文献类型：期刊文献

中文题名：基于特异性响应基因介导的级联反应通路探讨针刺干预视觉剥夺的效应机制

英文题名：Mechanism of cascade response pathway mediated by specific response gene in acupuncture intervention on visual deprivation

作者：王觉[1];刘安国[1];马重兵[1];贾静[1];魏玉婷[1];孙燕[1];严兴科[1]

第一作者：王觉

机构：[1]甘肃中医药大学针灸推拿学院,兰州730101

第一机构：甘肃中医药大学针灸推拿学院

年份：2021

卷号：36

期号：7

起止页码：3939

中文期刊名：中华中医药杂志

外文期刊名：China Journal of Traditional Chinese Medicine and Pharmacy

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金项目(No.81660816,No.81860879);甘肃省高等学校创新能力项目(No.2019A-073)。

语种：中文

中文关键词：视觉剥夺;弱视;全转录组测序;针刺响应基因;Grm2基因;mGluRs代谢通路;生物学功能

外文关键词：Visual deprivation;Amblyopia;RNA-Seq;Acupuncture response gene;Grm2 gene;mGluRs metabolic pathway;Biological function

摘要：目的:利用全转录组学测序技术(RNA-Seq)及组间差异基因比对韦恩分析法钓取针刺干预视觉剥夺效应的特异性响应基因,基于该基因的生物学功能分析初步阐明针刺对视觉剥夺效应的干预机制。方法:随机将72只14d龄健康SD幼鼠分为空白组、空白捆绑组、空白针刺组、模型组、模型捆绑组及模型针刺组,每组12只。模型捆绑组与模型针刺组在模型复制成功3d后,分别给予捆绑和针刺刺激。14d后行图形视觉诱发电位(P-VEP)检测,通过组间差异表达基因分析,钓取视皮质特异性针刺响应基因,并描述其生物学功能。采用SYBR Green荧光定量技术对基因表达进行扩增验证。结果:(1)与空白组P-VEP比较,模型组P100潜伏期显著延长,振幅显著降低(P<0.05);与模型组P-VEP比较,模型针刺组P100潜伏期显著缩短,振幅显著升高(P<0.05)。(2)共钓取左侧视皮质特异性针刺响应基因38个,其中上调表达18个,下调20个。右侧视皮质特异性针刺响应基因63个,上调表达20个,下调43个。(3)SYBR Green荧光定量对Grm2基因的验证结果表明其扩增效率较高,特异性较好。(4)基于GO分析和KEGG通路筛选,阐释了Grm2基因调控代谢型谷氨酸受体(mGluRs)信号通路在针刺干预视觉剥夺效应中的生物学机制。结论:Grm2基因介导的mGluRs信号通路激活可以改善视皮层神经元突触可塑性,从而减轻视觉剥夺性损害,这可能是针刺干预视觉剥夺效应的重要分子机制。
Objective: To extract the specific response gene of acupuncture on visual deprivation effect by using complete transcriptological sequencing(RNA-Seq) and intergroup differential gene comparison, and to preliminarily clarify the mechanism of acupuncture on visual deprivation based on the analysis of the gene’s biological feature. Methods: A total of 72 healthy SD mice aged 14 days were randomly divided into blank group, blank binding group, blank acupuncture group, model group, model binding group and model acupuncture group, with 12 rats in each group. The model binding group and the model acupuncture group were given binding and acupuncture stimulation respectively 3 days after successful model replication. After 14 days, we detected the the visual evoked potential(P-VEP), obtained the specific genes in visual cortex of acupuncture response through the analysis of the differentially expressed genes between groups, and described the genes biological functions at same time. Then, SYBR Green fluorescence quantitative technique was used to amplify and verify gene expression. Results:(1)Compared to the blank group, the incubation period of P100 in the model group was significantly prolonged and the amplitude was significantly decreased(P<0.05). Compared to the P-VEP in the model group, the incubation period of P100 in the model acupuncture group was significantly shortened and the amplitude was significantly increased(P<0.05).(2)A total of 38 left visual cortex specific acupuncture response genes were selected, among which 18 were up-regulated and 20 were down-regulated. There were 63 right visual cortex specific acupuncture response genes, which included 20 up-regulated and 43 down-regulated genes.(3)The results of SYBR Green fluorescence quantification on Grm2 gene showed high-level amplification efficiency and better specificity.(4)Based on GO analysis and KEGG pathway screening, the biological mechanism of Grm2 gene regulation of mGluRs signal in acupuncture intervention visual deprivation effect was explained. Conclusion: The activation of the mGluRs pathway mediated by Grm2 gene can improve the synaptic plasticity of the visual cortex neurons, so as to reduce the visual deprivation damage, which may be an important molecular mechanism for acupuncture on visual deprivation.