發(fā)布時間：2018-11-13 13:54

【摘要】：血管活性腸肽(vasoactive intestinal peptides，VIP)是肺內(nèi)重要的感覺神經(jīng)肽，主要由非腎上腺素能非膽堿能神經(jīng)纖維釋放。VIP陽性神經(jīng)纖維廣泛分布于氣道上皮細胞、支氣管平滑肌層、肺血管和支氣管血管的管壁以及粘膜下的腺體周圍。VIP的受體則主要分布于肺血管平滑肌、氣道平滑肌、氣道上皮層、粘膜下的腺體、 巨噬細胞、肺泡上皮和淋巴細胞等。VIP除擴張血管外，還具有舒張氣道平滑肌、減少肺炎性介質(zhì)釋放、清除氧自由基和抗細胞凋亡等生物學(xué)作用，其在機體內(nèi)的多種效應(yīng)日益受到重視。 支氣管上皮細胞是炎癥和物理刺激因子的靶細胞，在哮喘、支氣管炎等氣道高反應(yīng)性疾病中，常伴有不同程度氣道上皮結(jié)構(gòu)破壞和功能受損。氣道上皮的修復(fù)包括損傷邊緣細胞變平、鋪展、向損傷部位遷移和基細胞增殖分化成纖毛上皮細胞以及功能的恢復(fù)等環(huán)節(jié)。作為肺內(nèi)重要的神經(jīng)肽，VIP對氣道上皮損傷后的結(jié)構(gòu)及功能的恢復(fù)有何影響尚未見報道。因此，探討VIP在氣道上皮損傷修復(fù)中的作用及其分子機制，有助于揭示VIP在維持氣道穩(wěn)態(tài)中的作用，尋找促進肺損傷修復(fù)的可能途徑，具有重要的學(xué)術(shù)價值和臨床應(yīng)用前景。 目的：研究血管活性腸肽(VIP)對氣道上皮損傷后修復(fù)過程的影響，并探討其可能的信號轉(zhuǎn)導(dǎo)途徑。 方法：(1) VIP對人氣道上皮損傷修復(fù)過程的影響：以人支氣管上皮細胞(human bronchial epithelial cells，HBECs)株為研究對象，觀察VIP對HBECs損傷修復(fù)指數(shù)的影響；采用blind-well Boyden chamber法測定VIP對HBECs趨化性的影響；采用免疫細胞化學(xué)方法檢測HBECs增殖相關(guān)抗原Ki67的表達；流式細胞儀檢測VIP對HBECs S期細胞百分率(SPF)和細胞增殖指數(shù)(PI)的影響；采用免疫細胞化學(xué)、RT-PCR法檢測人支氣管上皮細胞鈣粘素(E-cd)蛋白及其mRNA的表達。(2)信號轉(zhuǎn)導(dǎo)機制的研究：以HBECs為研究對象，，雙察PKC抑制劑H-7、CaM抑制劑W-7、PKA抑制劑H-89和ERK抑制劑PD98059對VIP促損傷修復(fù)作用的影響。(3)CREB在VIP促氣道損傷
[Abstract]:Vasoactive intestinal peptide (vasoactive intestinal peptides,VIP) is an important sensory neuropeptide in the lung, which is mainly released by non-adrenergic non-cholinergic nerve fibers. VIP positive nerve fibers are widely distributed in airway epithelial cells and bronchial smooth muscle cells. VIP receptors are mainly distributed in pulmonary vascular smooth muscle, airway epithelium, submucosal glands, macrophages, and so on. Alveolar epithelium and lymphocytes. VIP not only dilates blood vessels, but also relaxes airway smooth muscle, reduces the release of pneumonic mediators, scavenges oxygen free radicals and inhibits apoptosis. Bronchial epithelial cells (BECs) are the target cells of inflammatory and physical stimulating factors. In airway hyperresponsiveness diseases such as asthma and bronchitis, bronchial epithelial cells are often accompanied with various degree of destruction of airway epithelial structure and impairment of function. The repair of airway epithelium includes flattening and spreading of injured edge cells, migration to injury site, proliferation and differentiation of basal cells into ciliated epithelial cells, and functional recovery. As an important neuropeptide in lung, the effect of VIP on the recovery of structure and function after airway epithelial injury has not been reported. Therefore, to explore the role of VIP in the repair of airway epithelial injury and its molecular mechanism is helpful to reveal the role of VIP in maintaining airway homeostasis and to find possible ways to promote the repair of lung injury, which has important academic value and clinical application prospect. Aim: to investigate the effect of vasoactive intestinal peptide (VIP) on the repair of airway epithelium after injury and to explore its possible signal transduction pathway. Methods: (1) the effect of VIP on the repair process of human airway epithelial injury: the effect of VIP on the repair index of HBECs injury was observed in human bronchial epithelial cell line (human bronchial epithelial cells,HBECs. The effect of VIP on chemotaxis of HBECs was determined by blind-well Boyden chamber assay, and the expression of HBECs proliferation-associated antigen Ki67 was detected by immunocytochemistry. The effect of VIP on the percentage of HBECs S phase (SPF) and cell proliferation index (PI) was detected by flow cytometry. The expression of cadherin (E-cd) protein and its mRNA in human bronchial epithelial cells were detected by immunocytochemistry and RT-PCR method. (2) the mechanism of signal transduction. The effects of CaM inhibitor W-7 PKA inhibitor H-89 and ERK inhibitor PD98059 on VIP induced injury repair. (3) the effect of CREB on VIP induced airway injury
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2006
【分類號】：R363

【參考文獻】

相關(guān)期刊論文 前10條

1 吳昌歸,毛寶齡,孫濱;神經(jīng)肽與哮喘關(guān)系的基礎(chǔ)研究[J];第三軍醫(yī)大學(xué)學(xué)報;1998年01期

2 張丙芳,林允信,戚好文,李志超,薛軍,賈斌,趙淑華;降鈣素基因相關(guān)肽對油酸型急性肺損傷家兔細胞因子的影響[J];第四軍醫(yī)大學(xué)學(xué)報;2001年18期

3 王曦,吳人亮,郝春榮,唐長生,熊密;吸煙小鼠呼吸道上皮細胞上皮鈣粘附素表達的研究[J];中國組織化學(xué)與細胞化學(xué)雜志;1998年03期

4 史育紅,譚天秩;血管活性腸肽受體研究現(xiàn)狀[J];國外醫(yī)學(xué)(放射醫(yī)學(xué)核醫(yī)學(xué)分冊);1998年04期

5 徐海燕;血管活性腸肽的免疫調(diào)節(jié)作用[J];國外醫(yī)學(xué)(免疫學(xué)分冊);2002年02期

6 管茶香,周伏文,羅自強,秦曉群,張長青,孫秀泓;血管活性腸肽對肺組織CTP:磷酸膽堿二胞苷酰轉(zhuǎn)移酶活性的影響[J];湖南醫(yī)科大學(xué)學(xué)報;2002年02期

7 秦曉群,孫秀泓,羅自強,管茶香,張長青;表皮生長因子對氣道上皮細胞VIP分泌及VIPR表達的影響[J];湖南醫(yī)科大學(xué)學(xué)報;1999年02期

8 李健,李曉玫,王海燕;低密度脂蛋白誘導(dǎo)腎小管上皮細胞增殖的信號通路[J];基礎(chǔ)醫(yī)學(xué)與臨床;2000年01期

9 秦曉群;氣道上皮細胞的抗氧化損傷保護及微環(huán)境調(diào)控[J];生理科學(xué)進展;1999年02期

10 秦曉群,孫秀泓,羅自強;血管活性腸肽、表皮生長因子上調(diào)支氣管上皮細胞bcl-2基因表達[J];生理學(xué)報;1999年04期

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