BQ123对大鼠肢体缺血-再灌注后肺损伤的影响

BQ123 Alleviates Lung Injury Following Limb Ischemia-Reperfusion in Rats

目的观察BQ123对大鼠肢体缺血-再灌注(I-R)后肺损伤的影响。方法雄性SD大鼠30只,随机分为对照组、I-R组和BQ123组,各10只。于缺血4 h再灌注4 h后,采用放射免疫法测定肺组织中内皮素(ET)-1的含量,用生物化学法测定髓过氧化物酶(MPO)活性和丙二醛(MDA)含量,采用酶联免疫吸附法检测P-选择素的浓度,免疫组织化学方法检测肺组织中凋亡相关因子Fas、Bcl-2和凋亡蛋白酶(caspase-3)的表达情况,利用原位末端标记法检测肺细胞凋亡情况。结果与对照组比较,I-R组肺组织ET-1、MPO、MDA和P-选择素均明显增加。肺组织Fas、Bcl-2和Caspase-3蛋白表达明显上调,其吸光度值分别为0.294±0.003,0.108±0.005,0.174±0.003,凋亡细胞百分率明显增高[(18.83±2.86)%](P<0.01);与I-R组比较,BQ123组ET-1、MPO、MDA、P-选择素含量明显下降。Bcl-2表达明显升高(0.128±0.005),Fas和Caspase-3的表达明显减弱,分别为0.115±0.007,0.159±0.006,细胞凋亡率下降[(10.67±2.16)%](P<0.01)。结论 BQ123可能通过抑制肺细胞凋亡相关蛋白的表达对肢体I-R后肺损伤产生保护作用。

Objective To observe BQ123 on lung tissue injury following ischemia-reperfusion of hind limbs in rats.Methods A total of 30 male SD rats were randomly divided into control,I-R,and BQ123 groups,with 10 rats in each group. After 4- hour of ischemia and 4-hour of reperfusion to the hind limbs, ET-1 was measured by radioimmunoassay, levels of myeloperoxidase （MPO） and malondialehyde （MDA） were examined by biochemical method,and the content of P-selectin was examined by ELISA. Immunohistochemical method was used to detect the expressions of Fas,Bcl-2 and Caspase-3 in lung tissues. Pulmonary apoptosis was examined by means of TdT-mediated dUTP nick end labeling （ TUNEL） . Results Compared with the control group,the levels of ET-1,MPO,MDA and P-selectin in lung tissues were all increased significantly in I-R group （ P〈0. 01）. The expressions of Fas,Bcl-2 and Caspase-3 were 0. 294±0. 003,0. 108±0. 005,and 0. 174±0. 003,significantly up-regulated in the I-R group. The apoptosis rate [（18. 83±2. 86）%] was significantly increased in the I-R group （P〈0. 01）. Compared with I-R group,the tissue contents of ET-1,MPO,MDA and P-selectin were significantly lowered in BQ123 group （P＆lt;0. 01）. The expression levels of Fas （0. 115±0. 007） and Caspase-3 （0. 159±0. 006） were decreased,but the expression of Bcl-2 was increased （0. 128±0. 005）. The apoptosis rate in BQ123 group was significantly lower [（10. 67±2. 16）%,P〈0. 01].Conclusion BQ123 may have protective effects on lung tissue after limb ischemia-reperfusion in rats by means of improving neutrophil aggregation and reducing the expression of proteins related to cell apoptosis.