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Title:: Effect of immune tolerance induced by donor rat splenocytes or bone marrow cells combined with asarinin on graft survival during rat heart transplantation

作者:: 张丽丽¹; 于波¹; 杨宝峰²; 哈尔滨医科大学： 1.附属第二医院心内科，2.药理学教研室，黑龙江哈尔滨 150086

Author(s):: ZHANG Li-li¹; YU Bo¹; YANG Bao-feng²; 1.Department of Cardiology, Second Affiliated Hospital, 2.Department of Pharmacology, Harbin Medical University, Harbin 150086, Heilongjiang, China

关键词:: 细辛脂素; 同种心脏移植; 免疫耐受; 大鼠

Keywords:: asarinin; cardiac allograft; immune tolerance; rat

分类号:: R654.2

DOI:: -

文献标识码:: A

摘要:: 目的: 探讨心脏移植前1周，输注供鼠脾/骨髓细胞于接受鼠胸腺内，同时灌服细辛脂素诱导免疫耐受对移植物的影响。方法: 提取中药细辛中的细辛脂素成分。以健康雄性Wistar大鼠为供者和SD大鼠为受者，建立大鼠腹部异位心脏移植模型。将接受心脏移植的80只SD大鼠分成5组，每组16只。阳性对照组：施行腹部异位心脏移植，不给任何处置；环孢霉素A（CsA）组：术前7 d开始灌服CsA；细辛脂素组：术前7 d开始灌服细辛脂素25mg/(kg·d)；脾细胞诱导组：术前7d开始输注供者脾细胞于受者胸腺内，术前7d开始灌服细辛脂素25mg/(kg·d)；骨髓细胞诱导组：术前7 d开始输注供者骨髓细胞于受体胸腺内，术前7 d开始灌服细辛脂素25 mg/(kg·d)。于术后7 d，每组各处死8只，另外8只用于观察移植物生存的天数。用MTT比色法检测供受体鼠混合淋巴细胞反应；应用ELISA法检测受体鼠血清中IL-2，IFN-γ，IL-4和IL-10的水平。结果: CsA组和免疫诱导组移植心的存活时间较阳性对照组显著延长（P<0.01）。供受体鼠的混合淋巴细胞反应免疫诱导组较阳性对照组差异显著（P<0.01）。免疫诱导组血清IL-4和IL-10的含量较CsA组显著升高（P<0.01）。CsA组和免疫诱导组血清IL-2和IFN-γ的含量较阳性对照组显著降低（P <0.01）。结论: 细辛脂素联合供者的脾细胞和骨髓细胞可成功地诱导受体鼠对心脏移植物的免疫耐受。输注供体鼠的脾细胞和骨髓细胞可起到相似的诱导免疫耐受的作用。

Abstract:: AIM: To explore the effect of immune tolerance in rat cardiac allograft induced by donor splenocytes or bone marrow combined with asarinin 7 days before surgery. METHODS: Asarinin was extracted from herba asari. Eighty Sprague Dawley rats (recipients) were randomly divided into five groups and were treated 7 days preoperatively: control group (no treatment); cyclosporin group, asarinin group, splenocyte-inducing group (splenocytes and asarinin), and bone marrow-inducing group (bone marrow and asarinin). Eight rats in each group were sacrificed and the other eight rats were left in order to observe survival time. Mixed lymphocyte reaction (MLR) between donor and recipient was examined with MTT and levels of serum IL-2, IFN-γ, IL-4 and IL-10 were detected with ELISA 7 days postoperatively (n=8). RESULTS: Allograft survival time was prolonged in cyclosporin group, asarinin group, splenocyte-inducing group and bone marrow-inducing group (P<0.01). Lymphocyte proliferation was alleviated in asarinin group, splenocyte-inducing group and bone marrow-inducing group compared with control group (P<0.01). Level of IL-4 and IL-10 significantly increased in splenocyte-inducing group and bone marrow-inducing group compared with the cyclosporin group (P<0.01). CONCLUSION: Immune tolerance, which is specific to the donor, can be induced by donor splenocytes and bone marrow cells. Immune tolerance induced by donor rat splenocytes or bone marrow cells combined with asarinin prolongs survival time of allografts and protects the transplanted organ.

参考文献/References

［1］张丽丽，关振中，张竞逵，等. 细辛脂素体外免疫抑制作用的实验研究［J］. 中华心血管病杂志， 2003， 31（6）：444-447.

［2］杨云，冯卫生. 中药化学成分提取分离手册［M］. 北京：中国中医药出版社， 1998:189-192.

［3］ van Denderen B, Peche H, Gagne K, et al. Identification of immunodominant fonor MHC peptides following rejection and donor strain transfusion-induced tolerance of heart allografts in adult rats［J］. Eur J Immunol, 2001, 31(5):1333-1339.

［4］ Mottram PL, Han WR, Purcell LJ, et al. Increased expression of IL-4 and IL-10 and decreased expression of IL-2 and interfer-on-gamma in long-survivinng mouse heart allografts after brief CD+4 monoclonal antibody therapy［J］. Transplantation, 1995, 59(4):559-565.

［5］ Goss JA, Nakafusa Y, Flye MW. MHC class II presenting cells are necessary for the induction of intrathymic tolerance［J］. Ann Surg, 1993, 217(5):492-501.

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细辛脂素+供鼠脾/骨髓细胞诱导的免疫耐受对大鼠心脏移植存活的影响

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

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