The Effect of Shenyuan Granules(肾元颗粒)on Diabetic Kidney Disease Inflammation Based on the hnRNPF-Mediated TLR4/NF-κB Signaling Pathway
Objective:Based on the hnRNPF-regulated TLR4/NF-κB signaling pathway,to explore the effect of Shenyuan granules on inflammation in db/db diabetic kidney disease mice and its underlying mechanisms.Methods:A total of 60 SPF-grade db/db mice were randomly assigned to model group and Shenyuan granules group,receiving 0.2％adenine model feed.Additionally,eight wild-type mice served as blank group and received regular feed.Mice were administered Shenyuan granules suspension by gavage in Shenyuan granules group,while an equivalent volume of double-distilled water by gavage in model group and blank group for 12 consecutive weeks.After 12 weeks of administration,metabolic cages were used to collect 24-hour urine from mice.After urine collection,serum and kidney tissues were obtained.Biochemical analysis included assessing serum creatinine(SCr),blood urea nitrogen(BUN)and alanine aminotransferase(ALT).ELISA was employed to measure neutrophil gelatinase-associated lipocalin(NGAL)levels in urine.Kidney tissue pathology was observed through PAS staining.Real-time PCR was used to detect expression levels of TLR4 mRNA,IL-6 mRNA,IKKαmRNA,NF-κB P65 mRNA,TNF-α mRNA and hnRNPF mRNA in mouse kidney tissues.Western blotting was performed to measure hnRNPF and TLR4 protein expression,as well as phosphorylation levels of IKKα and NF-κB P65.Results:The model group showed significantly higher levels of serum SCr,BUN and ALT than blank group(P＜0.05).In contrast,The Shenyuan granules group showed significantly lower levels of SCr,BUN and ALT than model group(P＜0.05).The model group showed significantly higher urinary NGAL level than blank group(P＜0.05).The Shenyuan granules group showed significantly lower urinary NGAL level than model group(P＜0.05).Compared to the blank group,mice in the model group showed severe kidney tissue damage,whereas the Shenyuan granules group showed some improvement compared to the model group.In terms of molecular markers,the model group showed higher relative expression levels of TLR4 mRNA,IL-6 mRNA,IKKαmRNA,NF-κB P65 mRNA and TNF-α mRNA than blank group(P＜0.05),while lower hnRNPF mRNA expression than blank group(P＜0.05).The Shenyuan granules group showed lower relative expression levels of TLR4 mRNA,IL-6 mRNA,IKKα mRNA,NF-κB P65 mRNA and TNF-α mRNA than model group(P＜0.05),while higher hnRNPF mRNA expression than model group(P＜0.05).The model group showed lower hnRNPF protein expression than blank group(P＜0.05),while higher TLR4 protein expression and phosphorylation levels of IKKα and NF-κB P65 than blank group(P＜0.05).The Shenyuan granules group showed higher hnRNPF protein expression than model group(P＜0.05),while lower TLR4 protein expression and phosphorylation levels of IKKα and NF-κB P65 than model group(P＜0.05).Conclusion:Shenyuan granules may potentially alleviate inflammation in db/db mice through the hnRNPF-mediated TLR4/NF-κB signaling pathway,improving relevant biochemical indicators and mitigating renal damage.
万方数据
维普
