本文選題：口腔扁平苔蘚 + 細(xì)胞免疫��； 參考：《鄭州大學(xué)》2014年博士論文

【摘要】：(1)口腔扁平苔蘚(oral lichen planus,OLP)是臨床上最為常見的慢性非感染性口腔黏膜病,是一種發(fā)生在皮膚及黏膜的慢性炎癥性疾病。本研究通過對(duì)65例口腔扁平苔蘚患者病變組織進(jìn)行組織病理學(xué)檢查證實(shí)其主要病理特征為:上皮過度角化或角化不全,棘層增生多見。上皮形狀不規(guī)則,呈鋸齒狀。基底細(xì)胞層液化、變性,導(dǎo)致基底層角質(zhì)細(xì)胞排列紊亂,基底膜界限不清,基底層角質(zhì)細(xì)胞液化壞死。固有層中有密集的淋巴細(xì)胞浸潤帶,該浸潤帶主要分布在固有層的淺層,部分還可深入到固有層的深層或者粘膜下層。本研究65例口腔扁平苔蘚患者中有23例在上皮的棘層、基底層或粘膜固有層可見圓形或卵圓形膠樣小體,為均質(zhì)性、嗜酸性,可能是屬于凋亡的產(chǎn)物。5/65例出現(xiàn)上皮輕度不典型增生,表現(xiàn)為上皮細(xì)胞異乎于常態(tài)的增生,具體表現(xiàn)為增生的細(xì)胞大小不一,形態(tài)多樣,核大而濃染,核漿比例增大,核分裂可增多但通常呈正常核分裂像。細(xì)胞排列較亂,細(xì)胞層次增多,極向消失。(2)應(yīng)用免疫組化S-ABC法檢測CD4+、CD8+T細(xì)胞在35例口腔扁平苔蘚組織及10例正常口腔粘膜中的表達(dá)水平,結(jié)果表明:正�？谇徽衬ど掀�(nèi)幾乎不表達(dá)CD4+、CD8+T細(xì)胞,僅基底層可見散在的CD4+、CD8+T細(xì)胞陽性表達(dá)�？谇槐馄教μ\固有層顯示中等量或大量淋巴細(xì)胞浸潤,92.3%(60/65)的病例出現(xiàn)淋巴細(xì)胞浸潤。口腔扁平苔蘚組與對(duì)照組比較CD8+表達(dá)水平顯著升高(P0.01),CD4+與對(duì)照組比較無顯著性差異(P0.05),表明口腔扁平苔蘚的發(fā)生可能與病變組織中細(xì)胞免疫改變有關(guān)。口腔扁平苔蘚組CD8+T細(xì)胞主要位于固有層淺層,接近基底膜破壞處表達(dá),CD4+T細(xì)胞主要于固有層深層表達(dá)。CD4/CD8比值下降,與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P0.0 5)。(3)應(yīng)用免疫組化S-ABC法檢測Bcl-2和Bax在口腔扁平苔蘚組及對(duì)照組中的表達(dá)水平。結(jié)果顯示,Bcl-2主要表達(dá)于上皮基底層及棘層細(xì)胞的胞漿和胞膜,在正�？谇火つぶ�,8/10例陽性(+)表達(dá),陽性表達(dá)率80%;在口腔扁平苔蘚的上皮組織中,17/35例陰性表達(dá),3/35例在基底層和棘層呈零星散在表達(dá),15/35例淋巴細(xì)胞浸潤帶處陽性表達(dá)增強(qiáng),陽性表達(dá)率51.4%;Bcl-2在口腔扁平苔蘚中陽性表達(dá)率與正常口腔黏膜比較無顯著性變化(P0.05),但在淋巴細(xì)胞浸潤帶處表達(dá)增強(qiáng)。35例口腔扁平苔蘚組織中,Bax陽性表達(dá)27例,陽性率77.1%,其中14例呈強(qiáng)陽性(+++)表達(dá),9例為中度陽性(++),4例弱陽性(+),陽性細(xì)胞以基底層和棘層中下部最明顯。10例正常人口腔黏膜中,3例弱陽性(+)表達(dá),其余病例均為陰性(-),陽性表達(dá)率為30.0%。Bax在口腔扁平苔蘚中陽性表達(dá)率和表達(dá)強(qiáng)度與正常口腔黏膜比較明顯增高(P0.01)。(4)用TUNEL法(原位末端轉(zhuǎn)移酶標(biāo)記法)檢測口腔扁平苔蘚患者病變組織及對(duì)照組中細(xì)胞凋亡的情況,結(jié)果表明,65例口腔扁平苔蘚病變組織口腔粘膜上皮層及固有層均可見凋亡細(xì)胞表達(dá),核輪廓清楚、致密,呈棕褐色。口腔扁平苔蘚病變組織的上皮細(xì)胞凋亡指數(shù)明顯較正常對(duì)照組高(P0.01),凋亡細(xì)胞多位于基底層液化壞死處,表達(dá)率為85.7%(30/35)。而對(duì)照組中的凋亡細(xì)胞多位于表層、棘層及基底層,表達(dá)率為80%(8/10)。(5)雙抗夾心酶聯(lián)免疫吸附試驗(yàn)檢測外周血血清TNF-a和IFN-γ含量,結(jié)果表明口腔扁平苔蘚患者外周血血清IFN-γ和TNF-α含量均顯著高于對(duì)照組(P0.05)。(6)采用單克隆抗體花環(huán)法對(duì)T細(xì)胞亞群進(jìn)行識(shí)別與測定,結(jié)果發(fā)現(xiàn),與健康組比較,20例口腔扁平苔蘚患者CD8+顯著升高(P0.05),CD4+與對(duì)照組比較無顯著性差異(P0.05),CD4+/CD8+與對(duì)照組比較顯著降低(P0.01),和免疫組織化學(xué)染色分析結(jié)果一致。(7)采用透射電鏡技術(shù)觀察發(fā)現(xiàn)口腔扁平苔蘚患者口腔黏膜棘層胞質(zhì)內(nèi)充滿張力原纖維,并變粗增多,排列紊亂,內(nèi)織網(wǎng)擴(kuò)張,內(nèi)含不定形物,線粒體腫大及峭消失,游離核糖體增多。多數(shù)細(xì)胞器減少,細(xì)胞核團(tuán)塊狀異染色質(zhì)增多,核周間隙加寬,有的核旁空泡,細(xì)胞變形,橋粒結(jié)構(gòu)破壞,細(xì)胞間隙明顯擴(kuò)大。基底細(xì)胞與基底膜半橋粒及細(xì)胞間橋粒破壞,部份細(xì)胞變形,排列紊亂,錯(cuò)位,水腫細(xì)胞間隙加大,空泡,變性,胞質(zhì)內(nèi)線粒體腫脹,峭明顯減少,內(nèi)質(zhì)網(wǎng)擴(kuò)張。細(xì)胞核增大,異染色質(zhì)增多,凋亡細(xì)胞增多。凋亡細(xì)胞出現(xiàn)頻率高于健康組。凋亡主要表現(xiàn)為染色質(zhì)沿核膜邊聚,且呈“C”形、新月形或團(tuán)塊狀,并可見凋亡小體。結(jié)論1.口腔扁平苔蘚的病理組織學(xué)特征為基底層角質(zhì)形成細(xì)胞的損傷,不同程度的變性、液化壞死,以及固有層T淋巴細(xì)胞的帶狀浸潤。2.口腔扁平苔蘚患者病損區(qū)組織固有層及外周血中T淋巴細(xì)胞亞群的顯著性改變,表明細(xì)胞免疫功能的紊亂在口腔扁平苔蘚發(fā)病中發(fā)揮著重要作用。3.口腔黏膜上皮組織中角質(zhì)形成細(xì)胞凋亡及凋亡抑制因子Bcl-2在病損區(qū)淋巴細(xì)胞浸潤帶處和凋亡促進(jìn)因子Bax在上皮基底層角質(zhì)形成細(xì)胞中表達(dá)增強(qiáng),提示細(xì)胞凋亡異常與口腔扁平苔蘚的發(fā)生、發(fā)展有重要關(guān)系,病損區(qū)組織超微結(jié)構(gòu)觀察結(jié)果也與之相符。4.TNF-α、IFN-γ在口腔扁平苔蘚患者外周血中異常高表達(dá),提示TNF-α、IFN-γ可能是造成口腔扁平苔蘚病損的重要因子之一。
[Abstract]:(1) oral lichen planus (oral lichen planus, OLP) is the most common chronic noninfectious oral mucous membrane disease in clinic. It is a chronic inflammatory disease occurring in the skin and mucous membrane. By histopathological examination of 65 cases of oral lichen planus, the main pathological features are epithelial hyperkeratosis. The epithelium is irregular and accretion. The epithelium is irregular and serrated. The basal cell layer is liquefied and denatured, resulting in the disorder of the basal layer keratinocytes, the indistinct basement membrane boundaries, and the liquefaction and necrosis of the basal layer keratinocytes. The lamina is densely infiltrated with lymphocytes, the infiltrating zone is mainly distributed in the shallow layer of the lamina and part is also available. In 65 cases of oral lichen planus, 23 of the 65 cases of oral lichen planus were in the epithelia, and the basal layer or the lamina propria showed circular or oval shaped corpuscle. It was homogeneous, eosinophilic, and may be the product of apoptosis. The epithelia is mild atypical hyperplasia, showing epithelial cell differentiation. The cell size is different, the cell size is different, the morphology is different, the nucleus is large and dense, the proportion of the nuclear plasma is increased, the nuclear division can be increased but usually the normal nuclear division. The cell arrangement is disorderly, the cell level increases and the pole disappears. (2) CD4+, CD8+T cells in the oral lichen planus tissue and the CD8+T cells in 35 cases of oral lichen planus tissue and The expression level in the normal oral mucosa of 10 cases showed that the normal oral mucosa almost did not express CD4+, CD8+T cells, only the scattered CD4+, and positive expression of CD8+T cells in the basal layer. The oral lichen lamina propria showed an equal or large number of lymphocyte infiltration, and 92.3% (60/65) cases had lymphocytic infiltration. Oral squamous cell was flat. The expression level of CD8+ in the moss group was significantly higher than that in the control group (P0.01), and there was no significant difference between the CD4+ and the control group (P0.05), indicating that the occurrence of oral lichen planus may be related to the cellular immune changes in the pathological tissue. The CD8+T cells in the oral lichen planus group are mainly located in the superficial layer of the lamina propria, close to the expression of the basement membrane destruction, and the CD4+T cell master. There was a significant difference in the expression of.CD4/CD8 ratio in the deep lamina of the lamina propria (P0.0 5). (3) the expression level of Bcl-2 and Bax in the oral lichen planus and the control group was detected by immunohistochemical S-ABC. The results showed that Bcl-2 was mainly expressed in the cytoplasm and membrane of the upper layer and the acanthosis cells in the normal oral cavity. In the mucosa, 8/10 positive (+) expression, positive expression rate was 80%, in the epithelial tissue of oral lichen planus, 17/35 negative expression, 3/35 cases in the basal layer and spinous layer were scattered in the expression, 15/35 cases of lymphocyte infiltration zone positive expression increased, positive expression rate of 51.4%; Bcl-2 in oral lichen planus positive expression rate and normal oral mucus There was no significant change in the membrane (P0.05), but in the lymphocytic infiltration zone, there were 27 cases of.35 positive oral lichen planus. The positive rate was 27, the positive rate was 77.1%, of which 14 were positive (+ + +), 9 were moderately positive (+ +) and 4 were weak positive (+). The positive cells were the most obvious normal human oral mucus in the basal layer and the middle and lower part of the spine. In the membrane, 3 cases were weakly positive (+), and the other cases were negative (-). The positive expression rate of 30.0%.Bax in oral lichen planus was significantly higher than that in normal oral mucosa (P0.01). (4) the detection of pathological tissue in oral lichen planus and the cell withering in the control group by TUNEL (in situ terminal transferase labeling) The results showed that the apoptotic cells were expressed in the oral mucosa epithelium and lamina propria of 65 cases of oral lichen planus, with clear, dense, brown brown outline of the nucleus. The apoptotic index of the epithelial cells of the oral lichen planus was higher than that of the normal control group (P0.01), and the apoptotic cells were mostly located in the liquefied necrosis of the basal layer. The rate of arrival was 85.7% (30/35). The apoptotic cells in the control group were mostly located in the surface layer, spinous layer and basal layer, and the expression rate was 80% (8/10). (5) double anti sandwich enzyme-linked immunosorbent assay was used to detect the content of TNF-a and IFN- gamma in peripheral blood serum. The results showed that the content of IFN- gamma and TNF- alpha in the peripheral blood serum of the oral lichen planus patients was significantly higher than that of the control group (P0.05) (6). The T cell subgroup was identified and measured by the monoclonal antibody ring method. The results showed that compared with the healthy group, the CD8+ of 20 patients with oral lichen planus increased significantly (P0.05). There was no significant difference between the CD4+ and the control group (P0.05), and the CD4+/CD8+ decreased significantly (P0.01) compared with the control group (P0.01), and the results were consistent with the immunohistochemical staining analysis. (7) By transmission electron microscopy, it was found that the oral mucosa of oral lichen planus was filled with tension fibrils in the acanthosis of oral mucous membrane, and it became more and more coarse, arranged in disorder, the inner webs were dilated, there were unshaped objects, the mitochondria were enlarged and the crurtosis disappeared, the free ribosome increased, the majority of the organelles were reduced, the nucleus mass heterochromatin increased, and the internuclear gap was added. Wide, some paratype vacuoles, cell deformations, destruction of pbridge structure and obvious enlargement of intercellular space. Basal cells and basal membrane half bridge particles and intercellular pbridges, partial cell deformation, disorder, dislocation, enlargement of edema cell gap, vacuoles, denaturation, cytoplasmic swelling of endoplasmic reticulum, dilatation of endoplasmic reticulum, nucleus enlargement, heterostaining The number of apoptotic cells increased. The frequency of apoptotic cells was higher than that in the healthy group. Apoptosis was mainly manifested by the accumulation of chromatin along the border of the nuclear membrane, which was "C", crescent shape or mass, and apoptotic body. Conclusion the histopathological characteristics of 1. oral lichen planus are the damage of basal stratum corneum cells, varying degrees of degeneration and liquefaction. Death and zonal infiltration of the lamina propria T lymphocyte in.2. oral lichen planus patients the significant changes in the lamina propria and the T lymphocyte subsets in the peripheral blood, indicating that the dysfunction of the cellular immune function plays an important role in the pathogenesis of oral lichen planus and the apoptosis and apoptosis of keratinocytes in the oral mucosa of.3. oral mucosa The inhibitory factor Bcl-2 increased in the epithelial cell infiltration zone and the apoptotic factor Bax in the epithelial basal layer keratinocytes, suggesting that the abnormal apoptosis has an important relationship with the occurrence of oral lichen planus. The ultrastructural observation results of the lesion area also correspond to.4.TNF- a, IFN- gamma is in the oral lichen planus Abnormal high expression in peripheral blood indicates that TNF- alpha and IFN- gamma may be one of the important factors causing oral lichen planus lesions.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R781.5

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