The elucidation of pathogenesis with the experimental model of dry eye by IL-12 and IL-18 simultaneous administration

同时滴入IL-12和IL-18滴眼液观察小鼠泪腺是否出现慢性炎症。在少数小鼠中证实了炎症细胞的侵袭，但在几乎所有滴眼液的小鼠中，几乎没有引起泪腺炎症和细胞凋亡。因此不能制备干眼、GVHD和干燥综合征模型小鼠。测定小鼠血清中细胞因子的含量。IL-18在给药后6天内升高，IL-12在开始给药后持续升高。在腹腔注射IL-12和IL-18后，血液中细胞因子的移动量等于1倍。虽然注射小鼠的血清细胞因子在注射小鼠身上的运动也相同，但从组织的变化没有产生相似的角度来看，这是有趣的。为了进一步研究，我们采用腹腔注射的方法，并以急性小鼠模型进行实验。采用Tunel染色法观察急性模型小鼠泪腺细胞凋亡。腹腔注射IL-12、IL-18后第2天泪腺腺上皮细胞全部凋亡，第3天开始出现少量腺上皮细胞坏死。此外，大约2周后，上皮细胞在正常腺体系统中恢复。采用ELISA法测定这段时间的血清细胞因子。血清中ifn浓度在2 ~ 3 d出现峰值，NO在1 ~ 2 d出现峰值，而IL-1、IL-6、IL-10和tnf均不能识别其变化。从敲除小鼠的结果来看，泪腺的凋亡似乎不是由Fas-FasR引起的，而是通过NO引起的。

Whether the chronic inflammation appeared by applyingg eye drops of IL-12 and IL-18 simultaneously in the lacrimal gland of the mouse was examined. The invasion of the inflammatory cell was confirmed in a few mouse but in almost of the mouse of applying eye drops, the inflammation and apoptosis in the lacrimal gland was not hardly caused. Thus model mouse of dry eye, GVHD and Sjoegren syndrome could not be made.The cytokine in mouse serum was measured. IL-18 was rosed within 6 days after, though IL-12 continuously rose after the instillation start. The movement of the blood cytokine equal to the one time in the peritoneal injection of IL-12 and IL-18. Though the movement of the serum cytokine is also same on the mouse which also injected the mouse which carried out the instillation, in the point that the change of the organization is not generated similarly, it was considered interesting. For more investigation we used the methos of peritoneal injection, and the experiment was carried out to be an acute mouse model.We observed apoptosis in the lacrimal gland of the mouse of the acute model by the Tunel staining. All glandular epithelium cells of the lacrimal gland caused the apoptosis next day from IL-12 and IL-18 peritoneal injection, and small number of glandular epithelium cells caused the necrosis from the third day. Furthermore, epithelium cells returned in the normal glandular system proven after about 2 weeks. Serum cytokine of this time was measured by the ELISA method. The serum concentration of IFN-showed the peak value in 2-3 day, NO in 1-2 day, but IL-1, IL-6, IL-10 and TNF-could not recognize the change the peak. From the result using knockout-mouse, apoptosis of the lacrimal gland seem to be not system by Fas-FasR but apoptosis through NO.