CAJ | 학술논문

目的：研究矽肺患者肺泡巨噬细胞（ AM）内质网应激（ ERS）和自噬相关蛋白的表达规律。方法采用单纯随机抽样方法，选择矽肺壹期、贰期、叁期男性患者20、28、22例分别设为矽肺壹期、贰期和叁期组，以6例诊断为观察对象的男性矽尘作业工人为对照组。收集研究对象的支气管肺泡灌洗液，分离、纯化和培养AM，经裂解后提取蛋白，采用免疫印迹法检测AM中葡萄糖调节蛋白78（ GRP78）、Beclin1和微管相关蛋白1轻链3（ LC3）蛋白的相对表达水平。结果矽肺壹期、贰期和叁期患者AM中GRP78、Beclin1和LC3的蛋白相对表达水平均高于对照组（P＜0．05），矽肺贰期和叁期组患者上述3个指标均高于矽肺壹期组（P＜0．05），矽肺叁期组患者AM中GRP78蛋白相对表达水平低于矽肺贰期组（P＜0．05）；矽肺叁期组患者AM中Beclin1和LC3的蛋白相对表达水平均高于矽肺贰期组，但差异均无统计学意义（ P＞0．05）。多重逐步线性回归分析结果显示，矽肺患者AM 中GRP78、Beclin1和LC3的蛋白相对表达水平均与矽肺期别呈正相关（P＜0．01）。结论不同期别矽肺患者AM中ERS和自噬的相关蛋白表达程度不同，AM的ERS和自噬在矽肺纤维化发生发展中可能发挥重要作用。
목적：연구석폐환자폐포거서세포（ AM）내질망응격（ ERS）화자서상관단백적표체규률。방법채용단순수궤추양방법，선택석폐일기、이기、참기남성환자20、28、22례분별설위석폐일기、이기화참기조，이6례진단위관찰대상적남성석진작업공인위대조조。수집연구대상적지기관폐포관세액，분리、순화화배양AM，경렬해후제취단백，채용면역인적법검측AM중포도당조절단백78（ GRP78）、Beclin1화미관상관단백1경련3（ LC3）단백적상대표체수평。결과석폐일기、이기화참기환자AM중GRP78、Beclin1화LC3적단백상대표체수평균고우대조조（P＜0．05），석폐이기화참기조환자상술3개지표균고우석폐일기조（P＜0．05），석폐참기조환자AM중GRP78단백상대표체수평저우석폐이기조（P＜0．05）；석폐참기조환자AM중Beclin1화LC3적단백상대표체수평균고우석폐이기조，단차이균무통계학의의（ P＞0．05）。다중축보선성회귀분석결과현시，석폐환자AM 중GRP78、Beclin1화LC3적단백상대표체수평균여석폐기별정정상관（P＜0．01）。결론불동기별석폐환자AM중ERS화자서적상관단백표체정도불동，AM적ERS화자서재석폐섬유화발생발전중가능발휘중요작용。
Objective To study the expression of endoplasmic reticulum stress (ERS) and autophagy-related proteins of alveolar macrophage (AM) in silicosis patients.Methods With simple random sampling, the male patients of silicosis at stage Ⅰ,Ⅱand Ⅲwere respectively chosen as the silicosis groupⅠ,ⅡandⅢwith 20, 28, 22 cases in each group.Six male observed subjects exposed to silica dust were selected as the control group .Bronchoalveolar lavage fluids of whole-lung lavage of all subjects were collected .After AM were separated , purified cultured and lysed , the relative protein expression levels of glucose regulated protein 78 ( GRP78 ) , Beclin1 and microtubule-associated protein 1 light chain 3 (LC3) were assayed by Western blot.Results Relative expression levels of GRP78, Beclin1 and LC3 proteins in AM of silicosis groupⅠ,Ⅱand Ⅲwere significantly higher than those of the control group respectively (P<0.05), the above three indicators of silicosis groupⅡandⅢwere significantly higher than those of the silicosis group Ⅰrespectively ( P<0.05), and the relative expression level of GRP78 protein in AM of silicosis group Ⅲwas significantly lower than that of the silicosis groupⅡ(P<0.05).Relative expression levels of Beclin1 and LC3 proteins in AM of silicosis group Ⅲwere higher than those of the silicosis group Ⅱ respectively, but there was no statistical significant difference (P >0.05). Analysis results of multiple stepwise linear regression indicated that there was a positive correlation between relative expression levels of GRP78, Beclin1 and LC3 proteins respectively in AM of silicosis patients and the stage of silicosis(P <0.01).Conclusion The related protein expression levels of ERS and autophagy in AM are different among silicosispatients at different silicotic stages.ERS and autophagy of AM may play an important role in the formation and developmentof silicosis.