The Study On The Apoptosis Of T Lymphocyte In Mouse Model Of Allergic Contact Dermatitis Induced By Tripterygium Glucoside

Objective To determine the effect of Tripterygium Glucoside to T cell subsetand apoptosis of T lymphocyte in mouse model of Allergic Contact Dermatitis(ACD) cultured in Vitro. and to identify the mechanism of lymphocyteapoptosis, then understand the part of action of the treatment to ACD byTripterygium Glucoside, and to provid some foundation about more rationaluse of the Tripterygium Glucoside to treat ACD.Methods The mouse model of ACD were induced by dinitrochlorobenzene(DNCB), and separate the lymphocyte from spleen, cultured in vitro for 48houses. The drug level in the control groups, Tripterygium Glucoside groupswere 0, 1, 2 and 4μmol/L.1 The items as below was performed using flow cytometer (FCM).1.1 The CD8, CD4 and CD3 subsets in the control groups, TripterygiumGlucoside groups;1.2 The T lymphocytes were treated by PI, and detect the ratio of G0/G1, S, G2/M and hypodiploid in cell cycle.1.3 Using the Annexin V-FITC Kit to detected the T lymphocytes apoptosisby FCM;1.4 Dyed the lymphocytes in vitro by PE anti-mouse CD95 (Fas/APO-1),APC-Cy7 anti-mouse CD4 fluorescence, and the expression of Fas was carriedout by FCM;1.5 Using the CaspGLOWTM Fluorescein Active Caspase-8 Staining Kit toexamine the concentration of Caspase-8 in cytoplasm.2 The Bcl-2 and Bax mRNA expression in the lymphocytes in vitro treated byTripterygium Glucoside were detected by RT-PCR. PCR products were detected by gel electrophoresis.Results1 The ratio of Th express on the ACD mouse models T lymphocyte in vitro inthe three group of Tripterygium Glucoside much lower than those in thecontrol group (P＜0.01), and the Th express in the 4.0 mol/L groupssignificantly lower than that in the 1 mol/L groups (P＜0.01). the Thexpress in the 4μmol/L groups lower than that in the 2μmol/L groups (P＜0.05) and the 2μmol/L groups lower than that in the 1μmol/L groups(P＜0.05).The ratio of Tc express on the ACD mouse models T lymphocyte in vitro inthe three group of Tripterygium Glucoside lower than those in the controlgroup. the Tc express in the 4.0 mol/L groups significantly lower than thatin the control groups (P＜0.01). the Tc express in the 2.0 mol/L groups and1.0 mol/L groups lower than that in the control groups (P＜0.05).The ratio of CD3+ express on the ACD mouse models T lymphocyte in vitroin the4.0 mol/L group of Tripterygium Glucoside significantly lower thanthat in the control groups (P＜0.01).2 The ratio of G1 phase in all the Tripterygium Glucoside treatment group atdifferent concentration compared with the control group had statisticalsignificance, and the ratio of G1-phase in the 4μmol/L group was muchhigher than that in the control group and 2μmol/L group (P＜0.01). theratio G1 in the2μmol/L gruop had statistic compared with those in thehigh concentration group and control group (P＜0.05).The ratio of S-phase in all the Tripterygium Glucoside treatment group atdifferent concentration compared with the control group had statistical significance (P＜0.01), and the ratio of S-phase in the 4μmol/L group wasmuch lower than that in the other concentration group (P＜0.01).The ratio of G2-phase in the Tripterygium Glucoside treatment group of 4μmol/L compared with other groups had statistical significance, and theratio of G2-phase in the 4μmol/L group was much lower than that in the1μmol/L group and control group (P＜0.01), the ratio of G2-phase in the4μmol/L group was lower than that in the 2μmol/L group (P＜0.05).The ratio hypo-diploid in the Tripterygium Glucoside high concentrationgruop had significante statistic compared with those in the control gruop (P＜0.01), and the ratio of hypo-diploid in the 4μmol/L TripterygiumGlucoside group higher than that in the 1μmol/L group (P＜0.05).3 The ratio of apoptosis cells in the Tripterygium Glucoside 4μmol/L and 1μmol/L gruop had significante statistic compared with those in the controlgruop (P＜0.01), and the ratio in the 4μmol/L and 2μmol/L TripterygiumGlucoside group higher than that in the 1μmol/L group (P＜0.05).4 The ratio of Fas express in all the Tripterygium Glucoside treatment groupshigher than that in the control group, and that in the 4 and 2μmol/Lgroups much higher than that in the control gruop (P＜0.01). The ratio ofFas express in the 4μmol/L groups higher than that in the 2 and 1μmol/Lgroup (P＜0.05).5 The concentration of Caspase-8 in the Tripterygium Glucoside treatmentgroup at 1.0μmol/L groups lower than that in the control group and 2mol/L group (P＜0.05). The concentration of Caspase-8 in the 1.0μmol/Lgroups much lower than that in the 4.0μmol/L group (P＜0.01).6 The express of Bax mRNA in the Tripterygium Glucoside treatment groupsof 4.0μmol/L much higher than that in the control group (P＜0.01). the difference between the 4.0μmol/L and 1.0μmol/L groups has statisticallysignificant (P＜0.05).Conclusion1 Tripterygium Glucoside may selectivity reduce the ratio of T cellsubset, including CD3+, Th, Tc.2 Tripterygium Glucoside could induce apoptosis in the ACD mouse modelsT lymphocyte in vitro, and it was one of possible mechanism to depress theactivated T lymphocyte.3 The mechanism of Tripterygium Glucoside to induce apoptosis was possiblerelatede to some factor as below, Tripterygium Glucoside could increase theFas express on the T lymphocyte in vitro, and could adjustment theCaspase-8 concentration, and could elevate the Bax mRNA express.