Study on berberine of reducing IL-6 and TNF-α secreted in HUVEC induced by visfatin through JNK signal pathway / 中草药

ABSTRACT

Objective: To study the regulation of c-Jun N-terminal kinase (JNK) pathway on interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) secreted in human umbilical vein endothelial cell (HUVEC) induced by visfatin and the intervention of berberine. Methods: HUVEC was cultured for the in vitro experiment, visfatin was added at different concentration (0, 50, 100, and 200 μg/L) and in different times (0, 6, 12, 24, and 48 h) to obserbe the effect on HUVEC. MTT assay was used to detect the proliferation of HUVEC, apoptosis rate was measured by Flow Cytometer, the contents of IL-6 and TNF-α in HUVEC supernatant were determined by enzyme-linked immuno-sorbent assay (ELISA) assay, and the protein expressions of p-JNK, Bax, and Bcl-2 in HUVEC lysate were determined by Western blotting. Berberine (50 μmol/L) and SP600125 (10 μmol/L) were added to interfere HUVEC and to detect the effect of berberine. Results: Compared with the control group, visfatin (100 μg/L) could significantly decrease the proliferation of HUVEC, induce HUVEC apoptosis, and increase the IL-6 and TNF-α contents in HUVEC supernatant. Meanwhile, it could increase p-JNK and Bax expression, decrease Bcl-2 expression after 24 h. Compared with visfatin group, berberine and SP600125 could increase the proliferation of HUVEC while decrease the IL-6 and TNF-α contents in HUVEC supernatant, restrain p-JNK and Bax expression while increase Bcl-2 expression in HUVEC. Conclusion: Berberine could decrease the contents of IL-6 and TNF-α in HUVEC supernatant and ease the injury induced by visfatin, the protective mechanism is related to the inhibition of JNK signal pathway activation.