Study On The Inhibitory Mechanism Of A Genistein Phenyl Piperazine Derivative On Oral Ulcers And Pigmentation

The present study was to investigate the inhibitory effect of a genistein phenyl piperazine derivative(GPD)on the oral ulcers and skin pigmentation of guinea pigs,and to elucidate the inhibition mechanism of the GPD on oral ulcers and skin pigmentation.By establishing a guinea pig oral ulcer model induced by 90% phenol,we studied the inhibitory effect and mechanism of GPD on oral ulcers.Watermelon frost was used as the positive control group.After the treatment of GPD,the pathological morphology of the oral ulcer tissues of guinea pigs were observed by hematoxylin eosin staining(HE).The levels of TNF-α,NF-κB,IL-10,SOD,ROS,MDA,AMPK,m TOR and NO were quantitatively analyzed by enzyme linked immunosorbent assay(ELISA).The positive expressions of COX-2 were analyzed by immunohistochemistry(IHC).Western blot was used to analyse the expression level of AMPK in total protein of cells.The results showed that the ulcer degree of GPD high dose group was significantly reduced compared with the model group(P < 0.01).The results of HE staining showed that the ulcer was restored to normal.The promotion rates of high dose GPD for IL-10,AMPK,SOD were 132%,84%,47%,respectively.And the inhibition rates for TNF-α,NF-κB,NO,m TOR,ROS,MDA were 33%,8%,20%,62%,36%and 40%,respectively.GPD could significantly increase the expression level of AMPK in the total cell protein(P < 0.01),and the high dose GPD had the best effect.And the positive expression of COX-2 in the high dose group of GPD decreased by 42%,and basically recovered to the normal.The research showed that high dose GPD has significant effect on the treatment of 90% phenol induced oral ulcer in guinea pigs by inhibiting inflammatory reaction,improving oxidative stress and regulating energy metabolism.In addition,by establishing a skin pigmentation model of guinea pig induced by UVB irradiation,we studied the inhibitory effect and mechanism of GPD on pigmentation.Arbutin was used as the positive control group.After GPD treatment,the changes in the contents of TYR,SOD,MDA,m TOR and AMPK in skin tissues were detected by ELISA.The morphology of skin tissues were observed by HE staining,and Masson-Fontana staining was used to observe the distribution of melanin in the skin epidermis of guinea pigs.The positive staining indexs of COX-2 and AMPK in guinea pig skin epidermis were detected by IHC.The results showed that compared with the model group,the damage of the skin tissues of the GPD medium dose group were improved,the relative density of the melanin was decreased.The promotion rates of the medium dose GPD for SOD and AMPK were 52% and 41%respectively,and the inhibition rates for TYR,MDA,m TOR were 22%,39%,32%,respectively.After GPD treatment,the positive staining index of AMPK was significantly increased(P < 0.01),and the effect of middle dose group was the best and the rate of promotion was 49%.Meanwhile,the positive staining index of COX-2 was significantly decreased(P<0.01),and the inhibition rate was 46%.The study showed that medium dose GPD could inhibit the skin pigmentation of guinea pig induced by UVB,by inhibiting the key enzyme of melanin-TYR,inhibiting the inflammatory reaction,improving the oxidative stress reaction and regulating energy metabolism.