Inhibition of NO Production in LPS-Stimulated Primary Rat Glial Cells by Gnidilatimonoein and Extract of Daphne mucronata.

Background: In the CNS, glial cells are involved in neuroinflammation and neuropathic pain. The glial cells are activated by a variety of pathological conditions and release pro-inflammatory mediators, including nitric oxide (NO). Overexpression of iNOS (inducible nitric oxide synthase) and extra NO is detrimental to neurophysiology and neuronal viability. Objectives: This study aimed to examine the effect of Gnidilatimonein isolated from D. mucronata and its leaves extract (as natural phytochemicals) on NO production in the LPS-induced primary glial cells. Materials and Methods: A preparative HPLC method was used to isolate gnidilatimonoein from leaves ethanolic extract. Various doses of Gnidilatimonoein, the ethanolic extract were applied to primary glial cells inflamed by lipopolysaccharide. A Colorimetric test, an MTT assay, and a RT-PCR analysis were then performed to analyze and compare NO production, cell viability, and iNOS expression. Results: Gnidilatimonoein treatment of pretreated primary glial cells significantly inhibited iNOS expression and decreased NO synthesis. Plant extracts also reduced NO production in inflamed microglial and glial at 0.1-3 mg.mL-1. At these concentrations, none of these compounds exerted a cytotoxic effect, suggesting that their anti-inflammatory effects were not due to the death of cells. Conclusion: This study indicates that D. mucronata and its active compound, Gnidilatimonoein, could have restrained effects on the expression of iNOS on the induced glial cells; however, further investigation is warranted.

Daphne mucronata enhances cell proliferation and protects human adipose stem cells against monosodium iodoacetate induced oxidative stress in vitro.

Mesenchymal stem cells (MSCs) are being used to treat many diseases as they exhibit great regenerative potential. However, MSC's transplantation sometimes does not yield the maximum regenerative outcome as they are unable to survive in inflammatory conditions. Several approaches including preconditioning are used to improve the survival rate of mesenchymal stem cells. One such recently reported approach is preconditioning MSCs with plant extracts. The present study was designed to evaluate the effect of Daphne mucronata extract on stressed human adipose-derived mesenchymal stem cells (hADMSCs). Isolated hADMSCs were preconditioned with different concentrations of Daphne muconata extract and the protective, proliferative, antioxidant and anti-inflammatory effect was assessed through various assays and expression analysis of inflammatory markers regulated through NF-κB pathway. Results suggest that preconditioning hADMSCs with Daphne mucronata increased the cell viability, proliferative and protective potential of hADMSCs with a concomitant reduction in LDH, ROS and elevation in SOD activity. Moreover, both the ELISA and gene expression analysis demonstrated down regulations of inflammatory markers (IL1-ß, TNF-α, p65, p50, MMP13) in Daphne mucronata preconditioned hADMSCs as compared to stress. This is the first study to report the use of MIA induced oxidative stress against hADMSC's and effect of Daphne mucronata on stressed hADMSCs. Results of these studies provided evidence that Daphne mucronata protects the hADMSCs during stress conditions by down regulating the inflammatory markers and hence increase the viability and proliferative potential of hADMSCs that is crucial for transplantation purposes.

Phytochemical screening, antioxidant and antibacterial properties of daphne mucronata.

OBJECTIVE: To aim at preliminary phytochemical screening of Daphne mucronata and evaluate its antioxidant and antibacterial activities. METHODS: Preliminary phytochemical screening was conducted for the crude extracts using standard methods. Antioxidant properties of crude methanolic extracts, n-hexane, chloroform, ethyl acetate and aqueous fractions of leaves, roots and bark were evaluated following standard procedures. The antibacterial activities were checked against Acinetobacter baumannii (A. baumannii), Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus), Morganella morganii (M. morganii) and vancomycin resistant S. aureus (VRSA). RESULTS: Carbohydrates, saponins, steroids, phenols/ tannins, flavonoids and glycosides were present in different parts of Daphne mucronata. The extracts showed good antioxidant activity with EC50 values of 157.82-361.61 µg/mL. Methanolic extracts of roots showed good activity against A. baumannii (86.95%), E. coli (85.18%) and S. aureus (84.61%). Methanolic extracts of bark were active against A. bumanni (65.21%), M. morganii (65.21%) and E. coli (62.96%). Methanolic extracts of leaves showed good activity against A. bumanni (78.26%), E. coli (77.78%), P. aeruginosa (74.07%), S. aureus (73.07%), M. morganii (69.56%), VRSA (68%) and Proteus vulgaris (60%). The n-hexane fraction of roots was effective against A. bumanni (78.26%). Chlorofom fraction of roots showed moderate activity against A. bumanni (60.86%) and S. aureus (61.53%). Ethyl acetate fraction of roots showed moderate activity against A. bumanii (69.56%), E. coli (62.96%) and S. aureus (69.23%). CONCLUSION: This study illustrates that Daphne mucronata possesses good antioxidant and antibacterial properties. The plant could be further exploited as potential natural antioxidant and as a new source of antimicrobials for treatment of various infections.