电针与润目灵治疗干眼的疗效及相关机理研究
发布时间：2019-02-08 18:27
【摘要】：目的：本研究拟通过活体动物实验,观察电针灸刺激眼周穴位及中药润目灵颗粒灌胃对干眼模型的疗效,探索电针及中药润目灵与兔泪腺抗凋亡作用及与M型胆碱受体及其介导的信号转导通路上关键因子的相关性,推断泪腺细胞主动分泌的可能作用及机制。方法：健康新西兰大白兔,实验分为以下六组：空白对照组、阿托品模型组、西药干预组、中药干预组、电针干预组、针药干预组,雌雄兼备,每组各6只。模型组和各治疗组均行1%硫酸阿托品滴眼液每日滴双眼4次(上午8：00、12：00,16：00、20：00)制备干眼模型,3天后水液缺乏性干眼模型形成并稳定后对各治疗组分别行毛果芸香碱滴眼液滴眼、给予润目灵灌胃、电针干预及润目灵灌胃结合电针干预治疗。实验标本采集和数据收集时间为：各组实验兔分别于实验前、实验第4天(模型稳定)、实验第11天(治疗第一周)、实验第18天(治疗第二周)检测泪流量、泪膜破裂时间、角膜染色分数；并在实验第18天取泪腺组织行病理形态学观察、Elisa法测乙酰胆碱含量、免疫组化及real-time PCR检测。结果：Schirmer I、泪膜破裂时间及角膜染色均证实西药、电针、中药、针药联合治疗均可促进泪液分泌、延长泪膜破裂时间、修复受损的角膜上皮,但针药干预组的效果最佳。实验18天,取各组的泪腺及角膜组织进行相关实验室检查,西药干预组、中药干预组及电针干预组的形态出现不同程度的改善,针药干预组的泪腺及角膜组织形态最接近于空白对照组。各组泪腺组织的乙酰胆碱含量较模型组有所上升,泪腺组织中的乙酰胆碱含量较模型组有统计学差异。针药干预组的凋亡因子Fas、FasL、Bax蛋白的半定量,与阿托品模型组比较有统计学差异。针药干预组中的泪腺组织中的M受体介导的的信号转导通路上关键因子的蛋白和蛋白及mRNA检测显示M3AchR及MAPK,与模型组比较有统计学差异,但IP3与模型组比较,无统计学差异。结论：电针和中药润目灵均可以抑制泪腺细胞凋亡,增加泪液分泌。电针增加泪液分泌与神经递质释放以及泪腺组织上M受体介导的信号转导通路关键因子也存在着一定的相关性。
[Abstract]:Objective: to observe the effect of electroacupuncture (EA) stimulation on acupoints around the eyes and Runmuling granule (RML) on dry eye model by living animal experiment. To explore the relationship between electroacupuncture and Runmuling in rabbit lacrimal gland and the key factors in M type choline receptor and its signal transduction pathway, the possible role and mechanism of active secretion of lacrimal gland cells were inferred. Methods: healthy New Zealand white rabbits were divided into the following six groups: blank control group, atropine model group, western medicine intervention group, traditional Chinese medicine intervention group, electroacupuncture intervention group, acupuncture and medicine intervention group, both sexes. The model group and the treatment group were treated with 1% atropine sulfate eye drops 4 times a day (8: 00: 12: 00: 16: 00 20: 00) to make dry eye model. After 3 days, the dry eye model of water deficiency was formed and stabilized. Each treatment group was treated with pilocarpine eye drops, treated with Runmuling, electroacupuncture and Runmuling combined with electroacupuncture intervention. The time of collecting samples and collecting data were: before experiment, 4 days (model stability), 11 days (the first week of treatment), 18 days (the second week of treatment) to detect the lacrimal flow and tear film rupture time. Corneal staining fraction; On the 18th day of the experiment, the lacrimal gland tissues were taken for histopathological observation, and the content of acetylcholine, immunohistochemistry and real-time PCR were measured by Elisa method. Results: Schirmer I, tear film rupture time and corneal staining confirmed that western medicine, electroacupuncture, Chinese medicine, acupuncture and medicine combined treatment can promote tear secretion, prolong tear film rupture time, repair damaged corneal epithelium, but acupuncture and medicine intervention group is the best. On the 18th day of the experiment, the lacrimal gland and corneal tissue of each group were taken for laboratory examination, and the morphology of the western medicine intervention group, the traditional Chinese medicine intervention group and the electroacupuncture intervention group were improved to some extent. The morphology of lacrimal gland and cornea in acupuncture and medicine intervention group was closest to that in blank control group. The content of acetylcholine in the lacrimal gland was higher than that in the model group, and the content of acetylcholine in the lacrimal gland was significantly higher than that in the model group. The semiquantitative expression of apoptotic factor Fas,FasL,Bax protein in acupuncture and drug intervention group was significantly different from that in atropine model group. The detection of protein, protein and mRNA of key factors in the signal transduction pathway mediated by M receptor in lacrimal gland tissue of acupuncture and drug intervention group showed that M3AchR and MAPK, were significantly different from model group, but IP3 had no statistical difference compared with model group. Conclusion: both electroacupuncture and Runmuling can inhibit the apoptosis of lacrimal gland cells and increase the secretion of tear. The increase of tear secretion by electroacupuncture is also related to the release of neurotransmitters and the key factors of signal transduction pathway mediated by M receptor in lacrimal gland.
【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R245
[Abstract]:Objective: to observe the effect of electroacupuncture (EA) stimulation on acupoints around the eyes and Runmuling granule (RML) on dry eye model by living animal experiment. To explore the relationship between electroacupuncture and Runmuling in rabbit lacrimal gland and the key factors in M type choline receptor and its signal transduction pathway, the possible role and mechanism of active secretion of lacrimal gland cells were inferred. Methods: healthy New Zealand white rabbits were divided into the following six groups: blank control group, atropine model group, western medicine intervention group, traditional Chinese medicine intervention group, electroacupuncture intervention group, acupuncture and medicine intervention group, both sexes. The model group and the treatment group were treated with 1% atropine sulfate eye drops 4 times a day (8: 00: 12: 00: 16: 00 20: 00) to make dry eye model. After 3 days, the dry eye model of water deficiency was formed and stabilized. Each treatment group was treated with pilocarpine eye drops, treated with Runmuling, electroacupuncture and Runmuling combined with electroacupuncture intervention. The time of collecting samples and collecting data were: before experiment, 4 days (model stability), 11 days (the first week of treatment), 18 days (the second week of treatment) to detect the lacrimal flow and tear film rupture time. Corneal staining fraction; On the 18th day of the experiment, the lacrimal gland tissues were taken for histopathological observation, and the content of acetylcholine, immunohistochemistry and real-time PCR were measured by Elisa method. Results: Schirmer I, tear film rupture time and corneal staining confirmed that western medicine, electroacupuncture, Chinese medicine, acupuncture and medicine combined treatment can promote tear secretion, prolong tear film rupture time, repair damaged corneal epithelium, but acupuncture and medicine intervention group is the best. On the 18th day of the experiment, the lacrimal gland and corneal tissue of each group were taken for laboratory examination, and the morphology of the western medicine intervention group, the traditional Chinese medicine intervention group and the electroacupuncture intervention group were improved to some extent. The morphology of lacrimal gland and cornea in acupuncture and medicine intervention group was closest to that in blank control group. The content of acetylcholine in the lacrimal gland was higher than that in the model group, and the content of acetylcholine in the lacrimal gland was significantly higher than that in the model group. The semiquantitative expression of apoptotic factor Fas,FasL,Bax protein in acupuncture and drug intervention group was significantly different from that in atropine model group. The detection of protein, protein and mRNA of key factors in the signal transduction pathway mediated by M receptor in lacrimal gland tissue of acupuncture and drug intervention group showed that M3AchR and MAPK, were significantly different from model group, but IP3 had no statistical difference compared with model group. Conclusion: both electroacupuncture and Runmuling can inhibit the apoptosis of lacrimal gland cells and increase the secretion of tear. The increase of tear secretion by electroacupuncture is also related to the release of neurotransmitters and the key factors of signal transduction pathway mediated by M receptor in lacrimal gland.
【学位授予单位】：南京中医药大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R245
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