钙敏感受体在大剂量瑞芬太尼诱导卵巢癌裸鼠心肌损伤中的作用机制研究

Mechanism of activation of Wnt/β-catenin pathway by calcium sensitive receptor in high-dose remifentanil-induced myocardial injury of nude mice with ovarian cancer

目的探讨钙敏感受体(CaSR)在大剂量瑞芬太尼诱导卵巢癌裸鼠心肌损伤中的作用机制及信号通路。方法将40只Nc-nu裸鼠按随机数字表法分为空白对照组(C组)、模型对照组(O组)、瑞芬太尼输注组(R组)和瑞芬太尼+CaSR拮抗剂Calhex231组(R+C组),每组10只。除C组外,其余3组裸鼠均进行卵巢癌模型构建。模型建立后,R组泵注瑞芬太尼20μg/(kg·min)持续1 h,R+C组尾静脉注射浓度为10-3 mol/L的Calhex231(每100 mg给予0.1μl),48 h后泵注瑞芬太尼20μg/(kg·min)持续1 h;C组、O组均输注等量0.9%氯化钠注射液。检测细胞上清液中肌酸激酶(CK)和乳酸脱氢酶(LDH)的变化情况,HE染色观察心肌细胞损伤情况,免疫组化检测CaSR表达分布,Western blot检测裸鼠心肌细胞中的CaSR、蓬乱蛋白同源物(Dvl-1)和β连环蛋白(β-catenin)表达情况。结果与C组和O组比较,R组CK和LDH水平均升高(均P<0.05),HE染色显示炎细胞浸润,心肌细胞肿胀,肌纤维排列紊乱,CaSR、Dvl-1及β-catenin表达明显升高(均P<0.05),免疫组化显示CaSR在心肌细胞膜及细胞质表达明显;而与R组比较,R+C组CaSR、Dvl-1和β-catenin表达均明显下降(均P<0.05)。结论CaSR通过Dvl-1介导的Wnt/β-catenin通路激活参与了大剂量瑞芬太尼诱导卵巢癌裸鼠心肌损伤的过程。

Objective To investigate the mechanism and signaling pathway of calcium sensitive receptor involved in myocardial injury induced by continuous high-dose infusion of remifentanil in nude mice with ovarian cancer.Methods Forty NC nude mice were randomly divided into 4 groups:control group(Group C),model group(Group O),remifentanil infusion group[20μg/(kg·min),Group R],remife ntanil+CaSR antagonists Calhex231 group(Calhex231100 mg per 0.1μl,Group R+C)with 10 rats in each group.The ovarian cancer model was induced in nude mice of Groups O,R and R+C.The CK and LDH levels in cell supernatant were tested,myocardial injury was observed by HE staining,and the expression of CaSR was detected by immunohistochemistry,the expression of CaSR,Dvl-1 andβ-catenin in cardiomyocytes was detected with Western blot analysis.Results Compared with Group C and Group O,the levels ofCKand LDH in Group R were higher(P<0.05),and HE staining showed inflammatory cells infilration,myocardial cell swelling,disordered muscle fibers arrangement in Group R.Meanwhile,Ca SR was significantly distributed in myocardial cell membrane and cytoplasm in Group R.Western blot showed the expression of CaSR,Dvl-1 andβ-catenin in cardiomyocytes was increased significantly(P<0.05)in Group R.Moreover,compared with R group,the expression of Ca SR,Dvl-1 andβ-catenin proteins was decreased significantly in Group R+C(P<0.05).Conclusion Activation of Dvl-1 mediated Wnt/β-catenin pathway by calcium sensitive receptor is involved in high-dose remifentanil-induced myocardial injury of nude mice with ovarian cancer.