摘要
目的观察电针(electroacupuncture,EA)GB30(环跳)、GB34(阳陵泉)对慢性压迫损伤(chronic compression injury,CCI)模型大鼠GABA能神经元功能的影响,分析电针镇痛与miR23b调控的GABA能神经元功能的关系。方法将52只SPF级SD雄性大鼠,随机均分为正常组、CCI组、CCI+EA组、CCI+miR23b组,每组13只。CCI组、CCI+EA组、CCI+miR23b组进行左侧坐骨神经结扎手术造成CCI模型,正常组除不结扎坐骨神经,其余步骤一样。术后CCI+miR23b组进行两次腰部硬脊膜注射miR23b,术后第4天进行第1次注射miR23b,第8天注射第2次miR23b,每次注射12.5μL,术后第14天取组织。CCI+EA组进行左侧GB30、GB34电针,每天电针1次,每次30 min,连续电针10天。实验结束后取动物脊髓腰段膨大组织及血清,进行实时定量PCR法检测GABA(A)、GABA(B)、miR23b的表达,ELISA法检测GABA的含量、原位杂交检测miR23b指标及免疫组化检测GABAAR beta-1,GABA B R2的表达。结果采用ELISA法检测大鼠血清GABA的含量的实验结果表明,与CCI组比,CCI+miR23b组、CCI+EA组大鼠血清GABA含量明显上调,差异具有显著性意义(P<0.05)。实时定量PCR的实验结果表明,与正常组比,CCI组大鼠的GABAA、GABAB及miR23b表达明显下调(P<0.05);与CCI组比,CCI+miR23b组、CCI+EA组GABAA、GABAB及miR23b表达明显上调(P<0.05)。原位杂交及免疫组化检测结果表明,与CCI组比,CCI+miR23b组、CCI+EA组GABA A R beta-1、GABA B R2、miR23b免疫阳性细胞表达灰度值明显下调,即表达上升,差异具有显著性意义(P<0.05)。结论电针可以调节疼痛引起的GABA A R beta-1、GABA B R2及miR23b指标的改变。说明电针通过调控miR23b的表达,改善疼痛模型大鼠脊髓背角GABA能神经元功能,达到镇痛的效果。
Objective To observe the effect of electroacupuncture(EA)Huantiao(GB30)and Yanglingquan(GB34)on the function of GABAergic neurons in CCI model rats,and to analyze the relationship between EA analgesia and the function of GABAergic neurons regulated by miR23b.Methods Fifty-two SPF SD male rats were randomly divided into control group,chronic compression injury(CCI)group,CCI+EA group and CCI+miR23b,with 13 rats in each group.In the CCI group,CCI+EA group and CCI+miR23b group,the left sciatic nerve ligation was performed to induce chronic compression injury(CCI)model.The control group did not ligated the sciatic nerve,and the other steps were the same.The CCI+miR23b group underwent two postoperative lumbar dural injections of miR23b;the first miR23b injection on postoperative day 4 and the second miR23b injection on postoperative day 8,each with 12.5μL;and the tissue was taken on postoperative day 14.The CCI+EA group underwent electroacupuncture of GB30 and GB34 on the left side,and electroacupuncture was performed once a day for 30 min for 10 consecutive days.At the end of the experiment,the expanded tissues and serum of the lumbar segment of the spinal cord were collected,and the expression of GABA(A),GABA(B)and miR23b were detected by real-time quantitative PCR,GABA content by ELISA,miR23b index by in situ hybridization and GABAAR beta-1 and GABA B R2 by immunohistochemistry.Results ELISA method was used to detect the content of serum GABA in rats.Compared with CCI group,the content of serum GABA in CCI+miR23b group and CCI+EA group was significantly increased(P<0.05).The results of real-time quantitative PCR showed that compared with the control group,the mRNA expressions of GABAA,GABAB and miR23b in the CCI group were significantly down-regulated(P<0.05).Compared with the CCI group,the mRNA expressions of GABAA,GABAB and miR23b in the CCI+miR23b group and the CCI+EA group were significantly up-regulated(P<0.05).The results of real-time PCR results displayed that the mRNA expression level of GABAA,GABAB and miR23b in CCI group,CCI+miR23b group and CCI+EA group were significantly increased(P<0.05).The in situ hybridization and immunohistochemical results showed that compared with the CCI group,the gray levels of GABA A R beta-1,GABA B R2 and miR23b immunopositive cells in the CCI+miR23b group and the CCI+EA group were significantly downregulated;namely,the expression was ascended,and the difference was remarkably significant(P<0.05).Conclusion Electroacupuncture can adjust the changes of GABA A R beta-1,GABA B R2 and miR23b indicators caused by pain.It shows that electroacupuncture may improve the function of GABAergic neurons in the spinal dorsal horn of pain model rats by regulating the expression of miR23b,and achieve analgesic effects.
作者
白凤媛
岳雯
赵冬梅
蔡仁军
宋素菲
刘涛
徐秋玲
Bai Fengyuan;Yue Wen;Zhao Dongmei;Cai Renjun;Song Sufei;Liu Tao;Xu Qiuling(The First Affiliated Hospital of Hainan Medical University,Haikou 570102,China;Hainan Medical University,Haikou 571199,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2022年第7期2852-2860,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
海南省科技厅海南省自然基金高层次人才项目(821RC571):基于脑源性神经营养因子启动子区甲基化研究电针镇痛的机理,负责人:徐秋玲
国家自然科学基金委员会地区科学基金项目(81660820):基于miR23b-NOX4调控氧化损伤研究电针治疗慢性疼痛的机理,负责人:徐秋玲
