神经肽因子对疱疹病毒性角膜基质炎小鼠眼表修复的影响及相关机制

目的探讨神经肽因子对疱疹病毒性角膜基质炎小鼠眼表修复的影响及相关机制。方法通过双眼角膜基质注射建立疱疹病毒性角膜基质炎小鼠模型,共42只,随机平分为模型组、阿昔洛韦组与神经肽因子组,各14只。3组分别在建模14 d分别多点皮下注射5μL的0.9%氯化钠溶液、5μL 10 mg/mL阿昔洛韦、5μL 10 mg/mL神经肽因子,1周1次,连续应用4次。采用角膜基质浑浊及新生血管评分标准对各组小鼠进行评分;采用酶联免疫试验法检测血清白细胞介素-6(IL-6)和白细胞介素-8(IL-8)含量;采用流式细胞仪检测CD4⁺T淋巴细胞比率;采用蛋白质印迹(Western blotting)法检测血管内皮生长因子(VEGF)蛋白表达水平。结果阿昔洛韦组与神经肽因子组治疗第2周与第4周的角膜基质浑浊及新生血管评分、血清IL-6和IL-8含量低于模型组(5.25±0.23)、(5.21±0.18)分;(56.33±3.14)、(56.98±1.58)μg/L;(78.28±3.22)、(78.19±2.22)μg/L],神经肽因子组(1.20±0.15)、(0.78±0.14)分;(10.38±2.11)、(7.09±1.29)μg/L;(10.76±0.87)、(8.17±0.82)μg/L]低于阿昔洛韦组(2.48±0.11)、(2.00±0.14)分;(23.09±1.22)、(18.09±2.15)μg/L;(34.09±3.76)、(24.98±2.76)μg/L],差异均有统计学意义(P<0.05)。阿昔洛韦组与神经肽因子组全血组织CD4⁺T淋巴细胞比率高于模型组(33.78±1.35)%],神经肽因子组(54.09±2.57)%]高于阿昔洛韦组(48.82±1.68)%],差异有统计学意义(P<0.05)。阿昔洛韦组(2.10±0.14)与神经肽因子组(1.09±0.21)治疗第4周的VEGF蛋白相对表达水平低于模型组(5.21±0.13),神经肽因子组低于阿昔洛韦组,差异有统计学意义(P<0.05)。结论神经肽因子对疱疹病毒性角膜基质炎小鼠的应用能促进眼表修复,抑制炎性因子的表达,改善小鼠的免疫功能,其作用机制可能与抑制VEGF蛋白表达有关。

Effect of neuropeptide factor on ocular surface repair in mice with herpes virus keratitis and related mechanisms

Objective To investigate the effects of neuropeptide factor on ocular surface repair in mice with herpes virus keratitis and related mechanisms.Methods A mouse model of herpesvirus-induced keratitis was established by two-eye corneal stroma injection.A total of 42 mice were randomly divided into model group,acyclovir group and neuropeptide factor group,14 each.The three groups were injected with 5μL of 0.9%sodium chloride solution,5μL of 10 mg/mL acyclovir,5μL 10 mg/mL neuropeptide factor at multiple points on the 14th day of modeling,once a week,4 consecutive applications.Corneal stromal turbidity and neovascularization scoring criteria were used to score rats in each group;Enzyme-linked immunoassay to detect serum interleukin-6(IL-6)and interleukin-8(IL-8)levels;Flow cytometry to detect the ratio of CD4⁺T lymphocytes;Western blotting was used to detect the expression level of vascular endothelial growth factor(VEGF)protein.Results The corneal stromal turbidity,neovascularization score,serum IL-6 and IL-8 levels in the acyclovir group and neuropeptide factor group were lower in the 2nd and 4th weeks of treatment than in the model group(5.25±0.23),(5.21±0.18)points;(56.33±3.14),(56.98±1.58)μg/L;(78.28±3.22),(78.19±2.22)μg/L](P<0.05),the neuropeptide factor group(1.20±0.15),(0.78±0.14)points;(10.38±2.11),(7.09±1.29)μg/L;(10.76±0.87),(8.17±0.82)μg/L]were lower than the acyclovir group(2.48±0.11),(2.00±0.14)points;(23.09±1.22),(18.09±2.15)μg/L;(34.09±3.76),(24.98±2.76)μg/L],the differences were statistically significant(P<0.05);the proportion of CD4⁺T lymphocytes in whole blood tissue were higher than that of the model group(33.78±1.35)%],and the neuropeptide factor group(54.09±2.57)%]were higher than the acyclovir group(48.82±1.68)%](P<0.05).The relative expression level of VEGF protein in the acyclovir group(2.10±0.14)and the neuropeptide factor group(1.09±0.21)were lower than that of the model group(5.21±0.13)at the 4th week of treatment,and the neuropeptide factor group were lower than the acyclovir group(P<0.05).Conclusion The application of neuropeptide factor to mice with herpes virus keratitis can promote ocular surface repair,inhibit the expression of inflammatory factors,and improve the immune function of mice.Its mechanism may be related to the inhibition of VEGF protein expression.