【摘要】： 目的檢測(cè)大鼠真菌性角膜潰瘍和細(xì)菌性角膜潰瘍中SP的表達(dá)變化過(guò)程及差異,探討SP在真菌性角膜潰瘍發(fā)病過(guò)程中的作用。 方法55只Wistar大鼠隨機(jī)分為空白對(duì)照組(A),實(shí)驗(yàn)一組(B),實(shí)驗(yàn)二組(C),右眼為實(shí)驗(yàn)眼,左眼為對(duì)照眼。A組5只大鼠不做任何處理；B組為真菌性角膜潰瘍組25只,接種鐮刀菌；C組為細(xì)菌性角膜潰瘍組25只,接種金黃色葡萄球菌；對(duì)照眼以生理鹽水代替菌株。每日裂隙燈觀察眼表情況,并刮取B組、C組潰瘍邊緣及底部組織涂片行革蘭染色和10%氫氧化鉀涂片,同時(shí)將刮取物接種于沙保弱培養(yǎng)基上25℃培養(yǎng)以確定真菌或細(xì)菌感染。詳細(xì)記錄各組大鼠角膜潰瘍生長(zhǎng)情況。分別于造模后1d、5d、8d、10d、14d隨機(jī)抽取各組大鼠取材。HE染色觀察角膜潰瘍組織學(xué)變化。免疫組織化學(xué)方法和RT-PCR法檢測(cè)SP的表達(dá)。 結(jié)果1、B組1d形成灰白色潰瘍?cè)?可見(jiàn)免疫環(huán)；5d為潰瘍高峰期,可出現(xiàn)穿孔；8d時(shí)潰瘍縮��；10d有新生血管生長(zhǎng),14d形成血管化角膜白斑。C組變化與B組時(shí)間變化類(lèi)似,但C組充血更嚴(yán)重,無(wú)免疫環(huán)。對(duì)照眼1d角膜水腫,5d角膜透明,未見(jiàn)感染。 2、HE染色示B組1d潰瘍形成,基質(zhì)淺層膠原纖維排列紊亂,中性粒細(xì)胞浸潤(rùn)。5d潰瘍加重,整個(gè)基質(zhì)層大量炎性細(xì)胞浸潤(rùn)；8d潰瘍縮小,基質(zhì)炎癥細(xì)胞減少,成纖維細(xì)胞增生,見(jiàn)新生血管形成；10d新生血管成熟；14d基質(zhì)層纖維呈瘢痕樣重構(gòu)。C組變化類(lèi)似,炎細(xì)胞浸潤(rùn)較多。 3、免疫組化示：(1)正常角膜上皮細(xì)胞中中等量表達(dá)SP,基質(zhì)層及內(nèi)皮有微弱表達(dá)(2)B組中角膜內(nèi)皮細(xì)胞、炎癥細(xì)胞、新形成的血管內(nèi)皮細(xì)胞,成纖維母細(xì)胞中SP表達(dá)均增強(qiáng),8d最強(qiáng),10d下降,14d降至低于正常,差異有統(tǒng)計(jì)學(xué)意義(P0.05)(3)C組其變化趨勢(shì)與B組中類(lèi)似,但相同時(shí)間點(diǎn)兩組間比較,B組低于C組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)(4)對(duì)照眼1d達(dá)到最高,5d降至正常水平。各時(shí)間點(diǎn)B組,C組實(shí)驗(yàn)眼與對(duì)照眼之間比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 4、RT-PCR與免疫組織化學(xué)檢測(cè)的變化趨勢(shì)一致。結(jié)論真菌性角膜潰瘍中SP表達(dá)早期與潰瘍進(jìn)展一致,晚期與潰瘍修復(fù)一致,提示SP可能參與了真菌性角膜潰瘍?cè)缙趽p傷和晚期修復(fù)過(guò)程；其表達(dá)低于同時(shí)間點(diǎn)的細(xì)菌性角膜潰瘍,提示其表達(dá)水平較低可能參與了真菌性角膜潰瘍疼痛較輕的機(jī)制。
[Abstract]:Objective to detect the expression of SP in fungal corneal ulcer and bacterial corneal ulcer in rats and to explore the role of SP in the pathogenesis of fungal corneal ulcer. Methods Fifty-five Wistar rats were randomly divided into control group (group B), experimental group 2 (group C), right eye (group C) and control group (group A) (n = 5). 25 rats in group B were inoculated with Fusarium in group B without any treatment. Group C was inoculated with Staphylococcus aureus in 25 rats of bacterial corneal ulcer group, and the control group was replaced by normal saline. Slit-lamp was used to observe the surface of the eye. Gram staining and 10% potassium hydroxide smear were performed on the edge and bottom tissue of ulcers in group B and C, and the scrape was inoculated at 25 鈩，

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