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目的探讨不同压力CO2缺血预处理对气腹模型大鼠肝脏缺血再灌注损伤的影响及可能机制。方法成年健康雄性SD大鼠30只,采用随机数字表法分为5组:空白对照组(C组)于麻醉后插入气腹针;气腹对照组(Pp组)以10 mm Hg气腹压力建立CO2气腹60 min;低压预处理组(L-IP组)、中压预处理组(M-IP组)、高压预处理组(H-IP组)在建立CO2气腹前,分别以5、10、15 mm Hg充气5 min放气5 min,连续3个循环。各组恢复灌注60 min后取肝组织,行HE染色光镜下观察肝组织病理学变化,分光光度法测定超氧化歧化酶(SOD)活性、总抗氧化物能力(T-AOC)及丙二醛(MDA)含量,免疫组化、Western blot法检测Bcl-2、Bax蛋白表达水平,RT-PCR法检测IL-1β、IL-6 m RNA表达水平。结果除C组外,Pp组与各IP组大鼠肝组织均出现病理学改变,其中,各IP组的损伤程度明显轻于Pp组;与Pp组比较,各IP组SOD活性、T-AOC升高及MDA含量下降,Bcl-2蛋白表达增加、Bax蛋白表达减少且Bcl-2/Bax比值升高,IL-1β及IL-6 m RNA表达下调(P均<0.05);与L-IP组、H-IP组比较,M-IP组SOD、TAOC、Bcl-2及Bcl-2/Bax比值均增加,MDA、Bax、IL-1β及IL-6 m RNA均减少。结论 CO2缺血预处理可减轻气腹模型大鼠肝脏缺血再灌注损伤,其中10 mm Hg压力的CO2预处理抗损伤作用更明显,其机制与减弱氧化应激反应,上调Bcl-2抗凋亡蛋白表达,下调Bax促凋亡蛋白表达及炎症细胞因子IL-1β、IL-6 m RNA的表达,减少细胞凋亡和降低炎性反应有关。
Objective To investigate the effects and mechanisms of CO2 preconditioning on hepatic ischemia-reperfusion injury in pneumopericardium model rats. Methods Thirty adult male Sprague-Dawley rats were randomly divided into five groups according to the random number table. The blank control group (C group) was inserted with the pneumoperitoneum after anesthesia. The pneumoperitoneum control group (Pp group) received pneumoperitoneum pressure of 10 mm Hg CO2 pneumoperitoneum was established for 60 min before the establishment of CO2 pneumoperitoneum in low pressure pretreatment group (L-IP group), medium pressure pretreatment group (M-IP group) and high pressure pretreatment group (H-IP group) , 10,15 mm Hg aeration 5 min deflated 5 min, continuous 3 cycles. After 60 min of reperfusion, the liver tissues were harvested and the pathological changes of liver were observed under light microscope. The activities of superoxide dismutase (SOD), total antioxidant capacity (T-AOC) The contents of MDA, Bcl-2 and Bax were detected by immunohistochemistry and Western blot. The expression of IL-1β and IL-6 mRNA was detected by RT-PCR. Results In addition to group C, pathological changes were observed in liver tissue of Pp group and each IP group, in which the damage degree of each IP group was lighter than that of Pp group. Compared with Pp group, SOD activity, T-AOC (P <0.05). Compared with L-IP (P <0.05), the expression of Bcl-2 protein increased, the expression of Bax protein decreased and the ratio of Bcl-2 / Compared with H-IP group, the ratios of SOD, TAOC, Bcl-2 and Bcl-2 / Bax in M-IP group increased and MDA, Bax, IL-1β and IL-6 m RNA decreased. Conclusions CO2 preconditioning can reduce the hepatic ischemia-reperfusion injury in pneumoperitoneum model rats. CO2 pretreatment with 10 mm Hg pressure has a more obvious anti-injury effect. Its mechanism is related to attenuating oxidative stress and up-regulating Bcl-2 anti-apoptosis The expression of proapoptotic protein Bax, the expression of pro-apoptotic protein Bax and the expression of inflammatory cytokines IL-1β and IL-6 m RNA were decreased, and apoptosis and inflammatory response were decreased.