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
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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
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Supported by the National Program on Key Basic Research(“973” Projects, 2005CB523304).

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    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。

    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.

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  • 收稿日期:2010-08-25
  • 最后修改日期:2011-01-07
  • 录用日期:2011-01-10
  • 在线发布日期: 2011-02-21
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