明目解毒方熏蒸对单纯疱疹病毒性角膜炎HSV-1及角膜CD4、CD14的影响

Effect of fuming and steaming with Mingmu Jiedu formula on HSV-1, CD4 and CD14 of herpes simplex keratitis

目的探讨明目解毒方熏蒸对单纯疱疹病毒性角膜炎(HSK)动物模型HSV-1及角膜CD4、CD14表达的影响。方法将20周龄新西兰大耳白兔60只随机分为4组:空白组6只,模型组、对照组、联合组各18只。除空白组外,其余各组实验兔皆予眼表单纯疱疹病毒Ⅰ型(HSV-1)接种造模。对照组予阿昔洛韦滴眼液点眼,联合组予以阿昔洛韦滴眼液点眼联合明目解毒方熏蒸治疗,空白组、模型组不予任何治疗。分别在治疗1 d、3 d、7 d后评价各组实验兔角膜上皮及基质损害的严重程度、记录角膜HSV-1含量。空白组分别在治疗的第3 d和第7 d,各随机抽取3只眼采用免疫组化SP法检测角膜CD4、CD14的表达结果,并随机抽取模型组、对照组、联合组每组6只眼,采用同样的方法检测角膜组织CD4、CD14、HSV-1的表达。结果(1)模型组的临床体征评分逐渐上升,对照组及联合组评分在治疗7 d后下降。(2)PCR检测结果,治疗1 d后,模型组、对照组、联合组均能检测出HSV-1表达;治疗3 d后,模型组6个样本皆可检测出HSV-1,对照组2个样本可检出,联合组未检出,差异有统计学意义(χ^2=12.600,P=0.002);治疗7 d后,模型组6个样本皆检出HSV-1,对照组、联合组均未检出,差异有统计学意义(χ^2=18.000,P=0.000)。(3)治疗后,模型组CD4表达较空白组、对照组及联合组表达均增强;治疗3 d后,对照组和联合组角膜CD4表达相当;治疗7 d后,联合组较对照组表达略增强。(4)治疗后,联合组的CD14表达较同期对照组增强。结论明目解毒方熏蒸联合阿昔洛韦滴眼液能有效地清除角膜组织HSV-1,提高角膜CD14的表达。

OBJECTIVE To explore the effect of Mingmu Jiedu decoction fuming and steaming on the expression of HSV-1,CD4 and CD14 in the cornea of herpes simplex keratitis(HSK)animal model.METHODS Twenty weeks old New Zealand white rabbits were randomly divided into 4 groups:6 in blank groups,18 in model group,18 in the control group and 18 in combination group.Except the blank group,all the other experimental rabbits were inoculated with HSV-1.The control group was given acyclovir eye drops,combined group was given acyclovir eye drops in addition to Mingmu Jiedu fuming and steaming treatment,blank group and model group were not given any treatment.One day,3 days and 7 days after treatment,the severity of corneal epithelial and stroma damage was evaluated and the content of corneal HSV-1 was recorded.On the 3rd and 7th day of treatment,three eyes of the blank group were randomly selected to detect the expression of corneal CD4 and CD14 by SP immunohistochemistry.Six eyes of model group,the control group and combination group were randomly selected respectively,and the expression of CD4,CD14 and HSV-1 in corneal tissue was detected by SP immunohistochemistry.RESULTS(1)The clinical signs scores of model group increased gradually,and that of the control group and combination group decreased 7 days after treatment.(2)One day after treatment,polymerase chain reaction(PCR)showed that HSV-1 expressions were detected in model group,the control group and combination group;after 3 days of treatment,HSV-1 were detected in 6 samples of the model group,2 samples of the control group,and none of the combined group,the difference was statistically significant(χ^2=12.600,P=0.002);after 7 days of treatment,HSV-1 was detected in 6 samples of the model group,none of the control group and the combination group,the difference was statistically significant(χ^2=18.000,P=0.000).(3)After treatment,the expression of CD4 in model group was higher than that in blank group,the control group and combination group;3 days after treatment,the expression of CD4 in cornea of the control group and combination group was similar,but after 7 days of treatment,the expression of CD4 in combination group was slightly higher than that in the control group.(4)After treatment,the expression of CD14 in the combination group was higher than that in the control group.CONCLUSIONS Mingmu Jiedu prescription fuming and steaming combined with acyclovir eye drops effectively removed HSV-1 from corneal tissue and improved the expression of CD14.