安脑片干预异体移植小鼠aGVHD效应的实验研究

Experimental study of ANNAO tablets on preventing acute graft-versus-host disease in bone marrow transplanted mice

目的观察中药安脑片对异体移植小鼠aGVHD的干预作用。方法 SPF级Balb/c雄性小鼠作为供鼠,以SPF级C57BL/6雌性小鼠为受鼠,建立aGVHD模型,分为安脑片组和空白组,每组10只。观察期间记录小鼠的一般状况及称质量、存活时间,给予临床aGVHD评价,移植后第30天杀鼠取材,制备肝脏、小肠、皮肤等标本,观察其组织学变化,以期评价其干预aGVHD的效果。结果空白组小鼠在移植后第15天全部死亡,死亡率100%,平均生存时间平均为11.9 d;安脑片组死亡3只,存活率70%,平均生存时间平均为27.3 d,差异显著(P=0.000)。安脑片组小鼠平均在移植后第12天精神状态开始恢复,喜动,饮食基本如常,体质量增加,毛发光泽,大便如常。安脑片组皮肤、腹泻、体质量减轻、姿势及活动度的临床评分分别为:0.10±0.32、0.20±0.42、0.12±0.00、0.10±0.10,总分为0.31±0.14,而空白组小鼠上述组织的临床评分分别为2.70±0.48、2.90±0.32、2.91±0.34、2.80±0.42,总评分为3.00±0.00,2组差异明显(P=0.026)。从组织病理学上,安脑片组小鼠肝脏、皮肤、小肠组织较空白组相比,均有不同程度的改善。结论安脑片可有效减轻异体移植小鼠aGVHD的症状和程度,延长小鼠存活时间,改善组织病理。

In order to observe the effect of ANNAO tablets on acute graft versus host disease, we set up an acute graft-versus-host disease mouse model. Hematopoietic stem cells of male Balb/c mice were transplanted to female C57BL/6 mice, which was irradiated by ^60Co X rays with dose 9.0 Gy before transplantation. Then recipient mice were randomly divided into two groups （ANNAO group and blank group）, and respectively administrated with ANNAO soup and 0.9% sodium chloride intragastrically. We observed and recorded clinical symptom, survival time and body weight of mice everyday during observation time. At day 15 posttransplantation, three mice died and seven mice survived in ANNAO group, and their mean survival time was 27.3 days, which was much longer than blank group （their mean survival time was 11.9 days）. The majority of ANNAO group demonstrated good mental state, good appetite, no weigh loss, normal stool, and burnish hairs after day 12. Their clinical scores of skin, weight, diarrhea, posture and activity were respectively 0.10±0.32, 0.20±0.42, 0.12±0.00, and 0.10±0.10, with the whole score of ANNAO group being 0.31±0.14, which was distinctly different as compared with that of blank group （P = 0.000）. 30 days later, survived mice were sacrificed and the samples of liver, intestine and skin were taken to observe histological features. We found there some improvements in liver, skin, and intestine tissue of ANNAO group as compared with blank control group. In conclusion, ANNAO tablets can relieve symptoms of acute GVHD mice effectively and prolong their survival time, and improve histopathological manifestations.