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Abstract：
The anti-inflammatory activity of cirsilineol in in vivo condition was assessed by measuring the relative organ weight, lung dry/wet weight ratio, protein concentration, and infiltration of inflammatory cells in bronchoalveolar lavage fluid. We estimated the myeloperoxidase activity and levels of cytokines, chemokines, and inflammatory markers to analyze the efficacy of cirsilineol against lipopolysaccharide (LPS)-induced lung inflammation. Furthermore, we quantified the gene expression of NFkB/IKK signaling molecules in cirsilineol-treated and untreated acute lung injury mice to confirm the anti-inflammatory property of cirsilineol. The lung histology was assessed with hematoxylin and eosin staining. Apart from in vivo experiments, in vitro tests with LPS-stimulated RAW 264.7 macrophages were also performed. Cell viability assay was performed in the presence and absence of LPS in RAW 264.7 macrophages to determine the cytotoxic effect of cirsilineol against macrophages. Reverse-transcription polymerase chain reaction (RT-PCR) analysis was done to analyze the gene expression of inflammatory markers in LPS-treated RAW 264.7 macrophages to prove that cirsilineol effectively inhibits inflammation in vitro. The results of our study prove that cirsilineol effectively inhibits inflammation in both in vivo and in vitro conditions. RT-PCR analysis results of NFkB/IKK signaling molecules clearly illustrate that cirsilineol inhibited the expression of NFkB/IKK signaling protein and thereby prevented inflammation in in vivo condition, and it is further confirmed with the results of inflammatory protein expression in vitro model. The lung histopathological studies authentically confirm that cirsilineol potentially prevented the mice from LPS-induced lung inflammation.
摘要：
通过测量相对器官重量来评估cirsilineol在体内条件下的抗炎活性，肺干/湿重比，蛋白质浓度，支气管肺泡灌洗液中炎性细胞浸润。我们估计了髓过氧化物酶的活性和细胞因子的水平，趋化因子,和炎症标志物来分析cirsilineol对抗脂多糖（LPS）诱导的肺部炎症的功效。此外，我们量化了cirsilineol治疗和未治疗的急性肺损伤小鼠中NFkB/IKK信号分子的基因表达,以证实cirsilineol的抗炎特性.用苏木精和曙红染色评估肺组织学。除了体内实验,还进行了LPS刺激的RAW264.7巨噬细胞的体外试验.在存在和不存在LPS的情况下,在RAW264.7巨噬细胞中进行细胞活力测定,以确定cirsilineol对巨噬细胞的细胞毒性作用。进行逆转录聚合酶链反应（RT-PCR）分析，以分析LPS处理的RAW264.7巨噬细胞中炎症标志物的基因表达，以证明cirsilineol在体外有效抑制炎症。我们的研究结果证明，cirsilineol在体内和体外条件下都能有效抑制炎症。NFkB/IKK信号分子的RT-PCR分析结果清楚地表明，cirsilineol抑制NFkB/IKK信号蛋白的表达，从而在体内条件下预防炎症。并与体外模型的炎性蛋白表达结果进一步证实。肺组织病理学研究真实地证实了cirsilineol潜在地预防了小鼠的LPS诱导的肺部炎症。
