目的：观察小檗碱(BBR)对阿霉素(DOX)诱导心肌损伤的影响及保护效应,并进一步探究BBR是否可以通过调控Wnt/β-catenin通路影响DOX诱导的心肌损伤。方法：以大鼠心肌细胞株H9C2为研究对象,分为Control组、模型组、BBR组、LiCl激动剂组。对照组细胞不作处理,模型组、BBR组、LiCl激动剂组细胞建立DOX诱导的心肌损伤模型,并使用BBR对BBR组进行干预,使用LiCl对LiCl激动剂组进行干预。采用CCK-8法检测心肌细胞活力,流式细胞仪检测细胞凋亡情况,Western blot检测Wnt/β-catenin通路蛋白β-catenin,细胞凋亡相关蛋白Bcl-2、Bax表达情况。结果：(1)CCK-8法:模型组心肌细胞活力低于Control组(P＜0.05),而BBR组和LiCl激动剂组细胞活力高于模型组(P＜0.05)。(2)流式细胞术:模型组细胞凋亡率高于Control组(P＜0.05);而BBR组和LiCl激动剂组细胞凋亡率低于模型组(P＜0.05)。(3)Western blot:与Control组相比,模型组心肌细胞Bax蛋白表达增加(P＜0.05),Bcl-2、β-catenin蛋白表达降低(P＜0.05),Bax/Bcl-2比值亦上升(P＜0.05);与模型组相比,BBR组和LiCl激动剂组Bax蛋白表达降低(P＜0.05),Bcl-2蛋白表达升高(P＜0.05),β-catenin蛋白表达升高(P＜0.05),Bax/Bcl-2比值下降(P＜0.05)。结论：BBR可能通过促进受损心肌细胞中β-catenin的表达,上调Wnt/β-catenin信号通路的活化程度,抑制心肌细胞凋亡,从而发挥其心肌细胞保护作用。
Objective: To observe the effect of berberine (BBR) on doxorubicin (DOX)-induced myocardial injury and its protective effect, and to further explore whether BBR can affect DOX-induced myocardial injury by regulating Wnt/β-catenin pathway. Methods: The rat myocardial cell line H9C2 was used as the research object and divided into Control group, model group, BBR group and LiCl agonist group. The cells in the control group were not treated. The DOX-induced myocardial injury model was established in the model group, BBR group and LiCl agonist group. The BBR group was intervened by BBR, and the LiCl agonist group was intervened by LiCl. CCK-8 method was used to detect the viability of cardiomyocytes. Flow cytometry was used to detect apoptosis. Western blot was used to detect the expression of Wnt/β-catenin pathway protein β-catenin, apoptosis-related proteins Bcl-2 and Bax. Results: (1)CCK-8 method : The cell viability of the model group was lower than that of the Control group (P＜0.05), while the cell viability of the BBR group and the LiCl agonist group was higher than that of the model group (P＜0.05). (2)Flow cytometry: The apoptosis rate of the model group was higher than that of the control group (P＜0.05). The apoptosis rate of BBR group and LiCl agonist group was lower than that of model group (P＜0.05). (3)Western blot: Compared with the Control group, the expression of Bax protein in the model group increased (P＜0.05), the expression of Bcl-2 and β-catenin protein decreased (P＜0.05), and the ratio of Bax/Bcl-2 also increased (P＜0.05). Compared with the model group, the expression of Bax protein in BBR group and LiCl agonist group decreased (P＜0.05), the expression of Bcl-2 protein increased (P＜0.05), the expression of β-catenin protein increased (P＜0.05), and the ratio of Bax/Bcl-2 decreased (P＜0.05). Conclusion: BBR may play a protective role in cardiomyocytes by promoting the expression of β-catenin in damaged cardiomyocytes, up-regulating the activation of Wnt/β-catenin signaling pathway and inhibiting cardiomyocyte apoptosis.
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