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本研究采用全细胞膜片钳技术、实时荧光定量PCR技术及Western blot技术检测自发性高血压大鼠(spontaneously hypertensive rat,SHR)和Wistar大鼠外周血淋巴细胞电压依赖性钾通道(voltage dependent potassium channel,Kv)电流密度及Kv1.3mRNA和蛋白表达水平,探讨淋巴细胞Kv通道在高血压病中的变化,为高血压病淋巴细胞激活提供证据。结果显示:(1)SHR淋巴细胞Kv峰值(取自阶跃电压中+60mV)电流密度为(119±10)pA/pF(n=30),Wistar大鼠为(56±9)pA/pF(n=40),SHR淋巴细胞Kv峰值电流密度明显高于Wistar大鼠(P<0.05);(2)与Wistar大鼠相比,SHR淋巴细胞Kv1.3 mRNA表达增多(P<0.05);(3)SHR淋巴细胞Kv1.3蛋白表达水平高于Wistar大鼠(P<0.05)。结果提示,SHR淋巴细胞上有更多功能性Kv通道,Kv通道可能与SHR淋巴细胞激活有关。
In this study, whole-cell patch clamp technique, real-time fluorescence quantitative PCR and Western blot were used to detect the voltage-dependent potassium channel in peripheral blood lymphocytes of spontaneously hypertensive rats (SHR) and Wistar rats , Kv) current density and Kv1.3 mRNA and protein expression levels, to explore changes in lymphocyte Kv channel in hypertension, provide evidence for hypertensive disease activation of lymphocytes. The results showed that: (1) The peak current of Kv of SHR lymphocytes (+60 mV from the step voltage) was (119 ± 10) pA / pF (n = 30) and Wistar rats was (56 ± 9) pA / pF (P <0.05); (2) Compared with Wistar rats, the expression of Kv1.3 mRNA in SHR lymphocytes increased (P <0.05); (3) The expression of Kv1.3 protein in SHR lymphocytes was higher than that in Wistar rats (P <0.05). The results suggest that SHR lymphocytes have more functional Kv channels, Kv channels may be related to SHR lymphocyte activation.