摘要
目的探讨依托泊苷对pristane诱导的弥漫性肺泡出血(diffuse alveolar hemorrhage, DAH)小鼠肺组织细胞凋亡的影响。方法 24只小鼠随机分为对照组、模型组和依托泊苷组,每组8只。模型组和依托泊苷组腹腔单次注射0.5 mL pristane制备DAH模型,对照组给予等量PBS。造模成功后,依托泊苷组腹腔注射10 mg/kg依托泊苷,对照组和模型组腹腔注射等量PBS,均1次/d,连续14 d。14 d后,取3组小鼠肺组织进行组织病理检查,HE染色观察小鼠肺组织出血情况和炎症细胞浸润情况,采用TUNEL染色试剂盒检测3组小鼠肺组织细胞凋亡率,采用Western blot法检测3组小鼠肺组织caspase-3、Bax、Bcl-2蛋白相对表达量。结果对照组小鼠肺组织大体呈正常粉红色,模型组为暗红色,依托泊苷组可见散在出血点。对照组小鼠肺泡结构完好,肺泡腔清晰,肺组织炎症细胞浸润不明显;模型组小鼠肺泡腔出现大量红细胞,伴大量中性粒细胞、巨噬细胞等炎症细胞浸润;依托泊苷组肺泡腔无明显红细胞,肺泡壁增厚。模型组小鼠肺组织细胞凋亡率[(7.87±0.79)%]及Bax、caspase-3蛋白相对表达量(0.87±0.01、2.11±0.17)高于对照组[(0.78±0.13)%、0.24±0.02、0.19±0.01]和依托泊苷组[(1.49±0.24)%、0.32±0.01、0.83±0.06](P<0.05),依托泊苷组高于对照组(P<0.05);模型组小鼠Bcl-2蛋白相对表达量(0.75±0.07)低于对照组(2.86±0.19)和依托泊苷组(5.62±0.21)(P<0.05),依托泊苷组高于对照组(P<0.05)。结论依托泊苷对DAH小鼠有治疗作用,可明显改善小鼠肺组织肺泡出血和炎症细胞浸润,降低肺组织细胞凋亡率,其机制可能与调节Bax、Bcl-2和caspase-3蛋白表达有关。
Objective To investigate the effect of etoposide on lung tissue cells apoptosis in mice with diffuse alveolar hemorrhage induced by pristane. Methods Twenty-four mice were randomly divided into control group, model group and etoposide group, with 8 mice in each group. The mice in model group and etoposide group were given a single intraperitoneal injection of 0.5 mL of pristane to prepare DAH models, and the mice in control group were given the same amount of PBS buffer. After successful modeling, the mice in etoposide group were intraperitoneally injected with 10 mg/kg etoposide, and the mice in control group and model group were given the same amount of PBS buffer, once a day for 14 consecutive days. After 14 days, the lung tissues were obtained for histopathological examination. HE staining was used to observe the hemorrhage and inflammatory cell infiltration in the lung tissues, TUNEL staining kit was used to detect the apoptosis rate, and Western blot method was used to detect the relative expressions of caspase-3, Bax and Bcl-2 proteins in the lung tissues of three groups. Results The lung tissues were generally in normal pink in control group, were in dark red in model group, and were found scattered bleeding points in etoposide group. The mice in control group had intact alveolar structure, clear alveolar cavity, and unobvious infiltration of inflammatory cells in the lung tissues. The mice in model group had a large number of red blood cells in the alveolar cavity, accompanied by a large number of neutrophils, macrophages and other inflammatory cells infiltration. The mice in etoposide group had no obvious red blood cells in the alveolar cavity, and the alveolar walls were thickened. The apoptosis rate of lung cells and the relative expressions of Bax and caspase-3 proteins were higher in model group((7.87±0.79)%, 0.87±0.01, 2.11±0.17) than those in control group((0.78±0.13)%, 0.24±0.02, 0.19±0.01) and etoposide group((1.49±0.24)%, 0.32±0.01, 0.83±0.06)(P<0.05), and higher in etoposide group than those in control group(P<0.05). The relative expression of Bcl-2 protein was lower in model group(0.75±0.07) than that in control group(2.86±0.19) and etoposide group(5.62±0.21)(P<0.05),and lower in etoposide group than that in control group(P<0.05).Conclusion Etoposide has a therapeutic effect on DAH mice.It can significantly alliviate alveolar hemorrhage and inflammatory cell infiltration in the lung tissues of mice,and reduce the apoptosis rate of the lung cells.The mechanism might be correlated with regulating the expressions of Bax,Bcl-2 and caspase-3 proteins.
作者
李向南
阎磊
孙恺
邵凤民
LI Xiang-nan;YAN Lei;SUN Kai;SHAO Feng-min(Department of Nephrology,Zhengzhou University People’s Hospital,Henan Provincial People's Hospital,Henan Provincial Key Laboratory of Kidney Disease and Immunology,Zhengzhou,Henan 450003,China;Department of Hematology,Zhengzhou University People’s Hospital,Henan Provincial People’s Hospital,Zhengzhou,Henan 450003,China)
出处
《中华实用诊断与治疗杂志》
2021年第4期325-328,共4页
Journal of Chinese Practical Diagnosis and Therapy
基金
河南省自然科学基金(182300410296)。