本文選題：真菌性角膜炎 + 血管活性腸肽��； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的:研究血管活性腸肽(Vasoactive intestinal peptide,VIP)在易感型C57BL/6小鼠和抵抗型BALB/c小鼠煙曲霉菌感染角膜中的表達(dá),明確VIP在BALB/c小鼠煙曲霉菌感染性角膜炎中的作用。方法:(1)實驗用抵抗型BALB/c及易感型C57BL/6兩種類型的小鼠。用煙曲霉菌感染小鼠角膜,建立小鼠真菌性角膜炎動物模型,采用RT-PCR技術(shù)方法檢測兩種小鼠正常角膜和煙曲霉菌感染后分別于第1天、3天、5天角膜中VIP m RNA的表達(dá)情況。(2)選用抵抗型BALB/c小鼠作為接下來實驗用的動物模型。給予BALB/c小鼠VIP拮抗劑預(yù)處理,用可拍照的裂隙燈顯微鏡定期觀察小鼠角膜感染情況,分別于感染的第1天,3天和第5天根據(jù)小鼠角膜的感染情況進(jìn)行臨床評分。利用RT-PCR技術(shù)檢測預(yù)處理前后正常角膜和感染后第3天小鼠角膜中促炎細(xì)胞因子MIP-2、IL-1β、TNF-α以及抗炎細(xì)胞因子IL-10、TGF-βm RNA的表達(dá);利用ELISA技術(shù)檢測小鼠正常角膜和煙曲霉菌感染后第3天及第5天角膜中促炎因子MIP-2、IL-1β、TNF-α蛋白和抗炎細(xì)胞因子IL-10及TGF-β蛋白的表達(dá)情況。結(jié)果:(1)BALB/c小鼠角膜在煙曲霉菌感染后第1天和第3天VIP m RNA的表達(dá)明顯高于C57BL/6小鼠煙曲霉菌感染角膜中的表達(dá),而在煙曲霉菌感染角膜后的第5天,兩種類型小鼠的角膜中VIP m RNA的表達(dá)未見明顯差異。正常兩種小鼠角膜中也可見VIP的表達(dá)。(2)BALB/c小鼠經(jīng)VIP拮抗劑處理后,與PBS處理對照組相比,VIP拮抗劑預(yù)處理小鼠角膜,在煙曲霉菌感染后的第3天和第5天可以增加小鼠角膜中炎癥細(xì)胞因子MIP-2、IL-1β、TNF-αm RNA及蛋白質(zhì)的表達(dá),明顯降低小鼠角膜中抗炎細(xì)胞因子IL-10、TGF-βm RNA及蛋白質(zhì)的表達(dá)水平。BALB/c小鼠角膜的炎癥反應(yīng)加重,真菌感染性角膜炎的臨床評分升高。結(jié)論:(1)煙曲霉菌感染性角膜炎中,抵抗型BALB/c小鼠VIP的表達(dá)明顯高于易感型C57BL/6小鼠的表達(dá)。(2)VIP拮抗劑處理加重BALB/c小鼠角膜炎癥反應(yīng)。(3)VIP通過降低促炎因子表達(dá)和增加抗炎因子表達(dá)來減輕煙曲霉菌感染性角膜炎的炎癥反應(yīng)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- 偽 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- 偽, 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- 偽 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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R772.21

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