藏药七十味珍珠丸灌胃给予脑缺血再灌注损伤模型大鼠的量-时-效关系研究
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摘要
目的:研究七十味珍珠丸治疗大鼠脑缺血再灌注损伤的量-时-效关系。方法:将大鼠随机分为假手术组(生理盐水,10mL/kg)、模型对照组(生理盐水,10 mL/kg)、阳性对照组(尼莫地平,30 mg/kg)、七十味珍珠丸不同剂量组(0.52、1.04、2.08、4.17、8.33、16.67、33.34、66.68、133.36、266.72、533.44 mg/kg),每组18只,各组大鼠灌胃相应药物1次;灌胃25 min后,除假手术组外,其余各组大鼠均用线栓法制造脑缺血再灌注损伤模型。脑缺血24、48、72 h后,对各组大鼠进行神经行为学评级,检测大鼠脑梗死率以评价脑缺血各时段七十味珍珠丸的最佳有效时间点、最佳给药剂量(D_(max))和最大效应(最小脑梗死率,E_(max)),然后采用Thermo Kinetica 5.1软件对七十味珍珠丸给药剂量与脑梗死率进行量-时-效关系拟合,计算量效曲线下面积(AUClast)、滞留剂量(MRTlast);并检测各组大鼠血清中超氧化物歧化酶(SOD)、丙二醛(MDA)的水平。结果:与假手术组比较,模型组大鼠神经行为显著异常(P<0.05或P<0.01),脑梗死率明显增加(P<0.01),大鼠血清中SOD水平显著降低(P<0.01,48 h),MDA水平显著升高(P<0.05,48 h)。与模型对照组比较,尼莫地平组神经行为异常无显著变化(P>0.05),脑梗死率显著降低(P<0.01,24、48 h),大鼠血清中SOD水平显著升高(P<0.01,48 h),MDA水平显著降低(P<0.05,48 h);七十味珍珠丸2.08～33.34 mg/kg剂量组神经行为异常显著改善(P<0.05,24 h);脑缺血24 h,七十味珍珠丸4.17～133.36 mg/kg剂量组脑梗死率均显著降低(以33.34 mg/kg剂量组最低,P<0.05或P<0.01),33.34～533.44 mg/kg剂量组大鼠血清中SOD水平均显著升高(P<0.05),0.52～2.08、8.33、33.34、266.72、533.44 mg/kg剂量组MDA水平均显著降低(P<0.05),D_(max)为33.34 mg/kg、E_(max)为3.02%、AUClast为5 141.76 mg/kg、MRTlast为329.161 mg/kg;脑缺血48 h,七十味珍珠丸2.08～133.36 mg/kg剂量组脑梗死率均显著降低(以66.68 mg/kg剂量组最低,P<0.05或P<0.01),1.04～533.44(除4.17外)mg/kg剂量组大鼠血清中SOD水平均显著升高(P<0.05),16.67～66.68、533.44 mg/kg剂量组大鼠血清中MDA水平均显著降低(P<0.05),D_(max)为66.68 mg/kg、E_(max)为2.13%、AUClast为5 219.36 mg/kg、MRTlast为340.521 mg/kg;脑缺血72 h,七十味珍珠丸各剂量组脑梗死率、MDA水平均无显著降低(P>0.05),SOD水平均无显著升高(除0.52 mg/kg剂量组外,P>0.05)。结论:七十味珍珠丸治疗大鼠脑缺血再灌注损伤的最佳有效时间点为48 h、D_(max)为66.68 mg/kg,其改善机制可能与增强SOD水平、降低MDA水平有关。
OBJECTIVE:To study the dose-time-effect relationship of Tibetan medicine Rannasangpei in cerebral ischemicreperfusion injury model rats with intragastric administration. METHODS:The rats were randomly divided into sham operation group(normal saline,10 mL/kg),model control group(normal saline,10 mL/kg),positive control group(nimodipine,30mg/kg),Rannasangpei different dose groups(0.52,1.04,2.08,4.17,8.33,16.67,33.34,66.68,133.36,266.72 and 533.44mg/kg),with 18 rats in each group. Each group was given relevant medicine intragastrically once;25 min after intragastric administration,cerebral ischemic-reperfusion injury model was established with suture-occluded method in those groups except for sham operation group. 24,48,72 h after cerebral ischemia,neuroethology of rats were graded in each group. The rate of cerebral infraction was detected to evaluate the optimal effective time,the optimal dose(D_(max)) and maximal effect(the rate of minimum cerebral infraction,E_(max))of Ratnasampil at different periods of cerebral ischemia. Dose-time-effect relationship of Rannasangpei dose with the rate of cerebral infraction was fitted with Thermo Kinetica 5.1 software. The area under curve(AUClast)and retention dose(MRTlast)of dose-effect curve were calculated,and detect the levels of SOD and MDA. RESULTS:Compared with sham operation group,the neurobehavior of model group was significantly abnormal(P<0.05 or P<0.01), and the rate of cerebral infarction was significantly increased(P<0.01); the level of SOD was decreased significantly(P<0.01,48 h),and the level MDA was increased significantly(P<0.05,48 h). Compared with model control group,there was no significant change in neurobehavioral abnormalities in the nimodipine group(P>0.05),and the rate of cerebral infraction was decreased significantly(P<0.01,24,48 h). The level of SOD in rats were increased significantly(P<0.01,48 h),while the level MDA decreased significantly(P<0.05,48 h). Rannasangpei 2.08-33.34 mg/kg could significantly improved neurobehavioral abnormalities(P<0.05,24 h);24 h after cerebral ischemia,the rate of cerebral infraction was decreased significantly in Rannasangpei 4.17-133.36 mg/kg group(the lowest is 33.34 mg/kg group,P<0.05 or P<0.01). The level of SOD in rats were increased significantly in 33.34-533.44 mg/kg groups(P<0.05). the level MDA was decreased significantly in 0.52-2.08,8.33,33.34,266.72 and 533.44 mg/kg groups(P<0.05). D_(max)was 33.34 mg/kg,E_(max)was 3.02%,AUClast was 5 141.76 mg/kg and MRTlast was 329.161 mg/kg. 48 h after cerebral ischemia,the rate of cerebral infraction was decreased significantly in Rannasangpei 2.08-133.36 mg/kg groups(the lowest is 66.68 mg/kg group,P<0.05 or P<0.01),while the level of SOD was increased significantly in 1.04-533.44(except for 4.17)mg/kg groups(P<0.05). The level of MDA was decreased significantly in 16.67-66.68,533.44 mg/kg groups(P<0.05),D_(max)was 66.68 mg/kg,E_(max)was 2.13%,AUClast was5 219.36 mg/kg and MRTlast was 340.521 mg/kg. 72 h after cerebral ischemia,the rate of cerebral infraction and the level of MDA had no significant decreased in Rannasangpei groups(P>0.05),and the levels of SOD had no significant increase(except for 0.52mg/kg group, P>0.05). CONCLUSIONS: The optimal effective time of Rannasangpei for the treatment of cerebral ischemia-reperfusion injury in rats is 48 h,and the D_(max)is 66.68 mg/kg. The improvement mechanism may be related to increase the level of SOD and decrease the level of MDA.
OBJECTIVE:To study the dose-time-effect relationship of Tibetan medicine Rannasangpei in cerebral ischemicreperfusion injury model rats with intragastric administration. METHODS:The rats were randomly divided into sham operation group(normal saline,10 mL/kg),model control group(normal saline,10 mL/kg),positive control group(nimodipine,30mg/kg),Rannasangpei different dose groups(0.52,1.04,2.08,4.17,8.33,16.67,33.34,66.68,133.36,266.72 and 533.44mg/kg),with 18 rats in each group. Each group was given relevant medicine intragastrically once;25 min after intragastric administration,cerebral ischemic-reperfusion injury model was established with suture-occluded method in those groups except for sham operation group. 24,48,72 h after cerebral ischemia,neuroethology of rats were graded in each group. The rate of cerebral infraction was detected to evaluate the optimal effective time,the optimal dose(D_(max)) and maximal effect(the rate of minimum cerebral infraction,E_(max))of Ratnasampil at different periods of cerebral ischemia. Dose-time-effect relationship of Rannasangpei dose with the rate of cerebral infraction was fitted with Thermo Kinetica 5.1 software. The area under curve(AUClast)and retention dose(MRTlast)of dose-effect curve were calculated,and detect the levels of SOD and MDA. RESULTS:Compared with sham operation group,the neurobehavior of model group was significantly abnormal(P<0.05 or P<0.01), and the rate of cerebral infarction was significantly increased(P<0.01); the level of SOD was decreased significantly(P<0.01,48 h),and the level MDA was increased significantly(P<0.05,48 h). Compared with model control group,there was no significant change in neurobehavioral abnormalities in the nimodipine group(P>0.05),and the rate of cerebral infraction was decreased significantly(P<0.01,24,48 h). The level of SOD in rats were increased significantly(P<0.01,48 h),while the level MDA decreased significantly(P<0.05,48 h). Rannasangpei 2.08-33.34 mg/kg could significantly improved neurobehavioral abnormalities(P<0.05,24 h);24 h after cerebral ischemia,the rate of cerebral infraction was decreased significantly in Rannasangpei 4.17-133.36 mg/kg group(the lowest is 33.34 mg/kg group,P<0.05 or P<0.01). The level of SOD in rats were increased significantly in 33.34-533.44 mg/kg groups(P<0.05). the level MDA was decreased significantly in 0.52-2.08,8.33,33.34,266.72 and 533.44 mg/kg groups(P<0.05). D_(max)was 33.34 mg/kg,E_(max)was 3.02%,AUClast was 5 141.76 mg/kg and MRTlast was 329.161 mg/kg. 48 h after cerebral ischemia,the rate of cerebral infraction was decreased significantly in Rannasangpei 2.08-133.36 mg/kg groups(the lowest is 66.68 mg/kg group,P<0.05 or P<0.01),while the level of SOD was increased significantly in 1.04-533.44(except for 4.17)mg/kg groups(P<0.05). The level of MDA was decreased significantly in 16.67-66.68,533.44 mg/kg groups(P<0.05),D_(max)was 66.68 mg/kg,E_(max)was 2.13%,AUClast was5 219.36 mg/kg and MRTlast was 340.521 mg/kg. 72 h after cerebral ischemia,the rate of cerebral infraction and the level of MDA had no significant decreased in Rannasangpei groups(P>0.05),and the levels of SOD had no significant increase(except for 0.52mg/kg group, P>0.05). CONCLUSIONS: The optimal effective time of Rannasangpei for the treatment of cerebral ischemia-reperfusion injury in rats is 48 h,and the D_(max)is 66.68 mg/kg. The improvement mechanism may be related to increase the level of SOD and decrease the level of MDA.
引文
[1]HU X,DE SILVA TM,CHEN J,et al.Genetic cerebral vascular disease and neurovascular injury in ischemic stroke[J].Circ Res,2017,120(3):449-471.
[2]汤海波,章剑,朱爱萍,等.2012-2015年江阴市脑卒中发病率及类型分析[J].江苏预防医学,2017,28(4):414-416.
[3]华俊.白果内酯对缺血性脑损伤的神经保护作用及其机制研究[D].南京:南京医科大学,2017.
[4]于秀华,周鸣,邓毅峰.脑心舒胶囊抗脑缺血作用研究[J].中国药房,2010,21(35):3282-3284.
[5]徐文龙,孙位军,王张,等.七十味珍珠丸抗大鼠脑缺血再灌注损伤的量效关系研究[J].现代药物与临床,2017,32(1):10-15.
[6]SCHMIDBUCHI S,HALFENS RJ,MULLER M,et al.Factors associated with supportive care needs of patients under treatment for breast cancer[J].Eur J Oncol Nurs,2013,17(1):22-29.
[7]颜娟,郑茂东,崔玉环,等.大黄酚脂质体对脑缺血再灌注损伤小鼠体内抗氧化作用的影响[J].中国老年学杂志,2018,38(3):703-704.
[8]洪森荣,尹明华.银杏叶提取物对缺血/再灌注小鼠脑细胞凋亡的保护作用[J].中草药,2007,38(12):1864-1867.
[9]毛继祖.藏医诊疗秘诀[M].兰州:甘肃人民出版社,2000:184.
[10]梁未雯,黄春娟,王张,等.藏药七十味珍珠丸对线栓致大鼠局灶性脑缺血再灌注损伤的时效关系初步研究[J].中药药理与临床,2015,31(1):182-187.
[11]王张,张艺,赵启鹏,等.灯盏细辛治疗脑缺血再灌注损伤的时-效关系初步研究[J].中药药理与临床,2012,28(2):62-64.
[12]LONGA EZ,WEINSTEIN PR,CARLSON S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[13]BEDERSON JB,PITTS LH,TSUJI M,et al.Rat middle cerebral artery occlusion:evaluation of the model and development of a neurologic examination[J].Stroke,1986,17(3):472-476.
[14]闫安,谢云亮.当归多糖对脑缺血再灌注损伤大鼠脑组织氧化应激水平及炎症因子表达的影响[J].中国实验方剂学杂志,2018,24(2):123-127.
[15]FISZER C,DOLBEAULT S,SULTAN S,et al.Prevalence,intensity,and predictors of the supportive care needs of women diagnosed with breast cancer:a systematic review[J].Psycho-Oncol,2014,23(4):361-374.
[16]CHEN SH,LIN MT,CHANG CP.Ischemic and oxidative damage to the hypothalamus may be responsible for heat stroke[J].Curr Neuropharmacol,2013,11(2):129-140.
[2]汤海波,章剑,朱爱萍,等.2012-2015年江阴市脑卒中发病率及类型分析[J].江苏预防医学,2017,28(4):414-416.
[3]华俊.白果内酯对缺血性脑损伤的神经保护作用及其机制研究[D].南京:南京医科大学,2017.
[4]于秀华,周鸣,邓毅峰.脑心舒胶囊抗脑缺血作用研究[J].中国药房,2010,21(35):3282-3284.
[5]徐文龙,孙位军,王张,等.七十味珍珠丸抗大鼠脑缺血再灌注损伤的量效关系研究[J].现代药物与临床,2017,32(1):10-15.
[6]SCHMIDBUCHI S,HALFENS RJ,MULLER M,et al.Factors associated with supportive care needs of patients under treatment for breast cancer[J].Eur J Oncol Nurs,2013,17(1):22-29.
[7]颜娟,郑茂东,崔玉环,等.大黄酚脂质体对脑缺血再灌注损伤小鼠体内抗氧化作用的影响[J].中国老年学杂志,2018,38(3):703-704.
[8]洪森荣,尹明华.银杏叶提取物对缺血/再灌注小鼠脑细胞凋亡的保护作用[J].中草药,2007,38(12):1864-1867.
[9]毛继祖.藏医诊疗秘诀[M].兰州:甘肃人民出版社,2000:184.
[10]梁未雯,黄春娟,王张,等.藏药七十味珍珠丸对线栓致大鼠局灶性脑缺血再灌注损伤的时效关系初步研究[J].中药药理与临床,2015,31(1):182-187.
[11]王张,张艺,赵启鹏,等.灯盏细辛治疗脑缺血再灌注损伤的时-效关系初步研究[J].中药药理与临床,2012,28(2):62-64.
[12]LONGA EZ,WEINSTEIN PR,CARLSON S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[13]BEDERSON JB,PITTS LH,TSUJI M,et al.Rat middle cerebral artery occlusion:evaluation of the model and development of a neurologic examination[J].Stroke,1986,17(3):472-476.
[14]闫安,谢云亮.当归多糖对脑缺血再灌注损伤大鼠脑组织氧化应激水平及炎症因子表达的影响[J].中国实验方剂学杂志,2018,24(2):123-127.
[15]FISZER C,DOLBEAULT S,SULTAN S,et al.Prevalence,intensity,and predictors of the supportive care needs of women diagnosed with breast cancer:a systematic review[J].Psycho-Oncol,2014,23(4):361-374.
[16]CHEN SH,LIN MT,CHANG CP.Ischemic and oxidative damage to the hypothalamus may be responsible for heat stroke[J].Curr Neuropharmacol,2013,11(2):129-140.
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