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通络方剂对实验性糖尿病大鼠胸腺CD4+CD25+调节性T细胞的影响
张兰予,冯晓云,邹俊杰,刘志民*
0
(ZHANG Lan-yu, FENG Xiao-yun, ZOU Jun-jie, LIU Zhi-min*)
摘要:
\[摘要\]目的探讨通络方剂对实验性糖尿病大鼠胸腺CD4+CD25+调节性T细胞(Treg)的影响。方法112只SD大鼠随机分为3组:正常对照组、糖尿病模型组、通络方剂干预组。后两组一次性腹腔注射链脲佐菌素60 mg/kg制备1型糖尿病大鼠模型。通络方剂干预组再分为2组:分别于糖尿病造模成功后当天及普通饲养12周后0.4 g·kg-1·d-1)灌胃给药,即预防性给药组(TL-1)和治疗性给药组(TL-2)。灌胃时间均为12周。实验周期6个月,不同时间点(对照组和模型组:4、8、12、16、20、24周;通络方剂组:12和24周)处死大鼠,流式细胞仪检测胸腺CD4+CD25+Treg数量及其占CD4+T细胞比例,H-E染色观察胸腺形态,免疫组化检测Foxp3蛋白表达。结果随糖尿病病程发展,与正常对照组相比,糖尿病大鼠胸腺逐渐萎缩,Treg占CD4+T细胞比例逐渐减少,Foxp3表达减少。两组通络方剂组较糖尿病组Treg比例升高(P<0.01),胸腺指数升高(P<0.05),Foxp3有表达。结论通络方剂可显著提高Treg比例,提高胸腺指数,提高Foxp3表达,可能对实验性糖尿病大鼠的胸腺Treg有保护作用。
关键词:  CD4+CD25+调节性T淋巴细胞  实验性糖尿病  通络方剂  胸腺  Foxp3
DOI:10.3724/SP.J.1008.2011.0167
投稿时间:2010-08-25修订日期:2011-01-07
基金项目:国家重点基础研究发展计划(“973”计划)项目(2005CB523304).
Effect of Tongluo recipe on CD4+CD25+ regulatory T cells in the thymus of experimental diabetic rats
第二军医大学长征医院内分泌科,上海 200003
(Department of Endocrinology, Changzheng Hospital,Second Military Medical University,Shanghai 200003, China)
Abstract:
\[Abstract\]ObjectiveTo observe the effect of Tongluo recipe(TLR) on CD4+CD25+ regulatory T cell in the thymus of experimental diabetic rats. MethodsTotally 112 SD rats were randomly divided into three groups: control group, diabetes mellitus(DM) group, and TLR treatment group. Rat diabetic model was induced by intraperitoneal injection of streptozotocin (60 mg/kg). The TLR treatment group was further divided into two subgroups: prevention group was given intragastric TLR (0.4 g·kg-1·d-1) for 12 weeks starting from the day of streptozotocin injection (TL-1 group), and the treatment group was given intragastric TLR (0.4 g·kg-1·d-1) for 12 weeks starting from 12 weeks after streptozotocin injection (TL-2). The animals were sacrificed at different time points (The control and DM group: 4, 8, 12, 16, 20, and 24 weeks; TLR group: 12 and 24 weeks after administration of TLR). Flow cytometry, H-E staining, and immunochemistry were performed to investigate the change of CD4+CD25+ regulatory T cell ratio in CD4+ T cells, the thymus morphology, and the expression of Foxp3 protein in the thymus.ResultsCompared with the normal control group, diabetic group had a gradually shrunk thymus, a decreased ratio of CD4+CD25+ regulatory T cells in CD4+ T cells, and a decreased expression of Foxp3. The ratios of CD4+CD25+ regulatory T cells and the thymus indices in the two TLR groups were significantly higher than those in the diabetic model group(P<0.05).ConclusionTongluo recipe can greatly increase the ratio of CD4+CD25+ regulatory T cells, thymus index, and Foxp3 expression in the thymus, and may protect the thymus CD4+CD25+ regulatory T cells in experimental diabetic rats.
Key words:  CD4+CD25+ regulatory T-lymphocytes  experimental diabetes mellitus  Tongluo recipe  thymus gland  Foxp3