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目的探讨磷脂酰肌醇-3-激酶/丝氨酸-苏氨酸激酶(PI3K-Akt)信号转导通路在舒芬太尼后处理减轻大鼠心肌缺血再灌注损伤中的作用。方法健康雄性SD大鼠50只,体质量230~280 g,2~3月龄,采用随机数字表法,将其随机分为5组(n=10):假手术组(S组)、心肌缺血再灌注损伤组(I/R组)、舒芬太尼后处理组(PO组)、舒芬太尼后处理组+PI3K特异性抑制剂LY294002组(PO+L组)、PI3K特异性抑制剂LY294002组(L组)。I/R组,PO组,PO+L组,L组采用结扎左冠状动脉前降支30 min,再灌注120 min的方法来建造心肌缺血再灌注的模型。I/R组在再灌注前5 min时尾静脉注射0.9%氯化钠注射液1 ml,PO组在再灌注前5 min时尾静脉注射1ml舒芬太尼稀释液1μg/kg,PO+L组尾静脉注射1 ml舒芬太尼稀释液1μg/kg+LY2940020抑制剂0.3mg/kg,L组尾静脉注射1 ml LY2940020抑制剂0.3 mg/kg。于再灌注120 min时处死大鼠,取缺血部位心肌组织,用10%甲醛固定,常规石蜡包埋,切片。光镜下观察病理学结果,采用免疫组织化学方法测定心肌细胞Akt及磷酸化Akt(p-Akt)的表达,计算p-Akt平均吸光度值与Akt平均吸光度值的比值(p-Akt/Akt),采用TUNEL染色法检测心肌细胞凋亡,计算心肌细胞凋亡指数(AI)。结果与S组比较,其余4组AI、p-Akt/Akt的表达均升高(P<0.05);PO组比I/R组、PO+L组、L组AI降低,p-Akt/Akt的表达升高(P<0.05);PO+L组比L组AI降低,p-Akt/Akt的表达升高(P<0.05);I/R组与PO+L组及L组上述指标比较差异无统计学意义(P>0.05)。结论 PI3K-Akt信号转导通路参与了舒芬太尼后处理减轻大鼠心肌缺血再灌注损伤的作用。
Objective To investigate the role of PI3K-Akt signal transduction pathway in sufentanil postconditioning in attenuating myocardial ischemia-reperfusion injury in rats. Methods Fifty healthy male Sprague-Dawley rats were randomly divided into 5 groups (n = 10): sham operation group (S group), myocardium (I / R group), sufentanil postconditioning group (PO group), sufentanil postconditioning group and PI3K specific inhibitor LY294002 group (PO + L group), PI3K-specific Inhibitor LY294002 group (L group). I / R group, PO group, PO + L group and L group were established myocardial ischemia-reperfusion model by ligating left anterior descending artery of coronary artery for 30 min and reperfusion for 120 min. In the I / R group, 1 ml of 0.9% sodium chloride injection was injected into the caudal vein 5 minutes before reperfusion and 1 ml / kg of sufentanil diluted into the caudal vein 5 minutes before reperfusion in PO group. PO + One ml of sufentanil diluted 1 μg / kg + LY2940020 inhibitor 0.3 mg / kg was injected into the tail vein of the tail vein, and 1 ml of the LY2940020 inhibitor 0.3 mg / kg was injected into the tail vein of the L group. Rats were sacrificed at 120 min after reperfusion. Myocardial tissue was removed from the ischemic area, fixed in 10% formaldehyde, embedded in paraffin and sectioned. The pathological results were observed under light microscope. The expression of Akt and phosphorylated Akt (p-Akt) was measured by immunohistochemistry. The ratio of the average absorbance of Akt and Akt (p-Akt / Akt) The cardiomyocyte apoptosis was detected by TUNEL staining and the cardiomyocyte apoptosis index (AI) was calculated. Results Compared with S group, the expression of AI and p-Akt / Akt in the other four groups were significantly increased (P <0.05); AI in PO group was lower than that in I / R group, PO + L group and L group (P <0.05). The AI in PO + L group was lower than that in L group, while the expression of p-Akt / Akt was increased in PO + L group (P <0.05) The difference was not statistically significant (P> 0.05). Conclusion The PI3K-Akt signaling pathway is involved in the effect of sufentanil to reduce the myocardial ischemia-reperfusion injury in rats.