[关键词]
[摘要]
目的 探讨炎调方对脓毒症急性肺损伤(ALI)大鼠肺组织热休克蛋白70(HSP70)mRNA和p38丝裂原活化蛋白激酶(p38 MAPK)的调控作用。方法 将清洁级健康雄性SD大鼠随机分为假手术组、模型组、炎调方(生药量9.9 g/kg)组、谷氨酰胺组、槲皮素组,采用盲肠结扎穿孔术法(CLP)建立脓毒症ALI模型。观察大鼠一般状况,各组分别于术后4、6、8、10、12、18、24 h采集肺组织标本,HE染色观察肺组织病理学改变;实时荧光定量PCR法检测肺组织HSP70 mRNA的表达水平;Western Blotting法检测肺组织p38 MAPK的蛋白表达水平。结果 炎调方组、谷氨酰胺组大鼠与模型组比较,术后不同时间点的精神状态、活动量等明显好转,可见少量稀便;肺组织损伤程度明显减轻,腔内出血较少,肺实变较轻。炎调方组不同时间点的肺组织HSP70 mRNA表达水平显著高于假手术组、模型组(除24 h时间点外)、槲皮素组(P<0.01),在8、12、18、24 h时间点显著低于谷氨酰胺组(P<0.01);炎调方组不同时间点的肺组织p38 MAPK的蛋白表达量显著低于模型组、槲皮素组(P<0.01),在4、6、10 h时间点明显低于谷氨酰胺组(P<0.05、0.01)。结论 炎调方可通过上调肺组织HSP70 mRNA的表达,下调肺组织p38 MAPK的蛋白表达,改善肺组织损伤,对脓毒症ALI发挥保护作用。
[Key word]
[Abstract]
Objective To investigate the regulation effect of Yantiao Prescription (YP) on the expression of heat shock protein 70 mRNA (HSP70) and p38 mitogen activated protein kinase (p38 MAPK) in lung tissue of rats with acute lung injury induced by sepsis. Methods Healthy male Sprague Dawley rats were randomly divided into sham group, model group, YP group, glutamine group, and quercetin group. Acute lung injury model induced by sepsis was established in rats by cecal ligation and puncture (CLP). At the corresponding time points, ten rats of each group were sacrificed and lung tissue samples were collected. The pathological changes of lung tissue were observed by HE staining. The expression of HSP70 mRNA in lung tissue was detected by real-time PCR. The expression of p38 MAPK in lung tissue was detected by Western Blotting. Results Compared with model group, the mental state and activity of the rats at different time points were obviously improved after the operation, and a small amount of loose stools was observed. The injury degree of lung tissue was obviously reduced, the hemorrhage in the cavity was less, and the pulmonary consolidation was lighter. The expression levels of HSP 70 mRNA in YP group were significantly higher than sham group, model group (except the 24 h time point), and quercetin group (P<0.01), lower than that of glutamine group in 8, 12, 18, and 24 h time point (P<0.01). The relative expression of p38 MAPK in lung tissue of rats in YP group was lower than that in model and quercetin group (P<0.01), and were lower than that of glutamine at the time points of 4, 6, and 10 h (P<0.05). Conclusion YP can improve the expression of lung tissue HSP70 mRNA, reduce the expression of p38 MAPK in lung tissue, improve lung histopathological changes, and plays a protective role against acute lung injury induced by sepsis.
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[基金项目]
国家自然科学基金资助项目(81303105)