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Chronic salt-loading downregulates large-conductance Ca^(2+)-activated potassium channel in mesenteric arterial smooth muscle cells from SD rats

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Journal of Pharmaceutical Analysis 2009年第4期21卷 215-221页

作者： Zhao-xia Zhou,Chao-feng Sun,Ai-qun Ma,Fang-yuan Chen,Na Wei,Fu-qiang Liu Department of cardiovascular Medicine,the First Affiliated Hospital,Medical School of xi’an Jiaotong University institute of cardiovascular channelopathy,key laboratory of environment and genes related to diseases(xi’an Jiaotong University),ministry of education,xi’an 71.061.china DepartmentofcardiovascularMedicine theFirstAffiliatedHospitalMedicalSchoolofXi'anJiaotongUniversity instituteofcardiovascularChannelopath KeyLaboratoryofEnvironmentandGenesRelatedtoDiseasesMinistryofEducationXi'an710061

Objective Large-conductance calcium-activated potassium(BKCa)channel modulates vascular smooth muscle *** the present study,we tested the hypothesis that salt,one of the factors which significantly influence blood pressure(BP),can regulate BKCa activity and then elevate blood *** Male Sprague-Dawley rats aged 6 weeks were randomized into high salt diet group(HS)and control group,fed with high salt diet(containing 5% NaCl)and standard rat chow(containing 0.4% NaCl)respectively for 16 *** systolic blood pressure(SBP),body weight(BW)and 24-hour urinary output were tested every 4 *** of urinary Na+ was detected using flame spectrophotometrical *** the end of 16 weeks,all the rats were killed,the mesenteric arteries were obtained,and single mesenteric smooth muscle cells were isolated at *** resting membrane potential(Em),the total potassium currents and the currents after perfusion with TEA solution of the cells were all recorded by whole cell patch *** transcriptions of BKCa channel α and β1 subunits in mesenteric arterial vascular smooth muscle cells(VSMC)of each group were calculated by real-time *** There was no difference in SBP and BW at each stage between control group and HS group;the urinary Na+ level in HS animals was elevated significantly after 4 *** negative values of Em in HS group VSMCs were reduced compared with those in the control *** of β1 subunit of BKCa channels were decreased in HS group,but α subunit transcriptions did not differ between the two *** cell potassium currents did not differ between HS and control groups,but BKCa currents of HS group VSMCs were lower than those of control group *** Even without elevating SBP,salt-loading can still modulate the expression and activity of BKCa channel in the mesenteric arterial VSMC and elevate vascular tone.

关键词： chronic salt-loading large-conductance calcium-activated potassium channel blood pressure

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多质粒共转染建立Cav1.3 L-型钙通道的研究模型

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陕西医学杂志 2009年第6期38卷 643-644,647页

作者：强华魏峰张爱峰史瑞明孙超峰马爱群西安交通大学医学院第一附属医院心内科教育部环境与疾病相关基因重点实验室离子通道病室西安710061

目的:提高多质粒转染效率,建立钙离子通道的研究模型。方法:在传统的脂质体转染过程中,通过不断调整细胞密度、转染质粒DNA的量,将含Cav1.3L-型钙通道相关亚基的质粒DNA转染至HEK293T细胞,采用全细胞膜片钳技术检测Cav1.3L-型钙通道的... 详细信息

目的:提高多质粒转染效率,建立钙离子通道的研究模型。方法:在传统的脂质体转染过程中,通过不断调整细胞密度、转染质粒DNA的量,将含Cav1.3L-型钙通道相关亚基的质粒DNA转染至HEK293T细胞,采用全细胞膜片钳技术检测Cav1.3L-型钙通道的电流表达。结果:转染后的HEK293T细胞在荧光显微镜下可见明显的绿色荧光;全细胞膜片钳技术检测出Cav1.3 L-型钙通道电流的表达。结论:通过调整细胞密度和转染质粒DNA的量,可以提高脂质体介导多质粒共转染HEK293T细胞的效率,建立钙离子通道的研究模型。

关键词：钙通道,L型基因脂质体 @人胚胎肾 293T细胞

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