基于肾小球内皮细胞内质网应激探索当归芍药散对db/db小鼠糖尿病肾病的保护作用及机制OA

Objective:To investigate the therapeutic efficacy of Danggui Shaoyaosan(DSS)on renal injury in db/db mice and its impact on endoplasmic reticulum stress(ERS)in renal tissues.Methods:Thirty 8-week-old male db/db mice and six db/m mice were acclimated for one week,after which urinary microalbumin and blood glucose levels were monitored to establish a diabetic kidney disease(DKD)model.The model mice were randomly divided into a model group,an irbesartan group,and three DSS treatment groups with different doses(16.77,33.54,and 67.08 g·kg-1·d-1).A normal group was set as control.Each group was intragastrically administered with the corresponding drugs or saline for 8 weeks.After the intervention,general conditions were observed.Serum cystatin C(Cys-C),24-hour urinary total protein(24 h-UTP),24-hour urinary microalbumin(24 h-UMA),urinary creatinine(Ucr),and urea nitrogen(UUN)were measured.Transmission electron microscopy(TEM)was used to observe glomerular basement membrane(GBM)and ultrastructural changes of the endoplasmic reticulum(ER)in glomerular endothelial cells.Western blot,real-time fluorescence quantitative polymerase chain reaction(Real-time PCR),and immunohistochemistry were used to analyze renal tissue structure and the expression of GRP78,CHOP,and related markers.Results:Compared with the normal group,the mice in the model group showed curled posture,sluggish response,poor fur condition,increased levels of Cys-C,24 h-UTP,24 h-UMA,and UUN(P＜0.05),while Ucr decreased(P＜0.05).The GBM was significantly thickened,with podocyte and foot process fusion.The protein expressions of GRP78,CHOP,and ATF6 were significantly upregulated(P＜0.05),the mRNA levels of GRP78 and CHOP increased(P＜0.05),and immunohistochemistry showed an enhanced GRP78 signal(P＜0.05).After treatment,the mice exhibited improved behavior,normalized GBM and podocyte structure,improved ER morphology and markedly better biochemical indicators.Western blot,Real-time PCR,and immunohistochemistry indicated that the ERS-related markers were downregulated in the DSS treatment groups(P＜0.05),suggesting alleviated ERS and improved renal function.Conclusion:DSS can effectively ameliorate renal pathological damage in db/db mice,possibly by regulating ERS in glomerular endothelial cells,although the underlying signaling mechanisms require further investigation.