CAJ | 학술논문

目的探讨缺氧诱导因子-1α(HIF-1α)在百草枯(PQ)中毒大鼠肺组织急性肺损伤早期肺纤维化中的作用.方法将48只健康SD大鼠随机分为对照组(6只)和PQ染毒组(42只).对照组大鼠予生理盐水1 ml,一次性灌胃；将质量分数为20％的PQ用生理盐水稀释,对染毒组大鼠一次性灌胃染毒1 ml(500 mg/kg),按灌胃后2、6、12、24、48、72及120 h分为7个亚组,每组6只.分别测定各组大鼠的动脉血氧分压(PaO2);进行肺组织HE染色、Mason染色,观察病理学变化；肺组织荧光染色观察肺组织变化及HIF1-a蛋白表达；用免疫蛋白印迹(Western blot)法检测HIF-1α蛋白表达.结果染毒72h组大鼠动脉血PaO2为(62.33±0.22) mm Hg,与对照组[(96.00±5.20) mm Hg]比较,差异有统计学意义(P＜0.05).HE染色可见,染毒后2h大鼠肺泡毛细血管内皮细胞、肺泡上皮细胞和肺间质的急性弥漫性损害,广泛肺间质炎性细胞浸润,增厚；染毒后12h肺间质水肿减轻,肺泡间距变窄；染毒后120 h,肺泡结构紊乱,较多量片状分布瘢痕条索,成纤维细胞明显增多,外周炎症吸收.Masson染色可见,染毒后2h大鼠肺泡上皮出现明显胶原沉积；24、48 h蓝染的胶原纤维逐渐增加；染毒后120 h肺泡组织明显破坏,大片纤维结缔组织沉积.免疫荧光及Westem blot法均显示,染毒组大鼠染毒后2h肺组织中HIF1-a蛋白表达明显升高,至12h达峰值,24、48 h逐渐下降,但仍高于对照组,染毒组各时间点HIF-1α蛋白表达与对照组的差异有统计学意义(P＜0.05).染毒组各时间点HIF-1α蛋白表达比较,差异无统计学意义(P＞0.05).结论 PQ中毒大鼠肺组织早期即出现肺纤维化及HIF-1α蛋白的表达,HIF-1α可能参与了肺纤维化的形成.
목적 탐토결양유도인자-1α(HIF-1α)재백초고(PQ)중독대서폐조직급성폐손상조기폐섬유화중적작용.방법 장48지건강SD대서수궤분위대조조(6지)화PQ염독조(42지).대조조대서여생리염수1 ml,일차성관위；장질량분수위20％적PQ용생리염수희석,대염독조대서일차성관위염독1 ml(500 mg/kg),안관위후2、6、12、24、48、72급120 h분위7개아조,매조6지.분별측정각조대서적동맥혈양분압(PaO2);진행폐조직HE염색、Mason염색,관찰병이학변화；폐조직형광염색관찰폐조직변화급HIF1-a단백표체；용면역단백인적(Western blot)법검측HIF-1α단백표체.결과 염독72h조대서동맥혈PaO2위(62.33±0.22) mm Hg,여대조조[(96.00±5.20) mm Hg]비교,차이유통계학의의(P＜0.05).HE염색가견,염독후2h대서폐포모세혈관내피세포、폐포상피세포화폐간질적급성미만성손해,엄범폐간질염성세포침윤,증후；염독후12h폐간질수종감경,폐포간거변착；염독후120 h,폐포결구문란,교다량편상분포반흔조색,성섬유세포명현증다,외주염증흡수.Masson염색가견,염독후2h대서폐포상피출현명현효원침적；24、48 h람염적효원섬유축점증가；염독후120 h폐포조직명현파배,대편섬유결체조직침적.면역형광급Westem blot법균현시,염독조대서염독후2h폐조직중HIF1-a단백표체명현승고,지12h체봉치,24、48 h축점하강,단잉고우대조조,염독조각시간점HIF-1α단백표체여대조조적차이유통계학의의(P＜0.05).염독조각시간점HIF-1α단백표체비교,차이무통계학의의(P＞0.05).결론 PQ중독대서폐조직조기즉출현폐섬유화급HIF-1α단백적표체,HIF-1α가능삼여료폐섬유화적형성.
Objective To explore the role of hypoxia-inducible factor lalpha (HIF-1α) in early lung fibrosis of rats with acute paraquat (PQ) poisoning.Methods Forty eight healthy SD rats were randomly divided into control group (6 rats) and paraquat poisoning group (42 rats).Control group was exposed to 1 ml normal solution by gastric garage.The paraquat group was exposed to 1 ml paraquat solution (50 mg/kg) by gastric gavage for 2,6,12,48,72 and 120 h,respectively.The arterial blood gas analysis (PaO2) was detected.The pathological examinations of lung tissues were performed by HE and Mason staining.HIF-1α in lung tissues were measured by immunofluorescence.Western blot assay was used to detect the expression levels of HIF-1α protein in lung tissues.Results PaO2 of rats exposed to paraquat for 72 h was (62.33±0.22) mm Hg,which was significantly lower than that (96.00±5.20) of control group (P＜0.05).Pathological examination by HE staining indicated that the acute diffuse lesion appeared in the lveolar capillary endothelium,epithelia and interstitial tissues,and there was the inflammatory cell infiltration in the alveolar of rats exposed to paraquat at 2 h after exposure.At 12 h after exposure,the interstitial edema in lung tissues of rats decreased and the lveolar space became narrow.At 120 h after exposure,there were the alveolar structure derangement,abundaut cicatrix,more fibroblasts and peripheral inflammation absorption.Pathological examination by Masson staining showed that there was obvious collagen deposition in the lveolar epithelia at 2h after exposure,the increased collagen fibrosis at 24 and 48 h after exposure and the obvious damage of alveolar tissues or much more fibrous connective tissue deposition at 120 h after exposure.The results of western blot and immunolfluorence assays exhibited that the expression levels of HIF-1α in lung tissues at 2,24 and 48 h after exposure significantly increased,as compared with control group (P＜0.05),but there were no significant differences of HIF-1α expression among sub-groups at different time points after exposure.Conclusion The results of present study shown that there were the pulmonary fibrosis and increased expression of HIF-1α in acute PQ poisoning rats at the early stage,and HIF-1α may be associated with pulmonary fibrosis.