本文選題：BALB/C + C57BL/6�。� 參考：《浙江大學(xué)》2010年碩士論文

【摘要】： 目的：(1)建立BALB/C和C57BL／6兩種不同品系小鼠的鐮刀菌和曲霉菌性的角膜炎模型；(2)對比兩種品系小鼠真菌性角膜炎的臨床表現(xiàn)差異；(3)測定感染眼球及同側(cè)頸部淋巴結(jié)的Th1相關(guān)細(xì)胞因子IFN-γ及Th2相關(guān)細(xì)胞因子IL-4的變化情況。 方法：(1)小鼠角膜真菌感染模型的制作：用2.0mm環(huán)轉(zhuǎn)在小鼠角膜中央印痕,自制基質(zhì)劃傷鉤于印痕范圍內(nèi)作線形劃傷至淺基質(zhì)層,共5道,板層刀去除損傷區(qū)域內(nèi)殘余的角膜上皮,然后將5μl的真菌孢子混懸液(濃度為108CFU／ml)滴入結(jié)膜囊,自然干燥。(2)接種后用裂隙燈顯微鏡觀察角膜浸潤病灶及眼部炎癥反應(yīng)的變化,每天1次。(3)接種后24h,48h和72h處死小鼠,摘除模型眼及相應(yīng)側(cè)頸部淋巴結(jié),分別制作成組織勻漿,ELISA法檢測Th1相關(guān)IFN-γ及Th2相關(guān)IL-4的變化。 結(jié)果：(1)BALB／c小鼠在接種鐮刀菌和曲霉菌后僅出現(xiàn)輕、中度角膜浸潤混濁和炎癥反應(yīng),無組織化膿壞死,5-7d內(nèi)所有角膜浸潤消失而痊愈,角膜恢復(fù)透明或僅出現(xiàn)少量疤痕；而C57BL／6小鼠在接種真菌后1-2d內(nèi)即出現(xiàn)濃密的角膜浸潤混濁,重度基質(zhì)水腫和前房積膿,3d后組織發(fā)生化膿壞死,角膜穿孔,眼內(nèi)容突出,最終以嚴(yán)重的角膜疤痕混濁而愈合,部分小鼠出現(xiàn)眼球萎縮。(2)BALB／c小鼠在感染鐮刀菌和曲霉菌后眼球IFN-γ均明顯升高,高峰值分別為正常的2.9和2.6倍；同時,頸淋巴結(jié)IFN-γ也明顯升高,但低于感染局部的升高幅度,高峰值分別為正常的2.0和1.5倍。而C57BL／6在感染鐮刀菌和曲霉菌后眼球IFN-γ僅輕度升高,且短時間內(nèi)下降至正常,高峰值僅為正常的1.3和1.1倍,但頸淋巴結(jié)IFN-γ則明顯升高,在72h分別達(dá)到正常的1.6和1.5倍。(3)BALB/c在感染鐮刀菌和曲霉菌后眼球IL-4均明顯降低,低谷水平僅為正常的0.6和0.5；而頸淋巴結(jié)IL-4則逐漸升高,高峰值為正常的1.2和1.5倍。而C57BL／6在感染鐮刀菌和曲霉菌后眼球IL-4也明顯降低,低谷水平為正常的0.5和0.6,且持續(xù)至72h仍維持低水平；但頸淋巴結(jié)IL-4也逐漸升高,高峰值為正常的1.9和1.6倍。 結(jié)論：(1)真菌性角膜炎是以Th1介導(dǎo)為主的免疫炎癥反應(yīng)過程；(2)感染局部Th1反應(yīng)的強(qiáng)弱是真菌性角膜炎預(yù)后的重要因素；(3)BALB/c小鼠感染局部的Th1反應(yīng)明顯強(qiáng)于C57BL／6小鼠；(4)在角膜真菌感染過程中,Th1極化在感染局部和頸淋巴結(jié)同步出現(xiàn),而Th2極化則在感染局部和頸淋巴結(jié)出現(xiàn)分離現(xiàn)象。
[Abstract]:Objective: (1) to establish Fusarium and aspergillus keratitis models of BALB / C and C57BL / 6 mice, (2) to compare the clinical manifestations of fungal keratitis between the two strains. (3) the changes of Th1-associated cytokine IFN- 緯 and Th2-associated cytokine IL-4 in infected eyeball and ipsilateral cervical lymph nodes were measured. Methods: (1) the model of corneal fungal infection in mice: the 2.0mm loop was applied to the central impression of the cornea of the mice, and the self-made matrix laceration hook was used for linear scratching to the superficial stromal layer in the range of the mark, for a total of 5 passages. The residual corneal epithelium in the injured area was removed by lamellar knife, and then 5 渭 l fungal spore suspension (concentration 108 CFU / ml) was dripped into conjunctival sac. (2) the corneal infiltrating focus and ocular inflammatory reaction were observed by slit lamp microscope after inoculation. (3) the mice were killed at 24 h, 48 h and 72 h after inoculation, the model eyes and the corresponding cervical lymph nodes were removed, and the changes of Th1-related IFN- 緯 and Th2-related IL-4 were detected by Elisa. Results: (1) after inoculation with Fusarium and Aspergillus, BALB / c mice only showed mild and moderate corneal infiltration, turbid and inflammatory reaction. All corneal infiltration disappeared and recovered within 5-7 days without tissue purulent necrosis, and the cornea recovered transparently or only a few scars appeared. However, C57BL / 6 mice had dense corneal infiltration and opacity within 1 to 2 days after inoculation of fungi, severe stromal edema and anterior chamber pyosis occurred 3 days later, corneal perforation, prominent eye content, and finally healed with severe corneal scar opacity. (2) after infection with Fusarium and Aspergillus, IFN- 緯 of BALB / c mice increased significantly, the peak value of IFN- 緯 was 2.9 and 2.6 times of normal value, meanwhile, IFN- 緯 of cervical lymph nodes was also significantly increased, but lower than that of local infection. The peak values were 2.0 and 1.5 times of normal values, respectively. After infection with Fusarium and Aspergillus, IFN- 緯 increased slightly and decreased to normal in a short time. The peak value of IFN- 緯 in C57BL / 6 was only 1.3 and 1.1 times of normal, but IFN- 緯 in cervical lymph nodes was significantly increased. (3) BALB / c decreased significantly after infection with Fusarium and Aspergillus, and the low level was only 0.6 and 0.5, while the level of IL-4 in cervical lymph nodes increased gradually, with a peak value of 1.2 and 1.5 times higher than that of normal. After infection with Fusarium and Aspergillus, C57BL / 6 decreased the level of IL-4 in eyeball, the low level of IL-4 was 0.5 and 0.6 of normal, and maintained a low level until 72 hours, but the IL-4 of cervical lymph node increased gradually, the peak value of IL-4 was 1.9 and 1.6 times of normal. Conclusion: (1) fungal keratitis is a Th1-mediated immune inflammatory process, (2) the intensity of local Th1 response is an important factor in prognosis of fungal keratitis, (3) the Th1 response in BALB / c mice is significantly stronger than that in C57BL / 6 mice. (4) Th1 polarization appeared simultaneously in infected area and cervical lymph node during corneal fungal infection, while Th2 polarization appeared separation phenomenon in infected area and cervical lymph node.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R772.21

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