School of Chinese Material Medicine, Yunnan University of Chinese Medicine, Kunming, 650500, China.
Key Laboratory of External Drug Delivery System and Preparation Technology in University of Yunnan, Kunming, 650500, China.
BMC Complement Med Ther. 2021 Nov 20;21(1):284. doi: 10.1186/s12906-021-03458-0.
Stephania yunnanensis H. S. Lo is widely used as an antipyretic, analgesic and anti-inflammatory herbal medicine in SouthWest China. In this study, we investigated the anti-inflammatory activity and mechanism of sinoacutine (sino), one of the primary components extracted from this plant.
A RAW264.7 cell model was established using lipopolysaccharide (LPS) induced for estimation of cytokines in vitro, qPCR was used to estimate gene expression, western blot analysis was used to estimate protein level and investigate the regulation of NF- κB, JNK and MAPK signal pathway. In addition, an acute lung injury model was established to determine lung index and levels of influencing factors.
Using the RAW264.7 model, we found that sino reduced levels of nitric oxide (NO), tumour necrosis factor-α (TNF-α), interleukin (IL)-1β and prostaglandin E (PGE) but increased levels of IL-6. qPCR analysis revealed that sino (50, 25 μg/ml) inhibited gene expression of nitric oxide synthase (iNOS). western blot analysis showed that sino significantly inhibited protein levels of both iNOS and COX-2. Further signalling pathway analysis validated that sino also inhibited phosphorylation of p65 in the NF-κB and c-Jun NH2 terminal kinase (JNK) signalling pathways but promoted the phosphorylation of extracellular signal regulated kinase (ERK) and p38 in the MAPK signalling pathway. In addition, in a mouse model induced by LPS, we determined that sino reduced the lung index and the levels of myeloperoxidase (MPO), NO, IL-6 and TNF-α in lung tissues and bronchoalveolar lavage fluid (BALF) in acute lung injury (ALI).
Taken together, our results demonstrate that sino is a promising drug to alleviate LPS-induced inflammatory reactions.
云南千金藤（Stephania yunnanensis H. S. Lo）作为一种解热、镇痛和抗炎草药，在中国西南部被广泛使用。在这项研究中，我们研究了从该植物中提取的主要成分之一——头花千金藤碱（sino）的抗炎活性和机制。
采用脂多糖（LPS）诱导 RAW264.7 细胞建立体外细胞模型，用于估计细胞因子，qPCR 用于估计基因表达，western blot 分析用于估计蛋白质水平并研究 NF-κB、JNK 和 MAPK 信号通路的调节。此外，建立急性肺损伤模型以确定肺指数和影响因素水平。
在 RAW264.7 模型中，我们发现头花千金藤碱降低了一氧化氮（NO）、肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β和前列腺素 E（PGE）的水平，但增加了 IL-6 的水平。qPCR 分析显示头花千金藤碱（50、25μg/ml）抑制了一氧化氮合酶（iNOS）的基因表达。western blot 分析表明，头花千金藤碱显著抑制了 iNOS 和 COX-2 的蛋白水平。进一步的信号通路分析验证了头花千金藤碱还抑制了 NF-κB 和 c-Jun NH2 末端激酶（JNK）信号通路中 p65 的磷酸化，但促进了 MAPK 信号通路中细胞外信号调节激酶（ERK）和 p38 的磷酸化。此外，在 LPS 诱导的小鼠模型中，我们确定头花千金藤碱降低了肺指数以及急性肺损伤（ALI）中肺组织和支气管肺泡灌洗液（BALF）中髓过氧化物酶（MPO）、NO、IL-6 和 TNF-α的水平。
综上所述，我们的研究结果表明头花千金藤碱是一种有希望的缓解 LPS 诱导的炎症反应的药物。
