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针刺干预对创伤性脑损伤大鼠神经功能、脑小胶质细胞活化及继发性神经损伤的影响

[Effect of acupuncture intervention on neurological function, cerebral microglia activation and secondary nerve damage in traumatic brain injury rats].

作者信息

Lin Shu-Jun, Zhang Yu-Juan, Lin Ji-Huan, Zhang Yi-Min, Zhu Ming-Min, Zhang Yu-Qing, Pu Liu, Chen Shu-Lian, Chen Wei-Hao, Cheng Shao-Bing

机构信息

College of Traditional Chinese Medicine of Jinan University, Guangzhou 510632, China.

Shi Shan Tang Clinic of Traditional Chinese Medicine, New Taipei City, Taiwan 23454, China.

出版信息

Zhen Ci Yan Jiu. 2019 Jan 25;44(1):19-24. doi: 10.13702/j.1000-0607.170475.

Abstract

OBJECTIVE

To observe the effect of acupuncture on activities of microglia in traumatic brain injury (TBI) rats.

METHODS

Fifty-four male SD rats were randomly and equally divided into normal control, model and acupuncture groups according to the random number table (=18 rats in each group). The TBI model was established by using a free fall brain injury striking device after exposing the local cranial bone (to induce the left parietal cerebral contusion). Acupoints "Baihui" (GV20), "Shuigou" (GV26), "Fengfu" (GV16), "Yamen" (GV15) and bilateral "Hegu" (LII4) were stimulated intensively by twirling the filiform needles with force at a range of >360° and a frequency of 160-180 cycles/min for 10 sec in every acupoint, once every 5 min during the 15 minutes' needle retaining. The treatment was given once every day for successive 14 days. The rats of the normal and model groups were grabbed and fixed with the same procedure. The behavioral changes were tested using modified neurological severity score (mNSS). The histopathological changes of the injured cerebral cortex tissues were observed by using hematoxylin-eosin (H.E.) staining, and the fluorescence intensity of Iba-1 (marker of microglia) positive products in the surrounding tissue of the cerebral focus was displayed by immunofluorescence staining, and the contents of neuron specific enolate (NSE) and neurite outgrowth inhibitor-A (Nogo-A) in serum (indicating a secondary nerve damage) were assayed by ELISA.

RESULTS

The mNSS scores were significantly increased on day 1, 3, 7 and 14 in the model group in comparison with the normal group (<0.01) and considerably decreased at the 4 time-points after acupuncture intervention relevant to the model group (<0.05, <0.01). H.E. staining showed that modeling induced pathological changes such as the excursion of cell nucleus, cellular swel-ling, vacuole-like change, neuron death, karyopyknosis dissolution, and proliferation of fibrous tissue were relatively milder in the acupuncture group. The average fluorescence intensity values of Iba-1-positive products, serum NSE and Nogo-A contents on day 3, 7 and 14 were significantly higher in the model group than in the normal group (<0.05, <0.01), and notably down-regulated in the acupuncture group than in the model group (<0.05, <0.01, except Nogo-A on day 3).

CONCLUSION

Acupuncture intervention may accelerate neurological function recovery in TBI rats, which is closely related to its effects in inhibiting the activation of microglia and secondary nerve damage.

摘要

目的

观察针刺对创伤性脑损伤(TBI)大鼠小胶质细胞活性的影响。

方法

54只雄性SD大鼠按随机数字表随机等分为正常对照组、模型组和针刺组(每组18只)。采用自由落体脑损伤打击装置暴露局部颅骨后建立TBI模型(致左侧顶叶脑挫伤)。用毫针强力捻转,幅度>360°,频率160~180次/分钟,在“百会”(GV20)、“水沟”(GV26)、“风府”(GV16)、“哑门”(GV15)及双侧“合谷”(LI4)穴位各强刺激10秒,留针15分钟期间每间隔5分钟行针1次。每天治疗1次,连续14天。正常组和模型组大鼠按同样操作抓取固定。采用改良神经功能缺损评分(mNSS)检测行为学变化。用苏木精-伊红(H.E.)染色观察损伤大脑皮质组织的组织病理学变化,用免疫荧光染色显示脑病灶周围组织中Iba-1(小胶质细胞标志物)阳性产物的荧光强度,并用酶联免疫吸附测定(ELISA)法检测血清中神经元特异性烯醇化酶(NSE)和神经突生长抑制因子-A(Nogo-A)含量(提示继发性神经损伤)。

结果

与正常组比较,模型组在第1、3、7和14天mNSS评分显著升高(<0.01),针刺干预后4个时间点与模型组比较显著降低(<0.05,<0.01)。H.E.染色显示,建模诱导的细胞核偏移细胞肿胀、空泡样改变、神经元死亡、核固缩溶解和纤维组织增生等病理变化在针刺组相对较轻。模型组第3、7和14天Iba-1阳性产物平均荧光强度值、血清NSE和Nogo-A含量显著高于正常组(<0.05,<0.01),针刺组与模型组比较显著下调(<0.0

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