血红素加氧酶1在白藜芦醇抑制阿霉素致心肌细胞凋亡中的作用

Effect of heme oxygenase-1 on protection of myocardial cells by resveratrol against doxorubicin-induced apoptosis

目的探讨血红素加氧酶1(HO-1)在白藜芦醇(RES)抑制阿霉素(DOX)致心肌细胞凋亡中的作用。方法 60只雄性C57BL/6J小鼠随机分为对照组、DOX组、DOX+RES组和DOX+RES+HO-1抑制剂锡原卟啉(ZnPP)组,每组15只;除对照组外,均采用经腹腔注射DOX(总剂量12 mg/kg)建立心肌损伤模型。苏木精-伊红(HE)染色后光学显微镜下观察心肌组织病理学改变。采用化学比色法测定血清肌酸激酶(CK)和乳酸脱氢酶(LDH)的活性;分光光度法检测心肌组织HO-1和半胱氨酸蛋白酶3(Caspase-3)的活性;Western blotting法检测心肌组织HO-1蛋白表达;免疫组织化学法检测心肌组织凋亡相关蛋白Bcl-2和Bax的表达;TUNEL法检测心肌细胞凋亡率。结果与对照组比较,DOX组小鼠的CK、LDH和Caspase-3活性、Bax蛋白表达及心肌细胞凋亡率均显著升高(P<0.01),而HO-1活性及HO-1、Bcl-2蛋白表达均显著降低(P<0.01)。与DOX组比较,DOX+RES组小鼠的CK、LDH和Caspase-3活性、Bax蛋白表达及心肌细胞凋亡率均显著降低(P<0.01),而HO-1活性及HO-1、Bcl-2蛋白表达均显著升高(P<0.01);与DOX+RES组比较,DOX+RES+ZnPP组小鼠的CK、LDH和Caspase-3活性、Bax蛋白表达和心肌细胞凋亡率均显著升高(P<0.01),而HO-1活性及HO-1、Bcl-2蛋白表达均显著降低(P<0.05)。结论 HO-1参与RES对DOX致心肌细胞凋亡的抑制作用。

Objective To investigate the effect of heme oxygenase-1 （HO-1） on protection of myocardial cells by resveratrol （RES） against doxorubicin （DOX）-induced apoptosis. Methods Sixty male C57BL/6J mice were randomly divided into control group, DOX group, DOX ＋ RES group and DOX ＋ RES ＋ HO-1 inhibitor ZnPP group, with 15 mice in each group. Except for control group, model of myocardial damage was established in the other groups by intraperitoneal injection of DOX （total dosage, 12 mg/kg）. Histopathological changes of myocardial tissues were observed by light microscopy with HE staining. The activity of serum creatine kinase （CK） and lactate dehydrogenase （LDH） was determined by chemiluminescent method, the activity of HO-I and Caspase 3 in myocardial tissues was detected by spectrophotometry, the expression of HO-1 protein in myocardial tissues was determined by Western blotting, the expression of apoptosis-related proteins Bcl-2 and Bax in myocardial tissues was detected by immunohistochemistry, and the apoptosis of myocardial cells was evaluated by TUNEL assay. Results Compared with control group, the activity of CK, LDH and Caspase-3, the expression of Bax protein and the apoptosis rate of myocardial cells were significantly higher （P 〈 0.01）, and the activity of HO-I activity as well as the expression of HO-1 and Bcl-2 protein were significantly lower in DOX group （P 〈 0.01）. Compared with DOX group, the activity of CK, LDH and Caspase-3, the expression of Bax protein and the apoptosis rate of myocardial cells were significantly lower （P 〈 0.01）, and the activity of HO-1 and the expression of HO-1 and Bcl-2 Droteinwere significantly higher in DOX ＋ RES group （P 〈0.01）. Compared with DOX-RES group, the activity of CK, LDH and Caspase-3, the expression of Bax protein and the apoptosis rate of myocardial ceils were significantly higher （ P 〈 0.01 ）, and the activity of HO-1 and the expression of HO-1 and Bcl-2 protein were significantly lower in DOX ＋ RES ＋ ZnPP group （P 〈 0.05）. Conclusion HO-1 participates in the inhibition effect of RES on myocardial apoptosis induced by DOX.