川芎嗪-丹参-当归配伍对大鼠脑缺血治疗时间窗的影响

Affect therapeutic time window of combined ligustrazine with salvia miltiorrhiza and angelica sinensis diels on cerebral ischemia/reperfusion in rats

目的研究川芎嗪-丹参-当归配伍对大鼠脑缺血治疗时间窗影响。方法应用线栓法制作大鼠MCAO模型。尾静脉给药对大鼠脑缺血模型进行干预。将60只成年雄性Wistar大鼠随机分成中药组和对照组。再将中药组和对照组随机分成5个小组,即MCAO2h,3h,4h,5h和6h组。中药组:川芎嗪(0.35mg/100g体重)+丹参(200mg/100g体重)+当归(165mg/100g体重)溶解于总体积1.5ml的等渗盐水中。对照组:等渗盐水1.5ml。给药时间:再灌注前20min、再灌注后12h和36h。再灌注48h后计算大鼠存活率、行神经功能缺损评分、脑血流量和脑梗死体积测定。结果大鼠存活率:中药组总存活率70%;各小组存活率分别为100.0%、83.3%、83.3%、66.7%和16.7%。对照组总存活率40%;各小组存活率分别为83.3%、50.0%、33.3%、16.7%和16.7%。神经功能评分:中药组2.12±0.38、2.52±0.45、2.69±0.51、3.39±0.51和4.63±0.71;对照组3.81±0.22、3.82±0.33、4.13±0.12、4.31±0.42和4.51±0.31。脑血流量变化相对值(%):中药组(155±31.42)、(139±39.53)、(129±31.69)、(113±29.98)和(83±41.63);对照组(93±519.12)、(87±29.22)、(81±29,.20)、(73±21.19)和(79±39.53)。大鼠脑梗死体积(mm3):中药组(128.2±65.2)、(183.1±78.2)、(221.5±41.3)、(347.1±37.7)和(403.12±61.8);对照组(183.2±71.3)、(249.6±37.2)、(347.5±14.4)、(381.3±12.0)和(415.61±32.7)。所有观测指标,各中药组与相应对照组(除MCAO6h组)比较,差异均有统计学意义(P<0.05或P<0.01)。结论川芎嗪-丹参-当归对大鼠脑缺血-再灌注损伤模型的治疗时间窗影响在2h～5h范围内。

Objective To investigate the therapeutic time window of Ligustrazine with Salvia miltiorrhiza and Angelica sinensis Diels in reperfusion injury after focal cerebral ischemia in rats. Methods Sixty adult male Wistar rats were randomly divided into two groups,the medicine group and saline control group. The two groups were once more divided into middle cerebral artery occlusion （MCAO） 2h,3h,4h,5h,and 6h groups （6/group）. A rat model of MCAO was induced by the intraluminal suture method;reperfusion was performed after cerebral ischemia 2h,3h,4h,5h,and 6h,respectively. Following success of MCAO model,each medicine group was administered at 20 minutes before reperfusion after cerebral ischemia,12h and 36h after reperfusion with Ligustrazine 0.35mg/100g plus Salvia miltiorrhiza 200mg/100g plus Angelica sinensis Diels 165mg/ 100g and isotonic saline solution 1.5mL,and the control group was administered isotonic saline solution 1.5mL.The survival rate of rats was calculated. Meanwhile,the volumes of cerebral infarction by TTC staining,neurological functional deficits were evaluated and cerebral blood flow was measured at 48h after the reperfusion. Results The survival rates of rats in the two group rats were 70% and 40%. The survival rates of rats in the medicine group for five MCAO times were： 100.0%,83.3%,83.3%,66.7% and 16.7%. And the control group：83.3%,50.0%,33.3%,16.7%,and 16.7%,respectively. Neurological deficit scores in the medicine group rats were： 2.12± 0.38,2.52± 0.45,2.69± 0.51,3.39±0.51,and 4.63± 0.71. And control group were： 3.81± 0.22,3.82± 0.33,4.13± 0.12,4.31±0.42,and 4.51±0.31,respectively. The relative magnitude of cerebral blood flow in the medicine group rats were： （155±31.42）,（139±39.53）,（129±31.69）,（113±29.98）,and （83±41.63） %. And the control group were： （93± 519.12）,（87± 29.22）,（81±29.20）,（73± 21.19） and （79±39.53） %. The volumes of cerebral infarction in the medicine group rats were： （128.2±65.2）,（183.1± 78.2）,（221.5±41.3）,（347.1±37.7）,and （403.12±61.8） mm3. And the control group were： （183.2±71.3）,（249.6± 37.2）,（347.5±14.4）,（381.3±12.0） and （415.61±32.7） mm3. There was statistical significance of all surveying between the medicine and control group （P0.05 or P0.01） in different times of MCAO （except MCAO 6h group）. Conclusion The neuroprotective effect of combined Ligustrazine with Salvia miltiorrhiza and Angelica sinensis Diels on focal cerebral ischemia/reperfusion injury in rats were 2h to 5h.