GSK-3β在脂肪乳抑制布比卡因诱导大鼠心肌细胞凋亡中的作用：RNA干扰腺病毒感染法  

Role of GSK.3β in lipid emulsion-induced inhibition of bupivacaine.induced apoptosis in cardiomyo-cytes of rats： evaluation using RNA interference adenovirus infection method

作　　者：马富强 吕丹妮 摆志霞 陈学新 李晓慧 Danni;Bai Zhixia;Chen Xuexin;Li Xiaohui(Department of Anesthesiology, Tumor Hospital, General Hospital of Ningxia Medical University, Yinchuan 750004, Chin;Department of Anesthesiology, First Affiliated Hospital of Xi＇ an Medical University, Xi＇an 710077, Chin)

机构地区：[1]宁夏医科大学总医院肿瘤医院麻醉科,银川市750004 [2]西安医学院第一附属医院麻醉科,710077

出　　处：《中华麻醉学杂志》2018年第2期155-158,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81241140）

摘　　要：目的采用RNA干扰（RNAi）腺病毒感染法评价糖原合成酶激酶3p（GSK-3p）在脂肪乳抑制布比卡因诱导大鼠心肌细胞凋亡中的作用。方法H9C2大鼠心肌细胞复苏培养后，以1×10^5个／ml细胞密度接种于96孔板上，采用随机数字表法分为8组（n=10）：空白对照组（c组）、布比卡因组（B组）、脂肪乳组（LE组）、布比卡因＋脂肪乳组（B＋LE组）、空白对照＋GSK-3[3RNAi腺病毒（GSK-313i）组（C＋GSK．313i组）、布比卡因＋GSK．313i组（B＋GSK-313i组）、脂肪乳＋GSK-313i组（LE＋GSK-313i组）、布比卡因＋脂肪乳＋GSK-3131组（B＋LE＋GSK．313i组）。B组、LE组、B＋LE组分别用终浓度为1mmol／L布比卡因、1％脂肪乳、lmmol／L布比卡因＋1％脂肪乳的培养基孵育。C＋GSK-313i组、B＋GSK．313i组、LE＋GSK-313i组、B＋LE＋GSK．313i组经GSK-313i腺病毒感染后第2天给予以上药物孵育。药物孵育24h后采用Westernblot法检测Bax和Bcl-2的表达，采用DAPI法确定细胞凋亡率。结果与c组比较．B组Bax表达上调，Bcl-2表达下调，细胞凋亡率升高（P〈0．05）。与B组比较，B＋LE组Bax表达下调，Bcl-2表达上调，细胞凋亡率降低（P〈0．05）。与B＋LE组比较，B＋LE＋GSK-313i组Bax表达上调，Bcl-2表达下调，细胞凋亡率升高（P〈0．05）。结论脂肪乳抑制布比卡因诱导大鼠心肌细胞凋亡的机制与GSK-313有关。Objective To evaluate the role of glycogen synthase kinase 3 beta （GSK-313） in lipid emulsion-induced inhibition of bupivacaine-induced apoptosis in cardiomyocytes of rats using RNA interfer-ence （RNAi） adenovirus infection method. Methods H9C2 cells were transferred into 96-well cell plates at a density of 1 ×10^5 cells/ml after culture and then divided into 8 groups （n = 10 each） using a random number table： control group （group C）, bupivacaine group （group B), lipid emulsion group （group LE）, bupivacaine plus lipid emulsion group （group B＋LE）, control plus GSK-313RNAi adenovirus （GSK-3βi） group （group C＋GSK-3βi）, hupivaeaine plus GSK-3βi group （group B＋GSK-3βi）, lipid emulsion plus GSK-3βi group （group LE＋GSK-3βi） and bupivaeaine plus lipid emulsion plus GSK-3[3i group （group B＋LE＋GSK-3βi）. In B, LE and B＋LE groups, the cells were incubated with culture medium containing 1 mmol/L bupivacaine, 1% lipid emulsion and 1 mmol/L bupivacaine plus 1% lipid emulsion, respectively.In C＋GSK-3βi, B＋GSK-3βi, LE＋GSK-3βi and B＋LE＋GSK-3βi groups, the cells were incubated with the drugs mentioned above on 2nd day after being infected by adenovirus. At 24 h after incubation with drugs, the expression of Bax and Bel-2 was determined by Western blot, and the apoptosis rate was calcu-lated using DAPI staining. Results Compared with group C, the expression of Bax was significantly up-regulated, the expression of Bei-2 was down-regulated, and the apoptosis rate was increased in group B （P〈0.05）. Compared with group B, the expression of Bax was significantly down-regulated, the expres-sion of Bel-2 was up-regulated, and the apoptosis rate was decreased in group B＋LE （P〈0. 05）. Compared with group B＋LE, the expression of Bax was significantly up-regulated, the expression of Bel-2 was down-regulated and the apoptosis rate was increased in group B ＋ LE ＋ GSK-3[3i （P〈 0. 05). Conclusion The mechanism by which lipid emulsion inhibit

关 键 词：脂肪乳剂 静脉注射用 布比卡因 细胞凋亡 肌细胞 心脏 

分 类 号：R614[医药卫生—麻醉学]