去卵巢小鼠血清性激素水平对泪腺炎症因子表达的影响

目的　研究去卵巢小鼠血清性激素水平对泪腺炎症因子表达的影响。方法　取雌性成年昆明小鼠60只，随机分为实验组和对照组，每组各30只。实验组摘除双侧卵巢，对照组模拟手术不摘除卵巢。于术前及术后1个月、3个月、6个月采用放射免疫法检测小鼠血清雌二醇、睾酮浓度，角膜荧光素染色及酚红棉线试验行眼表评估。于术后1个月、3个月、6个月行免疫组织化学法、ELISA、RT-PCR检测泪腺中白细胞介素（interleukin，IL）-1β、IL-6、肿瘤坏死因子（tumor necrosis factor，TNF）-α的表达。结果　术前实验组血清雌二醇、血清睾酮浓度与对照组之间相比，差异无统计学意义（均为P>0.05），术后1个月、3个月、6个月，实验组血清雌二醇、血清睾酮浓度均低于对照组，差异均有统计学意义（均为P<0.05）。术前实验组与对照组角膜荧光素染色评分均为0分，术后1个月、3个月实验组角膜荧光素染色评分与对照组相比，差异均无统计学意义（均为P>0.05），术后6个月实验组角膜荧光素染色评分高于对照组，差异有统计学意义（P<0.05）。术前及术后1个月、3个月实验组泪液分泌量与对照组相比，差异均无统计学意义（均为P>0.05），术后6个月实验组泪液分泌量明显少于对照组，差异有统计学意义（P<0.01）。泪腺免疫组织化学、ELISA、RT-PCR检测结果显示，术后1个月、3个月、6个月实验组IL-1β、IL-6、TNF-α的表达均高于对照组，差异均有统计学意义（均为P<0.05）。结论　小鼠去卵巢后可见泪腺组织炎症因子表达增加、泪液分泌量减少、角膜荧光素染色阳性，提示性激素水平病理性失调是导致干眼发生和发展的原因之一。

The influence of serum sex hormone levels on the expression of inflammatory cytokines in the lacrimal glands of ovariectomized mice

Objective　To study the effect of serum sex hormone levels on the expression of inflammatory factors in lacrimal glands of ovariectomized mice.Methods　Sixty female adult Kunming mice were randomly divided into experimental group and control group,30 mice in each group.Bilateral ovariectomy was performed in the experimental group and sham operation was performed on control animals with their ovaries reserved.At pre-operation,1,3 and 6 months after operation,the concentrations of serum estradiol and testosterone were detected by radioimmunoassay,ocular surface condition was assessed by corneal fluorescein staining and phenol red cotton thread test.The expression levels of Interleukin-1β (IL-1β),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α) in the lacrimal glands were detected by immunohistochemistry,enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative polymerase chain reaction (RT-PCR) at 1,3 and 6 months after surgery.Results　Serum estradiol and testosterone levels were no statistically significant differences in the experimental group compared with control group before surgery (all P>0.05).At 1,3 and 6 months after surgery,serum estradiol and testosterone levels in the experimental group were significantly lower than those of control group (all P<0.05).Corneal fluorescein staining showed grade 0 for experimental and control group before surgery,there were no statistically significant differences in the corneal fluorescein staining score between the experimental group and control group at 1 and 3 months after surgery (all P>0.05).At 6 months after surgery,corneal fluorescein staining score in the experimental group was significantly higher than that in the control group (P<0.05).There was no statistically significant difference in tear secretion between the experimental group and control group before surgery,at 1 and 3 months after surgery (all P>0.05).At 6 months after surgery,tear secretion in the experimental group was significantly lower than that in the control group (P<0.05).The results of immunohistochemistry,ELISA and RT-PCR of lacrimal glands showed that the expression of IL-1β,IL-6 and TNF-α in the experimental group was statistically higher than that in the control group at 1,3 and 6 months after surgery(all P<0.05).Conclusion　After ovariectomy in mice,the expression of inflammatory factors in lacrimal glands are increased,and tear secretion is decreased,accompanying with positive fluorescein staining in the cornea,which suggesting that the pathological sexual hormone imbalance is one reason for the occurrence and development of dry eye.