本文選題：高遷移率族蛋白B + 慢性阻塞性肺疾病��； 參考：《醫(yī)學(xué)研究生學(xué)報》2014年12期

【摘要】：目的肺動脈高壓與慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)密切相關(guān)。研究COPD繼發(fā)肺動脈高壓的機制,探討COPD模型大鼠遠(yuǎn)端肺動脈平滑肌細(xì)胞(pulmonary arterial smooth muscle cells,PASMCs)高遷移率族蛋白B1(high mobility group protein B1,HMGB1)表達(dá)水平與其合成分泌干擾素-γ(interferon-γ,IFN-γ)的相關(guān)性及臨床意義。方法建立COPD大鼠模型,原代培養(yǎng)PASMCs并鑒定,檢測香煙煙霧提取物(CSE)及脂多糖(LPS)誘導(dǎo)下PASMCs合成分泌細(xì)胞因子的功能,實驗分組:對照組、CSE組、LPS組、CSE+LPS組,同時4組另設(shè)瓶并分別加入1∶1000抗HMGB1抗體0.2 m L培養(yǎng)作為干預(yù)組;分別于培養(yǎng)12、24、48、72 h后,采用Western blot檢測各組PASMCs中HMGB1蛋白表達(dá)變化,ELISA檢測細(xì)胞培養(yǎng)上清液HMGB1、IFN-γ濃度,并進(jìn)行相關(guān)分析。結(jié)果光鏡下見細(xì)胞呈長梭形或"峰、谷"狀生長,α-actin免疫組化染色及細(xì)胞免疫熒光鑒定為PASMCs;Western blot及ELISA法檢測結(jié)果顯示,4實驗組組間培養(yǎng)12 h后HMGB1表達(dá)水平及細(xì)胞上清液HMGB1、IFN-γ,兩兩比較差異均有統(tǒng)計學(xué)意義(P0.05);HMGB1蛋白表達(dá)水平與IFN-γ濃度變化呈正相關(guān)(r=0.91);加入抗HMGB1抗體干預(yù)培養(yǎng)12 h后,干預(yù)組IFN-γ濃度均較對應(yīng)實驗組減弱(P0.05);同組內(nèi),于細(xì)胞培養(yǎng)12、24、48、72 h,Western blot及ELISA法檢測結(jié)果差異均無統(tǒng)計學(xué)意義(P0.05)。結(jié)論 HMGB1表達(dá)水平與COPD模型大鼠遠(yuǎn)端PASMCs合成分泌IFN-γ呈正相關(guān),抗HMGB1抗體為臨床治療COPD、抑制其全身炎癥反應(yīng)提供了新的干預(yù)靶點。
[Abstract]:Objective Pulmonary hypertension is closely related to chronic obstructive pulmonary disease (obstructive pulmonary). To investigate the mechanism of pulmonary hypertension, and to investigate the relationship between the expression of high mobility group protein B1 (high mobility group protein B1HMGB1) and the synthesis and secretion of interferon- 緯 -interferon- 緯 (IFN- 緯) in pulmonary smooth muscle cells of distal pulmonary artery of COPD model rats. Methods the rat model of COPD was established, PASMCs were cultured in vitro and identified. The synthesis and secretion of cytokines of PASMCs induced by cigarette smoke extract (CSE) and lipopolysaccharide (LPSs) were detected. The rats were divided into two groups: control group (CSE group) and LPS group (LPS group). At the same time, the 4 groups were cultured with 1: 1000 anti-HMGB1 antibody 0.2 mL respectively as the intervention group, the expression of HMGB1 protein in PASMCs of each group was detected by Western blot and the concentration of HMGB1 IFN- 緯 in the supernatant of cell culture was detected by Elisa. And carry on the correlation analysis. Results under the light microscope, the cells appeared to be fusiform or "peak". The expression of HMGB1 and the expression of HMGB1 in the supernatant of HMGB1 and HMGB1 in cell supernatant were detected by Western blot and Elisa after 12 h culture. There were significant differences between the two groups in the expression of HMGB1 and the expression of HMGB1 in the supernatant of HMGB1. The expression of HMGB1 and the expression of HMGB1 in the cell supernatant were significantly higher than those in the control group. The level of protein expression was positively correlated with the change of IFN- 緯 concentration, and the anti-HMGB1 antibody was added to culture for 12 h. The concentration of IFN- 緯 in the intervention group was lower than that in the corresponding experimental group, while in the same group, there was no significant difference in the results of Western blot and Elisa in the same group. Conclusion the level of HMGB1 expression is positively correlated with the synthesis and secretion of IFN- 緯 by distal PASMCs in COPD model rats. Anti-HMGB1 antibody provides a new intervention target for clinical treatment of COPDs and inhibition of systemic inflammatory response.
【作者單位】： 桂林醫(yī)學(xué)院附屬醫(yī)院呼吸內(nèi)科;
【基金】：國家自然科學(xué)基金(81360010) 廣西壯族自治區(qū)醫(yī)療衛(wèi)生重點科研課題(2012004)
【分類號】：R563.9

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本文編號：2035157