In vitro antioxidant activity of crude extracts of Harpagophytum zeyheri and their anti-inflammatory and cytotoxicity activity compared with diclofenac.

BACKGROUND: This study evaluated the in vitro antioxidant activity and comparison of anti-inflammatory and cytotoxic activity of Harpagopytum zeyheri with diclofenac. METHODS: In vitro assays were conducted using water, ethanol, and ethyl acetate extracts of H.zeyheri. The antioxidant activity was evaluated using the 2,2'-diphenyl-1-picrylhydrazy (DPPH) and 2,2'- azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays. The anti-inflammatory activity was determined by measuring the inhibition of nitric oxide (NO) on lipopolysaccharide (LPS)-induced RAW 264.7 mouse macrophages as well as cytokine (TNF-α and IL-10) expression on LPS-induced U937 human macrophages. For cytotoxicity, cell viability was determined using the 3-(4, 5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The ethyl acetate extract had the lowest IC50 values in the DPPH (5.91 µg/ml) and ABTS (20.5 µg/ml) assay compared to other extracts. Furthermore, the ethyl acetate extracts effectively inhibited NO and TNF-α and proved to be comparable to diclofenac at some concentrations. All extracts of H. zeyheri displayed dose-dependent activity and were associated with low levels of human-IL-10 expression compared to quercetin. Furthermore, all extracts displayed low toxicity relative to diclofenac. CONCLUSIONS: These findings show that H. zeyheri has significant antioxidant activity. Additionally, similarities exist in the inflammatory activity of H. zeyheri to diclofenac at some concentrations as well as low toxicity in comparison to diclofenac.

Natural Polyketides Isolated from the Endophytic Fungus Phomopsis sp. CAM212 with a Semisynthetic Derivative Downregulating the ERK/IκBα Signaling Pathways.

Three previously undescribed natural products, phomopsinin A - C (1:  - 3: ), together with three known compounds, namely, cis-hydroxymellein (4: ), phomoxanthone A (5: ) and cytochalasin L-696,474 (6: ), were isolated from the solid culture of Phomopsis sp. CAM212, an endophytic fungus obtained from Garcinia xanthochymus. Their structures were determined on the basis of spectroscopic data, including IR, NMR, and MS. The absolute configurations of 1: and 2: were assigned by comparing their experimental and calculated ECD spectra. Acetylation of compound 1: yielded 1A: , a new natural product derivative that was tested together with other isolated compounds on lipopolysaccharide-stimulated RAW 264.7 cells. Cytochalasin L-696,474 (6: ) was found to significantly inhibit nitric oxide production, but was highly cytotoxic to the treated cells, whereas compound 1: slightly inhibited nitric oxide production, which was not significantly different compared to lipopolysaccharide-treated cells. Remarkably, the acetylated derivative of 1: , compound 1A: , significantly inhibited nitric oxide production with an IC50 value of 14.8 µM and no cytotoxic effect on treated cells, thereby showing the importance of the acetyl group in the anti-inflammatory activity of 1A: . The study of the mechanism of action revealed that 1A: decreases the expression of inducible nitric oxide synthase, cyclooxygenase 2, and proinflammatory cytokine IL-6 without an effect on IL-1ß expression. Moreover, it was found that 1A: exerts its anti-inflammatory activity in lipopolysaccharide-stimulated RAW 264.7 macrophage cells by downregulating the activation of ERK1/2 and by preventing the translocation of nuclear factor κB. Thus, derivatives of phomopsinin A (1: ), such as compound 1A: , could provide new anti-inflammatory leads.

Anti-inflammatory and antioxidant properties of leaf extracts of eleven South African medicinal plants used traditionally to treat inflammation.

ETHNOPHARMACOLOGICAL RELEVANCE: Inflammation is a complex mechanism employed by the body to promote healing and restoration to normal function in the event of injury. Eleven plant species were selected in this study based on their use in traditional medicine against inflammation in South Africa. METHODS: Hexane, acetone, ethanol, methanol and water extracts of the powdered plants were prepared and a total of fifty-five extracts were tested for their anti-inflammatory and antioxidant activities. The anti-inflammatory activity of extracts was evaluated via the 15-lipoxygenase (15-LOX) inhibitory and the nitric oxide (NO) inhibition assays using lipopolysaccharide (LPS)-activated RAW 264.7 murine macrophages. Total flavonoid and total phenolic contents were determined. The antioxidant activity of the extracts was performed using radical scavenging DPPH (2, 2-diphenyl-1-picrylhydrazyl) and electron reducing ABTS (2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) assays. RESULTS: The hexane extract of Typha capensis (TC) had good lipoxygenase inhibitory activity with IC50 of 4.65 µg/mL, significantly (p < 0.05) higher than that of the positive control quercetin (IC50 = 24.60). The same extract also had good nitric oxide inhibitory activity with 86% NO inhibition and cell viability of 97% at 50 µg/mL. The TC acetone extract had the best antioxidant activity with IC50 of 7.11 and 1.91 µg/mL respectively in the DPPH and ABTS assays. Following fractionation of the TC plant material, the ethyl acetate fraction had interesting antioxidant activity and the methanol/water (35%) and hexane fractions had good 15-LOX inhibitory activity. The antioxidant and anti-inflammatory activities therefore resided in both polar and more non-polar fractions. CONCLUSION: The acetone extract of Typha capensis and its fractions had good anti-inflammatory and antioxidant activities, supporting the medicinal use of this species against inflammation. Other species including Ficus elastica, Carpobrotus edulis, Cotyledon orbiculata and Senna italica also had good activity worthy of further investigation.