除翳解毒方对复发单纯疱疹病毒性角膜炎大鼠模型血清CD4~+、CD8~+T细胞的影响
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摘要
目的观察除翳解毒方对单纯疱疹病毒性角膜炎(HSK)大鼠模型血清中CD4~+、CD8~+T细胞及其比值的影响,从细胞免疫水平探讨除翳解毒方对HSK的作用机制。方法将SD大鼠80只,随机分为正常组、模型组、中西药治疗组、西药对照组,每组20只;中西药治疗组给予除翳解毒方~+更昔洛韦眼凝胶治疗、西药对照组给予更昔洛韦眼凝胶治疗。各组大鼠行裂隙灯显微镜检查、眼前节照相,泪液细胞学培养,Biw;第6 w末,模型组、中西药治疗组及西药对照组大鼠行紫外线照射,制造复发感染模型;第10 w末,处死全部大鼠,行角膜组织氯化金染色检测角膜神经纤维,流式细胞仪检测CD4~+、CD8~+T细胞及其比值。结果本实验成功建立了原发、复发感染HSK大鼠模型;氯化金染色模型组大鼠角膜神经纤维束稀疏、变细、变短;中西药组大鼠治疗后角膜神经纤维较丰富,呈网络结构;西药组神经纤维束较稀疏,大致呈网络结构;模型组与正常组比较,CD4~+T细胞减少,CD8~+T细胞增加,CD4~+/CD8~+降低,差异有统计学意义(P<0.01);中西药组、西药组与模型组比较,CD4~+T细胞增加,CD8~+T细胞减少,CD~+4/CD~+8升高,差异有统计学意义(P<0.01);中西药组与西药组比较,CD4~+T细胞增加, CD8~+T细胞减少, CD4~+/CD8~+升高,差异有统计学意义(P<0.05)。结论除翳解毒方能够增加CD4~+、减少CD8~+T淋巴细胞数量,可以调节复发性HSK大鼠模型的免疫功能,减少HSK复发。
OBJECTIVE To investigate the effect of Chuyi Jiedu decoction on the serum levels of CD4~+, CD8~+T cells and their ratio in the herpes simplex keratitis(HSK) rats and to reveal its mechanism at cell immunological level. METHODS Eighty Sprague-Dawley rats were randomly divided into four groups: normal group, model group, Chuyi Jiedu decoction combined ganciclovir ophthalmic gel group, and ganciclovir ophthalmic gel group. There were 20 rats in each group. Rats in Chuyi Jiedu decoction and ganciclovir ophthalmic gel group were treated by Chuyi Jiedu decoction and ganciclovir ophthalmic gel. The ganciclovir ophthalmic gel group was given ganciclovir ophthalmic gel. Slit lamp microscopy, anterior segment photography, and tear fluid cytology culture were used in each group to test rats twice a week. At the end of the sixth week, rats in the model group, the Chuyi Jiedu Decoction and ganciclovir ophthalmic gel group, and ganciclovir ophthalmic gel group were irradiated with ultraviolet rays to inducerecurrence infection. At the end of the 10 thweek, corneal nerve fibers were examined by Gold chloride staining, and CD4~+, CD8~+T cells and their ratios were measured by flow cytometry.RESULTS The primary and recurrent HSK rat model was successfully established in this experiment. The corneal nerve fiber bundles in the gold chloride staining model group were sparse,thinning, and shortening. The corneal nerve fibers were rich and in network structure in Chuyi Jiedu decoction and ganciclovir ophthalmic gel group. The nerve fiber bundles in the ganciclovir ophthalmic gel group were sparse and roughly in a network structure; Compared with the normal group, the CD4~+T cells decreased, the CD8~+T cells increased, and the CD4~+/CD8~+decreased in model group(P <0.01); the Chuyi Jiedu decoction and ganciclovir ophthalmic gel group, and ganciclovir ophthalmic gel group compared with the model group, CD4~+T cells increased, CD8~+T cells decreased, CD4~+/CD8~+increased, the difference was statistically significant(P <0.01); In contrast to the ganciclovir ophthalmic gel group, CD4~+T cells increased, CD8~+T cells decreased,and CD4~+/CD8~+increased in Chuyi Jiedu decoction and ganciclovir ophthalmic gel group, and the difference was statistically significant(P <0.05). CONCLUSIONS The Chuyi Jiedu decoction increased CD4~+T lymphocyte and reduced the expression of CD8~+T lymphocyte subsets, which modulated the immune function of recurrent HSK rat models and therefore reduced the recurrence of HSK.
OBJECTIVE To investigate the effect of Chuyi Jiedu decoction on the serum levels of CD4~+, CD8~+T cells and their ratio in the herpes simplex keratitis(HSK) rats and to reveal its mechanism at cell immunological level. METHODS Eighty Sprague-Dawley rats were randomly divided into four groups: normal group, model group, Chuyi Jiedu decoction combined ganciclovir ophthalmic gel group, and ganciclovir ophthalmic gel group. There were 20 rats in each group. Rats in Chuyi Jiedu decoction and ganciclovir ophthalmic gel group were treated by Chuyi Jiedu decoction and ganciclovir ophthalmic gel. The ganciclovir ophthalmic gel group was given ganciclovir ophthalmic gel. Slit lamp microscopy, anterior segment photography, and tear fluid cytology culture were used in each group to test rats twice a week. At the end of the sixth week, rats in the model group, the Chuyi Jiedu Decoction and ganciclovir ophthalmic gel group, and ganciclovir ophthalmic gel group were irradiated with ultraviolet rays to inducerecurrence infection. At the end of the 10 thweek, corneal nerve fibers were examined by Gold chloride staining, and CD4~+, CD8~+T cells and their ratios were measured by flow cytometry.RESULTS The primary and recurrent HSK rat model was successfully established in this experiment. The corneal nerve fiber bundles in the gold chloride staining model group were sparse,thinning, and shortening. The corneal nerve fibers were rich and in network structure in Chuyi Jiedu decoction and ganciclovir ophthalmic gel group. The nerve fiber bundles in the ganciclovir ophthalmic gel group were sparse and roughly in a network structure; Compared with the normal group, the CD4~+T cells decreased, the CD8~+T cells increased, and the CD4~+/CD8~+decreased in model group(P <0.01); the Chuyi Jiedu decoction and ganciclovir ophthalmic gel group, and ganciclovir ophthalmic gel group compared with the model group, CD4~+T cells increased, CD8~+T cells decreased, CD4~+/CD8~+increased, the difference was statistically significant(P <0.01); In contrast to the ganciclovir ophthalmic gel group, CD4~+T cells increased, CD8~+T cells decreased,and CD4~+/CD8~+increased in Chuyi Jiedu decoction and ganciclovir ophthalmic gel group, and the difference was statistically significant(P <0.05). CONCLUSIONS The Chuyi Jiedu decoction increased CD4~+T lymphocyte and reduced the expression of CD8~+T lymphocyte subsets, which modulated the immune function of recurrent HSK rat models and therefore reduced the recurrence of HSK.
引文
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[3]陈伟.单疱病毒性角膜炎中医治疗的疗效观察[J].临床医药文献杂志,2015,2(1):37.
[4]黄俊珺,董学梅.益气养血祛风法治疗复发性单纯疱疹病毒性角膜炎的临床观察[J].国际眼科杂志,2017,17(12):2318-2320.
[5]Young RC,Hodge DO,Liesegang TJ,et a1.Incidence,recurrence and outcomes of herpes simplex virus eye disease in Olmsted County,Minnesota,1976-2007:the effect of oral antiviral prophylaxis[J].Arch Ophthalmol,2010,128:1178-1183.
[6]俞莹.单纯疱疹病毒性角膜炎发病机制的研究进展[J].中华实验眼科杂志,2013,31(2):201-203.
[7]王玉中,徐艳,王利群,等.益气解毒冲剂联合西药治疗单纯疱疹病毒性角膜炎107例临床研究[J].中医杂志,2014,55(2):133-136.
[8]陈丽平.《证治准绳·杂病·七窍门目》学术源流研究[J].河南中医,2014,34(8):1475-1477.
[9]陈子燕,李庆生.中药联合无环鸟苷治疗单纯疱疹病毒性角膜炎的临床疗效分析[J].世界中医药,2017,12(4):825-828,832.
[10]关瑞娟,亢泽峰,李凌,等.益气解毒中药减少单纯疱疹病毒性角膜炎复发的免疫机理研究[J].中国中医眼科杂志,2013,23(1):17-20.
[2]葛坚.眼科学(8年制)[M].北京:人民卫生出版社,2010:244.
[3]陈伟.单疱病毒性角膜炎中医治疗的疗效观察[J].临床医药文献杂志,2015,2(1):37.
[4]黄俊珺,董学梅.益气养血祛风法治疗复发性单纯疱疹病毒性角膜炎的临床观察[J].国际眼科杂志,2017,17(12):2318-2320.
[5]Young RC,Hodge DO,Liesegang TJ,et a1.Incidence,recurrence and outcomes of herpes simplex virus eye disease in Olmsted County,Minnesota,1976-2007:the effect of oral antiviral prophylaxis[J].Arch Ophthalmol,2010,128:1178-1183.
[6]俞莹.单纯疱疹病毒性角膜炎发病机制的研究进展[J].中华实验眼科杂志,2013,31(2):201-203.
[7]王玉中,徐艳,王利群,等.益气解毒冲剂联合西药治疗单纯疱疹病毒性角膜炎107例临床研究[J].中医杂志,2014,55(2):133-136.
[8]陈丽平.《证治准绳·杂病·七窍门目》学术源流研究[J].河南中医,2014,34(8):1475-1477.
[9]陈子燕,李庆生.中药联合无环鸟苷治疗单纯疱疹病毒性角膜炎的临床疗效分析[J].世界中医药,2017,12(4):825-828,832.
[10]关瑞娟,亢泽峰,李凌,等.益气解毒中药减少单纯疱疹病毒性角膜炎复发的免疫机理研究[J].中国中医眼科杂志,2013,23(1):17-20.