Effects Of YI FA Oral Liquid Combined With Glucocorticoid On Expression Of Glucocorticoid Receptor MRNA Level Of Severe Alopecia Areata Patients

ObjectiveTo analyze the pharmacological mechanism of YIFA oral liquid combined whth glucocorticoid(GC) through the observation of its clinical treatment effect on severe alopecia areata patients.To investigate the pathogenesis of severe alopecia areata by detecting the level of glucocorticoid receptors(GR)mRNA in peripheral blood mononuclear cells(PBMCs) from patients with severe alopecia areata.Analyze both the relationship between the expression level of GR and pathogenetic condition,and between the the expression level of GR and clinical effect.Further more to explore the relationship between GR and Chinese medicine combined with GC.MethodsAccording to the third album《guide line of Chinese herbal medicine clinical research》,32 cases clinical diagnosed as severe alopecia areata and deficiency of liver and kidney as their TCM syndrome were selected.And 20 cases as normal control group.32 cases of severe alopecia areata patients were randomly divided into integrated Chinese and western medicine group(17 cases) and western medicine group(15 cases).Integrated Chinese and western medicine group was given YIFA oral liquid,20ml each time and three times a day;prednisone 0.25mg/kg,every 8 a.m.;famotidine 20mg each time,twice a day.Western medicine group was given prednisone and famotidine only,usage and dosage with integrated Chinese and western medicine group.Evaluting the treatment effect of two groups after six months treatment.The therapeutic indexes including:the root of hair loss during treatment,the new hair area during treatment,the color,density,thickness of new hair and the whole situation of the patients.The criteria for treatment effect also reffered to the third album《guide line of Chinese herbal medicine clinical research》. The patients of two groups were observed blood routine,urine routine,soil routine,cardiac function(EKG),liver function(ALT,AST),renal function(BUN, Cr) and adverse reactions before and after treatment.Before treament,serum cortisol levels was measured by Chemiluminescence from 32 cases of severe alopeeia areata patients and 20 normal controls.GR was measured by reverse transcription-real time fluorescence quantitative polymerase chain reaction (RT-FQRealTime-PCR) from severe alopecia areata patients(before and after treatment) and normal controls.The data were inputted into a computer and were analyzed by statistical package for the social science(SPSS) 13.0.Results(A).Comparison of serum cortisol level:Serum cortisol level from severe alopecia areata patients had no significant difference compared to the normal controls(P＞0.05);Serum cortisol level between integrated Chinese and western medicine group and western medicine group alos had no significant difference(P＞0.05).(B).Comparison of the GRmRNA expression level1.The normal control group compared with severe alopecia areata patients group:GRmRNA was detected in PBMCs from all severe alopecia areata patients and the normal controls.The expression of GRmRNA before and after treatment of severe alopecia areata patients was significantly lower than that of the normal controls(P＜0.05);And the expression of GRmRNA after treatment of alopecia areata patients was significantly lower than that before treatment(P＜0.01).2.Integrated Chinese and western medicine group compared with western medicine group1:(1) Comparison within the groupsGRmRNA expression level after treatment of two groups was significantly lower than that before treatment by two pairs sample comparison.And western medicine group dropped greater than integrated Chinese and western medicine group.Integrated Chinese and western medicine group(P＜0.05),western medicine group(P＜0.01).(2) Comparison between the groups Before treatment,GRmRNA expression level of integrated Chinese and western medicine group had no significant difference compared to western medicine group(P＞0.05).After treatment,GRmRNA expression of integrated Chinese and western medicine group was significantly higher than that of western medicine group(P＜0.05).(C).Comparison of treatment effectAnalyzed the testers with PP analysis who had good compliance and complete treatmeat test.Analyzed all the testers with ITT analysis.PP analysis:the cure rate of integrated Chinese and western medicine group was 58.82%,while the total effective rate was 94.1%.Tthe curative rate of western medicine group was 40%,while the total effective rate was 80%.The cure rate and the total effective rate of integrated Chinese and western medicine group was significantly higher than that of western medicine group(P＜0.05).The results of ITT analysis was the same to the results of PP analysis.The adverse reactions between integrated Chinese and western medicine group and western medicine group had no significant difference(P＞0.01).(D).The patients were divided into alopecia universalis group,alopecia totalis group and patiching group according to the bald area.As the result, the GRmRNA expression lever of patiching group was significantly higher than that of alopecia universalis group and alopecia capitis totalis group.(E).According to the treatment effect,the patients were divided into recovery group,excellence group,effective group and ineffective group.GRmRNA expression level before treatment of four groups had no statistically difference(P＞0.05).But after treatment,GRmRNA expression level of four groups had significant difference(P＜0.05).Comparison within the groups: Before and after treatment,the differential value of GRmRNA expression level in recovery group,excellence group and effective group had significant difference(P＜0.05);While the differential value of GRmRNA expression level in ineffective group had no significant difference(P＞0.05).ConclusionWe can draw a conclusion as the following:1.We firstly reported that serum cortisol level from severe alopecia areata patients had no significant difference compared to the normal controls. However,the expression of GRmRNA of severe alopecia areata patients was significantly lower than that of the normal controls.It suggested that the disorder of GC-GR exist in alopecia areata patients.And the decrease of GRmRNA in peripheral blood mononuclear cells may be invoved in the pathogenesis of severe alopecia areata2.We firstly reported that the expression of GRmRNA in peripheral blood mononuclear cells of severe alopecia areata patients and deficiency of liver and kidney as their TCM syndrome was significantly lower than that of the normal controls(P＜0.05).This maybe the pathological change on receptor and gene level in severe alopecia areata patients and deficiency of liver and kidney as their TCM syndrome.It depends on further research that whether GRmRNA expression level can be the mordern pathology quantitative basis or not.3.We firstly reported that GRmRNA expression of integrated Chinese and western medicine group after treatment was significantly higher than that of western medicine group.The treatment effect of integrated Chinese and western medicine group was better that that of western medicine group.It indicated that YIFA oral liquid combined with glucocorticoid can raise GRmRNA expression of severe alopecia areata patients,or decrease the down-regulation of glucocorticoid towards its receptor,which can increase the sensibility of glucocorticoid and guarantee the binding level between GC and GR.It pointed out that the treatment effect of YIFA oral liquid combined with glucocorticoid was related to its regulation towards GR.This maybe one of the action mechanism of YIFA oral liquid to alopecia areata.4.We firstly reported that the ligher pathogenetic condition was,the higher GRmRNA level expressed.On the contrary,the heavier pathogenetic condition was,the lower GRmRNA level expressed.For this reason,examining GRmRNA expression level is useful in judging the prognosis of severe alopecia areata.5.We firstly reported that the better the treatment effect was the higher GRmRNA expressed in PBMCs of severe alopecia areata.On the contrary,the worse the treatment effect was the lower GRmRNA expressed.It indicated it would increases adverse reactions if we blindly increase the dosage of GC to the patients who had bad response to GC.Disadvantage would exceed disadvantage. It is supposed that examining GRmRNA expression level of severe alopecia areata before treatment maybe contribute to the effect prediction which the patients towards GC.Moreover it maybe one of the reference index to direct treatment and prognosis.