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الفعالية الحياتية لبعض مستخلصات النباتات الطبية على بعض انواع البكتريا والتعبير الجيني لانزيمات الكيراتينز المنتج من قبل Trichophyton rubrum == Biological Activity of some Medicinal Plant Extracts on some Bacteria and Gene Expression of Keratinases Produced from Trichophyton rubrum
Author name:
علي جليل عبيد حسن عينون
Supervisor name:
جواد كاظم عبود علي الجنابي | وجدان رضا محمود محمد تاج الدين
General topic:
Biology
Specific topic:
Microbiology
Degree:
Doctorate
University:
University of Babylon - College Of Science - Department Of Biology
Language:
English
University location:
Babylon
First pages:
24T3507 - p.pdf
Abstract:
The present study aimed to search for safe and effective antifungal agents through investigation of the phytochemical characterization and bioactive properties of medicinal plants (Pimpinella anisum essential oil and Moringa oleifera and Malva sylvestris leaf extracts) and their effect on gene expression of keratinases produced by T. rubrum using Real Time PCR.Phytochemical analysis of active materials of P. anisum essential oil and M. oleifera leaf extracts were carried out, through TLC accompanied autobiography, high performance liquid chromatography (HPLC) and GC - MS analyses were proceeded to relate the chemical profile with their antifungal activity. Also, the cytotoxic effects of active compound and crude extract for both plant extracts were determined using MTT (3 - (4,5 - dimethylthiazol - 2 - yl) - 2,5 - diphenyltetrazolium bromide dye) assay. Screening the antifungal activity of Pimpinella anisum essential oil, Moringa oleifera leaf extracts and four traditional antifungal agents against growth and development of two strains of T. rubrum differed in their virulence were conducted. MIC for crud extracts and antifungal agent were also determined using microdilution plate, moreover, the effects of crud extracts and active compounds on gene expression was evaluated.Qualitative TLC, HPLC analysis for the anise essential oil extract revealed the presence of anethole which was identified as a main volatile compound in this extract which was identified as anethole. GC - MS analysis also revealed that trans - anethole is the major compound (96.8%), then Himachalene (1.84%), other components were also present but less than 1% e.g. Estragole, cyclohexene and 2 - Methoxy - 5 - (1 - propenyl) phenol. On the other hand, LC - MS analysis were showed the existence of that keratinase inhibitor (extracted from Moringa oleifera leaf)IICryptochlorogenicacid (C16H18O9), Astragalin (C21H20O11), so the presences of these compounds reflected the inhibitory effects on keratinase enzyme.The results upon screening, revealed that the anise oil inhibited both strains of T. rubrum, since the growth inhibition for first strain were 21, 30 and 39 mm at 15.6, 31.2 and 62.5 μl/ml respectively, while in second strain were 24, 32 and 39 mm at 15.6, 31.2 and 62.5 μl/ml respectively. The tested strains of this fungus revealed substantial sensitivity pattern toward crude extract of essential oil and anethole as well. Results showed that there was considerable growth inhibition for both strains of T. rubrum at minimal inhibitory concentrations (MIC) between 15.6 μg/mL for crude oil and 7.5 μg/mL for anethole. The inhibition in radial growth due to the effect of anise essential oil increased from 56% at 3rd day to 86% at the 7th day. Sporulation was substantially decreased by anise oil treatment. Anise oil was significantly inhibited conidial germination for both strains. Treatment with anise oil induced morphological changes in the conidial group where the conidia were absent in treated sample. Protease inhibitor isolated from mature leaves of M. oleifera showed great ability to inhibit keratinase activity of T. rubrum.MTT assay revealed that anethole is safe as phototherapeutic agents, when it is having no effects on normal cell, but shown anticancer activity against cancer cell.Anise oil, anethole, M. oleifera crud inhibitor, Cryptochlorogenic acid and Astragalin showed high inhibitory effects on gene expression of keratinase enzyme in both strains of T. rubrum KP979791 and KP979787.
