摘要
目的:观察"百会"透"曲鬓"针刺法对脑出血大鼠侧脑室下区(SVZ)神经干细胞增殖的影响及Shh通路在此过程的作用。方法:SD大鼠随机分为空白组、模型组、针刺组、激动剂组,每组18只。采用自体血注入右侧大脑尾状核制备脑出血大鼠模型。针刺组予"百会"透"曲鬓"针刺法,激动剂组予腹腔注射Shh通路激动剂purmorphamine(1 mg/kg),两组均每天干预1次,连续7 d。观察各组治疗前后Bederson评分,HE染色法观察出血区脑组织病理损伤情况,免疫荧光法观察SVZ BrdU^+/Nestin^+细胞数,免疫组织化学法检测基底节部Shh通路相关因子Shh、Gli1的表达。结果:与同组治疗前比较,治疗后模型组、针刺组、激动剂组Bederson评分显著降低(P<0.05,P<0.001);与空白组比较,治疗前模型组、针刺组、激动剂组Bederson评分显著升高(P<0.001),治疗后模型组Bederson评分显著升高(P<0.001);与模型组比较,治疗后针刺组、激动剂组Bederson评分显著降低(P<0.05)。模型组脑出血组织周围红细胞浸润,有水肿带形成;针刺组和激动剂组可见陈旧性出血灶形成,组织结构排列紊乱。与空白组比较,模型组大鼠基底节部Shh、Gli1阳性表达显著升高(P<0.001);与模型组比较,针刺组和激动剂组SVZ BrdU^+/Nestin^+细胞数明显增多(P<0.001),基底节部Shh、Gli1阳性表达显著升高(P<0.001)。结论:"百会"透"曲鬓"针刺法可明显改善脑出血大鼠神经功能,这可能与其激活Shh通路,继而促进SVZ神经干细胞增殖有关。
Objective To observe the effect of penetrative needling from "Baihui"(GV20) to "Qubin"(GB7) on neural stem cell proliferation and sonic hedgehog(Shh) signaling in subventricular zone(SVZ) in intracerebral hemorrhage(ICH) rats so as to explore its mechanisms underlying improvement of ischemic injury of brain.Methods Male SD rats were randomly divided into blank control, model, acupuncture and agonist(Purmorphamine, an activator of Shh signaling pathway) groups(n=18 in each group, 6 for H.E. stain, 6 for examining neuronal cell proliferation, and 6 for immunohistochemistry). The ICH model was established by injecting autogenous blood(50 μL) into the right caudate nucleus. The neurological defect was scored with refe-rence to Bederson’s method. Penetrative needling from GV20 to GB7 was performed by manipulating the needle for 6 min(repeated 3 times in 30 min), once daily for 7 days. Intraperitoneal injection of Purmorphamine(1 mg/mL, 1 mg/kg) was performed, once daily for 7 days. Histopathological changes of the hemorrhagic penumbra region were observed under microscope after H.E. stain, the newborn neural stem cell proliferation(BrdU^+/Nestin^+ double labeled cells) in the SVZ was observed by immunofluorescence after intraperitoneal injection of BrdU(50 mg/kg), and the expression of Shh and glioma-associated hemolog-1(Gli1) detected by immunohistochemistry.Results After modeling, the neurological score and expression levels of Shh and Gil1 proteins were significantly increased in the model group relevant to the blank control group(P<0.001). Following the interventions, the neurological score was evidently decreased(P<0.05), while the number of BrdU^+/Nestin^+ double labeled cells and the expression levels of Shh and Gil1 proteins were significantly increased in both acupuncture and agonist groups in comparison with the model group(P<0.001). No significant differences were found between the acupuncture and agonist groups in down-regulating the neurological score and in up-regulating the number of BrdU^+/Nestin^+
作者
张菶
刘昊
李孟
孙晓伟
ZHANG Beng;LIU Hao;LI Meng;SUN Xiao-wei(Heilongjiang University of Chinese Medicine,Harbin 150040,China;Zhejiang Provincial Tongde Hospital,Hangzhou 310012;First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040)
出处
《针刺研究》
CAS
CSCD
北大核心
2020年第2期93-98,共6页
Acupuncture Research
基金
国家自然科学基金青年项目(No.81804168)
黑龙江中医药大学校科研基金项目(No.201825)
关键词
针刺
脑出血
神经干细胞增殖
SHH信号通路
侧脑室下区
Acupuncture
Intracerebral hemorrhage
Neural stem cell proliferation
Shh/Gil1 signaling
Subventricular zone
