فهرست مطالب somayeh babashpour

Most herbs play significant roles in the treatment of various diseases. Because dopamine functions in theanti-inflammatory process and the presence of this substance in Portulaca Oleracea L. native plant, investigating thisplant’s anti-inflammatory properties in treating neurological diseases is interesting. The objective of this study was to estimate the NO production and the expression level of inflammatory genesin lipopolysaccharide (LPS)-treated microglial cells affected by P. oleracea L. extraction. P. oleracea L. hairy root extract was isolated, and the primary microglial cell of the rat wasisolated from glial cells and confirmed by immunocytochemistry analysis. Microglial cells were pretreated with differentconcentrations of P. oleracea L. extract and then treated with 1 μg.mL-1 LPS. The control group did not receive anytreatment. The NO level in culture supernatants was measured by the Griess method. The mRNA expression levels ofiNOS (inducible Nitric oxide synthase) and TNF-α (tumor necrosis factor-alpha) in LPS-treated microglial cells wereevaluated using Real-Time PCR. The present study determined that 0.1 mg. mL-1 of the P. oleracea L. extract decreased the NO production in ratmicroglial cells. Different concentrations of the P. oleracea L. extract had no prominent effects on LPS-treated cell viability. The results of real-time PCR indicated that P. oleracea L extracts suppressed the mRNA expression levels of iNOS and TNF-α in LPS-treated cells. MTT assay determined that P. oleracea L. extract was not cytotoxic, and the anti-inflammatory P. oleracea L. extract effects observed were not because of cell death. P. oleracea L. extract might be helpful as an anti-inflammatory agent in treating inflammatory diseases.

Dopamine is one of the important medications of Portulaca oleracea L. To optimize the production of dopamine, one of the methods is the identification and engineering of metabolite pathways. To investigate the tyrosine decarboxylase (TDC) and tyrosinase, which seem to be the most important genes in dopamine synthesis pathway, hairy roots were produced from Portulaca oleracea using Agrobacterium rhizogenes and total RNA was extracted from hairy roots. A cDNA library was synthesized using RT-PCR. Then, the twogenes were amplified, isolated and cloned in a pTG 19-T vector. Bioinformatics' databases were used to predict the details of the structural, functional and biological characteristic of these genes. Nucleotide sequence analysis revealed that the cloned cDNAs expressed TDC and tyrosinase, and contained a single open reading frame of 1800 bp and 1750 bp, respectively. TDC has the most similarity with TDC of Arabidopsis thaliana (L.) Heynh.,but tyrosinase has 98% similarity withtyrosinase of Agaricus bisporus. Because of More negatively charged amino acids the TDC has hydrophobic properties, therefore affinity and hydrophilic chromatography can be used for purification of TDC. But tyrosinase has hydrophilic properties and hydrophobicity chromatography can be used for its purification. There were two peroxisomal signal peptide (KLAKEFEQL) and (KIEGRPLHL) in the TDC and tyrosinase, respectively. Therefore, they are biologically active in the peroxisomes, and included in biosynthesis dopamine through the transformation of L-lysine to L-dopa and finally to the dopamine. In conclusion, increasing the expression of TDC and tyrosinase through the genetic engineering can increase dopamine production in the Portolaca.