阿司匹林对脑缺血-再灌注大鼠脑能量代谢的影响

Effect of aspirin on brain energy metabolism in rats with cerebral ischemia-reperfusion injury

目的观察阿司匹林(ASA)对脑缺血-再灌注大鼠脑组织能量代谢的影响。方法取雄性SD大鼠50只,随机分为假手术组(12只)、溶剂组(26只)、ASA组(12只),根据再灌注时间将每组再分为24、72h两个亚组。线栓法制作大脑中动脉缺血2h,再灌24、72h模型。ASA组大鼠于再灌注同时灌胃给予ASA,60mg/kg(60mgASA加0.05ml无水乙醇助溶,用中性PBS液定容至10ml),1ml/100g。溶剂组大鼠灌胃给予等体积的0.5%无水乙醇PBS溶液。高效毛细管电泳分析法测定三磷酸腺苷酶(ATP)含量,无机磷法测定Na+-K+-ATP酶和Ca2+-ATP酶活性,比色法测定乳酸脱氢酶及乳酸含量。结果①ASA组大鼠再灌注24、72hATP含量为(27.7±3.5)、(29.7±2.5)μmol/g(湿重);Na+-K+-ATP酶为(8.6±0.9)、(7.8±0.6)μmol.mg-1.h-1;Ca2+-ATP酶为(6.1±0.8)、(6.6±0.7)μmol.mg-1.h-1,与溶剂组大鼠的(10.3±1.0)、(4.6±0.8)μmol/g(湿重),(4.5±0.7)、(5.8±0.8)及(4.5±0.4)、(2.5±0.5)μmol.mg-1.h-1比较,差异有统计学意义(P<0.01或0.05);与假手术组比较,除Ca2+-ATP酶含量高于假手术组[(4.3±0.5)、(4.6±0.3)μmol.mg-1·h-1]外,其他指标差异无统计学意义。②ASA组再灌注24、72h乳酸含量为(0.68±0.25)、(0.62±0.15)mmol/g(蛋白);乳酸脱氢酶活性为(9769±957)、(9677±532)U/g(蛋白);与溶剂组大鼠的(0.42±0.12)、(0.33±0.16)mmol/g(蛋白),(4033±723)、(5902±689)U/g(蛋白)比较,差异有统计学意义(P<0.01或0.05);与假手术组大鼠的(0.58±0.19)、(0.61±0.23)mmol/g(蛋白),(9430±273)、(9382±375)U/g(蛋白)比较,差异无统计学意义。结论阿司匹林对脑缺血-再灌损伤24和72h大鼠脑组织均有明显的保护作用,其机制可能与改善脑组织能量代谢有关。

Abstract： Objective To observe the effect of aspirin （ASA） on brain energy metabolism in rats with focal cerebral ischemia-reperfusion injuries. Methods Fifty male SD rats were randomly allocated to sham-operation （ n = 12 ）, vehicle （ n = 26） and ASA （ n = 12 ） groups. Each group was redivided into 2 subgroups according to the time of reperfusion（ 24 h and 72 h） . The models of focal middle cerebral artery ischemia for 2 h and reperfusion for 24 and 72 h were established by suture method. The ASA group rats were given ASA, 60 mg/kg by gavage at the same time with reperfusion. The vehicle group rats were given 0.5% absolute alcohol PBS. The level of adenosine triphosphate （ATP） was determined by capillary elec- trophoresis. The Na ＋ -K ＋ -ATPase and Ca2＋ -ATPase activity were determined by inorganic phosphate as- say, and the levels of lactate dehydrogenase and lactic acid were determined by colorimetric method. Results （1）The levels of ATP after 24 h and 72 h reperfusion in the ASA group were 27. 7 ± 3. 5 and 29. 7 ± 2. 5 μmol/g wet weight ; The Na ＋ -K ＋ -ATPase were 8.6 ± O. 9 and 7.8 ± 0. 6 μmol·mg-1·h-1 ; The Ca2＋ -ATPase were 6. 1 ±0. 8 and 6.6 ±0. 7 μmol·mg-1·h-1. As compared with 10. 3 ± 1.0 and 4. 6 ± 0. 8μmol/g wet weight, 4. 5 ±0. 7 and 5. 8 ±0. 8 μmo·mg-1·h-1, and 4. 5 ＋0. 4 and 2. 5±0. 5 μmol·mg-1·h-1 in the vehicle group, the differences were statistically significant. As compare with the sham- operation group, apart from the level of Ca2＋ -ATPase was higher than that of the sham-operation group （4. 3±0. 5 and 4. 6 ±0. 3 μmol·mg-1·h-1 ）, the differences of other indexes were not statistically sig- nificant. （2）The levels of lactic acid 24 h and 72 h after reperfusion were 0. 68 ± 0. 25 and 0. 62 ± 0. 15 mmol/g protein; the lactate dehydrogenase activities were 9769 ＋ 957 and 9677 ± 532 U/g protein; As compared with 0. 33 ±0. 16 and 0. 42 ＋0. 12 mmol/g protein, and 4033 ＋723 and 5902 ＋689 U/g pro- tein in the vehicle group, the differences were statistically significant. As compared with 0. 58 ± 0. 19 and 0. 61 ± 0. 23 mmol/g protein, and 9430±273 and 9382 ± 375 U/g protein in the sham-operation group, the differences were not statistically significant. Conclusion ASA has significant neuroprotective effect on 24 h and 72 h after focal cerebral ischemia-reperfusion injury, and its mechanism may be associated with the improvement of the energy metabolism of brain tissue.