淫羊藿总黄酮含药血浆对干眼症泪腺上皮细胞模型IL-6表达的影响

Effect of Total Flavonoids of Yinyanghuo( Herba Epimedii) on IL-6 Expression in Dry Eye Lacrimal Gland Epithelial Cell Model

目的研究淫羊藿总黄酮含药血浆对干眼症泪腺上皮细胞模型中IL-6表达的影响。方法取健康1月龄雄性新西兰大耳白兔61只,按随机的方法取1只雄兔泪腺,用于培养泪腺上皮细胞,取第3代泪腺上皮细胞用于实验。按随机的方法将60只雄兔分为空白组、淫羊藿总黄酮2.5倍组、淫羊藿总黄酮5倍组以及淫羊藿总黄酮10倍组。利用MTT法决定后面干预细胞含药血浆的浓度。将生长良好的泪腺上皮细胞,随机分成空白组、淫羊藿总黄酮组和雄激素组,分别添加空白血浆、淫羊藿总黄酮血浆、丙酸睾酮进行干预。干预48 h后,再分别加入H2O2继续培养1 h,以建立干眼症细胞模型。通过H2O2诱导雄兔泪腺上皮细胞凋亡,以建立干眼症细胞凋亡模型。用流式细胞术检测各组IL-6的表达。结果根据MTT实验结果选取干预细胞含药血浆的浓度为淫羊藿总黄酮2.5倍组。空白组中IL-6的表达均高于雄激素组、淫羊藿总黄酮组,差异具有统计学意义(P <0.05)。雄激素组中IL-6的表达均高于淫羊藿总黄酮组,差异具有统计学意义(P <0.05)。结论淫羊藿总黄酮含药血浆作用于干眼症泪腺上皮细胞模型中可抑制IL-6表达,从而使IL-6的表达下调。含淫羊藿总黄酮含药血浆可能是抑制IL-6表达途径从而达到抑制炎症作用,这可能是它治疗干眼症的关键机理之一。

Objective To study the effect of total flavonoids of Yinyanghuo( Herba Epimedii) on the expression of IL-6 in dry eye lacrimal gland epithelial cell model. Methods Sixty-one healthy 1-month-old male New Zealand white rabbits were randomly selected from the lacrimal gland of 1 male rabbit for the culture of lacrimal gland epithelial cells and the third generation of lacrimal gland epithelial cells for the experiment. According to the random method,60 male rabbits were randomly divided into blank group,total flavonoids of Yinyanghuo( Herba Epimedii) 2. 5 times group,total flavonoids of Yinyanghuo( Herba Epimedii)5 times group and total flavonoids of Yinyanghuo( Herba Epimedii) 10 times group. MTT assay was used to determine the concentration of drug-containing serum in the intervention cells. The well-growing lacrimal gland epithelial cells were randomly divided into blank group,total flavonoids of Yinyanghuo( Herba Epimedii) group and androgen group. Blank serum,total flavonoids of Yinyanghuo( Herba Epimedii) serum and testosterone propionate were added respectively. 48 hours after intervention,H2 O2 was added for 1 h to establish a dry eye model. H2 O2 induced apoptosis of lacrimal glandular epithelial cells in male rabbits. IL-6 expression in each group was detected by flow cytometry. Results According to the results of MTT test,the serum concentration of total flavonoids of Yinyanghuo( Herba Epimedii) was 2. 5 times. The expression of IL-6 in blank group was significantly higher than that in androgen group and total flavonoids group( P < 0. 05). The expression of IL-6 in androgen group was significantly higher than that in total flavonoids group( P < 0. 05). Conclusion Total flavonoids of Yinyanghuo( Herba Epimedii) can down-regulate the expression of IL-6 in dry eye lacrimal gland epithelial cell model. The serum containing total flavonoids of Yinyanghuo( Herba Epimedii) may inhibit the expression of IL-6 in order to inhibit inflammation,which may be one of the key mechanisms of Yinyanghuo( Herba Epimedii) in the treatment of dry eye.