Center for Cancer and Inflammation Research, School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China; Department of Animal Science, Patuakhali Science and Technology University, Dumki, Patuakhali, 8602, Bangladesh.
Center for Cancer and Inflammation Research, School of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China.
J Ethnopharmacol. 2021 Sep 15;277:114183. doi: 10.1016/j.jep.2021.114183. Epub 2021 May 13.
The dried rhizome of Atractylodes lancea (Thumb.) DC. (Compositae) has been prescribed in folk medicine for the management of various inflammatory conditions such as rheumatic diseases, gastritis and hepatitis. However, the molecular mechanisms underlying the beneficial properties of this herb remain elusive.
In this study, we investigated the anti-gastritis activities of Al-EE (an ethanolic extract of the herb) and explored the mechanism of action.
An ethanolic extract of the Atractylodes lancea (Thumb.) DC. (Compositae) rhizome, Al-EE, was prepared with ethanol (95%) and quality controlled using HPLC analysis. To determine the in vivo effects of this extract, we utilised a HCl/EtOH-induced gastritis rat model. In vitro assays were carried out using a lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cell model. MTT assays were used to examine cell viability, while Griess assays were carried out to measure nitric oxide (NO) production. Messenger RNA expression was examined by real-time PCR. Prostaglandin E (PGE) production was examined using ELISA assays. To examine protein expression and enzymatic activities, we employed western blot analysis. Nuclear transcription factor (NF)-κB activity was determined by Luciferase reporter assays.
The content of atractylenolide (AT)-1 and AT-2 in Al-EE was 0.45% and 5.07% (w/w), respectively (Supplementary Fig. 1). Al-EE treatment suppressed the production of NO and PGE, reduced the mRNA expression of inducible NO synthase (iNOS), cyclooxygenase (COX)-2 and tumor necrosis factor (TNF)-α, while also reducing the protein levels of iNOS and COX-2 in RAW264.7 macrophage cells. Furthermore, Al-EE inhibited the nuclear protein levels of NF-κB (p65) and NF-κB-driven luciferase reporter gene activity in RAW264.7 macrophage cells. Critically, intra-gastric injection of Al-EE (25 mg/kg) attenuated HCl/EtOH-induced gastric damage in SD rats, while the phosphorylation of Akt and IκBα was suppressed by Al-EE in vitro and in vivo.
In summary, Al-EE has significant anti-gastritis effects in vivo and in vitro, which can be associated with the inhibition of the Akt/IκBα/NF-κB signalling pathway. This mechanistic finding provides a pharmacological basis for the use of the A. lancea rhizome in the clinical treatment of various inflammatory conditions.
白术(Thumb。)DC 的干根茎(菊科)已在民间医学中用于治疗各种炎症性疾病,如风湿性疾病、胃炎和肝炎。然而,这种草药的有益特性的分子机制仍不清楚。
在这项研究中,我们研究了 Al-EE(草药的乙醇提取物)的抗胃炎活性,并探讨了其作用机制。
采用乙醇(95%)制备白术(Thumb。)DC.(菊科)根茎的乙醇提取物 Al-EE,并通过 HPLC 分析进行质量控制。为了确定该提取物的体内作用,我们利用了盐酸/乙醇诱导的胃炎大鼠模型。在体外试验中,我们使用脂多糖(LPS)刺激的 RAW 264.7 巨噬细胞细胞模型进行试验。MTT 试验用于检测细胞活力,而格里斯试验用于测量一氧化氮(NO)的产生。通过实时 PCR 检查信使 RNA 表达。使用 ELISA 测定法检查前列腺素 E(PGE)的产生。通过 Western blot 分析检查蛋白质表达和酶活性。通过荧光素酶报告测定法测定核转录因子(NF)-κB 活性。
Al-EE 中 AT-1 和 AT-2 的含量分别为 0.45%和 5.07%(w/w)(补充图 1)。Al-EE 处理抑制了 NO 和 PGE 的产生,降低了诱导型一氧化氮合酶(iNOS)、环氧化酶(COX)-2 和肿瘤坏死因子(TNF)-α的 mRNA 表达,同时还降低了 RAW264.7 巨噬细胞中的 iNOS 和 COX-2 蛋白水平。此外,Al-EE 抑制了 RAW264.7 巨噬细胞中核蛋白水平的 NF-κB(p65)和 NF-κB 驱动的荧光素酶报告基因活性。至关重要的是,Al-EE(25mg/kg)胃内注射可减轻 SD 大鼠盐酸/乙醇诱导的胃损伤,而 Al-EE 可在体外和体内抑制 Akt 和 IκBα的磷酸化。
总之,Al-EE 在体内和体外均具有显著的抗胃炎作用,这可能与抑制 Akt/IκBα/NF-κB 信号通路有关。该机制研究为白术根茎在临床治疗各种炎症性疾病中的应用提供了药理学依据。