杏芎氯化钠注射液对大鼠局灶性脑缺血再灌注损伤的保护作用

Neuroprotective effects of Floium Ginkgo extract and Tertram Ethypyrazine sodium chloride injection against cerebral ischemia-reperfusion injury in rats

目的研究杏芎氯化钠注射液对大鼠局灶性脑缺血再灌注损伤后脑组织中血小板α-颗粒膜蛋白-140(GMP-140)、组织型纤维酶原激活物(t PA)、自由基、氨基酸动态平衡和黏附分子的影响。方法健康雄性SD大鼠随机分为假手术组、模型组、对照组和实验组。各组均制备大鼠局灶性脑缺血模型。假手术组除不插尼龙线阻塞血管外,其余步骤与手术组相同。模型组与假手术组大鼠再灌注后腹腔注射生理盐水3 m L·kg^(-1),每天1次;对照组腹腔注射金纳多注射液15 mg·kg^(-1),每天1次;实验组腹腔注射杏芎氯化钠注射液41.75m L·kg^(-1),每天1次。用发色底物法测定GMP-140、t PA活性;用黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性;用硫代巴比妥酸法测定细胞脂质过氧化物丙二醛(MDA)含量;用高效液相色谱法测定脑组织中左旋谷氨酸(L-Glu)、左旋甘氨酸(L-Gly)、左旋天门冬氨酸(L-Asp)、γ-氨基丁酸(GABA)浓度;用免疫组化法测定黏附分子表达。结果处理后7 d,对照组和实验组的GMP-140含量分别为(7.40±2.90),(6.00±2.90)ng·m L^(-1),模型组为(20.40±5.80)ng·m L^(-1),差异均有统计学意义(均P<0.05)。处理后7 d,实验组与对照组的t PA的表达分别为(12.40±2.10),(6.60±1.40)ng·m L^(-1),差异有统计学意义(均P<0.05)。与假手术组、模型组相比,实验组与对照组SOD活性均显著升高、MDA含量均明显降低(均P<0.01);但实验组与对照组比较,差异均无统计学意义(均P>0.05)。实验组L-Asp、L-Glu表达与模型组相比,差异均有统计学意义(均P<0.05)。处理后3 d,实验组的GABA、L-Gly分别为(15.30±3.62),(23.63±6.22)mg·L^(-1),模型组分别为(12.68±1.63),(18.21±4.91)mg·L^(-1),差异均有统计学意义(均P<0.05)。处理后3 d,实验组ICAM、VCAM^(-1)水平与模型组相比明显降低。结论杏芎氯化钠注射液能够改善缺血再灌注大鼠脑内血小板活性,提高血液中t PA含量,提高脑缺血组织总抗氧化活力,降低自由基水平,抑制脂质过氧化,维持兴奋/抑制性氨基酸的动态平衡,降低黏附分子的表达,从而保护受损的神经元。

Objective To observe the effects of Floium Ginkgo Extract and Tertram Ethypyrazine Sodium Chloride injection on granule membrane protein 140（ GMP-140）,tissue-type plasminogen activator（ t PA）,free radicals,amino acid homeostasis and effects of adhesion molecules in rats with focal cerebral ischemia-reperfusion injury.Methods Adult male Sprague-Dawley rats were randomly divided into sham-operation group,model group,control group and experimentalgroup. Enrolled rats were subjected to middle cerebral artery occlusion（ MCAO）. The sham-operation group was given the same operation steps as model group except not blocking the blood vessels. Both groups were given normal saline 3 m L·kg^（-1）; control group was given Ginaton solution 15 mg·kg^（-1）; experiential group was given Floium Ginkgo Extract and Tertram Ethypyrazine Sodium Chloride injection 41. 75 m L·kg^（-1）,once a day. The activity of GMP-140 and t PA were determined by the chromogenic substrate method. The superoxide dismutase（ SOD） activity was measured by xanthine oxidase method. The content of malondialdehyde（ MDA） was determined by thiobarbituric acid method.The concentrations of amino acids L-glutamic acid（ L-Glu）,L-glycine（ L-Gly）,L-aminosuccinic acid（ L-Asp）,γ-aminobutyric acid（ GABA） in brain tissue were determined by liquid chromatography. Results The levels of GMP-140 in control group and experiential group on 7 d after treatment were（ 7. 40 ± 2. 90） and（ 6. 00 ± 2. 90）ng·m L^（-1）,the levels of GMP-140 in model group was（ 20. 40 ± 5. 80） ng · m L^（-1）,with significant difference（ P 0. 05）. The expressions of t PA in control group and experiential group were（ 12. 40 ± 2. 10） and（ 6. 60 ± 1. 40）ng·m L^（-1） on 7 d after treatment,with significant difference（ P 0. 05）. Compared with sham-operation group and model group,the activity of SOD significantly increased in experimental group and control group,while the content of MDA significantly reduced（ P 0. 01）. There is no statistical difference between experimental group and control group（ P 0. 05）. Compared with model group,the levels of L-Glu and L-Asp were significantly reduced（ P 0. 05）.The production and release of γ-aminobutyric acid（ GABA） and L-Gly were（ 15. 30 ± 3. 62） and（ 23. 63 ± 6. 22）mg·L^（-1） in experimental group,and were（ 12. 68 ± 1. 63）,（ 18. 21 ± 4. 91） mg·L^（-1） in model group,with significant difference（ P 0. 05）. Compared with model groups,the expressions of intercellular adhesion molecule^（-1）（ ICAM^（-1））,vascular cell adhesion molecule^（-1）（ VCAM^（-1）） were significantly reduced in experimental group on 3 d after treatment. Conclusion Floium Ginkgo Extract and Tertram Ethypyrazine Sodium Chloride injection can improve the platelet activity in rat brain after ischemia-reperfusion,increase the content of t PA in blood,increase the total antioxidant activity of cerebral ischemia,decrease the level of free radicals and inhibit the lipid peroxidation,maintain the excitatory/inhibitory amino acid homeostasis,and reduce the expression of adhesion molecules,thus protecting damaged neurons.