机构地区:[1]浙江中医药大学附属杭州市中医院,杭州310007
出 处:《浙江中医药大学学报》2024年第6期655-663,共9页Journal of Zhejiang Chinese Medical University
基 金:浙江省中医药科技计划项目(2022ZA110)。
摘 要:[目的]探究舒肌汤治疗帕金森病(Parkinson’s disease,PD)大鼠肌张力障碍的最佳配伍比例。[方法]采用6-羟基多巴胺诱导制备大鼠PD肌张力障碍模型,以正交试验法设计实验,随机分为模型组(等体积0.9%氯化钠溶液)、舒肌汤1-16组[按L;16;(2;15;)正交试验表配伍比例给药],同时设假手术组(等体积0.9%氯化钠溶液)作为对照,给药1次/d,连续28 d。采用步进试验、悬挂试验和异常不自主运动量(abnormal involuntary movement,AIM)评分检测行为学表现;透射电镜观察膈肌神经接头超微结构变化;免疫组织化学检测脑组织酪氨酸羟化酶(tyrosine hydroxylase,TH)表达水平。[结果]正交试验结果显示,舒肌汤最佳配伍组方为舒肌汤4组即厚朴:青风藤:粉防己:毛叶轮环藤:山梗菜(1:1:2:2:1)。与假手术组比较,模型组大鼠调整步数显著下降,潜伏期和AIM评分显著升高(P<0.01);与模型组比较,舒肌汤组大鼠调整步数均显著升高、潜伏期和AIM评分均显著降低(P<0.01),其中舒肌汤4组作用效果最明显(P<0.01)。透射电镜观察显示,舒肌汤可减少突触前膜囊泡、线粒体数目和突触后膜褶皱,减轻神经元结构损伤。免疫组化显示,与模型组比较,舒肌汤组大鼠TH蛋白表达水平均显著升高(P<0.05),其中舒肌汤4组的TH蛋白表达水平最高(P<0.01)。[结论]舒肌汤治疗PD大鼠肌张力障碍的最佳配伍比例是厚朴:青风藤:粉防己:毛叶轮环藤:山梗菜=1:1:2:2:1。[Objective]To investigate the optimal ratio of Shuji Decoction for the treatment of dystonia in Parkinson's disease(PD)rats.[Methods]The model of PD dystonia was prepared by induction of 6-hydroxydopamine,and the experiment was designed by the orthogonal experimental design,and randomly divided into model group(equal volume of 0.9%sodium chloride solution),Shuji Decoction group 1~16[dosing ratio according to L_(16)(2~(15))orthogonal test table],and sham operation group(equal volume of 0.9%sodium chloride solution)as control,which was administered once/d for 28 d.The stepping test,suspension test and abnormal involuntary movement(AIM)were used to detect the degree of muscular rigidity.Ultrastructural changes of the neuromuscular junctions of the phrenic nerve were observed by transmission electron microscope.The expression level of tyrosine hydroxylase(TH)in brain tissue was measured by immunohistochemistry.[Results]The results of the orthogonal test showed that the best combination of Shuji Decoction was Shuji Decoction group 4,i.e.,Magnoliae Officinalis Cortex:Clematis chinensis Osbeck:Stephania tetrandra S.Moore:Cyclea barbata Miers:Lobelia sessilifolia Lamb in the ratio of 1:1:2:2:1.Compared with sham operation group,there was significantly decreased number of adjusting steps whereas increased latency and AIM scores of rats in model group(P<0.01);compared with model group,the number of adjusting steps was significantly increased,while latency and AIM scores of rats in Shuji Decoction groups were significantly reduced(P<0.01),among which the effect of No.4 Shuji Decoction group was the most obvious(P<0.01).The electron microscopic examination revealed that Shuji Decoction could reduce the number of presynaptic membrane vesicles,mitochondria and postsynaptic membrane folds,and relieve the structural damage of neurons.Immunohistochemistry showed that compared with model group,the expression level of TH was significantly increased in Shuji Decoction groups(P<0.05),and the expression level of TH in Shuji Decoction
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