摘要
目的探讨双秦眼用凝胶对实验性小鼠复发性单疱病毒性角膜炎(HSK)在角膜组织结构及炎症因子的表达方面的干预作用。方法用病毒接种法给BALB/c小鼠角膜接种单纯疱疹病毒I型(HSV-I)SM44株,饲养6周后用紫外线光照射小鼠的右眼角膜,诱导复发,随机分为凝胶基质组(A组)、双秦眼用凝胶组(B组)、无环鸟苷滴眼液(C组)、无环鸟苷+可的松滴眼液组(D组),分别给予凝胶基质、双秦眼用凝胶、无环鸟苷滴眼液、无环鸟苷+可的松滴眼液点眼,连续给药14 d,于造模后第1、3、5、7、10、12、14 d行裂隙灯检查,观察角膜、结膜病变情况并评分,14 d后处死所有小鼠,行石蜡切片、HE染色、免疫组化、ELISA检测。结果(1)组织病理学:角膜病变变化结果显示,从第3 d开始,B组评分均低于A组,差异有统计学意义(t_(3d)=2.273,P=0.029;t_(5d)=5.874,P=0.000;t_(7d)=-2.459,P=0.020;t_(10d)=2.142,P=0.039;t_(12d)=8.607,P=0.000;t_(14d)=6.576,P=0.000);结膜病变变化结果显示,从第3 d开始,B组评分均低于A组,差异均有统计学意义(t_(3d)=5.620,P=0.000;t5 d=3.299,P=0.002;t_(7d)=8.426,P=0.000;t_(10d)=12.515,P=0.000;t_(12d)=8.318,P=0.000;t_(14d)=16.353,P=0.000)。且从第10 d开始,B组评分均低于C组,差异均有统计学意义(t_(10d)=2.657,P=0.012;t_(12d)=4.102,P=0.000;t_(14d)=6.869,P=0.000);角膜病理切片HE染色结果显示,B组基质层轻度水肿,各层结构大致正常。(2)角膜内白细胞介素-17(IL-17)抗体:与A组比较,B组的IL-17的表达量降低,差异有统计学意义(t=6.379,P=0.000),且B组表达量低于C组,差异有统计学意义(t=3.889,P=0.001)。(3)角膜内肿瘤坏死因子-α(TNF-α)的含量:ELISA方法检测角膜内TNF-α的含量结果显示,与A组比较,B组角膜内TNF-α的含量较低,差异有统计学意义(t=2.652,P=0.016)。结论双秦眼用凝胶不仅可以保护角膜组织结构的完整,而且可以抑制角膜内IL-17的表达,降低角膜内TNF-α的含量,从而抑制细胞凋亡,发挥对复发性HSK的治疗作用。
OBJECTIVE To investigate the effect of Shuangqin eye gel on the corneal structure and expression of inflammatory factors in experimental mice with recurrent herpes simplex keratitis(HSK).METHODS BALB/c mice were inoculated with herpes simplex virus type I(HSV-I)SM44 strain by corneal virus inoculation method.After six weeks of feeding,the right corneas of the mice were irradiated with ultraviolet light to induce relapse.They were randomly divided into gel matric group(Group A),Shuangqin eye gel group(Group B),acyclovir(ACV)eye drops group(Group C),and ACV+cortisone eye drops group(Group D).Gel matric,Shuangqin eye gel,ACV eye drops,and ACV+cortisone eye drops were given to the eyes for consecutive two weeks.Slit lamp examination was performed on the 1^(st),3^(rd),5^(th),7^(th),10^(th),12^(th),and 14^(th) days after the establishment of the model.Corneal and conjunctival lesions were observed and scored.After fourteen days,all mice were sacrificed for paraffin section,HE staining,immunohistochemistry and ELISA analysis.RESULTS(1)Histopathology:The results of corneal pathological changes showed that from the third day,the scores of Group B were lower than those of Group A,and the differences were statistically significant(t_(3d)=2.273,P=0.029;t_(5d)=5.874,P=0.000;t_(7d)=-2.459,P=0.020;t_(10d)=2.142,P=0.039;t_(12d)=8.607,P=0.000;t_(14d)=6.576,P=0.000).The results of conjunctival pathological changes showed that from the third day,the scores of Group B were lower than those of Group A,and the differences were statistically significant(t_(3d)=5.620,P=0.000;t_(5d)=3.299,P=0.002;t_(7d)=8.426,P=0.000;t_(10d)=12.515,P=0.000;t_(12d)=8.318,P=0.000;t_(14d)=16.353,P=0.000).And from the 10^(th) day,the scores of Group B were lower than those of Group C,with statistical significance(t_(10d)=2.657,P=0.012;t_(12d)=4.102,P=0.000;t_(14d)=6.869,P=0.000).The results of HE staining of corneal pathological sections showed that the stromal layer in Group B was slightly edematous,and the structure of each layer was mostly normal.(2)Expression of intracorneal IL-17 antibody:Compared with Group A,the expression of IL-17 in Group B was lower with statistical significance(t=6.379,P=0.000),and the expression of IL-17 in Group B was lower than that in Group C,with statistical significance(t=3.889,P=0.001).(3)TNF-αin the cornea:The content of TNF-αin the cornea was detected by ELISA analysis.The results showed that compared with Group A,the content of TNF-αin the cornea of Group B was lower with statistical significance(t=2.652,P=0.016).CONCLUSIONS Shuangqin eye gel can not only protect the integrity of the corneal structure,but also inhibit the expression of IL-17 and reduce the content of TNF-αin the cornea,thus inhibit cell apoptosis,and play a therapeutic role on recurrent HSK.
作者
王珍
梁丽娜
马群英
秦虹
唐由之
WANG Zhen;LIANG Lina;MA Qunying;QIN Hong;TANG Youzhi(Eye Hospital,China Acad-emy of Chinese Medical Sciences,Beijing 100040,China)
出处
《中国中医眼科杂志》
2022年第9期684-690,共7页
China Journal of Chinese Ophthalmology
基金
国家“重大新药创制”科技专项(20092X09301-005)。
