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

同时应用IL-12和IL-18滴眼液于小鼠泪腺中，观察是否出现慢性炎症。在少数小鼠中证实了炎性细胞的侵入，但在大多数滴眼液的小鼠中，几乎没有引起泪腺的炎症和凋亡。因此，不能建立干眼症、移植物抗宿主病和干燥综合征小鼠模型。IL-18在滴注后6天内升高，IL-12在滴注开始后持续升高。腹腔注射IL-12和IL-18后，血液中细胞因子的移动量相当于腹腔注射IL-12和IL-18的一次。虽然血清细胞因子的运动在同样注射了进行滴注的小鼠的小鼠上也是相同的，但在组织的变化没有类似地产生的观点上，这被认为是有趣的。为了进一步研究，我们采用腹腔注射法，建立了小鼠急性泪腺炎模型，通过Tunel染色观察了急性模型小鼠泪腺细胞凋亡的情况。腹腔注射IL-12和IL-18后第2天，泪腺腺上皮细胞全部凋亡，第3天开始，少量腺上皮细胞坏死。此外，上皮细胞在约2周后恢复到正常腺系统中。采用ELISA法测定血清细胞因子水平。血清中IFN-α在发病后2-3天出现峰值，NO在发病后1-2天出现峰值，而IL-1、IL-6、IL-10和TNF-α在发病后2 - 3天无明显变化。从基因敲除小鼠的实验结果来看，泪腺细胞的凋亡可能不是通过Fas-FasR介导的，而是通过NO介导的。

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.