摘要
目的探讨益气活血方(脑络欣通)对MCAO-R气虚血瘀证大鼠神经再生的影响及可能的作用机制。方法随机数字表法将大鼠分成4组:正常组、模型组、脑络欣通组、通心络组,其中脑络欣通组和通心络组分别用脑络欣通溶液、通心络溶液灌胃。观察各组大鼠生命状态并进行神经功能缺损、气虚证和血瘀证评分,运用激光多普勒仪动态监测局部脑血流量(r CBF),TTC染色检测脑梗死体积,免疫组化结合免疫荧光染色检测脑组织Nestin和Brd U表达,荧光定量PCR检测mi RNA-124水平,Western blotting检测Wnt3a、GSK3β和β-catenin蛋白表达水平。结果脑络欣通和通心络都能改善模型大鼠神经功能,降低气虚证和血瘀证评分,减少脑梗死体积,r CBF提高(P<0.01);脑络欣通和通心络组Brd U表达增强,与正常组比较,其他3组阳性细胞数明显增多(P<0.01),与模型组比较,脑络欣通和通心络组显著增多,Nestin表达明显增强(P<0.01)。脑络欣通能抑制模型大鼠脑组织mi RNA-124和Wnt3a应激性的高表达,同时提高β-catenin的表达水平(P<0.01)。但脑络欣通和通心络对模型大鼠GSK3β蛋白表达均无明显影响(P>0.05)。结论脑络欣通可能通过影响脑组织mi RNA-124的表达而激活Wnt通路,从而发挥对气虚血瘀型脑缺血的神经保护以及神经再生的作用。
Objective To investigate the effect of Yiqihuoxue prescription (NLXT) on nerve regeneration in MCAO-R rat models of qi deficiency and blood stasis syndrome and explore the underlying mechanisms. Methods The rats were randomized into 4 groups, namely the control group, model group, NLXT group and TXL group. The rats in NLXT group and TXL group were treated with gavage of NLXT and TXL solutions, respectively. The NFDS, QDSS and BSSS of the rats were evaluated. The regional cerebral blood flow (rCBF) were dynamically monitored with laser Doppler scanning, and the volume of cerebral infarction was detected with TTC-dye;the expression levels of nestin and BrdU were assayed with immunohistochemistry and mmunofluorescent staining. The expressions of miRNA-124, Wnt3a, GSK3βandβ-catenin in the rat brain tissue were detected with PCR or Western blotting. Results NLXT and TXL both improved the neurological functions of the model rats, reduced NFDS, QDSS, and BSSS scores, decreased the volume of cerebral infarction, and promoted the recovery of the rCBF (P〈0.01). Nestin and BrdU expression levels were significantly increased in the rat brain tissue in NLXT group and TXL group. NLXT significantly inhibited high expressions of miRNA-124 and Wnt3a in response to stress, and increased β-catenin expression level (P〈0.01). NLXT and TXL produced no obvious effect on GSK3βexpression in the model rats (P〉0.05). Conclusion NLXT can activate Wnt signaling by affecting miRNA-124 expression to offer neuroprotection and promote nerve regeneration in rats with cerebral ischemia with qi deficiency and blood stasis syndrome.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2017年第8期1047-1053,共7页
Journal of Southern Medical University
基金
国家自然科学基金青年项目(81603523)
安徽省高校自然科学研究重点项目(KJ2016A404)
国家科技支撑计划(2012BAI26B00)~~