VIP在烟曲霉菌感染角膜中的表达及作用

发布时间：2018-05-17 08:39

  本文选题：真菌性角膜炎 + 血管活性肠肽　； 参考：《青岛大学》2017年硕士论文

【摘要】：目的:研究血管活性肠肽(Vasoactive intestinal peptide,VIP)在易感型C57BL/6小鼠和抵抗型BALB/c小鼠烟曲霉菌感染角膜中的表达,明确VIP在BALB/c小鼠烟曲霉菌感染性角膜炎中的作用。方法:(1)实验用抵抗型BALB/c及易感型C57BL/6两种类型的小鼠。用烟曲霉菌感染小鼠角膜,建立小鼠真菌性角膜炎动物模型,采用RT-PCR技术方法检测两种小鼠正常角膜和烟曲霉菌感染后分别于第1天、3天、5天角膜中VIP m RNA的表达情况。(2)选用抵抗型BALB/c小鼠作为接下来实验用的动物模型。给予BALB/c小鼠VIP拮抗剂预处理,用可拍照的裂隙灯显微镜定期观察小鼠角膜感染情况,分别于感染的第1天,3天和第5天根据小鼠角膜的感染情况进行临床评分。利用RT-PCR技术检测预处理前后正常角膜和感染后第3天小鼠角膜中促炎细胞因子MIP-2、IL-1β、TNF-α以及抗炎细胞因子IL-10、TGF-βm RNA的表达;利用ELISA技术检测小鼠正常角膜和烟曲霉菌感染后第3天及第5天角膜中促炎因子MIP-2、IL-1β、TNF-α蛋白和抗炎细胞因子IL-10及TGF-β蛋白的表达情况。结果:(1)BALB/c小鼠角膜在烟曲霉菌感染后第1天和第3天VIP m RNA的表达明显高于C57BL/6小鼠烟曲霉菌感染角膜中的表达,而在烟曲霉菌感染角膜后的第5天,两种类型小鼠的角膜中VIP m RNA的表达未见明显差异。正常两种小鼠角膜中也可见VIP的表达。(2)BALB/c小鼠经VIP拮抗剂处理后,与PBS处理对照组相比,VIP拮抗剂预处理小鼠角膜,在烟曲霉菌感染后的第3天和第5天可以增加小鼠角膜中炎症细胞因子MIP-2、IL-1β、TNF-αm RNA及蛋白质的表达,明显降低小鼠角膜中抗炎细胞因子IL-10、TGF-βm RNA及蛋白质的表达水平。BALB/c小鼠角膜的炎症反应加重,真菌感染性角膜炎的临床评分升高。结论:(1)烟曲霉菌感染性角膜炎中,抵抗型BALB/c小鼠VIP的表达明显高于易感型C57BL/6小鼠的表达。(2)VIP拮抗剂处理加重BALB/c小鼠角膜炎症反应。(3)VIP通过降低促炎因子表达和增加抗炎因子表达来减轻烟曲霉菌感染性角膜炎的炎症反应程度。
[Abstract]:Aim: to study the expression of vasoactive intestinal peptide (Vasoactive intestinal peptide) in the cornea of susceptible C57BL/6 mice and resistant BALB/c mice infected with Aspergillus fumigatus, and to clarify the role of VIP in BALB/c mice with aspergillus fumigatus infective keratitis. Methods two types of resistant BALB/c and susceptible C57BL/6 were used in the experiment. The animal model of fungal keratitis was established by infecting the cornea of mice with Aspergillus fumigatus. RT-PCR technique was used to detect the expression of VIP m RNA in normal cornea and aspergillus fumigatus of two kinds of mice on day 1, day 3 and day 5, respectively. The resistant BALB/c mice were selected as the next experimental animal model. BALB/c mice were pretreated with VIP antagonist. The corneal infection of mice was observed regularly by slit lamp microscope. The clinical scores were evaluated on the 1st day and 5th day according to the infection of the cornea of the mice. The expression of pro-inflammatory cytokine MIP-2IL-1 尾 -TNF- 伪 and anti-inflammatory cytokine IL-10 TGF- 尾 m RNA in normal cornea before and after preconditioning and 3 days after infection were detected by RT-PCR technique. The expression of proinflammatory factor MIP-2TNF- 伪, anti-inflammatory cytokine IL-10 and TGF- 尾 in normal cornea and mice cornea after aspergillus fumigatus infection was detected by ELISA technique on the 3rd and 5th day after infection. Results the expression of VIP m RNA in the cornea of BALB / c mice was significantly higher than that in C57BL/6 mice on the 1st and 3rd day after infection, but on the fifth day after aspergillus fumigatus infection, the expression of VIP m RNA in the cornea of BALB / c mice was significantly higher than that in the C57BL/6 mouse cornea infected with Aspergillus fumigatus. There was no significant difference in the expression of VIP m RNA between the two types of mice. The expression of VIP was also seen in the cornea of two normal mice. After treated with VIP antagonist, VIP antagonist was used to pretreat the cornea of mice compared with the control group treated with PBS. On the 3rd and 5th day after aspergillus fumigatus infection, the expression of inflammatory cytokine MIP-2IL-1 尾 -TNF- 伪 m RNA and protein in the cornea of mice was increased. The expression level of anti-inflammatory cytokine IL-10 TGF- 尾 m RNA and protein in the cornea of mice was significantly decreased. The inflammatory reaction of BALB / c mice was aggravated, and the clinical score of fungal infectious keratitis was increased. Conclusion 1) aspergillus fumigatus infective keratitis, The expression of VIP in the resistant BALB/c mice was significantly higher than that in the susceptible C57BL/6 mice. The VIP antagonist treatment aggravated the corneal inflammation in BALB/c mice by reducing the expression of proinflammatory factor and increasing the expression of anti-inflammatory factor to alleviate the infection of Aspergillus fumigatus. Degree of inflammation in keratitis.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R772.21

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