基于PI3K/Akt/mTOR信号通路探讨萆薢分清颗粒对抗单侧输尿管梗阻大鼠肾纤维化影响的研究OA

Objective:To establish a unilateral ureteral obstruction(UUO)rat model and investigate the effect of Bixie Fenqing Granules(Rhizoma Dioscoreae Granules for Clearing Turbid Urine)on the expression of renal fibrosis(RF)-related proteins in UUO rats,thereby exploring its mechanism against RF.Methods:Sixty SPF-grade male SD rats aged 8 weeks were randomly divided into six groups(n=10 per group):blank group,model group,losartan potassium group,and high-,medium-,and low-dose Bixie Fenqing Granules groups.The general condition of the rats was observed during drug intervention.At week 4,serum and urine samples were collected to measure 24-hour urine protein quantification(24 h-UPQ),serum creatinine(Scr),and blood urea nitrogen(BUN)levels.Urinary creatinine content was detected by the sar-cosine oxidase method,and urinary microalbumin was measured by ELISA to calculate the urinary microalbumin-to-creatinine ratio(ACR).Renal tissue specimens were stained with hematoxylin-eosin(HE)and Masson staining to observe the degree of renal fibrosis.The expression levels of PI3K,Akt,and mTOR proteins in renal tissues were detected by West-ern blot and immunohistochemistry.Results:(1)Compared with the blank group,the model group showed lethargy and poor general condition;24 h-UPQ,ACR,Scr,and BUN levels were significantly increased(P<0.05,P<0.01).HE stain-ing revealed renal tubular atrophy,structural damage with vacuolar degeneration,interstitial fibroblast proliferation,in-flammatory cell infiltration,and glomerular structural destruction and consolidation.Masson staining showed extensive blue collagen fiber deposition,indicating severe RF.Immunohistochemistry and Western blot results indicated significantly en-hanced expression of PI3K,Akt,and mTOR in the model group(P<0.05,P<0.01).(2)After 4 weeks of intervention,all treatment groups showed improved general conditions,with the losartan potassium and high-dose Bixie Fenqing Granules groups exhibiting the most significant improvement.HE staining indicated marked alleviation of pathological damage in the losartan potassium and high-dose Bixie Fenqing Granules groups compared to the model group.Masson staining showed reduced blue collagen fiber area and improved fibrosis in all intervention groups except the low-dose Bixie Fenqing Gran-ules group.Compared with the model group,the high-and medium-dose Bixie Fenqing Granules groups and the losartan potassium group had significantly lower 24 h-UPQ,ACR,Scr,and BUN levels(P<0.05,P<0.01),while the low-dose group showed reduced Scr and BUN levels(P<0.05).Immunohistochemistry and Western blot results demonstrated that the losartan potassium group and the high-and medium-dose Bixie Fenqing Granules groups had significantly lower expression of PI3K,Akt,and mTOR in renal tissues(P<0.05,P<0.01).The low-dose group showed reduced PI3K and Akt expression(P<0.05,P<0.01),and the high-dose group had significantly lower PI3K expression than the medium-and low-dose groups(P<0.05).(3)Compared with the losartan potassium group,the high-dose Bixie Fenqing Granules group showed equivalent effects in improving general condition,reducing 24 h-UPQ,ACR,Scr,and BUN levels,and alleviating renal pathological damage and fibrosis(P>0.05).Immunohistochemistry and Western blot results indicated that the two groups equally suppressed PI3K and Akt protein expression in UUO rat renal tissues(P>0.05),while the high-dose group had lower mTOR expression than the losartan potassium group(P<0.05).Conclusion:Bixie Fenqing Granules reduce protein-uria,alleviate renal pathological damage,improve renal function,and protect the kidneys by lowering 24 h-UPQ,ACR,Scr,and BUN levels in UUO rats.They delay RF progression by downregulating the expression of RF-related proteins PI3K,Akt,and mTOR in renal tissues.