Clinical Study Of Jilishouwu Recipe On The Treatment Of Vitiligo And Mechanism Research On GSTT For Melanocytes

Objectives:1. To observe the clinical effects and action mechanism research of Jilishouwu Recipein the treatment of vitiligo.2. To reveal the action mechanism of the effective monomer GSTT in principal drugtribulus terrestris in Jilishouwu Recipe on oxidative stress damaged melanocytes, and toestablish the theoretical basis for new clinical drug development and utilization.Methods:1. To collect100cases of clinically diagnosed vitiligo patients, clinical staging,typing and typing of TCM symptoms, and randomly divided them into two groups. Let thetreatment group take the decoction of Jilishouwu Recipe, and externally use the topicalmometasone furoate ointments; let the control group oral intake the Vitiligo pills, andexternally use the topical mometasone furoate ointments, to determine the curative effectafter the treatment course of3months. Take the venous blood from the treatment group,the control group before and after treatment and control group, to detect the CAT, GSH-PX,SOD, MDA levels, randomly take the samples of50healthy subjects samples from thehospital medical center during the same period, to act as the healthy control group.2. Take the normal melanocytes as research object to subculture. Add250μmol/LH2O2into melanocytes to culture for12hours, to serve as the melanocytes oxidative stressmodel. The experiment is divided into negative control group, drug treatment group, andpositive control group. Drug treatment group is divided into three drug concentrations of50mg/L,100mg/L, and150mg/L. Using the methods of MTT and Annexin-V/PI doublestaining, to observe the melanocytes vitality and apoptosis under the impact of GSTTwith different concentrations. Results:1. Clinical study results:(1) Effects comparison between the treatment group and the control groupThe recovery rates of the treating group and the control group were44%and12%, thecomparison result between two groups was statistically significant (χ2=6.349, P<0.05). theeffective rates were76%and40%respectively, the difference between two groups wasstatistically significant (χ2=6.650, P <0.05).(2) Effects comparison between the treatment group and the control group withdifferent syndrome typing:In the treatment group, the recovery rate of wind-heat syndrome was significantlyhigher than that of the control group, so there was statistically significant difference(χ2=4.773, P<0.05). The effective rate of gas stagnation and blood stasis in treatment groupwas significantly higher than that of the control group, so there was statistically significantdifference (χ2=4.000，P<0.05).(3) The serum antioxidase level comparison between two groups of patients beforetreatment and the health control groupCompared with the healthy control group, the CAT, GSH-PX, SOD levels of serum inthe treatment group and the control group of vitiligo patients decreased significantly, andMDA increased significantly, there was statistical significance (P<0.001).(4) The serum antioxidase level comparison between the treatment group before andafter treatment and the health control groupAfter the treatment, the CAT, GSH-PX and SOD levels of serum in the treatmentgroup increased significantly, the difference between them had statistical significance(P<0.001). Compared with the healthy control group, the GSH-PX and MDA levels ofserum in the treatment group after the treatment had no statistical difference (P>0.05). TheSOD level of serum was higher than that of the healthy control group, there was statisticalsignificance (P<0.05).(5) The serum antioxidase level comparison for the control group before and aftertreatmentAfter the treatment, the CAT, GSH-PX and SOD levels of serum of patientssignificantly increased, and MDA level of serum significantly decreased, the differencebefore and after the treatment had statistical significance (P<0.001). Compared with thehealthy control group, the MDA level in the treatment group had no statistical difference (P>0.05), the SOD level is higher than that of the healthy control group, there wasstatistical difference (P>0.05). After the treatment, the CAT and GSH-PX levels are stilllower than those of the healthy control group, there was statistical difference (P<0.001).(6) The serum antioxidase level comparison for the control group and control groupAfter the treatment, the CAT and GSH-PX levels of serum of patients is much higherthan that of the control group, the difference had statistical significance (P<0.001). Thecomparison of SOD and MDA had no statistical significance (P>0.05).2. The results of the experiment(1) Effects of GSTT with different concentrations for the activity of oxidativestress-induced damaged melanocytesThe comparison of medium concentration group and high concentration group in drugtreatment group with positive control group showed no statistical significance (P>0.05),the comparison of cell viability for medium concentration group and high concentrationgroup with negative control treated group, the difference was statistically significant(P<0.05), the comparison of cell viability for the medium concentration group and highconcentration group was statistical significance (P<0.05), the comparison of cell viabilityfor low concentration group with the negative control group had no statistical significance(P>0.05).(2) Effects of GSTT with different concentrations for the apoptosis of oxidativestress-induced damaged melanocytesThe comparison between the low concentration group and control group had nostatistical significance, while the melanocyte apoptosis rate comparison between the highconcentration drug treatment group and the control group had significant difference(P<0.05), the melanocyte apoptosis rate comparison between the high concentrations drugtreatment group and the medium concentration drug treated melanocytes apoptosis rate hadstatistically significant (P<0.05).Conclusions:1. The effect of Jilishouwu recipe in the treatment of vitiligo is exact, without adversereaction, antioxidant effect may be the effective mechanism of Jilishouwu Recipe in thetreatment of vitiligo.2. GSTT has anti-oxidation effect, can increase the oxidative stress-induced melanincell viability, decrease the apoptosis, the antioxidation effects play a protective role onmelanocytes..