益肾通络方调控TLR4/MyD88/NF-κB信号通路改善糖尿病肾病小鼠细胞焦亡的机制研究OA北大核心

Objective To investigate the mechanism of Yishen Tongluo Formula in ameliorating renal pyroptosis in diabetic nephropathy mice by regulating the toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway.Methods Sixty C57BL/6 male mice were randomly divided into control(10 mice)and intervention groups(50 mice)using random number table method.The diabetes nephropathy model was established by intraperitoneally injecting streptozotocin(50 mg/kg).After modeling,the intervention group was further divided into model,semaglutide(40 μg/kg),and high-,medium-,and low-dose Yishen Tongluo Formula groups(15.6,7.8,and 3.9 g/kg,respectively)using random number table method.The high-,medium-,and low-dose Yishen Tongluo Formula groups were administered corresponding doses of medication by gavage,the semaglutide group received a subcutaneous injection of semaglutide injection,and the control group and model groups were administered distilled water by gavage for 12 consecutive weeks.Random blood glucose levels of mice in each group were monitored,and the 24-h urinary protein content was measured using biochemical method every 4 weeks;after treatment,the serum creatinine and urea nitrogen levels were measured using biochemical method.The weight of the kidneys was measured,and the renal index was calculated.Hematoxylin and eosin,periodic acid-Schiff,periodic Schiff-methenamine,and Masson staining were used to observe the pathological changes in renal tissue.An enzyme-linked immunosorbent assay was used to detect urinary β2-microglobulin(β2-MG),neutrophil gelatinase-associated lipocalin(NGAL),and kidney injury molecule-1(KIM-1)levels.Western blotting and real-time fluorescence PCR were used to detect the relative protein and mRNA expression levels of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3),Caspase-1,gasdermin D(GSDMD),interleukin-1β(IL-1β),and interleukin-18(IL-18)in renal tissue.Immunohistochemistry was used to detect the proportion of protein staining area of the TLR4,MyD88,and NF-κB in renal tissue.Results Compared with the control group,the random blood glucose,24-h urinary protein,serum creatinine,urea nitrogen,and renal index of the model group increased,and the urine β2-MG,NGAL,and KIM-1 levels increased.The relative protein and mRNA expression levels of NLRP3,Caspase-1,GSDMD,IL-1β,and IL-18 in renal tissue increased,and the proportion of TLR4,MyD88,and NF-κB protein positive staining areas increased(P<0.05).Pathological changes such as glomerular hypertrophy were observed in the renal tissue of the model group.Compared with the model group,the Yishen Tongluo Formula high-dose group showed a decrease in random blood glucose after 12 weeks of treatment(P<0.05).The Yishen Tongluo Formula high-and medium-dose groups showed a decrease in 24-h urinary protein,creatinine,urea nitrogen,and renal index,as well as decreased β2-MG,NGAL,and KIM-1 levels.NLRP3,Caspase-1,GSDMD,IL-1 β,and IL-18 relative protein and mRNA expression levels were also reduced,and the proportion of TLR4,MyD88,and NF-κB protein positive staining areas was reduced(P<0.05).Pathological damage to renal tissue was ameliorated.Conclusion Yishen Tongluo Formula may exert protective renal effects by inhibiting renal pyroptosis and alleviating tubular interstitial injury in diabetic nephropathy mice by regulating the TLR4/MyD88/NF-κB signaling pathway.