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目的观察昆明山海棠(THH)对急性移植物抗宿主病(aGVHD)小鼠血浆 IFN-γ、IL-4、IL-10浓度的影响,探讨 THH 预防 aGVHD 的可能机制。方法 BALB/c 受鼠清髓性预处理后输注C57BL/6供鼠的骨髓细胞和脾细胞混合液,建立 aGVHD 模型。实验分为4组:对照组(A 组)、环孢素(CsA)预防组(B 组)、THH 预防组(C 组)、THH 和 CsA 联合预防组(D 组)。观察 aGVHD 发生情况和受鼠嵌合体植入情况,ELISA 法检测各组受鼠血浆 IFN-γ、IL-4、IL-10浓度。结果①B、C、D 组受鼠血浆 IFN-γ的浓度明显低于 A 组(P<0.05),D 组下降最明显;IL-4浓度无明显变化;IL-10浓度均高于 A组(P<0.05),D 组上升最明显;②A组受鼠移植后中位生存时间为9 d,B、C、D 组移植后中位生存时间均>30 d,较 A 组明显延长(P<0.05);③A组受鼠均有典型翘毛、弓背、腹泻、体重减轻表现,20 d内全部死于 GVHD;B、C、D 组仅有翘毛、弓背表现,腹泻、体重减轻不明显;B、C 组病理改变皮肤无异常,仅肝脏或小肠有少量淋巴细胞浸润;D 组存活小鼠病理检查未见明显异常,均比 A 组轻;④B、C、D组受鼠+30天骨髓细胞表而 H-2~b 分子百分率分别为(99.18±0.58)%、(97.68±0.59)%、(99.15±0.11)%。结论 THH 可以调节 aGVHD 小鼠细胞因子分泌,可用于 aGVHD 的预防,小剂量 THH 与CsA 联合应用有协同作用,能减少 CsA 的用量。
Objective To observe the effect of Kunming Sanguisorba (THH) on plasma IFN-γ, IL-4 and IL-10 concentrations in mice with acute graft-versus-host disease (aGVHD) and explore the possible mechanisms of THH in preventing aGVHD. Methods BALB/c mice were treated with myeloablative pretreatment and infused with a mixture of bone marrow cells and splenocytes from C57BL/6 donor mice to establish aGVHD model. The experiment was divided into 4 groups: control group (A group), cyclosporine (CsA) prevention group (B group), THH prevention group (C group), THH and CsA combined prevention group (D group). Observe the occurrence of aGVHD and the implantation of chimera-infected mice. Determine the concentrations of plasma IFN-γ, IL-4 and IL-10 in each group by ELISA. Results The levels of plasma IFN-γ in group 1B, C and D were significantly lower than those in group A (P<0.05). The decrease in group D was most obvious; the concentration of IL-4 did not change significantly; the concentration of IL-10 was higher than that of group A ( P<0.05), group D had the most significant increase; group 2A had a median survival time of 9 days after transplantation; group B, C, and D had median survival time >30 days after transplantation, which was significantly longer than group A (P< 0.05); 3A group had typical hair curling, arch back, diarrhea, weight loss, and all died within 20 days of GVHD; group B, C, and D had only hair curls, arches, diarrhea, weight loss Obviously, there was no abnormality in the pathological changes of skin in group B and C. Only a small amount of lymphocytes infiltrated in the liver or small intestine. No obvious abnormality was found in pathological examination of surviving mice in group D. All of them were lighter than those in group A; rats in groups 4B, C, and D were treated with mice+30. The percentage of H-2~b molecules on day bone marrow cells was (99.18±0.58)%, (97.68±0.59)%, and (99.15±0.11)%, respectively. Conclusion THH can regulate the secretion of cytokines in aGVHD mice and can be used for the prevention of aGVHD. The combination of low-dose THH and CsA has a synergistic effect and can reduce the amount of CsA.