目的 探讨酪氨酸蛋白激酶2/信号转导子和转录激活子3(JAK2/STAT3)信号通路在抵抗素诱导H9C2心肌细胞肥厚中的作用及其可能分子机制.方法 取生长同步化的H9C2心肌细胞随机分为对照组、抵抗素组、抵抗素+ Ag490组和Ag490组.对照组细胞用低血清培养基培养;抵抗素组细胞用含浓度为50μg· L-1抵抗素的低血清培养基培养;抵抗素+ Ag490组细胞先用含100μmol Ag490低血清培养基培养,然后用含抵抗素浓度50 μg· L-1的低血清培养基继续培养;Ag490组细胞先用含100 μmol Ag490低血清培养基培养,然后低血清培养;各组细胞均培养48 h(第1部分实验)或3h(第2部分实验).应用Image J软件测心肌细胞面积像素;二喹啉甲酸(BCA)法测蛋白含量;采用Western blot检测各组心肌细胞磷酸化酪氨酸激酶(P-JAK2)、磷酸化信号转导子和转录激活子3(P-STAT3)、α肌动蛋白(α-SMA)和C-MYC蛋白相对表达量.结果 抵抗素组心肌细胞表面积像素和单细胞蛋白含量均大于对照组、抵抗素+ Ag490组和Ag490组(P＜0.05);抵抗素+Ag490组心肌细胞表面积像素和单细胞蛋白含量大于对照组和Ag490组(P＜0.05);对照组和Ag490组心肌细胞表面积像素及单细胞蛋白含量比较差异无统计学意义(P＞0.05).抵抗素组心肌细胞α-SMA蛋白表达量较对照组、抵抗素+Ag490组、Ag490组均显著增加(P＜0.05);其余各组心肌细胞o-SMA蛋白表达量比较差异均无统计学意义(P＞0.05).抵抗素组心肌细胞C-MYC蛋白表达量较对照组、抵抗素+ Ag490组、Ag490组均显著增加(P＜0.05);抵抗素+Ag490组心肌细胞C-MYC蛋白表达量较对照组增加(P＜0.05);其余各组心肌细胞C-MYC蛋白表达量比较差异无统计学意义(P＞0.05).抵抗素组心肌细胞P-JAK2、P-STAT3蛋白表达量较对照组、抵抗素+Ag490组、Ag490组显著增加(P＜0.05);其余各组心肌细胞P-JAK2、P-STAT3蛋白表达量比较差异无统计学意义(P＞0.05).结论 抵抗素可诱导H9C2心肌细胞肥厚;JAK2/STAT3信号通路的激活在抵抗素诱导大鼠心肌肥大过程中发挥作用.%Objective To explore the role of janus kinase 2/signal transducer and activator of transcription 3 (JAK2/ STAT3) signaling pathway in resistin induced H9C2 cardiomyocytes hypertrophy and randomly its possible molecular mechanisms.Methods The synchronized grow H9C2 myocardial cells were selected and randomly divided into control group,resistin group,resistin + Ag490 group and Ag490 group.The myocardial cells in control group were cultured with low blood serum medium;the myocardial cells in resistin group were cultured with low blood serum medium which contained 50 μg · L-1 resistin;the myocardial cells in resistin + Ag490 group were first cultured with low blood serum medium which contained 100 μmol Ag490,then were cultured with low blood serum medium which contained 50 μg · L-1 resistin;the myocardial cells in Ag490 group were cultured with low blood serum medium which contained 100 μmol Ag490.After culturing for 48 h(the first part of the experiment) or 3 h (the second part of the experiment),the surface area picture element of myocardial cells were measured by Image J software;the content of protein was measured by bicinchoninic acid (BCA)method;the relative expression levels of alpha smooth muscle aorta (α-SMA),C-MYC,phosphorylated janus kinase 2 (P-JAK2),phosphorylated transducer and activator of transcription 3 (P-STAT3) were measured by Western blot.Results The surface area picture element of myocardial cells and protein content of single cell in resistin group were significantly greater than those in the control group,resistin +Ag490 group and Ag490 group(P ＜0.05);the surface area picture element of myocardial cells and protein content of single cell in resistin + Ag490 group were significantly greater than those in the control group and Ag490 group(P ＜ 0.05);there was no statistic difference in the surface area picture element of myocardial cells and protein content of single cell between the control group and Ag490 group(P ＞ 0.05).The expression of α-SMA protein in myocardial cells in resistin group was significantly higher than that in the control group,resistin + Ag490 group and Ag490 group(P ＜ 0.05);there was no statistic difference in the expression of α-SMA protein in myocardial cells among other groups(P ＞ 0.05).The expression of C-MYC protein in myocardial cells in resistin group was significantly higher than that in the control group,resistin + Ag490 group and Ag490 group (P ＜ 0.05);the expression of C-MYC protein in myocardial cells in resistin + Ag490 group was significantly higher than that in the control group (P ＜ 0.05);there was no statistic difference in the expression of C-MYC protein in myocardial cells among other groups(P ＞ 0.05).The expression of P-JAK2 and P-STAT3 protein in myocardial cells in resistin group was significantly higher than that in the control group,resistin + Ag490 group and Ag490 group (P ＜ 0.05);there was no statistic difference in the expression of P-JAK2 and P-STAT3 protein in myocardial cells among other groups (P ＞ 0.05).Conclusion Resistin can induce H9C2 cardiomyocytes hypertrophy;JAK2/STAT3 signaling pathways play a role in the process of resistin induced myocardial hypertrophy.
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