针刺对兔干眼模型眼表损害及泪腺细胞凋亡的影响

发布时间：2018-06-03 21:13

本文选题：针刺 + 干眼　；参考：《南京中医药大学》2017年硕士论文

【摘要】：目的:通过皮下注射氢溴酸东莨菪碱注射液建立水液缺乏性兔干眼模型,探讨针刺对此模型眼表及泪腺细胞的影响,探索针刺对干眼的作用机制。方法:健康新西兰白兔24只随机分为空白组、模型组、针刺组共3组,每组8只,空白组不作任何处理;模型组和针刺组皮下注射氢溴酸东莨菪碱,每日4次,每次2mg.kg-1,直至实验结束;针刺组在干眼模型建立成功后进行针刺,每日针1次,共14天。不同时段观察各组的SchirmerⅠ试验(SIT)、泪膜破裂时间(BUT)、角膜荧光素钠染色(CFS),光镜、电镜观察泪腺形态学变化,免疫组化检测泪腺组织中凋亡相关基因蛋白的表达,联酶免疫法检测泪腺组织中乙酰胆碱(ACh)的含量。结果:1.SIT:针刺第4、7、14天,针刺组与模型组相比,两组泪流量差异均有统计学意义(P0.05)。针刺第14天,针刺组与实验前相比,差异无统计学意义(P0.05)。2.BUT:针刺第7、14天,针刺组与模型组相比差异有统计学意义(P＜0.05)。针刺第14天,针刺组与实验前相比,差异无统计学意义(P0.05)。3.CFS:针刺第7、14天,针刺组与模型组相比差异有统计学意义(P0.05)。针刺组在针刺后的各时段与针刺前相比,差异显著(P0.05),且在针刺第4天时,CFS评分最高,此后即呈现逐渐下降的趋势。4.泪腺形态学观察:空白组的正常泪腺组织中未见明显细胞扩张及萎缩;模型组的泪腺上皮细胞萎缩,细胞内物质较少,可见大量淋巴细胞浸润;针刺组中泪腺柱状上皮细胞扩张,未见明显腺体萎缩,胞浆丰富,腺体间质少量淋巴细胞浸润,。5.泪腺组织凋亡相关基因蛋白的表达:空白组中偶见阳性细胞表达,模型组可见大量淡黄色至棕黄色细颗粒细胞,针刺组中见少量淡黄色至棕黄色细颗粒细胞;针刺组泪腺细胞凋亡因子相关基因蛋白Bax、Fas及FasL阳性表达的细胞数明显低于模型组,P0.05;针刺组的Bax/Bcl_2、Fas/FasL与空白组均无统计学差异,P0.05。6.泪腺中Ach含量测定:针刺组泪腺组织中Ach含量较模型组高,差异显著,有统计学意义,P0.05;且针刺组与空白组相比时,差异均无统计学意义,P0.05。结论:1皮下注射氢溴酸东莨菪碱可以成功制备兔干眼模型,且稳定持续。2光镜、电镜下观察模型组泪腺组织可见泪腺上皮细胞萎缩,细胞内物质较少,可见大量淋巴细胞浸润。针刺组的泪腺细胞胞浆丰富,排列紧密,活动良好,与空白组接近。3针刺对水液缺乏性兔干眼模型有良好干预作用,可以提高泪液分泌量,增加泪膜稳定性,改善角膜状况,其机制可能为通过促进Ach释放,抑制干眼兔泪腺组织的细胞凋亡和腺体萎缩,改善兔干眼模型的泪腺代谢,从而达到增加泪液分泌的作用。
[Abstract]:Objective: to study the effect of acupuncture on ocular surface and lacrimal gland cells in rabbits with water deficiency by subcutaneous injection of scopolamine hydrobromide, and to explore the mechanism of acupuncture on dry eye. Methods: Twenty-four healthy New Zealand white rabbits were randomly divided into three groups: model group and acupuncture group with 8 rabbits in each group without any treatment, the model group and acupuncture group were subcutaneously injected with scopolamine hydrobromide 4 times a day. The acupuncture group was treated with acupuncture once a day for 14 days after the establishment of the dry eye model. Schirmer I test, lacrimal film rupture time, corneal fluorescein sodium staining, morphological changes of lacrimal gland were observed by light microscope and electron microscope. The expression of apoptosis-related gene protein in lacrimal gland was detected by immunohistochemistry. The content of acetylcholine (ache) in lacrimal gland was detected by Elisa. Results 1. Sit: on the 14th day of acupuncture, there was a significant difference in lacrimal flow between the acupuncture group and the model group (P 0.05). On the 14th day of acupuncture, there was no significant difference between the acupuncture group and the experimental group (P < 0.05). 2.But: on the 714th day of acupuncture, the difference between the acupuncture group and the model group was statistically significant (P < 0.05). On the 14th day, there was no significant difference between the acupuncture group and the experimental group. 3. CFS: on the 714th day of acupuncture, the difference between the acupuncture group and the model group was statistically significant. The difference between the acupuncture group and the acupuncture group was significant (P 0.05), and the CFS score of the acupuncture group was the highest on the 4th day after acupuncture, and then decreased gradually. 4. Morphological observation of lacrimal gland: there was no obvious cell expansion and atrophy in normal lacrimal gland tissue in blank group, but the epithelial cells of lacrimal gland in model group were atrophied, the substance in the cells was less, and a large number of lymphocytes were infiltrated. In acupuncture group, the columnar epithelial cells of lacrimal gland were dilated, no obvious gland atrophy, abundant cytoplasm, and a small amount of lymphocyte infiltration in glandular stroma. The expression of apoptosis-related gene protein in lacrimal gland tissue: positive cells were occasionally found in blank group, a large number of light yellow to brown fine granular cells were found in model group, and a small amount of light yellow to brown fine granular cells were found in acupuncture group. The number of positive expression of Bax-FAS and FasL in lacrimal gland apoptosis-associated gene protein in acupuncture group was significantly lower than that in model group (P0.05), but there was no significant difference in Bax / Bcl2Fas-FasL between acupuncture group and blank group (P0.05.6). The content of Ach in lacrimal gland in acupuncture group was higher than that in model group (P 0.05), and there was no significant difference between acupuncture group and blank group (P 0.05). Conclusion Anisodamine hydrobromide subcutaneously can be used to establish rabbit dry eye model, and it is stable and persistent under light microscope. Under electron microscope, the lacrimal gland epithelial cells atrophy, the intracellular substance is less, and a large number of lymphocytes infiltrate can be seen in the lacrimal gland tissue of the model group. The cytoplasm of lacrimal gland cells in acupuncture group was rich, compact and active, which had a good intervention effect on dry eye model of rabbits with water deficiency, and could increase tear secretion, increase the stability of lacrimal film and improve the corneal condition. The mechanism may be to promote the release of Ach, inhibit the apoptosis of lacrimal gland and atrophy of gland in dry eye rabbit, improve the metabolism of lacrimal gland in rabbit dry eye model, so as to increase the secretion of lacrimal fluid.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245;R-332

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