【摘要】：目的 探討CLEC-2B (C-type lectin domain family 2, member B)基因在白癜風(fēng)患者外周血與皮損中的表達(dá)與疾病臨床類型、病程、病期、轉(zhuǎn)歸的相關(guān)性,并進(jìn)行皮損區(qū)CLEC-2B基因的蛋白表達(dá),以明確CLEC-2B基因表達(dá)的細(xì)胞定位。通過本實驗發(fā)現(xiàn)疾病多樣性及臨床轉(zhuǎn)歸與基因差異表達(dá)的相關(guān)性,據(jù)此預(yù)測白癜風(fēng)發(fā)生、臨床分型、分期及預(yù)后,從而對臨床起到一定的指導(dǎo)作用。 方法 應(yīng)用反轉(zhuǎn)錄-聚合酶鏈?zhǔn)椒磻?yīng)(reverse transcription-polymerase chain reaction, RT-PCR)方法,對白癜風(fēng)患者及正常對照者外周血及皮膚組織中CLEC-2B的表達(dá)量進(jìn)行分析,擴(kuò)增產(chǎn)物經(jīng)4.0%瓊脂糖溴乙錠電泳,凝膠成像儀下攝像證實。按照白癜風(fēng)的診斷標(biāo)準(zhǔn),利用統(tǒng)計軟件分析不同臨床類型、不同病期、不同病程、不同轉(zhuǎn)歸的白癜風(fēng)皮損區(qū)、外周血及正常人皮膚、外周血CLEC-2B基因表達(dá)的差異。并對白癜風(fēng)患者皮損組織與正常皮膚組織,用免疫組織化學(xué)方法確定皮膚組織中CLEC-2B基因的蛋白表達(dá)。 結(jié)果 1.白癜風(fēng)患者組皮損中CLEC-2BmRNA表達(dá)量高于正常對照組,差異有統(tǒng)計學(xué)意義(t=2.581,P0.05)。 2.白癜風(fēng)患者與正常對照組外周血中CLEC-2BmRNA的表達(dá)量比較差異無統(tǒng)計學(xué)意義(t=1.375,P0.05)。 3.不同性別白癜風(fēng)患者皮損組中CLEC-2BmRNA表達(dá)量比較無統(tǒng)計學(xué)差異(t=1.341,P0.05)。 4.不同病程白癜風(fēng)患者皮損中CLEC-2BmRNA的表達(dá)量比較差異無統(tǒng)計學(xué)意義(F=1.479,P0.05)。 5.尋常型白癜風(fēng)皮損組中CLEC-2BmRNA的表達(dá)量高于節(jié)段型白癜風(fēng)組,差異有統(tǒng)計學(xué)意義(t=2.432,P0.05)。 6.進(jìn)展期白癜風(fēng)患者組皮損中CLEC-2BmRNA的表達(dá)量高于穩(wěn)定期患者組,差異有統(tǒng)計學(xué)意義(t=2.204,P0.05)。 7.在不同轉(zhuǎn)歸的白癜風(fēng)患者中,各組皮損中CLEC-2B mRNA的表達(dá)量差別有統(tǒng)計學(xué)意義(F=5.218,P0.05)。 8.通過免疫組織化學(xué)方法證實在白癜風(fēng)皮損真皮層中的淋巴細(xì)胞中有CLEC-2B蛋白的表達(dá)。 結(jié)論 1.白癜風(fēng)患者皮損中CLEC-2B的表達(dá)水平的增高,提示CLEC-2B在白癜風(fēng)發(fā)病機(jī)制中具有一定的意義。 2.白癜風(fēng)患者進(jìn)展期CLEC-2B的表達(dá)水平高于穩(wěn)定期患者,提示CLEC-2B與白癜風(fēng)病期進(jìn)展有關(guān)。 3. CLEC-2B在尋常型白癜風(fēng)的表達(dá)水平高于節(jié)段型白癜風(fēng),提示CLEC-2B在決定白癜風(fēng)的分型中起一定的作用,同時推測CLEC-2B可能參與白癜風(fēng)的免疫機(jī)制。 4.不同轉(zhuǎn)歸的白癜風(fēng)患者CLEC-2B的表達(dá)水平差異有統(tǒng)計學(xué)意義,從而在基因水平揭示同一疾病不同轉(zhuǎn)歸的謎底,指導(dǎo)臨床診斷和預(yù)后。同時為尋找調(diào)控該基因的藥物,改善疾病轉(zhuǎn)歸奠定基礎(chǔ)。 5. CLEC-2B蛋白表達(dá)于白癜風(fēng)患者皮損真皮層淋巴細(xì)胞的細(xì)胞核中,進(jìn)一步加深了對本基因的認(rèn)識。
[Abstract]:Objective to investigate the expression of CLEC-2B (C-type lectin domain family-2, member B) gene in peripheral blood and lesions of patients with vitiligo and its correlation with clinical type, course of disease, stage of disease and prognosis. The protein expression of CLEC-2B gene was carried out to identify the cellular localization of CLEC-2B gene expression. In this study, we found the correlation between disease diversity and clinical outcome and gene differential expression, so as to predict the occurrence, clinical classification, staging and prognosis of vitiligo, and thus play a guiding role in clinical practice. Methods reverse transcription polymerase chain reaction (reverse transcription-polymerase chain reaction, RT-PCR) was used to analyze the expression of CLEC-2B in peripheral blood and skin of patients with vitiligo and normal controls. The amplified products were confirmed by 4.0% agarose bromide ethidium electrophoresis. According to the diagnostic criteria of vitiligo, statistical software was used to analyze the difference of CLEC-2B gene expression in different clinical types, different disease stages, different course of disease, different prognosis of vitiligo skin, peripheral blood and normal skin, and peripheral blood. The protein expression of CLEC-2B gene in the skin tissue of patients with vitiligo and normal skin tissue was determined by immunohistochemical method. Outcome 1. The expression of CLEC-2BmRNA in the lesions of patients with vitiligo was significantly higher than that of normal controls (t = 2.581, P0.05). 2. There was no significant difference in the expression of CLEC-2BmRNA in peripheral blood between vitiligo patients and normal controls (t = 1.375, P0.05). 3. There was no significant difference in the expression of CLEC-2BmRNA between male and female patients with vitiligo (t = 1.341, P0.05). 4. There was no significant difference in the expression of CLEC-2BmRNA in the lesions of patients with vitiligo at different stages (F = 1.479, P0.05). 5. The expression of CLEC-2BmRNA in vitiligo vulgaris group was higher than that in segmental vitiligo group (t = 2.432, P0.05). 6. The expression of CLEC-2BmRNA in skin lesions of patients with progressive vitiligo was significantly higher than that of patients with stable vitiligo (t = 2.204, P0.05). 7. In the patients with vitiligo with different outcomes, the expression of CLEC-2B mRNA in the lesions of each group was significantly different (F = 5.218, P0.05). 8. The expression of CLEC-2B protein was confirmed by immunohistochemical method in the lymphocytes of the dermis of vitiligo. Conclusion 1. The increased expression of CLEC-2B in the lesions of patients with vitiligo suggests that CLEC-2B may play an important role in the pathogenesis of vitiligo. 2. The expression level of CLEC-2B in patients with advanced vitiligo was higher than that in patients with stable vitiligo, suggesting that CLEC-2B was related to the progression of vitiligo. 3. The expression level of CLEC-2B in vitiligo vulgaris is higher than that in segmental vitiligo, suggesting that CLEC-2B may play a role in determining the classification of vitiligo, and that CLEC-2B may be involved in the immune mechanism of vitiligo. 4. The expression level of CLEC-2B in vitiligo patients with different prognosis has statistical significance, so as to reveal the mystery of different outcomes of the same disease at the gene level, and guide the clinical diagnosis and prognosis. At the same time, it lays a foundation for finding the drugs that regulate the gene and improving the prognosis of the disease. 5. The expression of CLEC-2B protein in the nuclei of dermis lymphocytes from patients with vitiligo further enhanced the understanding of this gene.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R758.41

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