Experimental Study Of CaN/NFAT Signaling Pathway Related Molecular Expression In The Peripheral Blood T Lymphocytes Of Kazakh Patients With Hypertension In Xinjiang

Objective:(1) To observe the inhibitory effect of drug intervention on the culture and proliferation of peripheral blood T lymphocytes in patients with essential hypertension(EH) in Xinjiang Kazakh.(2) To study whether calcineurin/nuclear factor of activated T lymphocytes(CaN/NFAT) signaling pathway related molecules are expressed in Kazakh patients with EH in Xinjiang and their expressional changes.(3) To explore the effects of telmisartan on Ca N/NFAT signaling pathway related molecular expression after T lymphocyte activation and proliferation of peripheral blood T lymphocytes in Xinjiang Kazakh patients with EH. To provide theoretical evidence and new ideas for the inflammatory theory and pathogenesis of hypertension, and to develop new target for the therapy of hypertension. Methods:(1) 90 EH patients without antihypertensive treatment in Xinjiang Kazakh were selected and randomly divided into control group, telmisartan group and potassium channel blocker group(4–Aminopyridine, 4-AP group)(n=30 in each group). The male to female ratio was 1:1 in each group. Peripheral blood T lymphocytes were isolated by immunomagnetic cell sorting method and cultured in vitro. The proliferative changes of T lymphocytes were detected by CCK analysis after 0h, 24 h, 48 h, 72 h of drug intervention.(2) 30 untreated Xinjiang Kazakh hypertensive patients and 30 Kazakh healthy subjects were randomly selected. The male to female ratio was 1:1 in each group. Peripheral blood T lymphocytes were isolated by immunomagnetic cell sorting method. The real-time fluorescent quantitative polymerase chain reaction(q RT-PCR) and western blot analysis were performed to detect the m RNA and relative protein expression levels of CaN/NFAT sinaling pathway related molecules, respectively.(3) 100 Xinjiang Kazakh hypertensive patients without antihypertensive drug therapy were randomly selected and the subjects were randomly divided into Control group, Telmisartan group, CaN blocker group(cyclosporin A, Cs A group), NFAT blocker group(VIVIT group) and 4-AP group(n=20 in each group). The male to female ratio was 1:1 in each group. Peripheral blood T lymphocytes isolated by immunomagnetic cell sorting method were firstly cultured for activation and proliferation in vitro, then incubated for 48 h after corresponding treatment. The qRT-PCR and western blot analysis were employed to detect the mRNA and relative protein expressional changes of CaN/NFAT signaling pathway related molecules, respectively. Results:(1) Under different treatment conditions, T lymphocyte proliferation was different. The T lymphocyte proliferation among three groups were significantly different(P<0.01). At the same time point, the cellular activity after T lymphocyte proliferation of telmisartan group and 4-AP group were decreased significantly than control group(all P<0.01). In each group, the cellular activity after T lymphocyte proliferation of different time point were different significantly(all P<0.01). The cellular activity of control group increased and cellular activity of telmisartan group and 4-AP group decreased along with the prolonged(all P<0.01). Telmisartan and 4-AP inhibited T lymphocyte proliferation, which were time-dependent within 72 h.(2) No significant differences in baseline data between these two groups were observed(P>0.05). Both m RNA and protein expression of NFATc1, interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) were found to be expressed in the peripheral blood T lymphocytes. Compared to the controls, the mRNA and relative protein expression level of NFATc1, IL-6 and TNF-α were significantly increased(P<0.05, respectively) in Xinjiang Kazakh patients with EH.(3) No significant differences in general conditions among these five groups were observed(P>0.05). Under different treatment conditions, the mRNA and relative protein of NFATc1(all P<0.05), IL-6(all P<0.05) and TNF-α(all P<0.05) in peripheral blood T lymphocytes were expressed in five groups. The mRNA and protein expression level of NFATc1, IL-6 and TNF-α in T lymphocytes of Telmisartan group, Cs A group, VIVIT group and 4-AP group decreased significantly compared with control group(P<0.05); m RNA and protein expression level of NFATc1, IL-6 and TNF-α in T lymphocytes of CsA group, VIVIT group and 4-AP group were lower than telmisartan group(P<0.05); there were no statistical differences among Cs A group, VIVIT group and 4-AP group in mRNA and protein expression level of NFATc1, IL-6 and TNF-α in T lymphocytes(P>0.05). In T lymphocytes after 48 h treatment with telmisartan, inhibitory rate of NFATc1, IL-6 and TNF-α mRNA expression were 40.8%, 56.6%, 70.6% and the inhibitory rate of protein expression were 47.5%, 47.9%, 21.1% respectively. Conclusion:(1) Telmisartan can significantly inhibit the activation and proliferation of peripheral blood T lymphocytes in patients with EH in Xinjiang Kazakh, which indicates telmisartan can regulate the inflammatory response of hypertension by inhibiting T lymphocytes proliferation.(2) The peripheral blood T lymphocytes expressed endogenous CaN/NFAT signaling pathway related molecules, and the expression of Ca N/NFAT and its downstream factors IL-6, TNF-α were significantly increased in Xinjiang Kazakh patients with EH.(3) Telmisartan can effectively inhibit NFATc1, IL-6, TNF-α mRNA and protein expression of peripheral blood T lymphocytes in hypertensive patients in Xinjiang Kazakh.