双秦眼用凝胶对小鼠单疱病毒性角膜炎的抗复发作用研究  

Research on the anti-relapse effect of Shuangqin Eye Gel for herpes simplex keratitis in mice

作　　者：王珍[1] 梁丽娜[1] 白昱旸[2] 唐由之[1] 周尚昆[1] 马群英[1] 

机构地区：[1]中国中医科学院眼科医院,北京100040 [2]广州中医药大学临床药理研究所,广州510405

出　　处：《中华中医药杂志》2015年第5期1761-1766,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家科技重大专项"重大新药创制"项目(No.20092X09301-005)~~

摘　　要：目的:探讨双秦眼用凝胶对小鼠单疱病毒性角膜炎(HSK)的抗复发作用。方法:用角膜病毒接种法给BALB/c小鼠接种单纯疱疹病毒I型(HSV-1)SM44株,随机分为对照组、无环鸟苷(ACV)组、双秦眼用凝胶组。连续给药2周,停止用药8周后紫外线B(UV-B)光照射,诱导HSK复发,观察各组小鼠泪液膜擦拭液的病毒检出率、UV-B光照射前后病变部位体征评分、角膜组织结构、聚合酶链反应(PCR)方法检测三叉神经节(TG)内HSV-1的潜伏,以及反转录聚合酶链反应(RT-PCR)方法检测TG内潜伏相关转录子(LAT)的表达情况。结果:UV-B光照射后,各组检出率均呈不同程度的升高;在照射后7d的泪液膜擦拭液里,除第3-5天双秦眼用凝胶组的泪液膜擦拭液里的病毒检出率高于ACV组外,其余几天,双秦眼用凝胶组的病毒检出率均低于其余两组。但是,比较各组间每天的病毒检出率,均无显著统计学差异;在病变部位体征评分方面,应用双秦眼用凝胶治疗后的小鼠在UV-B光照射诱导后病变程度轻于其余两组(P<0.05);HE染色观察角膜组织结构发现,UV-B光照射后,对照组及ACV组角膜均有不同程度的损伤,双秦眼用凝胶组角膜各层结构大致正常;双秦眼用凝胶组TG内HSV-1的潜伏及LAT的表达均受到抑制;用PCR技术检测各组小鼠TG内HSV-1 DNA的灰度值,结果显示,双秦眼用凝胶优于对照组(P<0.05)及ACV组(P<0.01);应用RT-PCR技术检测并比较各组小鼠TG内LAT m RNA的灰度值,双秦眼用凝胶亦优于对照组(P<0.01)及ACV组(P<0.05)。结论:双秦眼用凝胶在一定程度上可以降低HSK小鼠泪液膜擦拭液的病毒检出率,减轻UV-B光照射后角膜病变体征、保护角膜的组织结构、抑制TG内HSV-1的潜伏和LAT的表达,但双秦眼用凝胶对小鼠HSK的抗复发作用及机制仍需在增加样本量、排除外界因素影响的基础上进一步研究探讨。Objective： To evaluate the anti-relapse effect of Shuangqin Eye Gel intervention in mice with herpes simplex keratitis （HSK）. Methods： The corneas of BALB/c mice were inoculated with herpes simplex virus type 1 （HSV-1） SM44 strain. The mice were randomly divided into three groups： control group, acyclovir （ACV） group and Shuangqin Eye Gel group. Each group was treated by corresponding method for two weeks. After 8 weeks the corneas were irradiated by ultraviolet B （UV-B） light in order to induce the relapse of HSK, the virus detection rate of tear film cleaning fluid, lesion symptom were detected. The hiding of HSV-1 in the trigeminal ganglion （TG） and the expression of the latency-associated transcript （LAT） in the trigeminal ganglion were detected by polymerase chain reaction （PCR） and reverse transcription polymerase chain reaction （RT-PCR）. Results： Virus detection rate of tear film cleaning fluid of HSK mice in the three groups were increased with different degrees. At the 7 day after irradiating by UV-B light, the virus detection rates of Shuangqin eye gel had been lower than the other two groups expect the 3th day and the 5th day. while there was no statistical difference among the three groups. The corneal lesions of Shuangqin eye gel group were lighter than the other two groups （P〈0.05）, with significant difference; The corneas of the control group and the ACV group were damaged while the Shuangqin Eye Gel group was generally normal; The PCR gray value of the HSV-1 DNA in TG of Shuangqin Eye Gel group was superior to the control group （P〈0.05） and the ACV group （P〈0.05）. The RT-PCR gray value of the LAT mRNA in TG of Shuangqin Eye Gel group was superior to the control group （P〈0.01） and ACV group （P〈0.05）. Conclusion： Shuangqin Eye Gel could reduce the virus detection rate of HSV-1, alleviate the symptoms of HSK, and protect the normal tissue structure of corneas, but its anti-relapse and mechanism still need to be further explored.

关 键 词：双秦眼用凝胶 单疱病毒性角膜炎 抗复发作用 潜伏相关转录体 RT-PCR 

分 类 号：R285.5[医药卫生—中药学]