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- 2017
血管活性肠肽对自身免疫性脑脊髓炎大鼠CD4+CD25+Treg/CD4+T细胞比例及TGF-β1表达的影响
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Abstract:
摘要：目的 探讨血管活性肠肽(VIP)对实验性自身免疫性脑脊髓炎(EAE)大鼠CD4+CD25+Treg/CD4+T细胞比例及转化生长因子-β1(TGF-β1)表达的影响。方法 健康雌性Wistar大鼠随机分成EAE模型对照组和VIP低剂量防治组、VIP高剂量防治组，利用髓鞘碱性蛋白(MBP)+完全福氏佐剂(CFA)诱导建立EAE模型，另设正常对照组。自造模当日起，每隔1d分别对VIP低、高剂量防治组大鼠腹腔注射VIP 4nmol/kg(0.2mL)、16nmol/kg(0.8mL)，正常对照组及EAE对照组腹腔注射0.8mL生理盐水，连续10d。观察各组大鼠发病高峰期神经功能障碍评分情况；HE染色观察脑组织基本病理改变；免疫组化技术检测脑组织中星形胶质细胞活化情况；流式细胞术及ELISA法检测脾组织中CD4+CD25+Treg/ CD4+T细胞比例及脑组织匀浆中TGF-β1含量变化。结果 VIP各剂量防治组大鼠发病高峰期神经功能障碍评分(NDS)降低，脑组织中炎细胞浸润程度明显下降、活化的星形胶质细胞数量减少、脾组织中CD4+CD25+Treg/CD4+T细胞比例升高、脑组织匀浆中TGF-β1含量升高，且各低、高剂量组间存在一定剂量依赖关系。结论 VIP通过提高脾组织中CD4+CD25+Treg/CD4+T细胞比例、增加脑组织匀浆中TGF-β1含量，从而减轻脑组织炎症细胞的浸润程度，抑制星形胶质细胞活化，发挥对EAE的防治作用。
ABSTRACT: Objective To explore the effects of vasoactive intestinal peptide (VIP) on the ratio of CD4+CD25+Treg/CD4+T cell and the expression of TGF-β1 in experimental autoimmune encephalomyelitis (EAE) rats. Methods We randomly divided 60 healthy female Wistar rats into normal control group, EAE control group, VIP low-dose group and VIP high-dose group. We used myelin basic protein (MBP) + complete adjuvant (CFA) to establish EAE model. Since the day of model construction, the low- and high-dose VIP groups received intraperitoneal injection of 4nmol/kg (0.2mL) and 16nmol/kg (0.8mL) of VIP every other day, respectively; normal control group and EAE group received injection of saline of 0.8mL for 10 days in a row. We recorded the peak of neurological dysfunction score (NDS) changes in the rats, observed the pathological changes and GFAP？？+ astrocyte activation in the brain at the morbidity peak of rats with HE staining, and detected the ratio of CD4+CD25+Treg/CD4+T in the spleen with FACS and TGF-β1 cytokine level in brain tissue with ELISA. Results The peak nerve dysfunction score was decreased in each VIP dose group. In normal control group, there were decreased inflammatory cell infiltration and decreased number of active astrocytes in the brain tissue. The degree of infiltration of inflammatory cells and astrocyte activation in VIP control groups were significantly lower than those in EAE group. The CD4+CD25+Treg/CD4+T cell ratio of the spleen tissue in each dose VIP treated group rats was higher than that in EAE control group. The cytokine level of TGF-β1 in the brain tissue increased in each VIP dose group in the dose-dependent manner. Conclusion Through up-regulating the ratio of CD4+CD25+Treg/CD4+T cell in the spleen tissue, increasing TGF-β1 content in brain tissue, and inhibiting the infiltration of inflammatory cells and the astrocyte activation, VIP plays an important role in prevention and control of EAE
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