密蒙花总黄酮对去势雄鼠干眼症模型角膜和泪腺组织中TNF-α,IL-1β表达的影响

Effects of buddleja offcinalis flavone on dry eye syndrome and the expression of IL-β,TNF-α in castrated rats

目的:探讨密蒙花总黄酮对去势雄鼠干眼症动物模型角膜及泪腺组织炎症因子表达的影响与组织损伤的关系。方法:选取150只健康1月龄Wistar雄性大鼠随机分为5组(正常组、假手术组、手术对照组、雄激素治疗组、密蒙花总黄酮治疗组,分别用A,B,C,D,E组表示),每组30只。将C,D,E三组实验鼠切除双侧睾丸及附睾;B组只切开阴囊,不切除睾丸,作为假手术对照组,A组不作处理。自造模后1wk,待伤口基本愈合开始用药。A,B,C组以生理盐水灌胃,1次/d;D组用丙酸睾酮注射液大腿肌肉注射,3d1次;E组以密蒙花提取物生理盐水混悬液灌胃,1次/d。各组实验动物分别于用药1,3,5mo后随机取10只,行SchirmerⅠ试验、泪膜破裂时间(break-up time,BUT)及角膜荧光素染色检查并处死各组动物,取双眼泪腺、角膜组织采用免疫组织化学方法观察各组角膜和泪腺组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),白细胞介素1β(interleukin 1β,IL-1β)蛋白的表达。结果:A与B组泪腺、角膜上皮细胞中TNF-α,IL-1β阳性表达均不明显;C组泪腺、角膜上皮细胞中TNF-α,IL-1β阳性表达的细胞数明显高于D,E组(P<0.05);D,E各组间差异不显著(P>0.05)。结论:去势雄鼠干眼症角膜和泪腺组织局部炎症反应可能是角膜损伤和泪腺分泌功能丧失的原因之一,TNF-α,IL-1β表达增加与其炎症反应有关。密蒙花总黄酮能够下调去势雄鼠角膜和泪腺局部TNF-α,IL-1β蛋白的表达,可能是其治疗干眼症的关键机制之一。

AIM： To assess the relationships between the inflammatory factors and tissue injury in cornea and lacrimal gland of castrated rat eye models by intervention of buddlija offcinalis flavone. METHODS： One hundred and fifty healthy Wistar rats were divided randomly into 5 groups each with 30 rats..the normal group （A） ; the false operation group （B） ; the model（castrated） group （C）; the therapy group treated with testosterone（D）; the therapy group with buddleja offcinalis flavone （E）. Bilateral testis and epididymis of group C, D and E were resected; group B were performed scrotal incision only, not being removed the testicles, as sham-operated control group. Nothing was performed on group A. When the wounds were basic healing, the rats were treated with medicine 1 week after the model was established. Group A, B and C were administered saline one time a day; group D were treated with testosterone propionate by thigh intramuscular injection once per 3 days; group E were treated with buddleja officinalis for one time a day. Ten experimental animals in each group were randomly choosed after 1 month,3,5 months medication respectively,which were performed Schirmer I test, tear film break-up time and corneal fluorescein staining . And then the animals were killed; Expression of tumor necrosis factor-α （ TNF-α ） and interleukin 1β （ IL-1β ） protein in cornea and lacrimal glands was observed by immunohistochemical methods. RESULTS- Expression of TNF-α and IL-1β protein in group A and B were not obvious; Expression of TNF-α and IL-1β protein in group C was significantly higher than that of group D and E（P〈0.05） ;The difference between D and E was no significant （P〉0.05）. CONCLUSION, Inflammatory response of cornea and lacrimal gland tissue in male castration rats with dry eye may be the main reason for injury of cornea and the loss of secretory function, and the increased , expression of TNF-α and IL-1β were relevant to inflammatory response. Buddleja officinalis flavonoids can lower the local protein expression of TNF-α and IL-1β,which may be one of the key mechanisms for the treatment of dry eye.