催乳素对刀豆蛋白A诱导活化T淋巴细胞免疫应答的影响

Effection of Prolactin on Immunoresponsiveness of Activated T Lymphocytes Induced by Concanavalin A

目的研究催乳素(PRL)对刀豆蛋白A(ConA)诱导活化T淋巴细胞的影响,探讨PRL在T淋巴细胞活化中的作用。方法用25μg/L的PRL与500μg/L的溴隐亭(Brc)单独或联合刺激5 mg/LConA诱导活化的CD4+T淋巴细胞系JurkatE6-1细胞,实验设置空白对照组、ConA组、ConA和PRL联合刺激组(PRL组)、ConA和Brc联合刺激组(Brc组)、ConA和PRL及Brc联合刺激组(PRL-Brc组),药物刺激48 h后,Trizol法提取Jurkat E6-1细胞总RNA,反转录后,利用PCR技术检测TNF受体相关因子6(TRAF6)基因的表达,利用反转录酶PCR技术检测TNF超家族成员4(TNFSF4)和相对分子质量为37 000的杀伤特异性分泌蛋白(KSP37)基因的表达。结果与空白对照组、ConA组比较,PRL组和Brc组活化T淋巴细胞TRAF6、TNFSF4、KSP37基因的表达显著降低(Pa<0.05);与PRL组和Brc组比较,PRL-Brc组可明显逆转PRL对活化T淋巴细胞TRAF6、TNFSF4、KSP37基因表达的抑制(Pa<0.05);与空白对照组比较,PRL-Brc组可明显抑制活化T淋巴细胞TRAF6基因的表达(Pa<0.01),KSP37及TNFSF4的表达高于空白对照组但是无统计学意义(Pa>0.05);与ConA组比较,PRL-Brc组可明显抑制活化T淋巴细胞TRAF6、TNFSF4、KSP37基因的表达(Pa<0.01)。结论生理浓度的PRL可以通过抑制活化T淋巴细胞中TRAF6、TNFSF4、KSP37的表达参与T淋巴细胞应答;Brc可通过拮抗PRL对活化T淋巴细胞的TRAF6、TNFSF4、KSP37的抑制用于治疗PRL引起的疾病。

Objective To study the effect of prolactin（PRL） on the activation of T lymphocytes stmiulated by concanavalin A（ ConA）, and to explore the action of PRL in the activation of T lymphocytes. Methods After CD4 ＋ T cell line JurkatE6 - 1 cells were respectively st- miulated by 5 mg,/L ConA, 25μg/L PRL and 500 μg/L bromocriptine （Brc）. The blank control group, the ConA group, the PRL and ConA group（ PRL group）, the Bre and ConA group（ Brc group）, the PRL and Brc group（PRL - Brc group） were set in the experiment. The total RNA was extracted by Trizol after 48 hours and was reversed transcription immediately. The expression of tumor necrosis factor receptor asso- ciated factor 6 （TRAF6） mRNA of T lymphocytes was checked by PCR. The expressions of tumor necrosis factor （ligand） super family 4 （ TNFSF4 ） and Killer specific secretory protein of 37 000 （ KSP37 ） mRNA of T lymphocytes were detected by real - time polymerase chain reaction. Results The PRL group and the Brc group could inhibit the expressions of TRAF6, TNFSF4, and KSP37 mRNA of the activated T lymphocyte compared with the blank control group and the ConA group（Pa 〈 0.05） ; In PRL- Brc group, Brc could reverse significantly the inhibitions of TRAF6, TNFSF4, and KSP37 mRNA caused by PRL in the activated T lymphocyte compared with the PRL group（P 〈 0.01 ）. The PRL - Brc group could inhibit the expressions of TRAF6 mRNA of the activated T lymphocyte compared with the blank control group, the expressions of KSP37and TNFSF4 in the PRL -Brc group was higher compared with the blank control group,but the mean differences was not significant （ Pa 〉 0.05 ）. The PRL - Brc group could inhibit significantly the expressions of TRAF6, TNFSF4, and KSP37 mRNA of the activa- ted T lymphocyte compared with the ConA group（ P~ 〈 0.01 ）. Conclusions PRL at physiological concertration might inhibit the expressions of TRAF6, TNFSF4, and KSP37 mRNA to participate in T lymphoeytes response ; Brc can rivalry the inhibitions of TRAF6, KSP37 and TNFSF4 induced by PRL and treat some diseases caused by PRL.