Department of Biochemistry Faculty of Science, Burapha University, Saensook, Muang, Chonburi 20131, Thailand.
J Ethnopharmacol. 2011 Feb 16;133(3):1008-14. doi: 10.1016/j.jep.2010.11.029. Epub 2010 Nov 19.
Cissus quadrangularis (family: Vitaceae) has been widely used in traditional herbal medicine for the treatment of hemorrhoids, gastric ulcers and bone healing. In the present study, we determined the anti-inflammatory activity and the molecular mechanism of the ethyl acetate extract of Cissus quadrangularis stem (CQE) in LPS-stimulated RAW 264.7 macrophage cells.
The inhibitory effect of CQE on LPS-induced nitric oxide (NO) production was evaluated in conditioned media. Cell viability was monitored by MTT assay. Protein and mRNA expressions were determined by RT-PCR and Western blotting analysis, respectively.
CQE potently inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophage cells in a dose-dependent manner. The mRNA and protein expressions of inducible nitric oxide synthase (iNOS) were suppressed also by CQE as was p65 NF-κB nuclear translocation. Further study demonstrated that CQE by itself induced heme oxygenase-1 (HO-1) gene expression at the protein and mRNA levels in dose- and time-dependent manner. In addition, the inhibitory effects of CQE on NO production were abrogated by a HO-1 inhibitor, zinc protoporphyrin IX (ZnPP).
Collectively, these results suggest that CQE exerts an anti-inflammatory effect in macrophages, at least in part, through the induction of HO-1 expression. These findings provide the scientific rationale for anti-inflammatory therapeutic use of Cissus quadrangularis stem.
筋骨草（卫矛科）在传统草药中被广泛用于治疗痔疮、胃溃疡和骨折愈合。本研究旨在确定筋骨草茎乙酸乙酯提取物（CQE）在 LPS 刺激的 RAW 264.7 巨噬细胞中的抗炎活性和分子机制。
通过条件培养基评估 CQE 对 LPS 诱导的一氧化氮（NO）产生的抑制作用。通过 MTT 测定法监测细胞活力。通过 RT-PCR 和 Western blotting 分析分别测定蛋白和 mRNA 的表达。
CQE 以剂量依赖性方式强烈抑制 LPS 诱导的 RAW 264.7 巨噬细胞中一氧化氮（NO）的产生。CQE 还抑制诱导型一氧化氮合酶（iNOS）的 mRNA 和蛋白表达以及 p65 NF-κB 核易位。进一步的研究表明，CQE 本身以剂量和时间依赖性方式诱导血红素加氧酶-1（HO-1）基因在蛋白和 mRNA 水平上的表达。此外，HO-1 抑制剂锌原卟啉 IX（ZnPP）可消除 CQE 对 NO 产生的抑制作用。
总之，这些结果表明 CQE 通过诱导 HO-1 表达在巨噬细胞中发挥抗炎作用。这些发现为筋骨草茎的抗炎治疗用途提供了科学依据。
