宣肺通腑方对脂多糖致急性肺损伤大鼠肺组织髓样分化蛋白―2核因子―κB表达影响.docVIP

宣肺通腑方对脂多糖致急性肺损伤大鼠肺组织髓样分化蛋白―2核因子―κB表达影响 　　摘要：目的 观察宣肺通腑方对脂多糖（LPS）诱导的急性肺损伤（ALI）大鼠肺组织髓样分化蛋白-2（MD-2）、核因子-κB（NF-κB）蛋白及其基因表达的影响，并探讨其作用机制。方法 Wistar大鼠随机分为正常组、模型组、地塞米松组和宣肺通腑方大、中、小剂量组，每组8只。尾静脉注射LPS 6 mg/kg制备大鼠ALI模型。宣肺通腑方各组于造模前以15.12、7.56、3.78 g原药材/kg宣肺通腑方水煎液灌胃，1次/d，连续3 d；地塞米松组在造模前1 h按5 mg/kg腹腔注射地塞米松。造模后9 h麻醉取材，采用免疫组化ABC法和实时荧光定量聚合酶链反应检测MD-2、NF-κB蛋白及其基因表达，光镜下观察大鼠肺组织病理变化。结果 与正常组比较，模型组MD-2、NF-κB蛋白及其基因表达均明显上调（P0.05）。病理观察结果显示，与正常组比较，模型组大鼠肺组织出现大片出血及坏死，炎症细胞大量浸润；宣肺通腑方各剂量组和地塞米松组大鼠肺组织病理改变明显轻于模型组，肺细支气管偶见炎症改变，水肿较轻，炎性细胞渗出较少。结论 宣肺通腑方能减轻ALI大鼠肺组织损伤，对肺损伤有保护作用，其机制可能与其降低ALI大鼠MD-2、NF-κB蛋白及其基因表达有关。 　　关键词：宣肺通腑方；急性肺损伤；髓样分化蛋白-2；核因子-κB；蛋白表达；基因表达；大鼠 　　DOI：10.3969/j.issn.1005-5304.2014.06.021 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2014）06-0065-04 　　Abstract：Objective To investigate the effects of Xuanfeitongfufang Decoction on expressions of MD-2， NF-κB protein and its mRNA in lung of the rats with acute lung injury caused by LPS. Methods Wistar rats were randomly divided into normal group， model group， dexamethasone group and Xuanfeitongfufang Decoction large-， medium-， small-dose group， each group had eight rats. The ALI rat model was established by LPS tail-intravenous injection （6 mg/kg）. The rats in Xuanfeitongfufang Decoction groups were pretreated by Xuanfeitongfufang Decoction （15.12， 7.56， 3.78 g/kg） for 3 days before LPS induced ALI. The rats in dexamethasone group were pretreated by dexamethasone （5 mg/kg）. MD-2， NF-κB protein and its mRNA were measured by immunohistochemistry and PCR. The histopathology of the lung injury was observed by light microscope. Results Compared with normal group， the expression of MD-2， NF-κB protein and itsmRNA were obviously increased in model group （P 　　Key words：Xuanfeitongfufang Decoction；acute lung injury；MD-2；NF-κB；protein expression；mRNA expression；rats 　　髓样分化蛋白-2（MD-2）是结合在Toll样受体4 （TLR4）胞外区的一种髓样分化蛋白，与TLR4组成复合受体介导脂多糖（LPS）信号的跨膜转导，增强细胞经TLR4对LPS的反应，最终激活肺泡巨噬细胞核因子-κB （NF-κB）[1]。Schwartz等[2]研究表明，LPS激活NF-κB，进而诱导促炎因子大量生成是急性肺损伤（ALI）的关键发病环节，NF-κB激活可能是大量炎症反应基因表