uri-nary albumin-creatinine ratio(ACR)was computed.After 10 weeks,mice in each group were assayed for 24 h total urine protein,serum creatinine(Scr),urea nitrogen(BUN)levels;Western blotting analy-sis was conducted to detect the expression of p-AMPK,p-mTOR,and p-ULK1,as well as the expres-sion of autophagy related proteins homolog of yeast Atg6(Beclin-1),autophagy-related proteins microtubule-associated protein 1 light chain 3(LC3),P62 in renal tissue;Immunohistochemistry was used to measure the expression of podocyte la-cunar membrane proteins(Nephrin,Podocin)in re-nal tissues;The pathological morphology of renal tissue was observed by light microscopy and trans-mission electron microscopy.RESULTS:Compared with the model group,FBG,ACR,and 24 h total urine protein were reduced in the dapagliflozin group and DHT groups of mice,and there was no statistically significant difference in Scr and BUN;In renal tissues,there is increased expression of p-AMPK and p-ULK1,decreased expression of p-mTOR,increased expression of LC3Ⅱ/LC3Ⅰ and Be-clin-1,and decreased expression of P62(P＜0.01,P＜0.05);differentially upregulated in glomeruli are the podocyte lacunar membrane proteins Nephrin and Podocin(P＜0.01,P＜0.05);renal pathologic damage was reduced to varying degrees;transmis-sion electron microscopy showed an increase in the number of autophagic vesicles and autophagic lyso-somes.CONCLUSION:DHT can delay the develop-ment of DN by regulating the AMPK/mTOR/ULK1 signaling pathway,enhancing podocyte autophagy,and protecting glomeruli.