Mechanism of action of Jinghua Xiaozi Mixture in the treatment of Henoch Schonlein Purpura Nephritis revealed by Wnt/β-Catenin signaling pathway

The experimental HSPN rat model was established by using the co-administration of bovine serum albumin (BSA), lipopolysaccharide (LPS), and carbon tetrachloride (CCl4). Blood and proteinuria were detected in each group of rats. IgA deposition was detected by immunofluorescence, pathological changes of glomeruli and renal tubules under light microscope and changes of basement membrane, podocytes and mesangial area under electron microscope were observed, and whether to establish an effective HSPN rat model was comprehensively evaluated. Then the experimental rats were given different concentrations of JHXZ by gavage for 4 weeks, and western medicine piperazine ferulate tablets were used as positive control to observe the effects of JHXZ intervention on the pathological changes of renal tissue and the contents of urinary red blood cells and urinary protein in experimental rats. Finally, by measuring the protein expressions and mRNA levels of Wnt3a, Wnt4, Wnt9b, GSK-3β, β-catenin, TCF-4, MMP-7 and E-cadherin in renal tissues and mesangial cells, the correlation of Wnt/β-catenin signaling pathway in the pathogenesis of HSPN and its role in the treatment of HSPN diseases were evaluated.The results of the study showed that the results of urine testing, immunofluorescence, light microscope and electron microscope showed that there were increased red blood cells and proteins in urine, IgA deposition and mesangial proliferation, basement membrane proliferation, abnormal changes in podocyte morphology and quantity in all groups except the normal group, especially in the model group, which indicated that the HSPN rat model was successfully prepared. After high-dose JHXZ treatment, the IgA deposition in kidney, the proliferation degree of mesangial area and basement membrane, and the morphology and number of podocytes in HSPN rat model are close to normal, while the red blood cells, proteins in urine and IL-2, IL-4 and IFN-γ in blood are obviously decreased, and the effect is better with the increase of dose. The results of PCR and WB in vivo and in vitro were highly consistent. According to PCR and WB detection, it was found that high-dose JHXZ treatment could significantly reduce the gene and protein expression levels of Wnt3a, Wnt4, Wnt9b, GSK-3β, β-catenin, TCF-4, MMP-7 in model rats, and maintain the high expression of E-cadherin gene and protein. In addition, piperazine ferulate tablets are the first-line therapeutic drugs for HSPN, which was used as a positive control in this experiment. Our results show that compared with rats treated with piperazine ferulate tablets, the high-dose JHXZ group has better effects on improving the pathological changes of renal tissue, reducing urine red blood cells and reducing the expression of GSK-3β gene and protein. Our experiments proved that the activation of Wnt/β-catenin is involved in the pathogenesis of HSPN. JHXZ can regulate mesangial cells through Wnt/β-catenin to protect kidney.