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电针“夹脊”穴对急性脊髓损伤大鼠少突胶质前体细胞增殖与分化的影响

[Effect of electroacupuncture at "Jiaji" (EX-B 2) points on the proliferation and differentiation of oligodendrocyte precursor cells in rats with acute spinal cord injury].

作者信息

Hu Rong, Chen Yi, Xu Hai-Peng, He Ke-Lin, Sun Lian-Zhu, Wu Lei, Ma Rui-Jie

机构信息

Third Clinical Medical College of Zhejiang Chinese Medicine University, Hangzhou 310053, China.

Third Clinical Medical College of Zhejiang Chinese Medicine University, Hangzhou 310053, China; Department of Acupuncture and Moxibustion, Third Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou 310053.

出版信息

Zhongguo Zhen Jiu. 2020 May 12;40(5):519-25. doi: 10.13703/j.0255-2930.20190326-0008.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) at "Jiaji" (EX-B 2) points on the proliferation and differentiation of oligodendrocyte precursor cells in rats with acute incomplete spinal cord injury, and to explore the mechanism of EA on improving motor function of spinal cord injury.

METHODS

A total of 72 male SPF SD rats were randomly divided into a sham operation group, a model group, an EA group and a medication group, 18 rats in each group. Each group was further divided into 1-day subgroup, 7-day subgroup and 14-day subgroup, 6 rats in each subgroup. The T acute incomplete spinal cord injury model was established by modified Allen's method in the model group, EA group and medication group. The rats in each group received intraperitoneal injection of 5-bromodeoxyuridine (BrdU, 50 mg/kg), once a day, and each subgroup received continuous injection for 1, 7, 14 times for cell proliferation labeling. The rats in the EA group were treated with EA at "Jiaji" (EX-B 2) points 3-4 mm next the spinous process of the upper and lower segments of the injured spinal cord (T, T) with a frequency of 2 Hz/100 Hz and intensity of 1-2 mA. The muscle twitch at the treatment site was taken as the degree. The treatment was given 20 min each time, once a day. In the medication group, monosialogangliosides (GM1) was injected intraperitoneally (10 mg/kg), once a day. The subgroups of EA group and medication group were treated for 1, 7, 14 times. The score of Basso Beattie Bresnahan (BBB) was used to evaluate the motor function of hind limbs. The co-expression of BrdU/NG2 positive cells was detected by immunofluorescence, and the expression of Olig2 and Sox10 was detected by Western blot.

RESULTS

Compared with the sham operation group, the BBB score was decreased 1 day, 7 days and 14 days after operation in the model group (<0.05), the expression of Olig2 and Sox10 was increased (<0.05), and the co-expression of BrdU/NG2 positive cells was increased 7 days and 14 days after operation (<0.05). Seven days and 14 days after operation, the BBB score in the EA group and medication group was higher than that in the model group (<0.05), and the co-expression of BrdU/NG2 in the medication group was higher than that in the model group (<0.05). Fourteen days after operation, the co-expression of BrdU/NG2 in the EA group was higher than that in the model group (<0.05); 1 day, 7 days and 14 days after operation, the expression of Olig2 and Sox10 in the EA group and medication group was higher than that in the model group (<0.05). Compared with the medication group, the co-expression of BrdU/NG2 positive cells in the EA group 14 days after operation was decreased (<0.05); 1 day, 7 days and 14 days after operation, the expression of Olig2 and Sox10 in the EA group was decreased (<0.05).

CONCLUSION

EA at "Jiaji" (EX-B 2) points could promote the expression of Olig2 and Sox10 after spinal cord injury, which has similar effects with GM1. It could promote the proliferation and differentiation of oligodendrocyte precursor cells into oligodendrocytes, so as to promote the recovery of motor function of rats.

摘要

目的

观察电针“夹脊”(EX - B 2)穴对急性不完全性脊髓损伤大鼠少突胶质前体细胞增殖与分化的影响,探讨电针改善脊髓损伤后运动功能的机制。

方法

将72只雄性SPF级SD大鼠随机分为假手术组、模型组、电针组和药物组,每组18只。每组再分为1天亚组、7天亚组和14天亚组,每组6只。模型组、电针组和药物组采用改良Allen法建立T急性不完全性脊髓损伤模型。各组大鼠腹腔注射5-溴脱氧尿嘧啶核苷(BrdU,50 mg/kg),每日1次,各亚组连续注射1、7、14次进行细胞增殖标记。电针组大鼠于损伤脊髓上下节段棘突旁开3 - 4 mm处的“夹脊”(EX - B 2)穴进行电针治疗,频率为2 Hz/100 Hz,强度为1 - 2 mA,以治疗部位肌肉出现轻微抽动为度,每次治疗20 min,每日1次。药物组腹腔注射单唾液酸四己糖神经节苷脂(GM1,10 mg/kg),每日1次。电针组和药物组各亚组分别治疗1、7、14次。采用Basso Beattie Bresnahan(BBB)评分评价后肢运动功能。免疫荧光法检测BrdU/NG2阳性细胞的共表达情况,Western blot法检测Olig2和Sox10的表达。

结果

与假手术组比较,模型组术后1天、7天和14天BBB评分降低(<0.05),Olig2和Sox10表达升高(<0.05),术后7天和14天BrdU/NG2阳性细胞共表达增加(<0.05)。术后7天和14天,电针组和药物组BBB评分高于模型组(<0.05),药物组BrdU/NG2共表达高于模型组(<0.05)。术后14天,电针组BrdU/NG2共表达高于模型组(<0.05);术后1天、7天和14天,电针组和药物组Olig2和Sox10表达高于模型组(<0.05)。与药物组比较,术后14天电针组BrdU/NG2阳性细胞共表达降低(<0.05);术后1天、7天和14天,电针组Olig2和Sox10表达降低(<0.05)。

结论

电针“夹脊”(EX - B 2)穴可促进脊髓损伤后Olig2和Sox10的表达,与GM1作用相似,可促进少突胶质前体细胞增殖分化为少突胶质细胞,从而促进大鼠运动功能恢复。

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