【作者】 黄正桥；

【导师】 王飞；

【作者基本信息】 成都中医药大学 ， 中医内科学， 2017， 博士

【摘要】 【目的】建立脂多糖(LPS)诱导的急性肺损伤(ALI)SD大鼠动物模型,探讨中药复方痰热净对大鼠急性肺损伤的保护作用及其相关机制。【方法】健康雄性SD大鼠48只,随机分为正常组、模型组、痰热净高、中、低剂量组、地塞米松治疗组,每组各8只。除空白组外每组大鼠气管内滴入LPS(3mg/kg);空白组气道内滴入与LPS等量的生理盐水。痰热净组灌服痰热净(由桑白皮、黄芩等组成),高、中、低剂量分别为6.48g/kg2d、3.24g/kg2d、1.62g/kg2d。地塞米松治疗组按照0.001g/kg的剂量每只大鼠腹腔注射地塞米松磷酸钠溶液0.25ml(地塞米松磷酸钠注射液5mg稀释于10ml生理盐水中制成浓度为0.5mg/ml溶液);其余两组灌服生理盐水。观察各组大鼠症状体征改变。7天后处死各组大鼠,留取肺组织及肺灌洗液待检。检测肺组织湿/干重比(W/D),光镜下观察肺组织病理学改变,酶联免疫法检测肺灌洗液黏蛋白5AC(Muc5AC)、IL-6、IL-21;蛋白印迹法(Westen-blot)检测肺组织TGF-β1、Smad3的表达情况。【结果】(1)光镜下观察:与空白组比较,LPS模型组大鼠肺组织毛细血管扩张充血,肺泡间隔明显增宽,见大量炎性细胞浸润和大量红细胞漏出,肺泡腔和间质水肿明显,严重者可见部分肺泡腔萎陷不张、肺泡断裂。部分大鼠肺组织肺泡间隔断裂、结构破坏明显。痰热净治疗各组大鼠肺组织中性粒细胞浸润仍较明显,肺泡间隔有明显增宽,并可见毛细血管扩张充血,部分肺泡腔和间质水肿,严重者也存在部分肺泡腔萎陷和肺泡间隔断裂。但是与LPS模型组比较,痰热净高、中剂量组正常肺泡结构破坏较少,肺泡腔萎陷不张、肺泡断裂程度轻,肺组织毛细血管扩张充血、炎性细胞浸润及红细胞的漏出较轻,肺泡腔渗出和肺间质肿胀程度明显减轻,病理改变较LPS模型组明显减轻。提示LPS气道内滴注能诱发大鼠产生ALI的肺部病理学改变,痰热净可以减轻ALI大鼠肺组织炎症反应,其效应与剂量呈正相关。(2)与空白组相比,LPS模型组、地塞米松组、痰热净高中低剂量组大鼠肺组织的W/D比值显著增加(p<0.05);与模型组比较痰热净高、中剂量组W/D值低于模型组,有显著性差异(p<0.05)。提示痰热净可以显著改善LPS所致ALI大鼠的肺水肿程度。(3)模型组大鼠肺泡灌洗液中Muc5AC明显升高。与模型组比较,痰热净各组大鼠肺泡灌洗液中的Muc5AC的量均有降低,差异具有显著性(p<0.05)。(4)与空白组比较,模型组大鼠肺泡灌洗液中IL-6、IL-21明显升高。与模型组比较,痰热净各组大鼠肺泡灌洗液中的IL-6、IL-21均有降低,差异具有显著性(p<0.05)。(5)在LPS急性肺损伤大鼠模型肺组织中,TGF–β1、Smad3表达量升高,痰热净高剂量干预后肺组织中TGF–β1、Smad3表达明显低于模型组,差异有显著性(P<0.05)。提示痰热净能够抑制ALI大鼠肺组织内TGF–β1、Smad3的表达。【结论】1.痰热净可改善LPS致ALI大鼠肺组织病理学改变,减轻肺水肿,减少炎症因子IL-6、IL-21的释放和Muc5AC的生成,使肺组织中TGF-β1、Smad3水平下降,表明痰热净对LPS诱导的ALI有保护作用,而这种保护作用的潜在机制可能与痰热净抑制TGF-β1、Smad3等有关。2.基于中医肺脏生理病理理论与ALI发病之间的关系,认为ALI病位在肺,痰、热等病理产物骤生,壅阻肺络、气机逆乱是其发展加重的核心环节;以清热化痰法治疗ALI顺应肺臧生理、契合病机,可有效改善ALI时肺脏功能失调。更多还原

【Abstract】 [Purpose]The aim of this study is to explore the protective effect of Tanrejing Decoction and their potential mechanism in rat with lipopolysaccharides(LPS)-induced acute lung injury(ALI).[Methods]48 male SD rats were randomly divided normal group,model control group,dexamethasone-treated group,24 rats in Tanrejing-treated group.Intratracheal instillation of LPS(3mg/kg)of LPS group;Normal group intratracheal instillation of equivalent saline.And at 7d,lungs were to be seized after the animals were killed.The lung wet/dry weight ratio(W/D),light microscope observation of lung histopathology were simultaneously detected.Using the ELISA method to measure the content of MUC5 AC 、IL-6、IL-21 of serum;TGF-β1、Smad3 of lung was detected with Western-blot method.[Results]Light microscope: compared with the normal group,LPS group rat lung tissue shows alveolar septal thickening and capillary congestion significantly,a large number of inflammatory cell infiltration,a lot of red blood cells leakage,alveolar and interstitial edema.In severe case,alveolar collapse and alveolar septal rupture can be seen.Compared with the LPS group,Tanrejing-treated rats lung tissue shows less damage of normal alveolar structure,inflammatory cell infiltration and the leakage of erythrocytes was lighter,alveolar and interstitial lung exudates was significantly reduced(p<0.05).Prompts airway instillation of LPS can induce ALI pathological changes in rat lung tissues,Tanrejing Decoction can significantly reduce lung tissue inflammation,alveolar structures damage in rat with ALI.Compared with the normal group,the rat lung tissue W/D ratio of model control group,dexamethasone-treated group,and Tanrejingtreated group increased significantly(p<0.05).Tanrejing-treated group rat lung tissue W/D ratio were significantly lower than the model control group(p<0.05).Prompts Tanrejing Decoction can significantly improve the pulmonary edema of LPS-induced ALI in rat.In the model group,the Muc5 AC in BALF was significantly increased.Compared with the model group,Muc5 AC levels in rat BALF decreased significantly(p<0.05).Compared with the normal group,IL-6 and IL-21 levels in rat BALF increased significantly.Compared with the model group,IL-6 and IL-21 levels in Tanrejing-treated group decreased significantly(p<0.05).The TGF –β1 and Smad3 levels of LPS model group in rat lung tissue were significantly increased.Compared with the model group,IL-6 and IL-21 levels in rat BALF decreased significantly(p<0.05).Compared with the model group,TGF –β1 and Smad3 levels in Tanrejing-treated group decreased significantly(p<0.05).Prompts Tanrejing Decoction can significantly inhibit the expression of TGF –β1 and Smad3 of LPS-induced ALI in rat.[Conclusion]1.Tanrejing could significantly attenuate pulmonary histopathological changes including weaking pulmonary edema,diminishing inflammatory cytokines and protein in acute lung injury.The potential mechanism of this action may involve the inhibitions of IL-6 and IL-21 expression by regulating TGF-β1 and Smad3 activation..2.Purifying and descending function of the lung,phlegm heat obstructing the lung,phlegm heat is an important factor leading to ALI damage.Therefore,in ALI,clearing heat phlegm can recover the physiologic function of lung.更多还原