氧化苦参碱对肺纤维化大鼠肺微血管的影响

Effect of oxymatrine on lung micro-blood vessels in rats with pulmonary fibrosis

目的观察氧化苦参碱(OMT)对肺纤维化大鼠肺组织肿瘤坏死因子-α(TNF-α)表达及肺微血管变化的影响,并探讨OMT治疗肺纤维化的可能机制。方法气管注射博来霉素制作大鼠肺纤维化模型。将72只雄性SD大鼠分为正常组、模型组、氧化苦参碱治疗组(简称治疗组),每组24只。正常组不做任何处理,正常饲养;治疗组造模后第2天起腹腔注射氧化苦参碱注射液,直至处死;模型组造模后每天腹腔注射同等量的生理盐水。每组在造模后第3,7,14,28天随机处死6只,观察肺组织病理变化,检测肺组织羟脯氨酸(HYP)含量,计算机图形自动分析系统(CIAS)评估肺纤维化程度。通过双抗夹心酶联免疫吸附法(ELISA)测定肺组织匀浆TNF-α含量。肺组织切片行CD-31免疫组化标记,通过CIAS评估肺微血管动态改变。结果模型组第14,28天肺HYP含量均高于同期正常组(P均<0.05),治疗组第28天肺HYP含量明显低于同期模型组(P<0.05);病理学半定量分析显示治疗组肺纤维化程度较模型组减轻;模型组及治疗组肺微血管密度(MVD)呈动态改变,在第3,7,14天2组肺组织MVD均高于正常组,在第28天时则明显低于正常组,其中模型组肺MVD第3,7天较治疗组均更高(P均<0.05);ELISA检测示模型组和治疗组各时间点肺组织匀浆TNF-α均高于正常组(P均<0.05),治疗组第7,14天TNF-α较同期模型组均明显下调(P均<0.05)。结论在肺纤维化发展过程中存在肺组织微血管密度动态改变,早期大量新生血管形成,晚期微血管减少甚至消失,氧化苦参碱能一定程度阻止肺纤维化早期新生血管形成。氧化苦参碱治疗肺纤维化可能部分与抑制TNF-α升高有关。

Objective It is to observe the effects of oxymatrine（OMT） on the expression of TNF-α and the change of lung micro-blood vessels in rats with pulmonary fibrosis,and to explore its possible mechanism about the treatment with OMT for pulmonary fibrosis.Methods Pulmonary fibrosis rat models were established by bleomycin via single intratracheal perfusion.Seventy-two male SD rats were randomly divided into normal group,model group and treatment group（24 rats in each group）.Normal group wasn＇t treated with any drugs.Treatment group was treated with oxymatrine via intraperitoneal injection untill the animals were sacrificed.Model group was given the same dosage of normal saline after model established.Six rats from each group were sacrificed randomly at the 3rd,the 7th,the 14th and the 28th days after model established to observe their pathological changes and determine hydroxyproline（HYP） content in lung tissue.The fibrosis was judged by computer image analysis system（CIAS）.TNF-α in lung tissue homogenate were determined by ELISA.Anti-CD-31 immunohistochemisty was performed and the microvessel density（MVD） was measured by CIAS.Results The HYP of model group was both higher than that in normal group at the 14th and the 28th days（both P〈0.05）.The HYP of treatment group was lower than that in model group（P〈0.05）at the 28th day.Semi-quantitative pathological analysis showed that the fibrosis degree of treament group was extenuated than that in model group.The immunohistochemical stain showed a dynamic change of the lung tissue MVD in both model group and treatment group,the MVD in both groups was significantly higher than that in normal group at the 3th,the 7th and the 14th days,but was lower than that in normal group on the 28th day.TNF-α concentrations of lung tissue homogenate in both model group and treated group were significantly higher than that in nomal group during the whole course（both P〈0.05）.The TNF-α concentrations in treatment group was lower than that in model group at the 7th and the 14th days.Conclusion There is a dynamic change of the lung tissue MVD during the course of lung fibrosis,large amount of neovascularization in the lung tissue can be observed in the early stage of the disease,in the end stage the micro-vessel in lung tissue is decreased or even disappears.OMT can reduce neovascularization in the early stage.OMT can improve pulmonary fibrosis,while the mechanisms may be partly related to its inhibition on TNF-α heightening.