摘要
Ginsenosides, the main active constituents of Panax ginseng Meyer （P. ginseng）, have potential therapeutic effects. All tested ginsenosides except gin- senoside F1 have previously been reported in inflammation studies using the RAW 264.7 macrophage cell line. We ex- amined the anti-inflammatory effects of single sugar moiety ginsenosides such as compound K （CK）, Rh2, Rhl, and F1 that were isolated from P. ginseng through in silico docking studies. We investigated their biological activity predictions, including absorption, distribution, metabolism, excretion, and toxicity and PASS properties, on the suppression of NF- κB, followed by in vitro validation in lipopolysaccharide （LPS）-stimulated RAW 264.7 macrophage cells. The molecular docking results of our study showed that all treated ginsenosides are non-toxic and may be drug-like molecules. The molecular binding interactions of these ginsenosides with the active residues of NF-κB noticeably support their anti-inflammatory activity. CK and Rhl sig- nificantly reduced the production of nitric oxide, cyclooxy- genase-2 （COX-2）, and pro-inflammatory cytokines such as prostaglandin E2 and tumor necrosis factor alpha （TNF-α） in a dose-dependent manner. Real-time PCR and Western blot analyses further confirmed that protopanaxadiols （PPDs） and protopanaxatriols （PPTs） inhibitory effects may have been due to the down-regulation of TNF-α, inducible nitric oxide synthase, COX2, nuclear factor kappa B （NF-κB）, and I kappa B kinase. The expression of co-stimulatory molecules such as ROS was also inhibited by CK and Rhl in a dose- dependent manner. Furthermore, activation of NF-κB in LPS-stimulated RAW 264.7 macrophages was significantly suppressed by CK and Rhl. Taken together, these results provide evidence that PPD- and PPT-type ginsenosides in- cluding CK and Rhl may exhibit strong anti-inflammatory effects by inhibiting pro-inflammatory mediators through down-regulation of NF-κB.
通过in silico对接实验研究了CK,Rh1,Rh2及F1这4种单糖结构人参皂苷的抗炎作用.结果表明,所有测试的人参皂苷均为无毒的类药性分子.人参皂苷与NF-κB活性残基的相互作用显示出抗炎活性.其中CK和Rh1可呈剂量依赖性地减少NO,COX-2以及促炎性细胞因子PGE 2和TNF-α.通过RT-PCR和Western blot检测证实抑制效果可能来源于人参皂苷对TNF-α等因素的向下调节作用.活性氧的表达也可由CK和Rh1以剂量依赖性效应得到抑制.PPD,PPT型人参皂苷CK和Rh1能显著抑制LPS受激RAW264.7细胞内活化的NF-κB,通过对NF-κB的向下调节抑制炎症介质,显示很强的抗炎效果.
基金
supported by a post-doctoral fellowship grant from the Kyung Hee University in 20120351
