“调脏通络”电针对糖尿病周围神经病变大鼠坐骨神经细胞中GRP78表达的影响及其神经保护作用

doi:10.13481/j.1671-587x.20180603

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (06): 1124-1130.doi: 10.13481/j.1671-587x.20180603

“调脏通络”电针对糖尿病周围神经病变大鼠坐骨神经细胞中GRP78表达的影响及其神经保护作用

潘鸿^1,2, 王宇峰¹, 黄海鹏¹, 张丽颖¹, 钟祯¹, 马诗棋¹, 董理¹, 王洪峰³

1. 长春中医药大学研究生学院, 吉林长春 130117;
2. 北京市第一中西医结合医院儿科, 北京 100018;
3. 长春中医药大学针灸研究所, 吉林长春 130117

收稿日期:2018-03-31 出版日期:2018-11-28 发布日期:2018-11-28
通讯作者: 王洪峰,教授,博士研究生导师(Tel:0431-86045206,E-mail:ccwhf@126.com) E-mail:ccwhf@126.com
作者简介:潘鸿(1980-),女,吉林省长春市人,副主任医师,医学博士,主要从事针灸治疗脏腑病方面的研究。
基金资助:
国家自然科学基金资助课题（81674051）；吉林省科技厅自然科学基金资助课题（20160101217JC）；吉林省教育厅"十三五"科学技术研究规划项目资助课题（JJKH20170727KJ）；吉林省教育厅"十三五"科学技术研究规划项目资助课题（JJKH20181243KJ）

Influence of Adjust Zang Dredge Meridian electroacupuncture in expression of GRP78 in sciatic nerve cells of rats with diabetic peripheral neuropathy and its neuroprotective effect

PAN Hong^1,2, WANG Yufeng¹, HUANG Haipeng¹, ZHANG Liying¹, ZHONG Zhen¹, MA Shiqi¹, DONG Li¹, WANG Hongfeng³

1. Graduate School, Changchun University of Chinese Medicine, Changchun 130117, China;
2. Department of Pediatrics, Fist Hospital of Integrated Chinese and Western Medicine, Beijing City, Beijing 100018, China;
3. Institute of Acupuncture, Changchun University of Chinese Medicine, Changchun 130117, China

Received:2018-03-31 Online:2018-11-28 Published:2018-11-28

摘要/Abstract

摘要： 目的：探讨糖尿病周围神经病（DPN）大鼠坐骨神经中葡萄糖调节蛋白78（GRP78）的表达，阐明"调脏通络"电针治疗DPN的临床意义及其机制。方法：86只成年雄性SD大鼠随机分为正常对照组（n=5）和模型组（n=81）。模型组大鼠均采用链脲佐菌素（STZ）一次注射法诱导建立糖尿病大鼠模型。将造模成功的大鼠随机分为模型对照组（n=27）、药物干预组（n=26）和电针干预组（n=26），正常对照组和模型对照组大鼠维持饲料喂养，不给予任何特殊处置。药物干预组大鼠给予甲钴胺灌胃，20 mg·kg^-1·d^-1，每日1次。电针干预组大鼠取双侧肺俞、脾俞、肾俞、合谷、足三里、三阴交和太冲进行"调脏通络"电针治疗，每日1次，每次30 min。检测各组大鼠热敏感度值、胫神经的运动神经传导速度（MCV）和感觉神经传导速度（SCV），透射电镜观察各组大鼠坐骨神经超微结构，TUNEL法检测各组大鼠坐骨神经细胞凋亡率，免疫荧光法检测各组大鼠坐骨神经中GRP78表达情况，Western blotting法检测各组大鼠坐骨神经中GRP78表达水平。结果：与药物干预组比较，电针干预组大鼠热敏感度值升高（P<0.01）。干预12周后与模型对照组比较，药物干预组和电针干预组大鼠胫神经MCV和SCV升高（P<0.01）；与干预前比较，干预12周后各组大鼠胫神经MCV和SCV均明显降低（P<0.01）。透射电镜下模型对照组大鼠坐骨神经可见部分髓鞘板层疏松，呈不规则的膜样团块，较粗的有髓神经纤维受累严重；药物干预组和电针干预组上述改变均减轻。TUNEL法检测，与模型对照组比较，药物干预组和电针干预组大鼠坐骨神经细胞凋亡率降低（P<0.01）。Western blotting法检测，与模型对照组比较，药物干预组和电针干预组大鼠坐骨神经中GRP78表达水平明显降低（P<0.05）。结论："调脏通络"电针能使DPN大鼠坐骨神经中GRP78表达水平降低，从而抑制内质网应激（ERS）发生，降低细胞凋亡率，对神经起到保护作用。

关键词: 糖尿病周围神经病, "调脏通络"电针, 内质网应激, 葡萄糖调节蛋白78

Abstract: Objective: To investigate the expression of glucose regulating protein 78(GRP78) in the sciatic nerve cells of the rats with diabetic peripheral neuropathy (DPN), and to elucidate the clinical significance and mechanism of Adjust Zang Dredge Meridianelectroacupuncture in the treatment of DPN.Methods: A total of 86 adult male SD rats were divided into normal control group(n=5)and model group (n=81). The diabetic rat models were built through one-time injection of streptozocin(STZ), and then the diabetic rats were randomly divided into model control group(n=27),medicine intervention group(n=26), and electroacupuncture intervention group(n=26).The rats in normal control group and model control group were fed with the maintenance feed.During the course of experiment, no special treatment was done for the rats in normal control group and model control group.The rats in drug intervention group were treated with intragastric administration of mecobalamin,20 mg·kg^-1·d^-1,once per day. The rats in electroacupuncture intervention group were treated with Adjust Zang Dredge Meridian electroacupuncture in the bilateral Feishu, Pishu, Shenshu, Hegu, Zusanli,Sanyinjiao and Taichong, once per day, 30 min per time.The thermal sensitivity value, motor nerve conduction velocity(MCV)and sensory nerve conduction velocity(SCV)of the tibial nerve of the rats in various groups were measured, the ultrastructures of sciatic nerve of the rats in various groups were observed under transmission electron microscope, the apoptotic rates of sciatic nerve cells of the rats in various groups were detected by TUNEL method, the expressions of GRP78 in sciatic nerve cells of the rats in various groups were detected by immumofluorescence,and the expression levels of GRP78 in sciatic nerve of the rats in various groups were detected by Western blotting method.Results: Compared with medicine intervention group, the thermal sensitivity value of the rats in electroacupuncture intervention group was significantly decresed (P<0.01). Compared with model control group, the MCV and SCV of tibial nerve of the rats in medicine intervention group and electroacupuncture intervention group were increased after 12-week-intervention (P<0.01); compared with before intervention, the MCV and SCV of tibial nerve of the rats in various groups were decreased after 12-week-intervention (P<0.01).Under transmission electron microscope, some of the myelin sheath lamellae of sciatic nerve of the rats in model control group showed irregular membrane-like mass, and most of the thicker myelinated nerve fibers were seriously involved; the above changes were alleviated in medicine intervention group and electroacupuncture intervention group.The TUNEL assay results showed that compared with model control group, the apoptotic rates of sciatic nerve cells of the rats in medicine intervention group and electroacupuncture intervention group were significantly decreased (P<0.01). The Western blotting results showed that compared with model control group,the expression levels of GRP78 in sciatic nerve cells of the rats in medicine intervention group and electroacupuncture intervention group were significantly decreased (P<0.05).Conclusion: Adjust Zang Dredge Meridian electroacupuncture can reduce the expression level of GRP78 in sciatic nerve cells of the DPN rats, inhibit the occurrence of endoplasmic reticulum stress(ERS), decrease the apoptotic rate of cells, and protect the nerves.

Key words: diabetic peripheral neuropathy, Adjust Zang Dredge Meridian electroacupuncture, endoplasmic reticulum stress, glucose regulated protein 78

中图分类号:

R587.1

潘鸿, 王宇峰, 黄海鹏, 张丽颖, 钟祯, 马诗棋, 董理, 王洪峰. “调脏通络”电针对糖尿病周围神经病变大鼠坐骨神经细胞中GRP78表达的影响及其神经保护作用[J]. 吉林大学学报(医学版), 2018, 44(06): 1124-1130.

PAN Hong, WANG Yufeng, HUANG Haipeng, ZHANG Liying, ZHONG Zhen, MA Shiqi, DONG Li, WANG Hongfeng. Influence of Adjust Zang Dredge Meridian electroacupuncture in expression of GRP78 in sciatic nerve cells of rats with diabetic peripheral neuropathy and its neuroprotective effect[J]. Journal of Jilin University(Medicine Edition), 2018, 44(06): 1124-1130.

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“调脏通络”电针对糖尿病周围神经病变大鼠坐骨神经细胞中GRP78表达的影响及其神经保护作用

Influence of Adjust Zang Dredge Meridian electroacupuncture in expression of GRP78 in sciatic nerve cells of rats with diabetic peripheral neuropathy and its neuroprotective effect

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