Protection of Ang1-7 througth MKK/P38MAPKs inflammatory signal pathway on TNF-α stimulated mouse HL-1 cells

Abstract Background to explore the effect of Ang1-7 througth MKK/P38MAPKs inflammatory signaling pathway on TNF-α-stimulated mouse HL-1 cells. Methods Using TNF-α (100 µg/ml) to establish an inflammatory atrial fibrillation model in HL-1 cell, which derived from mouse atrial myocyte. treated HL-1 cells with different concentrations of Ang 1-7 (0.1, 1 and 10 mmol/L) and divided into 5 groups, namely A group(control group), B group(TNF ), C group(TNF + Ang 1-7 0.1 mmol/L), D group(TNF + Ang 1-7 1 mmol/L ) and E group(TNF + Ang 1-7 10 mmol/L ). Firstly, different concentrations of Ang 1-7 (0.1 mmol/L, 1 mmol/L and 10 mmol/L) were used to stimulate for half an hour, and then TNF-α (100 µg/ml) was added to stimulate for four hours. Both the cells and supernatant were collected. Cells were collected for Western Blotting to detect the protein expression of MKK3, MKK4, MKK6, PMMK4 and PP38. The supernatant was subjected to flow cytometry for detecting multi-inflammatory factors. Results Compared with the A group, the protein expression of MKK3, MKK4, MKK6, PMMK4 and PP38 was statistically significant increased after stimulation with inflammatory factors (TNF-α) (P < 0.05). After intervention with Ang 1-7, the protein expression of MKK3, MKK4, MKK6, PMMK4 and PP38 was statistically significant lower than that of B group (P < 0.05). There is no significant difference of the protein expression of P38 after stimulation with inflammatory factor (TNF-α). Compared with the A group, there was no significant difference in the protein expression of MAS after the stimulation of inflammatory factor (TNF-α). After the intervention of Ang 1-7, the protein expression of MAS was higher than that of the A group and B group, but there was no significant difference (P > 0.05). The expression of MAS protein had an increasing trend, but there was no significant difference (P > 0.05). TGF-β, TNF-α was significantly increased after stimulating factor (TNF-α) was given, but was decreased after the intervention of Ang 1-7, both there were statistically significant (P < 0.05). IL-6 also had the same trend, but there was no significant difference. Conclusion Ang1-7 througth MKK/P38MAPKs inflammatory signal pathway protected on TNF-α stimulated mouse HL-1 cells