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目的:评价低温缺氧复氧时大鼠心肌成纤维细胞(RCF)对H9c2细胞缝隙连接蛋白43(Cx43)表达的影响。方法:体外培养的H9c2细胞，采用随机数字表法分为4组(n n＝12)：对照组(C组)、低温缺氧复氧组(HHR组)、RCF共培养组(Co组)和RCF共培养+低温缺氧复氧组(Co+HHR组)。C组于37 ℃、5%COn 2+95%空气条件下培养5 h。HHR组于4 ℃、5%COn 2+95%Nn 2条件下培养1 h，然后于37 ℃、5%COn 2+95%空气条件下培养4 h。Co组和Co+HHR组将0.3×10n 5个/孔H9c2细胞接种于Transwelln ?小室下层培养皿、0.6×10n 5个/孔RCF接种于上层小室。Co组于37 ℃、5%COn 2+ 95%空气条件下培养5 h；Co+HHR组于4 ℃、5%COn 2+95%Nn 2条件下培养1 h后，于37 ℃、5%COn 2+95%空气条件下培养4 h。采用台盼蓝染色法测定H9c2细胞死亡率，免疫荧光法测定Cx43和细胞外调节蛋白激酶1/2(ERK1/2)的表达水平，Western blot法测定Cx43、磷酸化Cx43、ERK1/2及磷酸化ERK1/2的表达水平。n 结果:与C组相比，HHR组H9c2细胞死亡率升高，Cx43表达及磷酸化水平降低，ERK1/2表达及磷酸化水平升高(n P0.05)；与Co组相比，Co+HHR组H9c2细胞死亡率升高，Cx43及ERK1/2的表达和磷酸化水平降低(n P<0.05)；与HHR组相比，Co+HHR组H9c2细胞死亡率升高，Cx43及ERK1/2的表达和磷酸化水平降低(n P<0.05)。n 结论:低温缺氧复氧时RCF可降低H9c2细胞Cx43的表达水平和活性，其机制可能与下调ERK1/2的表达、抑制ERK1/2的活性有关。“,”Objective:To evaluate the effect of rat cardiac fibroblasts (RCF) on the expression of connexin43 (Cx43) in H9c2 cells during hypothermic hypoxia/reoxygenation.Methods:H9c2 cells cultured n in vitro were divided into 4 groups (n n=12 each) using the random number table method: control group (group C), hypothermic hypoxia/reoxygenation group (group HHR), RCF co-culture group (group Co) and RCF co-culture plus hypothermic hypoxia/reoxygenation group (group Co+ HHR). Group C was incubated at 37℃ in 5% COn 2 + 95% air for 5 h. Group HHR was incubated at 4 ℃ in 5% COn 2 + 95% Nn 2 for 1 h and then at 37 ℃ in 5% COn 2 + 95% air for 4 h. In group Co and group Co+ HHR, H9c2 cells 0.3×10n 5 cells/well were inoculated in the lower chamber and RCF 0.6×10n 5 cells/well in the the upper chamber of a transwelln ? culture dish.Group Co was incubated at 37 ℃ in 5% COn 2 + 95% air for 5 h. Group Co+ HHR was incubated at 4℃ in 95% Nn 2 + 5% COn 2 for 1 h, and then incubated at 37 ℃ in 5% COn 2 + 95% air for 4 h. The mortality rate of H9c2 cells was measured by trypan blue staining, the expression of Cx43 and extracellular signal-regulated protein kinases 1/2 (ERK1/2) by immunofluorescence, and the expression of Cx43, phosphorylated Cx43, ERK1/2 and phosphorylated ERK1/2 by Western blot.n Results:Compared with group C, the mortality rate of H9c2 cells was significantly increased, the expression and phosphorylation of Cx43 were decreased, and the expression and phosphorylation of ERK1/2 were increased in group HHR (n P0.05). Compared with group Co, the mortality rate of H9c2 cells was significantly increased, and the expression and phosphorylation of Cx43 and ERK1/2 were decreased in group Co+ HHR (n P<0.05). Compared with group HHR, the mortality rate of H9c2 cells was significantly increased, and the expression and phosphorylation of Cx43 and ERK1/2 were decreased in group Co+ HHR (n P<0.05).n Conclusions:RCFs can decrease the expression and activity of Cx43 in H9c2 cells during hypothermic hypoxia/reoxygenation, and the mechanism may be related to the down-regulation of ERK1/2 expression and inhibition of ERK1/2 activity.