本文選題：大皰性類天皰瘡 + 趨化作用��； 參考：《大連理工大學(xué)》2012年博士論文

【摘要】：大皰性類天皰瘡(Bullous Pemphigoid, BP)是一種皮膚水皰性自身免疫性疾病,其主要癥狀表現(xiàn)為張力性水皰的形成、表皮和真皮的分離、嚴(yán)重的炎癥細(xì)胞浸潤(rùn)及半橋粒和細(xì)胞外基質(zhì)的破壞,該病的發(fā)生和抗半橋粒蛋白BP180和BP230的自身性抗體有關(guān)。通過IgG被動(dòng)轉(zhuǎn)移動(dòng)物模型,我們了解到一些BP的發(fā)病機(jī)制。在BP的IgG被動(dòng)轉(zhuǎn)移動(dòng)物模型中,抗BP180的IgG誘導(dǎo)水皰的形成,而水皰的形成依賴于補(bǔ)體的激活、肥大細(xì)胞的脫顆粒和嗜中性粒細(xì)胞的浸潤(rùn)。此外,在BP病人和實(shí)驗(yàn)動(dòng)物的水皰液中還發(fā)現(xiàn)了許多蛋白水解酶,如血纖維蛋白酶、膠原蛋白酶、嗜中性粒細(xì)胞彈性蛋白酶(Neutrophil Elastase, NE)和基質(zhì)金屬蛋白酶(Matrix Metalloproteinase, MMP-9)的存在。由于這些蛋白水解酶能夠降解細(xì)胞外基質(zhì)組分,因此他們對(duì)于BP水皰的形成有重要的作用。如果能夠找到這些蛋白酶在BP中產(chǎn)生及參與病癥的途徑,并抑制這些途徑的發(fā)生,則對(duì)BP的治療具有很大的幫助。 本論文通過基因敲除和炎癥細(xì)胞重建的方法分析了NE和肥大細(xì)胞蛋白酶-4(Mouse mast cell protease-4, mMCP-4)在BP動(dòng)物模型中參與疾病發(fā)生的機(jī)制,更重要的是這一發(fā)現(xiàn)為BP的治療提供了潛在的靶點(diǎn)。 我們的實(shí)驗(yàn)結(jié)果顯示NE能夠降解鼠BP180重組蛋白,其酶切位點(diǎn)在BP180細(xì)胞外結(jié)構(gòu)域的第506和561號(hào)氨基酸上,產(chǎn)生的小肽分別命名為p506和p561。在體內(nèi)和體外的實(shí)驗(yàn)結(jié)果都表明,p561小肽對(duì)嗜中性粒細(xì)胞有趨化活性。向B6小鼠體內(nèi)局部注射NE能夠?qū)⑹戎行粤＜?xì)胞招募至皮膚,而如果向小鼠體內(nèi)同時(shí)注射NE的抑制劑α1-PI(al-proteinase inhibitor)則無嗜中性粒細(xì)胞的浸潤(rùn)。更重要的是,NE能夠直接降解人和鼠的BP180以及天然狀態(tài)存在的人BP180三聚體分子。這些結(jié)果表明：1)NE能夠直接造成細(xì)胞外基質(zhì)的損傷；2)NE降解BP180后產(chǎn)生具有嗜中性粒細(xì)胞趨化活性的小肽,這個(gè)小肽能夠加重發(fā)病初期的病癥強(qiáng)度。 除了NE外,mMCP-4也和許多自身免疫性疾病和炎癥反應(yīng)有關(guān),然而其參與疾病的機(jī)制尚不清楚。在實(shí)驗(yàn)中,我們發(fā)現(xiàn)mMCP-4對(duì)于自身免疫性疾病BP動(dòng)物模型中水皰的形成至關(guān)重要。mMCP-4缺陷型小鼠能夠抑制BP的發(fā)生,但mMCP-4-/-小鼠中補(bǔ)體的激活、肥大細(xì)胞的脫顆粒和嗜中性粒細(xì)胞的早期浸潤(rùn)都和野生型類似。而向mMCP-4-/-小鼠體內(nèi)注射致病性IgG后,由于mMCP-4的缺失,激活的MMP-9的量明顯低于野生型。野生型和mMCP-4-/-小鼠的肥大細(xì)胞重建不能完全恢復(fù)激活的MMP.9的數(shù)量。在mMCP-4-/-(?)勺小鼠體內(nèi)重建mMCP-4+/+小鼠的肥大細(xì)胞,能夠使機(jī)體產(chǎn)生水皰和嗜中性粒細(xì)胞的浸潤(rùn),而重建mMCP-4-/-小鼠的肥大細(xì)胞則不能。mMCP-4也能夠在體內(nèi)和體外降解半橋粒蛋白BP180,并造成表皮和真皮的分離。這些實(shí)驗(yàn)結(jié)果表明,mMCP-4在BP的發(fā)病機(jī)制中發(fā)揮兩個(gè)作用：它既能夠激活MMP-9,又能直接降解BP180,從而造成半橋粒和基底膜的細(xì)胞外基質(zhì)損傷。
[Abstract]:Bullous pemphigoid (BPP) is a skin blistering autoimmune disease characterized by the formation of tensional blisters, the separation of epidermis and dermis, severe inflammatory cell infiltration and the destruction of semidomere and extracellular matrix. The development of the disease is related to the autoantibodies against hemidopontin BP180 and BP230. Through the IgG passive transfer animal model, we know some BP pathogenesis. In BP IgG passive transfer animal model, the formation of vesicles was induced by anti-BP180 IgG, and the formation of vesicles depended on the activation of complement, degranulation of mast cells and infiltration of neutrophils. In addition, many proteolytic enzymes such as blood fibrinase, collagenase, neutrophil elastase (NEUtrophil) and matrix metalloproteinase (MMP-9) were found in the blister fluid of BP patients and laboratory animals. Because these proteolytic enzymes can degrade extracellular matrix components, they play an important role in the formation of BP blister. If we can find the pathway that these proteases produce and participate in the disease in BP, and inhibit the occurrence of these pathways, it will be of great help to the treatment of BP. In this paper, the mechanism of NE and mast cell protease-4mouse mast cell protease-4 (mMCP-4) involved in the development of disease in BP animal model was analyzed by gene knockout and inflammatory cell reconstruction. More importantly, this discovery provided a potential target for the treatment of BP. Our results showed that NE could degrade mouse BP180 recombinant protein, and its restriction site was on amino acids 506 and 561 of the extracellular domain of BP180, and the resulting peptides were named p506 and p561respectively. The results in vivo and in vitro showed that p561 small peptide was chemoattractant to neutrophils. Local injection of NE into B6 mice could recruit neutrophils into the skin, but no infiltration of neutrophils was observed if the inhibitor 偽 1-PI(al-proteinase inhibitor of NE was injected simultaneously into the skin of B6 mice. More importantly, it can directly degrade BP180 of human and mouse and human BP180 trimer in natural state. These results suggest that the small peptide with neutrophil chemotactic activity can be produced after the degradation of BP1 80 by 1: 1 ne, which can aggravate the intensity of the disease in the early stage of the disease. In addition to NE, mMCP-4 is also associated with many autoimmune diseases and inflammatory reactions, but the mechanism of its involvement in the disease is unclear. In our experiment, we found that mMCP-4 is very important to the formation of blisters in the animal model of autoimmune disease BP. MMCP-4 deficient mice can inhibit the occurrence of BP, but the complement activation in mMCP-4-r-mice. Degranulation of mast cells and early infiltration of neutrophils are similar to wild type. However, after injection of pathogenic IgG into mMCP-4-r-mice, the amount of activated MMP-9 was significantly lower than that of wild type due to the absence of mMCP-4. In wild type and mMCP-4-r-mice, mast cell remodeling did not fully restore the number of activated MMP.9. At mMCP-4-r-p-) Reconstructing mast cells in mMCP-4 / mice in spoons can cause blisters and neutrophils to infiltrate the body. But the mMCP-4-r-mouse mast cells could not. MMCP-4 could also degrade BP180 in vivo and in vitro, resulting in the separation of epidermis and dermis. These results suggest that mMCP-4 plays two roles in the pathogenesis of BP: it not only activates MMP-9, but also directly degrades BP180, thus causing damage to the extracellular matrix of the semidopontine and basement membrane.
【學(xué)位授予單位】：大連理工大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R758.6

【共引文獻(xiàn)】

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