黄连素合用环孢素A对小鼠皮肤移植排斥反应的影响

目的观察黄连素合用环孢素A抗同种异体小鼠皮肤移植排斥反应的影响,探讨其作用机制。方法以BALB/c小鼠为供体,以C57BL/6小鼠为受体,采用背-背皮肤移植手术法制备模型。将受体小鼠按随机数字表法分为模型组、黄连素组、环孢素A组、联合组,每组20只;另取20只健康C57BL/6小鼠作为假手术组。造模后黄连素组小鼠腹腔注射黄连素100 mg/kg,环孢素A组小鼠腹腔注射环孢素A注射液10 mg/kg,联合组腹腔注射黄连素100 mg/kg及环孢素A注射液5 mg/kg,假手术组和模型组小鼠腹腔注射等体积0.5%羧甲基纤维素钠。1次/d,连续10 d。记录受体小鼠移植皮片的存活时间;采用ELISA法检测血浆Th1类细胞因子[IL-2、γ-干扰素(interferon-γ,IFN-γ)]和Th2类细胞因子(IL-4、IL-10)水平;采用流式细胞术检测脾脏T淋巴细胞亚群CD4+CD25+比例。结果与模型组比较,各给药组受体小鼠移植皮片的存活时间延长(P<0.01);联合组移植皮片存活时间较黄连素组或环孢素A组延长(P<0.01)。与模型组比较,联合组血浆IL-2[(11.55±3.14)pg/ml比(19.85±2.42)pg/ml]、IFN-γ[(26.41±6.20)pg/ml比(57.23±10.15)pg/ml]水平降低,IL-4[(192.45±70.12)pg/ml比(61.09±21.61)pg/ml]、IL-10[(106.79±27.83)pg/ml比(40.08±11.23)pg/ml]水平升高(P<0.05),脾脏CD4+CD25+调节性T细胞比例[(7.65±2.42)%比(3.69±0.83)%]升高(P<0.01)。结论黄连素合用环孢素A通过促使Thl向Th2细胞的偏移、诱导免疫耐受、刺激CD4+CD25+调节性T细胞的表达等途径,抑制同种异体小鼠的皮肤移植排斥反应。

The anti-rejection effect of berberine combined with ciclosporin A on mice with xenogenic skin

Objective To investigate the anti-rejection effect of the berberine combined with ciclosporin A(CsA)against mice with xenogenic skin transplantation and explore the mechanisms.Methods The skin transplantation model was established by an operation,tergal skin flaps from BALB/c donor mice were transplanted to the back of C57BL/6 recipient mice.Then recipient mice were randomly divided into 5 groups(n=20),namely sham(0.5%CMC-Na),model(0.5%CMC-Na),berberine(100 mg/kg),CsA(10 mg/kg),berberine combined with CsA(berberine 100 mg/kg and CsA 5 mg/kg).All recipient mice were intraperitoneally administered once a day for 10 days.The skin graft survival time of all mice were noted everyday.At the end of the experiment,the levels of Th1 cytokines(IL-2 and IFN-g)and Th2 cytokines(IL-4 and IL-10)in plasma were detected by ELISA,and quantity of CD4+CD25+T cells were analysed by flow cytometry.Results Compared with model group,the survival days of skin graft of those in groups treated with drugs were all significantly prolonged(P<0.01).Berberine combined with CsA group showed a longer survival days than berberine or CsA group(P<0.01).Compared to the model group,the content of IL-2(11.55±3.14 pg/ml vs.19.85±2.42 pg/ml),IFN-g(26.41±6.20 pg/ml vs.57.23±10.15 pg/ml)of berberine combined with ciclosporin A group significantly decreased(P<0.05).The content of IL-4(192.45±70.12 pg/ml vs.61.09±21.61 pg/ml),IL-10(106.79±27.83 pg/ml vs.40.08±11.23 pg/ml)of berberine combined with ciclosporin A group significantly increased(P<0.05).The amount of CD4+CD25+T cells(7.65%±2.42%vs.3.69%±0.83%)in berberine combined with ciclosporin A group significantly increased(P<0.01).Conclusions The immunosuppressive effect of berberine combined with CsA maybe related with induction of immune tolerance,balance of Th1/Th2 cytokines and increasement of CD4+CD25+T cells expression.