益气活血中药对急性肺损伤大鼠炎症因子及Toll样受体4 mRNA表达的影响

Effect of Supplementing Qi and Activating Blood Circulation Herbs on Inflammatory Factors and Toll-like Receptor 4 mRNA Expression in Rats with Acute Lung Injury

目的:观察和探讨益气活血中药对脂多糖(LPS)诱导的急性肺损伤(ALI)大鼠支气管肺泡灌洗液中(BALF)炎症因子及肺组织中Toll样受体4(TLR-4)mRNA表达的影响。方法:SD雄性大鼠72只,随机分成正常组、模型组、地塞米松组、益气活血中药组4组,每组18只;除正常组气管内滴注生理盐水,其余各组滴注LPS(5 mg·kg^(-1))制备ALI模型,造模前后3 d各组予以相应药物或生理盐水灌胃;每组分别于造模后8,24,72 h各处死6只,苏木素-伊红(HE)染色光镜下观察各组大鼠肺组织病理学改变,酶联免疫(ELISA)法检测BALF中炎症因子肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL^(-1)β),IL-6的浓度,实时荧光定量PCR(Real-time PCR)测定肺组织中TLR-4 mRNA的表达。结果:肺组织病理学评分:与正常组比较,同时相的LPS各组均显著升高(P<0.01);与模型组比较,地塞米松组和中药组均降低(P<0.05,P<0.01),且72 h组降低最为明显。大鼠BALF中TNF-α,IL^(-1)β,IL-6浓度:LPS造模各组均明显高于同时相的正常组(P<0.01),模型组均高于中药组和地塞米松组(P<0.01),且随着时间的延长,模型组逐渐上升,而药物干预组逐渐降低(P<0.05,P<0.01)。肺组织TLR-4mRNA相对表达量:与正常组比较,LPS造模各组同时相肺组织TLR-4 mRNA的相对表达量均显著升高(P<0.01),以模型组8 h点为最高;与模型组比较,中药组各时间点表达均降低(P<0.05,P<0.01)。结论:益气活血中药能够降低急性肺损伤大鼠BALF中TNF-α,IL^(-1)β,IL-6的水平,减轻肺组织的病理损伤,其防治ALI的作用机制可能与抑制TLR-4 mRNA的表达有关。

Objective： To observe and investigate the effects of supplementing Qi and activating blood circulation herbs on inflammatory factors in bronchoalveolar lavage fluid（BALF） and Toll-like receptor 4 mRNA expression in lung tissues of rats with acute lung injury（ALI） induced by lipopolysaccharide（LPS）. Method：Seventy-two healthy male SD rats were randomly divided into normal group,model group,dexamethasone group and supplementing Qi and activating blood circulationherbs group,n = 18 in each group. ALI models were established by intratracheal instillation of LPS（5 mg·kg^-1）,while normal group received normal saline. Three days before and after making the models,the corresponding drugs or normal saline were given in each group by gavage. Six rats in each group were put to death at the end of the 8,24,72 h after modeling. Then the pathological changes of lung tissues were observed under light microscope; the concentrations of tumor necrosis factor-α（TNF-α）,interleukin1β（IL-1β）,and interleukin 6（IL-6） in rat bronchoalveolar lavage fluid（BALF） were measured by enzymelinked immune sorbent assay（ELISA）,and the TLR-4 mRNA expression levelsin lung tissues were determined by real-time polymerase chain reaction（Real-time PCR）. Result： For the pathological score of lung tissues,as compared with normal group,LPS in other groups exhibited a higher level（P〈0. 01）; as compared with model group,dexamethasone group and Chinese herbs group exhibited a lower level（P〈0. 05,P〈0. 01） and the level was lowest in 72 h. For the contents of TNF-α,IL-1β and IL-10 in BALF,LPS in other groups displayed an obvious increase as compared with normal group,but the levels in dexamethasone group and herbs group were decreased as compared with model group（P〈0. 01）; the level in model group was increased gradually over time in model group and the levels indrug intervention groups were decreased over time（P〈0. 05,P〈0. 01）. For the TLR4 mRNA expression level in lung tissues,as compared with normal group,LPS in other groups exhibited a higher level（P〈0. 01） and the level was highest in model group at 8 h; as compared with the model group,the expression levels in herbs group were decreased at various time points（P〈0. 05, P〈0. 01）. Conclusion：Supplementing Qi and activating blood circulation herbs could decrease the levels of TNF-α,IL-1β and IL-10 in BALF in rats with ALI induced by LPS,improve the pathological changes in lung tissues,and the prevention and treatment mechanism for ALI may be associated withinhibiting the TLR4 mRNA expression.