CaN/NFAT抑制剂对自发性高血压大鼠T淋巴细胞Kv1.3钾通道的影响

Effect of CaN/NFAT inhibitor on Kv1.3 potassium channel of T lymphocytes in spontaneously hypertensive rats

目的研究钙调神经磷酸酶(CaN)/活化T细胞核因子(NFAT)抑制剂环孢素A(CsA)和VIVIT对自发性高血压大鼠(SHR)的T淋巴细胞电压依赖性钾离子通道(Kv1.3)的抑制作用。方法 2017年1月至2018年12月,共设立5组,初始每组10只大鼠,进入实验后:外购的12周龄健康雄性Wistar-Kyoto(WKY)大鼠为WKY组(n=3),外购同龄雄性SHR分为四组,不作处理的为SHR组(n=6),给予安慰剂(PLA)生理盐水、CaN/NFAT抑制剂CsA和VIVIT的分别为PLA组(n=6)、CsA组(n=5)和VIVIT组(n=4)。予以相应处理后,分离淋巴细胞,利用qRT-PCR技术和Western blot技术检测T淋巴细胞Kv1.3和TNF-α、IL-6的表达情况。结果 Kv1.3、IL-6、TNF-α的mRNA相对表达量在SHR组(3.139±0.305、3.015±0.423、2.586±0.284)较WKY组(1.002±0.075、1.014±0.195、1.003±0.097)升高(P均<0.05);在CsA干预组(1.643±0.125、1.515±0.152、1.606±0.064)和VIVIT干预组(1.780±0.236、1.638±0.168、1.676±0.166)较PLA组(3.148±0.250、2.809±0.307、2.649±0.299)显著下降(P均<0.05)。Kv1.3、IL-6、TNF-α的蛋白相对表达量在SHR组(0.796±0.153、0.907±0.153、0.719±0.033)较WKY组(0.391±0.075、0.359±0.066、0.351±0.036)升高(P均<0.05);在CsA干预组(0.425±0.078、0.464±0.147、0.423±0.092)和VIVIT干预组(0.573±0.073、0.617±0.067、0.504±0.097)较PLA组(0.914±0.171、0.921±0.138、0.774±0.070)显著下降(P均<0.05)。结论 SHR T淋巴细胞上有很多被激活的Kv1.3钾离子通道;CsA、VIVIT两种CaN/NFAT抑制剂对Kv1.3钾离子通道具有抑制作用。

Objective To investigate the the inhibitory effect of calcineurin(CaN)/nuclear factor of activated T cells(NFAT) inhibitors--cyclosporine A(CsA) and VIVIT on Kv1.3 potassium channels of T lymphocytes in spontaneously hypertensive rats(SHR).Methods From January 2017 to December 2018,5 groups were set up with 10 rats in each group.The specific groups and the number of rats after entering the experiment were as follows: the purchased 12-week-old healthy male Wistar-Kyoto(WKY) rats were into WKY Group(n=3);purchased male SHR of the same age were divided into four groups, the ones without treatment were SHR group(n=6),the ones given with normal saline、CsA and VIVIT respectively were PLA group(n=6),CsA group(n=5) and VIVIT group(n=4).After corresponding intervention, lymphocytes were isolated, qRT-PCR and Western blot were used to detect the expression of Kv1.3 potassium channel, TNF-α and IL-6.Results The relative expression levels of Kv1.3,IL-6,TNF-α mRNA in SHR group(3.139±0.305,3.015±0.423,2.586±0.284) were higher than those in WKY group(1.002±0.075,1.014±0.195,1.003±0.097)(all P<0.05);but those in CsA intervention group(1.643±0.125,1.515±0.152,1.606±0.064) and VIVIT intervention group(1.780±0.236,1.638±0.168,1.676±0.166) were significantly decreased compared with PLA group(3.148 ±0.250,2.809±0.307,2.649±0.299)(all P<0.05).The relative expression levels of Kv1.3,IL-6,TNF-α protein in SHR group(0.796±0.153,0.907±0.153,0.719±0.033) were higher than those in WKY group(0.391±0.075,0.359±0.066,0.351±0.036)(all P<0.05);but those in CsA intervention group(0.425±0.078,0.464±0.147,0.423±0.092) and VIVIT intervention group(0.573±0.073,0.617±0.067,0.504±0.097) were significantly decreased compared with PLA group(0.914±0.171,0.921 ±0.138,0.774±0.070)(all P<0.05).Conclusions There are many activated Kv1.3 potassium channels in T lymphocytes of SHR.Two CaN/NFAT inhibitors--CSA and VIVIT can inhibit Kv1.3 potassium channels.