川芎嗪-灯盏乙素苷元孪药对脑缺血再灌注损伤模型大鼠的保护作用及机制研究

Study on protective effects of twin drugs of tetramethylpyrazine-scutellarein on cerebral ischemiareperfusion injury model rats and its mechanism

目的 研究川芎嗪-灯盏乙素苷元孪药(TMSC4)对脑缺血再灌注损伤(CIRI)模型大鼠的保护作用及机制。方法 将105只SD大鼠随机分为假手术组、模型组、灯盏乙素苷元组(0.7 mmol/kg)、川芎嗪组(0.7 mmol/kg)和TMSC4低、中、高剂量组(0.35、0.7、1.4 mmol/kg),每组15只。假手术组、模型组大鼠灌胃等体积生理盐水,其余各组大鼠灌胃相应药物,每天1次,连续14 d。除假手术组外,其余各组大鼠均采用线栓法建立CIRI模型。大鼠缺血2 h再灌注22 h后,测定大鼠脑指数及脑含水量;检测大鼠血清中白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)的水平,脑组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化氢酶(GSH-Px)、过氧化氢酶(CAT)的水平和大脑皮层神经细胞的凋亡情况以及B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化型胱天蛋白酶3(cleaved-caspase-3)蛋白表达水平。结果 与假手术组比较,模型组大鼠脑指数和脑含水量,血清中IL-1β、IL-6、TNF-α水平以及脑组织中MDA水平,神经细胞凋亡率,Bax、cleaved-caspase-3蛋白表达水平均显著升高(P<0.05);脑组织中SOD、 GSH-Px、CAT水平和Bcl-2蛋白表达水平均显著降低(P<0.05)。与模型组比较,各给药组大鼠上述指标水平均显著逆转(P<0.05),且TMSC4中、高剂量组的逆转作用显著优于灯盏乙素苷元组和川芎嗪组(P<0.05)。结论 TMSC4对CIRI模型大鼠具有保护作用,其作用机制可能与减轻炎症反应、氧化应激损伤,抑制神经细胞凋亡有关。

OBJECTIVE To study the protective effects of twin drugs of tetramethylpyrazine-scutellarein(TMSC4)on cerebral ischemia-reperfusion injury(CIRI)model rats and its mechanism.METHODS One hundred and five SD rats were randomly divided into sham operation group,model group,scutellarein group(0.7 mmol/kg),tetramethylpyrazine group(0.7 mmol/kg),and TMSC4 low-dose,medium-dose and high-dose groups(0.35,0.7,1.4 mmol/kg),with 15 rats in each group.Sham operation group and model group were given constant volume of normal saline intragastrically,and other groups were given relevant drug intragastrically,once a day,for consecutive 14 d.Except for sham operation group,all other groups were treated to establish the CIRI model using the thread occlusion method.After 2 hours of ischemia and 22 hours of reperfusion,the brain index and brain water content of the rats were measured.Serum levels of interleukin 1β(IL-1β),IL-6 and tumor necrosis factorα(TNF-α),the levels of superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px)and catalase(CAT)in brain tissues,the situation of neuronal cell apoptosis,and the protein expressions of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax)and cleaved-caspase-3 were evaluated.RESULTS Compared with sham operation group,the brain index,brain water content,the serum levels of IL-1β,IL-6 and TNF-α,the levels of MDA in brain tissues,the brain cell apoptosis and the protein expressions of Bax and cleaved-caspase-3 in model group were significantly increased(P<0.05);the levels of SOD,GSH-Px and CAT and the protein expression of Bcl-2 in brain tissues were significantly decreased(P<0.05).Compared with model group,the above indexes of rats were reversed significantly in administration groups(P<0.05),while the reverse effects of TMSC4 medium-dose and high-dose groups were significantly better than those of scutellarein group and tetramethylpyrazine group(P<0.05).CONCLUSIONS TMSC4 has a certain protective effect in CIRI model rats,the mechanism of which may be related to relieving inflammatory reaction and oxidative stress,inhibiting cell apoptosis.