目的 探討比索洛爾預處理對缺氧/復氧誘導的心肌細胞凋亡和纖維化的影響，并分析分子機制。方法 構建缺氧/復氧細胞模型，將H9c2細胞分為對照組、模型組、比索洛爾組、細胞外信號調(diào)節(jié)激酶（ERK1/2）通路激活劑（LM22B-10）組。采用酶聯(lián)免疫吸附測定法（ELISA）檢測細胞中乳酸脫氫酶（LDH）和肌酸激酶同工酶（CK-MB）水平，CCK-8法檢測細胞活力，流式細胞術檢測細胞凋亡水平，免疫印跡法（Western blotting）法檢測凋亡蛋白、纖維化蛋白和ERK1/2信號通路蛋白表達。結(jié)果 與模型組比較，比索洛爾組H9c2細胞中LDH和CK-MB水平、細胞凋亡率，切割型半胱氨酸天冬氨酸蛋白水解酶-3（cleaved Caspase-3）、B淋巴細胞瘤-2相關蛋白（Bax）、I型膠原（Col I）、III型膠原（Col III）、α-平滑肌肌動蛋白（α-SMA）、基質(zhì)金屬蛋白酶9（MMP-9）蛋白表達以及磷酸化細胞外信號調(diào)節(jié)激酶1/2（p-ERK1/2）/（ERK1/2）比值顯著降低；細胞活力，B淋巴細胞瘤2（Bcl2）、金屬蛋白酶組織抑制因子1（TIMP-1）蛋白表達顯著升高（P<0.05）。與比索洛爾組比較，LM22B-10組H9c2細胞中LDH和CK-MB水平、細胞凋亡率，Cleaved Caspase-3、Bax、Col I、Col III、α-SMA、MMP-9蛋白表達以及（p-ERK1/2）/（ERK1/2）比值均顯著升高，細胞活力，Bcl-2、TIMP-1蛋白表達均顯著降低（P<0.05）。結(jié)論 比索洛爾預處理通過抑制ERK1/2信號通路活化減輕缺氧/復氧誘導的心肌細胞凋亡和纖維化。

Objective To investigate the effects of bisoprolol pretreatment on cardiomyocyte apoptosis and fibrosis induced by hypoxia reoxygenation, and to analyze the its mechanisms. Methods Hypoxia/reoxygenation cell model were constructed, H9c2 cells were divided into control group, model group, bisoprolol group, and LM22B-10 group. LDH and CK-MB levels in cells were detected by enzyme-linked immunosorbent assay (ELISA), cell viability was detected by CCK-8 assay, and cell apoptosis was detected by flow cytometry. The expressions of apoptotic proteins, fibrotic proteins and ERK1/2 signaling pathway proteins were detected by Western blotting. Results Compared with model group, LDH and CK-MB levels and apoptosis rate of H9c2 cells, protein expression of cleaved Caspase-3, Bax, Col I, Col III, α-SMA, MMP-9, and (p-ERK1/2)/(ERK1/2) ratio were significantly decreased. Cell viability, Bcl-2, and TIMP-1 protein expressions were significantly increased in bisoprolol group (P < 0.05). Compared with bisoprolol group, LDH and CK-MB levels, apoptosis rate of H9c2 cells, protein expression of cleaved Caspase-3, Bax, Col I, Col III,α-SMA, MMP-9, and (p-ERK1/2)/(ERK1/2) ratio were significantly increased, and cell viability, Bcl-2 and TIMP-1 protein expression were significantly decreased in LM22B-10 group (P < 0.05). Conclusion Bisoprolol pretreatment attenuated hypoxia/reoxygenation induced cardiomyocyte apoptosis and fibrosis by inhibiting the activation of ERK1/2 signaling pathway.

R965

海南省衛(wèi)生健康行業(yè)科研項目（20A200060）