靶向CD40的小RNA干扰抑制大鼠异体肢体移植急性排斥反应及细胞凋亡表达的研究

Resistance to acute rejection and apoptosis in limbs alltransplantation of rats by RNA interference of CD40 costimulatory molecule

目的探讨靶向CD40的小RNA干扰对大鼠异体肢体移植急性排斥反应及细胞凋亡的影响。方法以纯系SD大鼠为供体,纯系Wistar大鼠为受体,行同种异体右后肢移植。27只大鼠肢体移植后随机分为3组:A组:注射梭华-Sofast(15μL)-siCD40-2/pSilencer(100μg)载体复合物600μL;B组:在肢体移植后,即注射Sofast(15μL)-pSilencer 4.1-CMV neo(100μg)空载体复合物600μL;C组:在肢体移植后注射生理盐水600μL。观察移植物排斥反应征象及存活情况,并于第7天对产生免疫耐受大鼠进行混合淋巴细胞反应(MLR),同时进行组织学检查。结果与其它实验组相比,A组移植物发生排斥反应的时间及存活时间均显著延长(P<0.01)(>13d),未见排斥反应征象,其它组均于术后近期发生排斥反应;A组大鼠对供体的淋巴细胞呈现低反应性,移植的供体同系大鼠的肢体得以存活。A组移植物细胞凋亡率低于其他组。A组表达CD40的B细胞低于B、C组。结论在术后不应用免疫抑制剂的情况下,靶向CD40的shRNA干扰可以抗大鼠异体肢体移植急性排斥反应。

Objective To study the effect on resistance to acute reaction of composite tissue allotransplantation by using CD40 costimulatory molecule on rat lymphocytes inhibited by shRNA.Methods The inbred SD rats were chosen as donors and inbred Wistar rats as recipients. 27 recipients were divided into 3 groups randomly after allogeneic leg transplantation. Rats in group A injection of Sofast（ 15 μL）-siCD40-2/pSilencer 600 μL the day after transplantation; group B rats received Sofast （15μL）-pSilencer 4.1-CMV neo（100μg） 600μL the day after transplantation; In group C, injection of sodium chloride 600 μL the day after transplantation. The condition of survival and rejection of grafts was observed. Rats which still survived 7 days after operation were received the mixed lymphocyte reaction （MIR）. Results Compared three groups, mean survival time of limb allografts was prolonged obviously in group A （ P 〈 0.01 ）. Rejection of grafts were never observed. Rejection of grafts was all emerged during short term after transplantation in the other groups. Rats in group A had a low reaction to the donors＇ lymphocyte. The expression of apoptosis in group A was lower than that in other groups.Conclusion Without any immnosuppressants after the operation, the method of using RNA interference of CD40 costimulatory molecule can resist acute rejection in limb allotransplantation in rats.