摘要
目的
观察清瘟败毒饮对脂多糖诱导急性肺损伤大鼠肺组织中核因子-κB表达的影响,以及对血清中细胞因子IL-1β和工L-13含量的影响,探讨清瘟败毒饮对ALI的治疗作用及干预机制。
方法
1.选取健康雄性SD大鼠144只,随机分为6组,每组24只,分别为正常对照组,模型组,泼尼松组和清瘟败毒饮大、中、小剂量组。脂多糖溶液(2mg/kg)气管内滴注复制急性肺损伤(ALI)模型,连续两天。各治疗组在第二次滴注完LPS溶液1小时后,用生理盐水将相应的药物稀释至4ml灌胃,一天两次。各组分别于灌胃后24h、48h处死6只大鼠,于72h处死余下所有大鼠,并收集所需标本；
2.通过测定支气管肺泡灌洗液(BALF)白细胞总数、肺组织干湿比、肺系数的变化、镜下观察HE染色肺组织病理变化,评价肺损伤的程度和清瘟败毒饮的治疗效果；
3.采用ELISA法检测血清中细胞因子工L-1β和IL-13的含量；
4.采用Western blot方法检测肺组织中核因子-κB的表达水平；
5.采用SPSS13.0统计软件分析处理数据。
结果
1.LPS致ALI大鼠肺组织病理检查结果显示肺泡结构破坏或实变,肺间质明显水肿,大量炎性细胞浸润,肺泡腔和肺间质内可见局灶性炎症细胞浸润和大量红细胞渗出,其BALF中白细胞总数均显著升高,肺组织干湿比和肺系数均明显增高,与正常对照组比较有显著性或非常显著性差异(P<0.05或P<0.01)；
2.清瘟败毒饮各剂量治疗组大鼠BALF中白细胞总数均下降,肺泡损伤、炎症细胞浸润和红细胞渗出情况有较大程度的改善,病理学炎症积分(除72h小剂量组外)均降低,大剂量组大鼠肺干湿比下降,各剂量组大鼠肺系数均下降；与同时相模型组相比均有显著性差异或非常显著性差异(P(0.05或P<0.01)；
3.清瘟败毒饮各剂量治疗组(除24h小剂量组外)大鼠血中IL-1β含量均下降,IL-13含量均升高,与同时相模型组相比均有显著性差异或非常显著性差异(P<0.05或P<0.01)：
4.清瘟败毒饮各剂量治疗组中48h清瘟败毒饮大、中剂量组和72h各剂量组大鼠肺组织中NF-K B p65的表达均降低,与同时相模型组相比均有显著性差异或非常显著性差异(P<0.05或P<0.01)；
5.清瘟败毒饮大剂量治疗组,除48h肺组织中NF-κBp65的表达显著高于泼尼松组(P<0.05)外,其余各项观测指标与泼尼松组相比均无显著性差异(P>0.05)。
结论
1.清瘟败毒饮能减轻脂多糖致急性肺损伤大鼠肺组织的损伤程度,在一定程度上减少炎症细胞在肺部积聚、浸润、渗出,起到比较明确的肺保护作用；
2.清瘟败毒饮可通过有效调节脂多糖致急性肺损伤大鼠血中炎症细胞因子IL-1β和抗炎症细胞因子IL-13表达水平,促使炎症和抗炎症细胞因子趋于动态平衡,从而减轻肺部炎症细胞浸润,起到修复保护损伤肺组织的作用；
3.清瘟败毒饮能有效减少脂多糖致急性肺损伤大鼠肺组织NF-κB的表达,通过抑制NF-κB的活化和炎性细胞因子的生成,对炎症反应起到了抑制作用；
4.在脂多糖致急性肺损伤早期使用清瘟败毒饮,可有效减轻肺组织的损伤程度,减轻肺部的炎症反应；
5.随着给药时间的延长大剂量应用清瘟败毒饮治疗脂多糖致急性肺损伤可达到与泼尼松治疗相近疗效。
Objective
To observe the effects of Qingwen Baidu decoction on NF-κB activation in rats lung tissue and the content of IL-1β,IL-13 in blood serum with acute lung injury induced by LPS, so to explore the treatment and the intervention mechanism for acute lung injury.
Methods
1.144 male rats were randomly divided into six groups, and each group have 24 rats: normal control group, model group, prednisone group, and three Qingwen Baidu decoction groups. Then instill LPS solution (2mg/Kg) into trachea to prepare for ALI model once a day and for two days. After one hours later, use 4 ml drug solution diluted by normal saline to intragastric administration twice a day. Kill 6 rats and collect specimen in each group at 24h, 48h, and kill the others at 72 hours after administration.
2. Through examine the total white cell in bronchoaleolar lavage fluid (BALF), wet/dry ratio of lung tissue and the lung index changes, and the pathological changes under light microscope with HE stained rats lung tissue to evaluate the degree of injuried lung and the therapeutic effect of Qingwen Baidu decoction to the ALI.
3. Using ELISA method to detect the content of IL-1βand IL-13 in rats serum.
4. Using Western Blot method to analyse the expression level of NF-κB in rats lung tissue.
5. Using Spssl3.0 statistics software to analyze and process data.
Results
1. The pathology of rats Lung tissue with LPS induced ALI results the alveolar structure damaged or consolidation, the lung interstitial edema obviously, the alveolar cavity and the lung interstitial were infiltrated by the focal inflammatory cells and red blood cells. And the total white cell in the BAIF, Wet/dry ratio of lung tissue and the lung index were increased significantly. Compared with the normal control group has significant or very significant difference (P<0.05, P<0.01);
2. The white cell in the BAIF, the pathological classification of inflammation (except low dose group of 72h) and the lung index were decreased in all doses of Qingwen Baidu decoction treatment groups. And the damaged alveolar, the infiltrated inflammatory cells and erythrocyte exudated were gradully improved. And the wet/dry ratio of large doses of Qingwen Baidu decoction was decreased obviously. Compared with the model group phase has significant or very significant difference (P<0.05, P<0.01);
3. The content of IL-1βin rats serum decreased (except low dose group of 24h), and the IL-13 in blood serum rised in all doses of Qingwen Baidu decoction treatment groups. Compared with the model group phase has significant or very significant difference (P<0.05, P<0.01);
4. The expression of NF-κB p65 in high and middle dose of Qingwen Baidu decoction treatment groups decreased in 48h, and all does of Qingwen Baidu decoction decreased in 72h. Compared with the model group phase has significant or very significant difference (P<0.05, P<0.01);
5. Except the total white cell in the BAIF in 24h and the the expression of NF-κB p65 in the lung tissue higher than prednisone group(P<0.05), Other observational indexes in large does of Qingwen Baidu decoction has no significant difference (P>0.05)
Conclusion
1. The Qingwen Baidu decoction can reduce the injured degree of ALI rats tissue, and can reduce the inflammatory cells gather, infiltrate and exudate in the lung tissue, so to play a clearer role in lung protection.
2. The Qingwen Baidu decoction can accommodate the express of inflammatory cytokine IL-1βand anti-inflammatory cytokine IL-13 to promote the inflammation and anti-inflammatory cytokines have a dynamic balance, reduce the lung inflammation so to have the function of repair and protect the injured lung tissue.
3. The Qingwen Baidu decoction can effectively reduced the express of NF-κB in LPS induced ALI rats tissue. By inhibit the activation of NF-κB and the generation of inflammatory cytokines can reduce the inflammatory response in rats lung tissue.
4. Using the Qingwen Baidu decoction early can effectively reduced lung tissue injured degree and can reduced the inflammatory response in ALI rats lung tissue induced by LPS.
5. Compared with the group of prednisone, applicating the high-dose Qingwen Baidu decoction can achieve similar efficacy with the time prolonged.
引文
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