The Intervention Study Of CaN/NFAT Inhibitors On CaN/NFAT Signaling Pathway, Potassium Channel And Inflammatory Factors In T SHR Lymphocytes

Objective: Previous studies have shown Ca N/NFAT pathway inhibitors prevent hypertension. However, the mechanism of CaN/NFAT inhibitors in preventing hypertension is unknown. The present study was designed to examine the current variation of Kv1.3 channels and the expression level of inflammatory cytokines KCCN4、IL-6 and TNF-α in lymphocytes derived from sponstaneously hypertensive rats(SHR) after the intervention of calcineurin-nuclear factor of activated T-cells(CaN-NFAT) signaling pathway inhibitors with cyclosporin A(CsA) and VIVIT(NFAT inhibitor). Then investigate the relationship among Kv channels-CaN/NFAT signal pathway-inflammation and hypertension, from intervention level of Ca/NFAT signal pathway inhibitor in vivo. Attempt to experiment to verify the hypothesis that Kv channels and inflammatory cytokinesmight be involved in the occurrence and development of hypertension via CaN/NFAT signal pathways activating lymphocytes to release inflammatory, so as to provide possible research evidence for the inflammation regulating mechanism in treatment of hypertension. Methods: Spontaneously hypertensive rats(SHR) were divided into five groups: Control group(WKY), Model group(SHR), Placebo group(PLA), CsA group(CsA), VIVIT group(VIVIT). The Kv1.3 channel electrophysiological characteristics of T cells before and after the intervention of the CsA and VIVIT were analyzed with patch-clamp techniques.Fluorescent immunohistochemical detect theactivity of CaN and NFAT in T lymphocytes.Real-Time PCR and Western Blot were applied to detect the influences of CsA and VIVIT on the expression of KCCN4, IL-6 and TNF-α. Results: The Kv1.3 channel current density of T lymphoeytes was blocked after treated with CaN/NFAT inhibitors. CaN/NFAT inhibitors(CsA and VIVIT) displayed a inhibition on Kv channel of T cells in SHR compared with the controls. The results demonstrated that CaN/NFAT inhibitors CsA and VIVIT significantly down regulated KCCN4, IL-6 and TNF-α expressionin spontaneously hypertensive rats compared to SHR group. Conclusion: CaN/NFAT inhibitor CsA, VIVIT could block Kv1.3 channel currents in SHR lymphocyte. CsA and VIVIT can significantly reduce the gene and protein expression level of Kv channel(KCCN4), IL-6, TNF-αin SHRT lymphocytes. CsA and VIVIT could reduce the activity of NFAT and CaN. CaN/NFAT pathway plays a crucial role in controlling Kv1.3 channel and inflammatory cytokines IL-6, TNF-α expression in spontaneously hypertensive rats.