Detection of some active compounds in Paliurus spina-christi plant :their histopathological effect and molecular mechanism on dermatophytes

Fifty five samples of patients with skin infections by dermatophytes have been studied during the period from June 2010 to September 2010.Eighteen isolates were isolated. The most common species were Trichophyton mentagrophytes, Trichophyton rubrum, and Microsporum gypsum .The relation between
dermatophytes infections and the age and gender of patients were studied .The first group age (20-40 years old)showed the highest rate of infections .Tinea corporis was the most common one. The percentage of infection with Trichophyton mentagrophytes was(13.63%) for male and(22.72%) for female ,Trichophyton rubrum percentage was (12.5%) for male only .This was followed by Microsporum gypsum(23.53%) detected in male and female .The clinical examination revealed that tinea corporis was most common among
other tinea types such as (tinea cruris,tinea capitis, tinea unguium, tinea manum). High performance liquid chromatography(HPLC) analysis of methanolic extracts of Paliurus spina-christi revealed the presence of Gallic acid,caffeic acid,syringic acid & epicotichen acid , while Gas chromatography( GC) revealed a wide range of chemical compounds (36).They were varied in types (sugar,amino acids ,phenols, fatty acids and esters).The Paliurus spina-christi methanolic extract was significantly the most effective in comparison with the ethanolic extract ,125μg/ml of Paliurus spina-christi methanolic extract was the most effective in inhibiting the mycelial growth of T.mentagrophytes .The mean diameter of T. mentagrophytes growth after incubation for 7 days in media treated with 125μg/ml of Paliurus spina-christi methanolic extract concentration was (1.5±0.03)cm. This effect was statistically significant(P<0.05),as compared with the control group whose mean diameter was (8.3 ±0.87)cm .On the other hand ,the mean diameter of T.mentagrophytes growth after incubation for 7 days in medium treated with125μg/ml of ethanolic Paliurus spina-christi extract was (2.4±0.09). This effect was statistically significant(P<0.05) as compared with the control group which recorded (8.3±0.87).The ethanolic extract of spina-christi had a moderate effect in
comparison with the methanolic extract at concentration 125 μg/ml on growth inhibiting .Clinical features of mice skin infected with T. mentagrophytes were characterized by redness ,swelling , regular margin with ulceration and loss of hair. The light microscopic study had shown certain degenerative changes in the
infected area. These changes included edema with heavy and acute inflammatory cells in the dermis area which may extend to hypodermis causing abscess .Infected mice treated with Paliurus spina-christi methanolic extract showed the disappearance of redness and hair regeneration after 7 days leaving
a white scar, usually slightly depressed along its entire length .The incision was covered with newly formed epidermis and hair was seen in comparison with treatment of Clotrnazole-P ,where healing was characterized by slower regenerative changes and the incision area was less in comparison with the area
treated with Paliurus spina-christi methanolic extract. Light microscopic study of the infected area after 14 days revealed that the moderate inflammatory cells were present in the dermis along with a normal
epidermis with its common structures in comparison to the other groups (+ve and vehicle)which emonstrated the moderate healing after21 days of treatment :keratin tissue ,sweat glands, hair follicle regeneration in the dermal layer. Healing progress was more observed in the treated skin section after 28 days of treatment ;complete healing was observed with the absence of inflammatory cells,edema in contrast with the abundance of collagen and fibroblast cells in the dermis layer with sweat glands and hair follicles .
The electron microscope study revealed that the normal shape of the epidermal layer with the dermal layer showed the melanocytes regeneration to its normal shape and size .The molecular assessment was characterized by evaluating the effect of methanolic extract on Tri m4 gene expression .The study found that the methanolic extract at concentration (125μg/ml) did nothave any effect on the expression of this gene; its expression was the same in the absence and presence of P.spina-christi extract by using PCR.