摘要
目的探讨黄芪注射液对急性肺损伤血清肿瘤坏死因子-α（TNF-α）、白细胞介素-α（IL-α）及肺组织热休克蛋白70（HSP70）、核转录因子-κBBp65（NF-κBp65）蛋白表达的影响。方法54只Sprague-Dawley（sD）雄性大鼠随机分为对照组、模型组、黄芪组，每组18只，对照组生理盐水5．0mlMkg尾静脉注射，模型组尾静脉注射脂多糖（LPS）5．0mg／kg复制急性肺损伤模型，30min后黄芪组给予黄芪注射液10∥kg，对照组和模型组给予等量生理盐水，各组分别于制模后2、6、12h处死大鼠，心脏取血、留取肺组织标本，采用苏木素一伊红（HE）染色、光镜下观察肺组织病理变化，检测肺组织湿干重比（W／D），酶联免疫吸附法（ELISA）检测血清TNF-α、IL-6浓度，免疫组织化学法检测HSPT0和NF-κBp65蛋白表达量。结果模型组肺间质及肺泡明显充血水肿，大量炎性细胞浸润；黄芪组肺间质及肺泡充血水肿明显减轻，少量炎性细胞浸润。模型组、黄芪组各时间点肺组织W／D明显高于对照组（P〈0．01）；模型组肺组织W／D12h达高峰；黄芪组肺组织W／D12h下降程度最大，明显低于模型组各时间点（P〈0．01）。模型组、黄芪组各时间点血清TNF-α、IL-6浓度明显高于对照组（P〈0．01）；模型组血清TNF-α、IL-6浓度6h达高峰；黄芪组血清TNF-α、IL-6浓度12h降至最低，明显低于模型组各时间点（P〈0．01）。模型组、黄芪组各时间点HSP70蛋白表达量明显高于对照组（P〈0．01）；模型组HSP70蛋白表达量6h达高峰；黄芪组HSP70蛋白表达量12h达高峰，明显高于模型组各时间点（P〈0．01）。模型组、黄芪组各时间点NF-KBp65蛋白表达量明显高于对照组（P〈0．01）；模型组NF-κB0p65蛋白表达量6h达高峰；黄芪组NF-κBp65蛋白表达量12h降至最低，明显低于模型组各时间点（P〈0．01）。结论黄芪注射液可能通过提高HSP70表达，从而抑制NF-κB活性，下调TNF-α、IL-6的合成，减轻炎症反应，发挥肺损伤的保护作用。
Objective To study the effects of astragalus membranous injection on TNFα and IL 6 level in the serum of rat and lung tissue expression of heat shock protein 70 （ HSP70）, nuclear factor KBp65 （NF KBp65 ） protein in acute lung injury （ALI） rat. Methods Fifty four healthy SI） rats were randomly divided into control group, lipopolysaccharides （LPS） group, astragalus membranous group and each group was consisted of 18 animals. Control group rats were injected intravenously with 5. 0 mL/kg saline （ NS）, LPS group rats were injected intravenously with 5 mg/kg LPS, astragalus membranous group rats were injected intravenously with 10 g/kg astragalus membranous injection 30 min later. Control and LPS group rats were injected respectively with same dose NS injection. Six rats in each group were killed randomly, the heart blood was extracted and lungs were taken out at 2, 6, 12 h respectively. Histopatbology changes of rigbt lung tissue were examined with HEstaining under light microscopy. Left lung tissue was detected for the W/D ratio of lung tissue. The serum levels of TNFαand IL 6 were detected by enzyme linked immunosorbent assay （ELISA）. The protein expression of HSP70 and NF KBp65 in the lung tissue were detected by immunohistochemistry. Results The patho structures of lung tissue in LPS group rats was congestive, dropsy and abundant edematous fluid and inflammatory cells soakage in lung stroma and alveolus, but which were more conspicuous in LPS group than those in astragalus membranous group. The W/D ratio of lung tissue in LPS and astragalus membranous groups was significant higher than that in control group （ P 〈 0. O1 ）. The W/D ratio in LPS group reached the peak at 12 h, however the drop extent of the W/D ratio in astragalus membranous group was larger at 12 h, which was significant lower than that in LPS group rats at all time points （ P 〈 0.01 ）. The serum concentration of TNF α, IL 6 in LPS and astragalus membranous groups rats were significant higher than those in control group rats （ P 〈 0.01 ）. Those in LPS group reached the peak at 6 h, however those in astragalus membranous group rats reduced to the minimum at 12 h, which were significant lower than those in LPS group rats at all time points （P 〈 O. 01 ）. The protein expression of HSP70 in LPS and astragalus membranous groups were significantly higher compared with control group （P 〈 0.01 ）. In LPS group, the protein expression of HSPT0 reached the peak at 6 h, however that in astragalus membranous group reached the peak at 12 h, which was significant higher than that in LPS group at all time points （P 〈0.01 ）. The protein expression of NF KBp65 in LPS and astragalus membranous groups were significantly higher than those in control group （P 〈0.01 ）. In LPS group, the protein expression of NF KBp65 reached the peak at 6 h, however that in astragalus membranous group reduced to the minimum at 12 h, which was significantly lower than that in LPS group rats at all time points （ P 〈 0.01 ）. Conclusion Astragalus membranous injection may improve the ALl by promoting HSP70 expression and inhibiting NF KB release to down regulate the level of TNFα, IL6 in ALI rat induced by LPS.
出处
《中国急救医学》
CAS
CSCD
北大核心
2012年第3期227-231,I0001,I0002,共7页
Chinese Journal of Critical Care Medicine
关键词
黄芪注射液
急性肺损伤
热休克蛋白70
核转录因子--κB
Astragalus membranous injection
Acute lung injury （ALI）
Heat shockprotein 70 （HSP70）
Nuclear factor κB （ NF - KB）
