Anti-inflammatory Mechanism of Wuwei Xiaoduyin on IgA Nephropathy Rats Based on NF-κB Inflammatory Signaling Pathway / 中国实验方剂学杂志

ABSTRACT

Objective:To observe the effect of Wuwei Xiaoduyin on the nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） signaling pathway in immunoglobulinA nephropathy（IgAN） rats， and to explore its mechanism of action in the treatment of IgA nephropathy. Method:The 40 SD rats were randomly divided into control group， model group， benazepril group （10 mg·kg·d^-1） and Wuwei Xiaoduyin group （2.75 g·kg·d^-1）， with 10 rats in each group. The IgA nephropathy rat model was established by intragastric administration of bovine serum albumin （BSA）， subcutaneous injection of carbon tetrachloride （CCl₄） and tail vein injection of lipopolysaccharide （LPS） for 9 weeks. The rats in each group were given corresponding doses of drugs by gavage， while the rats in the control group and model group were given the same amount of normal saline for successive 28 days. The levels of 24-hour urinary protein （UTP）， serum creatinine （SCr）， blood urea nitrogen （BUN） and serum albumin （ALB） were detected. The contents of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-1<italic>β</italic> （IL-1<italic>β</italic>） and interleukin-6 （IL-6） in serum were detected by enzyme-linked immunosorbent assay （ELISA）， the hematoxylineosin staining （HE）， immunofluorescence and transmission electron microscopy were used to observe the renal pathological changes， the expressions of IL-6， I<italic>κ</italic>B kinase <italic>β</italic> （IKK<italic>β</italic>）， phosphorylated I<italic>κ</italic>B kinase <italic>β</italic> （p-IKK<italic>β</italic>）， NF-<italic>κ</italic>B inhibitor protein <italic>α</italic> （I<italic>κ</italic>B<italic>α</italic>）， phosphorylated NF-<italic>κ</italic>B inhibitor protein <italic>α</italic> （p-I<italic>κ</italic>B<italic>α</italic>）， NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 were detected by immunohistochemistry （IHC）， real-time PCR （RT-PCR） and Western blot， respectively. Result:Compared with the control group， the level of UTP in the model group significantly increased （<italic>P</italic><0.01）， cultured glomerular mesangial cells proliferated， mesangial matrix and electronic dense deposit increased， mesentery thickened. A large amount of IgA was deposited in the glomerular mesangial area and showed irregular particles and mass distribution， the levels of TNF-<italic>α</italic>， IL-1<italic>β</italic>， IL-6 in serum significantly increased （<italic>P</italic><0.01）， the expression levels of IL-6， IKK<italic>β</italic>， p-IKK<italic>β， </italic>NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 in renal tissue significantly increased （<italic>P</italic><0.01）.Compared with the model group， the levels of UTP in each administration group significantly decreased （<italic>P</italic><0.01）， and the renal tissue injury alleviated， the levels of TNF-<italic>α</italic>， IL-1<italic>β</italic>， IL-6 in serum significantly decreased （<italic>P</italic><0.01）， the expressions of IL-6， IKK<italic>β</italic>， p-IKK<italic>β</italic>， I<italic>κ</italic>B<italic>α</italic>， p-I<italic>κ</italic>B<italic>α</italic>， NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 in the renal tissue significantly decreased （<italic>P</italic><0.01）. There were no significant differences in serum creatinine， blood urea nitrogen and serum albumin among the groups. Conclusion:Wuwei Xiaoduyin can reduce proteinuria in IgA nephropathy rats， and its mechanism may be related to the inhibition of NF-<italic>κ</italic>B signaling pathway and the over expression of related inflammatory factors.