针刺对干眼兔泪腺形态学及泪腺上皮细胞凋亡相关基因蛋白表达的影响

目的 探讨针刺对干眼兔泪腺形态学及泪腺上皮细胞凋亡相关基因蛋白表达的影响,分析针刺对干眼的作用机制.方法 取健康新西兰白兔18只随机分为空白组、模型组、针刺组,每组6只,空白组不作任何处理;模型组皮下注射氢溴酸东莨菪碱,每天4次,每次2 mg·kg-1共24 d;针刺组在与模型组同样处理的基础上进行针刺,每天1次,共14 d.不同时段观察各组泪液分泌试验检测泪液量的结果,光镜、电镜观察泪腺形态学变化,免疫组织化学检测泪腺组织中凋亡相关基因蛋白的表达.结果 泪液分泌试验检测泪液量结果显示在针刺第4、7、14天同时段针刺组与模型组相比明显升高,差异均有统计学意义(均为P＜0.05);在针刺第14天时,针刺组与实验前相比,差异无统计学意义(P＞0.05).光镜、电镜示针刺组泪腺上皮细胞胞浆丰富,排列紧密,活动良好,与空白组接近.免疫组织化学检测显示与模型组比较,针刺组泪腺导管及腺泡上皮细胞中Bax的阳性细胞半定量评分值明显低,差异有统计学意义(P ＜0.05);Bcl-2的阳性细胞半定量评分值亦低,但差异无统计学意义(P＞0.05).与空白组相比,针刺组Bax、Bcl-2评分值差异均无统计学意义(均为P＞0.05).与模型组比较,针刺组的泪腺导管及腺泡上皮细胞中Fas、FasL的阳性细胞半定量评分值明显低,差异均有统计学意义(均为P＜0.05).与空白组相比,针刺组Fas、FasL评分值差异均无统计学意义(均为P＞0.05).结论 针刺通过抑制干眼兔泪腺组织的细胞凋亡及腺体萎缩,促进兔干眼模型的泪腺代谢,从而达到增加泪液分泌的作用.

Effects of acupuncture on morphology and apoptosis related gene expression of lacrimal gland in dry eye model rabbits

Objective To investigate the effects of acupuncture on morphology and apoptosis related gene expression of lacrimal gland in dry eye model rabbit,and explore the possible mechanism of acupuncture on dry eye.Methods Eighteen healthy New Zealand white rabbits were randomly divided into three groups:blank group,model group,acupuncture group,6 rabbits in each group.Blank group did not accept any treatment.Model group and acupuncture group were treated with subcutaneous injections of scopolamine hydrobromide,4 times a day with the dose of 2.0 mg · kg-1,when the acupuncture group was also treated with acupuncture,once a day for fourteen days.Three groups were examined Schirmer Ⅰ test (S Ⅰt) at different stages,and morphological changes of lacrimal gland under light and electron microscope were observed.The immunohistochemical method was used to detect the protein levels of apoptosis related gene in lacrimal gland of all rabbits.Results Between the acupuncture group and model group,the S Ⅰt was significantly different on day 4,7,14 (all P ＜ 0.05).At the same time,compared with that before experiment,S Ⅰt had no significant difference on day 14 in acupuncture group (P ＞ 0.05).Optical and electron microscope showed the lacrimal gland epithelial cells of acupuncture group closely arranged with abundant cytoplasm,closed to that of the blank group.Meanwhile,the number of lacrimal epithelium cells with Bax,Fas and FasL positive expression was significantly less than that in the model group (all P ＜ 0.05).But when it compared with blank group,there was no significant difference of Bax/Bcl-2 and Fas/FasL positive expression in acupuncture group (all P ＞ 0.05).Conclusion Acupuncture can restrain the apoptosis and gland atrophy of lacrimal gland tissue of dry eye rabbit,and promote metabolism,thereby increase the tear secretion.