【摘要】：目的外周血單核細(xì)胞表型偏移失衡與肺纖維化病理進(jìn)展關(guān)系密切。文中通過(guò)闡述循環(huán)單核細(xì)胞亞群在實(shí)驗(yàn)性肺纖維化小鼠不同病理時(shí)期的動(dòng)態(tài)變化,探討其表型偏移與肺組織炎癥和纖維化的關(guān)系。方法 100只雄性C57BL/6J小鼠隨機(jī)數(shù)字表法分為等滲鹽水組和博萊霉素組,經(jīng)口咽吸入法分別給予等滲鹽水及博萊霉素A5,術(shù)后第1、3、7、14及21天處死小鼠,病理學(xué)方法計(jì)算各組小鼠肺組織炎癥評(píng)分(inflammation score,IS)及膠原容積分?jǐn)?shù)(collagen volume fraction,CVF),常規(guī)方法進(jìn)行支氣管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)細(xì)胞計(jì)數(shù)及分類計(jì)數(shù),熒光定量PCR法檢測(cè)各組小鼠肺組織膠原Ⅰ、膠原Ⅲ的mRNA表達(dá)量,氯胺T法檢測(cè)肺組織羥脯氨酸(hydroxyproline,HYP)含量,流式細(xì)胞術(shù)檢測(cè)循環(huán)單核細(xì)胞亞群的變化。結(jié)果博萊霉素組小鼠第3天炎性細(xì)胞浸潤(rùn)、肺泡壁水腫等炎癥程度(IS評(píng)價(jià))較等滲鹽水組顯著增加(P0.01),第7天達(dá)到峰值后呈下降趨勢(shì)。博萊霉素組小鼠第14、21天肺組織CVF較等滲鹽水組明顯增高(P0.01);與等滲鹽水組相應(yīng)時(shí)間點(diǎn)比較,博萊霉素組第1天細(xì)胞總數(shù)無(wú)明顯變化,第3天明顯增多(25.18±2.06 vs 8.82±1.59,P0.01),第7天升至高峰(56.56±5.03 vs 3.68±0.79,P0.01),后呈下降趨勢(shì),但第14~21天細(xì)胞總數(shù)仍高于等滲鹽水組(P0.01),肺泡巨噬細(xì)胞比例的變化趨勢(shì)與細(xì)胞總數(shù)相一致;而中性粒細(xì)胞數(shù)量第1、3、7天顯著高于等滲鹽水組(9.086±1.268 vs 1.108±0.229、5.551±0.511 vs 0.315±0.100、8.093±0.922 vs 0.249±0.074),差異均有統(tǒng)計(jì)學(xué)意義(P0.01);博萊霉素組膠原Ⅰ、ⅢmRNA表達(dá)水平在第14、21天顯著高于等滲鹽水組(P0.05);HYP檢測(cè)結(jié)果顯示博萊霉素組第7、14、21天高于等滲鹽水組(P0.01),其中第21天達(dá)最高值;與等滲鹽水組相比,博萊霉素組第1天Ly6Chi單核細(xì)胞亞群比例顯著升高,第3天達(dá)到高峰(P0.01),此后呈降低趨勢(shì),Ly6Clo亞群比例變化與之相反。各組小鼠Ly6Chi單核細(xì)胞比例與IS及CVF值均呈正相關(guān)(P0.01,P=0.001 3)。結(jié)論在博萊霉素A5誘導(dǎo)的小鼠肺纖維化模型中,不同病理生理階段外周血中Ly6Chi、Ly6Clo單核細(xì)胞亞群比例處于動(dòng)態(tài)變化,與肺組織炎性細(xì)胞滲出、浸潤(rùn)等病理進(jìn)程相比,Ly6Chi單核細(xì)胞亞群應(yīng)對(duì)肺組織損傷、炎癥反應(yīng)迅速,早期Ly6Chi單核細(xì)胞亞群比例的升高可能與肺組織炎癥反應(yīng)及后期纖維化程度存在密切的聯(lián)系。
[Abstract]:Objective the imbalance of phenotypic deviation of peripheral blood monocytes is closely related to the pathological progress of pulmonary fibrosis. In this paper, the dynamic changes of circulating monocyte subsets in different pathological stages of experimental pulmonary fibrosis mice were described, and the relationship between phenotypic deviation and inflammation and fibrosis of lung tissue was discussed. Methods 100 male C57BL/6J mice were randomly divided into isoosmotic saline group and bleomycin A5 group. The mice were sacrificed on the 1st, 3rd, 7th, 14th and 21st day after operation. The lung tissue inflammation score (inflammation score,IS) and collagen volume fraction (collagen volume fraction,CVF) were calculated by pathological method. Bronchoalveolar lavage fluid (bronchoalveolar lavage fluid,) was performed by routine method. The mice were killed on the 1st, 3rd, 7th, 14th and 21st day after operation. The lung tissue inflammation score (inflammation score,IS) and collagen volume fraction (collagen volume fraction,CVF) of mice in each group were calculated by pathological method. Bronchoalveolar lavage fluid (bronchoalveolar lavage fluid,) was performed by routine method. The mRNA expression of collagen 鈪，

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