SAT-456 Effect of FangjiHuangqi decoction on IL-6 and TGF-β1 secretion by HMC after intervention of inflammatory factors secreted by lymphocytes based on p38MAPK signaling pathway

Abstract

Based on the previous experimental research, we designed the experiment to select inflammatory factors secreted by peripheral blood lymphocytes to co-incubate with human mesangial cells, and to explore the effects of Fangji Huangqi Decoction on IL-6, TGF-β1 and p38MAPK signaling pathways affecting IgA nephropathy. Human mesangial cells (HMC) were induced by inflammatory factors secreted by HPBL from healthy people and IgA nephropathy patients for 24 hours. The cells were divided into five groups: (1) Healthy blank group: HMC culture medium + 10% healthy control HPBL culture supernatant + 10% blank healthy human serum; (2) Healthy dosing group: HMC culture medium + 10% healthy control HPBL culture supernatant + 10% medicated serum; (3) IgA + blank serum: HMC culture medium + 10% IgA nephropathy HPBL culture supernatant + 10% blank healthy human serum; (4) IgA + medicated serum group: HMC culture medium + 10% IgA nephropathy HPBL culture supernatant + 10% IgA nephropathy patients HPBL culture supernatant IgA + blocker group: HMC culture medium + 10% IgA nephropathy patients HPBL culture supernatant + P38MAPK specific blocker SB203580 + 10% drug-containing serum. After 48 hours of culture, the expression of TGF-β1, IL-6 and p38 MAPK was detected by RT-PCR, Western Blot and immunofluorescence. The expression of IL-6, TGF-β1 and p38MAPK in IgA+blank serum group was significantly higher than that in healthy blank group (P<0.05). The expression of IL-6, TGF-β1 and p38MAPK in IgA+blank serum group was lower than that in IgA+blank serum group (P<0.05). There was no significant difference in the expression of IL-6, TGF-β1 and p38MAPK between IgA+blocker group and healthy group (P>0.05). The expression of IL-6, TGF-β1 and p38MAPK in the dosage group was lower than that in the IgA + blank serum group (P < 0.05). _The expression of IL-6, TGF-β1, p38 and p-p38 protein in IgA+containing serum group was lower than that in IgA+blank serum group (P < 0.05). There was no significant difference in the expression of IL-6, p38 and p-p38 protein between IgA+blocker group and healthy group (P > 0.05). The expression of IL-6, TGF-β1, p38 and p-p38 protein in IgA+blocker group was lower than that in IgA+blank serum group (P < 0.05). The results of epidemic fluorescence showed that the secretion of IL-6, TGF-β1 and p38 in IgA + blank serum group was higher than that in healthy blank serum group. The same effect of IgA + drug-containing serum group and IgA + blocker group could reduce the levels of IL-6, TGF-β1 and p38. The activation level of p38 MAPK in HMC induced by inflammatory factors secreted by peripheral blood lymphocyte in IgA nephropathy patients is significantly increased, and the secreted inflammatory factors IL-6 and TGF-β1 are also increased. Fangji Huangqi Decoction can significantly reduce the activation level, suggesting that the inhibition of p38 MAPK activation may also be one of the mechanisms of Fangji Huangqi Decoction in inhibiting the secretion of inflammatory factors by HMC. Fangji Huangqi Decoction can delay the progress of IgA nephropathy by reducing the activation of p38 MAPK, reducing the secretion of TGF-β1 and IL-6 by mesangial cells, and reducing the inflammatory reaction of mesangial cells.