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)  rev-ealed 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 demonstrated  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.