Anti-Inflammatory Activity of Root Ethanolic Extract of Harpagophytum procumbens on Human Peripheral Blood Mononuclear Cells and Screening of its Bioactive Compounds by GC-MS Method
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Anti-Inflammatory, Peripheral Blood Mononuclear Cells, Bioactive Compounds, GC-MS Technique
Abstract
Background: A traditional African medicine consisting of an anti-inflammatory fame is the Devil s Claw, e.g. Harpagophytum procumbens. Its extracts have a broad application in Europe on the instances of musculoskeletal pain and low back pain as the therapy of such inflammatory issues. It has been found out that Harpagophytum procumbens is capable of alleviating pain and reducing the need of NSAIDs and possibly as analgesic and an anti-inflammatory agent, mainly in osteoarthritis.
Procedures: The powder was resieved with certified sieve. The volatility and polyphenols are the classes of phytochemicals whose biocompatibility and solubility dictated the choice of the solvents. An inert GCMS selective detector 6890N/5973 of Agilent Technologies, USA, was used to analyze by GC-MS. PBMCs isolation and harvesting of peripheral blood samples. The pretreatment and incubation of PBMCs with the three different doses of Harpagophytum procumbens was done after which the incubation time was permitted to elapse after two hours of which LPS (1.0 0g/mL) was added to stimulate the cells. The concentration of the pro-inflammatory mediators was measured after removing and storing at 70 25C the supernatants of the LPS-stimulated and the pretreatment PBMCs after 24 hours.
Results: GC-MS analysis belongs to the number of the common techniques that could be applied to study the chemical composition of the plant extracts, the one of H. procumbens included. The method is detectable to volatile and non-volatile root and plant extract constituents. Over 31 compounds are discovered as a result of the research. Speaking about the most frequent kind of chemicals we may point out the existence of p-Vinylguaiacol, Decahydronaphthalene, 1-methylcyclohexane-1-carboxamide, Tetradecanoic acid, 1-methylethyl ester, 6-methoxypyridine-3-carboxylic acid, Pentadecanoic acid, 15-amino, Octahydrophenanthrene, Oleic acid, trimethylsilyl ester, Cell viability of PBMCs after incubation with six concentrations (0, 10, 20, 40, 80, 160, and 320 ug/ml) of Harpagophytum procumbens at different time points (24, 48 and 72 h) was found to be 94.83+/-3.1, 91.13+/-3.7, 90.03+/-3.8, 87.03+/- The production of pro-inflammatory mediators (TNF-alpha, Interleukin-1b and Interleukin-6 Pg/mL respectively) was recorded as (3.96, 8.27, 4.91, 3.52, 30.46, 15.70, 12.50 and 9.58) Pg/mL respectively in the effect of Harpagophytum procumbens (0, 10, 60, 150, LPS But Interleukin-1 5.50, 10.49, 3.67, 1.50, 35.70, 16.88, 10.00 and 6.49 Pg/mL was recorded respectively. During the same time the Interleukin-6 levels were (35.00, 60.71, 53.98, 18.93, 180.01, 176.61, 144.00 and 133.58) Pg/mL respectively.
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