目的探讨不同时间给予17β-雌二醇(E2)替代治疗对apo E-/-雌鼠动脉粥样硬化(As)病变内效应性T细胞(Teff)和调节性T细胞(Treg)平衡和氧化应激的影响及其与As的关系。方法 5周龄apo E-/-鼠摘除卵巢后用高脂餐喂养,随机分为早期、延迟及持...目的探讨不同时间给予17β-雌二醇(E2)替代治疗对apo E-/-雌鼠动脉粥样硬化(As)病变内效应性T细胞(Teff)和调节性T细胞(Treg)平衡和氧化应激的影响及其与As的关系。方法 5周龄apo E-/-鼠摘除卵巢后用高脂餐喂养,随机分为早期、延迟及持续治疗组,给予E2皮下注射(5μg/天)8周或16周,检测主动脉根部As病变大小、组成改变以及硫氧还蛋白TRX1的表达,并检测As病变内Th1细胞的细胞因子IFN-γ、Treg细胞的转录因子Foxp3和TRX1 m RNA的表达。结果 (1)和对照组相比,早期E2治疗使apo E-/-鼠脂质条纹的面积显著减少(8.82%±0.98%vs.19.3%±1.26%in controls,P<0.01);而延迟E2治疗纤维脂质斑块的面积明显增加(30.7%±0.74%vs.25.1%±0.94%in controls,P<0.05);持续E2治疗纤维脂质斑块的面积亦明显降低(22.48%±0.92%vs.25.63%±1.04%in controls,P<0.05);(2)延迟E2治疗斑块内胶原和平滑肌细胞含量明显减少,而巨噬细胞数量显著增加;持续E2治疗斑块内胶原和平滑肌细胞含量明显增加,而巨噬细胞数量明显减少;(3)早期及持续E2治疗As病变内代表促炎作用的IFN-γm RNA的表达明显降低,而延迟E2治疗斑块内IFN-γm RNA的表达明显增加;(4)早期及持续E2治疗AS病变内代表抑炎作用的Foxp3 m RNA的表达明显增加,而延迟E2治疗斑块内Foxp3 m RNA的表达明显降低;(5)早期及持续E2治疗,apo E-/-鼠血清过氧化脂质(LPO)的含量明显减少,而延迟E2治疗血清LPO含量明显增加;(6)早期及持续E2治疗As病变内TRX1 m RNA及蛋白水平的表达明显降低;而延迟E2治疗斑块内TRX1 m RNA及蛋白水平的表达却明显增加。结论 apo E-/-鼠摘除卵巢后,早期E2替代治疗可抑制As病变的形成,而缺乏雌激素一定时间后,再给予E2治疗却加速了As病变的进展并使斑块趋于不稳定,其机制与调节Teff/Treg细胞平衡以及对氧化应激的影响有关。展开更多
目的:观察调脂方对动脉粥样硬化(AS)模型小鼠外周血白细胞端粒、端粒酶以及主动脉病变、血脂、炎症指标的影响,探讨其抗AS作用机制。方法: 30只8周龄雄性ApoE -/-小鼠随机分为模型组、阿托伐他汀组、中药干预组,每组10只。另取同周龄雄...目的:观察调脂方对动脉粥样硬化(AS)模型小鼠外周血白细胞端粒、端粒酶以及主动脉病变、血脂、炎症指标的影响,探讨其抗AS作用机制。方法: 30只8周龄雄性ApoE -/-小鼠随机分为模型组、阿托伐他汀组、中药干预组,每组10只。另取同周龄雄性C57BL/6小鼠10只做空白对照组。采用高脂饲料喂养方法建立AS模型,空白对照组、模型组小鼠用蒸馏水灌胃,中药干预组灌服中药“调脂方”,阿托伐他汀组阿托伐他汀钙灌胃给药,持续12周。运用生化仪检测脂代谢指标:总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)及炎症指标:超敏C反应蛋白(hs-CRP)、白介素-6(IL-6);采用油红“O”染色,观察动脉斑块形成及脂质堆积程度;荧光定量聚合酶链反应法检测外周血白细胞及血管细胞的端粒长度及端粒酶活性。结果:与正常空白对照组比较,模型组血清脂代谢指标和炎症指标均明显升高( P <0.01);与模型组比较,阿托伐他汀组和中药干预组均显著降低TC、LDL、HDL、hs-CRP( P <0.01),TG、IL-6( P <0.05);与阿托伐他汀组比较,中药干预组降低TC的作用较弱( P <0.05),而中药干预组在降低TG程度上优于阿托伐他汀组( P <0.05),在降低LDL、hs-CRP、IL-6及升高HDL上无明显差异。主动脉病理油红“O”染色显示模型组AS程度明显,中药干预组和阿托伐他汀组AS程度较模型组明显改善,阿托伐他汀组作用优于中药干预组。模型组外周血白细胞端粒长度明显短于空白对照组、外周血白细胞端粒酶活性低于空白对照组( P <0.05)。与空白对照组比较,中药干预组端粒酶活性、阿托伐他汀组端粒长度、端粒酶活性明显提高( P <0.01),中药干预组与阿托伐他汀组之间外周血白细胞端粒长度及端粒酶活性则无明显统计学差异。结论:调脂方具有抗小鼠AS作用,其机制可能与改善脂代谢、炎症指标及调控端粒长度及端粒酶活性相关。展开更多
Background Subtypes of T cells, called regulatory T cells （Treg cell）, play a critical role in limiting autoimmune processes and inflammatory responses, The aim of this study was to explore functional roles of FOXP3...Background Subtypes of T cells, called regulatory T cells （Treg cell）, play a critical role in limiting autoimmune processes and inflammatory responses, The aim of this study was to explore functional roles of FOXP3 in the manifestation of atherosclerosis in Apolipoprotein E deficient （ApoE）-/- mice. Methods Lentivirus-mediated （siRNA） was used to knock down FOXP3 and FOXP3high＋CD4＋ CD25＋ T cells adoptive transfer assays in high fat diet ApoE-/- mice were done. The resulting atherosclerotie lesions were assessed by determining FOXP3 transcript levels and investigating the expression of FOXP3 protein in different tissues. Results Animals treated with siRNA of FOXP3 showed a significant increase in atherosclerotic lesion formation and a reduction in the number of FOXP3＋CD4＋CD25＋ T cells compared with other groups. Transfer of FOXP3highCD4＋CD25＋ T cells significantly decreased atherosclerotic plaque formation and increased the number of FOXP3＋ CD4＋ CD25＋ T cells. FOXP3 protein levels and FOXP3 transcript levels were lowest in the siRNA group, and were highest in tissues from the Treg transfer group. Conclusion FOXP3 plays an important role in regulating the inflammatory response within the atherosclerotic lesion. It can inhibit significantly the progression of the atherosclerosis plaque in ApoE-/- mice.展开更多
Background Immune inflammatory response is throughout the entire process of atherosclerosis （AS）. It was unclear whether the mechanism of mesenchymal stem cells （MSCs） transplantation for treatment of AS is involv...Background Immune inflammatory response is throughout the entire process of atherosclerosis （AS）. It was unclear whether the mechanism of mesenchymal stem cells （MSCs） transplantation for treatment of AS is involved with inflammation regulation in the plaque area. The aim of this study was to explore the effects of MSCs in the formation of atherosclerosis plaque in hypercholesterolemic apoliprotein （apo）E-/- mice. Methods ApoE-/- mice MSCs were isolated and identified. At 8 weeks of age, 30 male ApoE-/- mice were randomly divided into negative control group （Neg, n = 10）, positive control group （Pos, n = 10） and mesenchymal stem cells group （MSCs, n = 10）. MSCs were injected through caudal vein into the body of Pos and MSCs group. The plaque area of all subjects were compared, the percentage of CD4^＋CD25^＋Tregs in different tissues were analyzed by fluorescence activated cell sorter （FACS）, proliferation response of splenocytes to MSCs was detected and cytokines in the supernatant were determined by enzyme linked immunosorbent assay （ELISA）. Results Compared with controls, MSCs resulted in a significant decrease of latherosclerotic plaques size （P 〈 0.05）, and a significant increase of CD4^＋CD25^＋ regulatory T cells in spleen （P 〈 0.05）. Specific proliferation response of CD4^＋CD25^＋ regulatory T cells in splenocytes to MCSs was significantly suppressed, the superanant level of TGF-[3 and IL-10 in MSCs group were increased while IFN-γ/ decreased significantly. Conclusion MSCs play an important role in regulating the inflammatory response and significantly inhibit the formation of the atherosclerosis plaque in ApoE-/-mice.展开更多
文摘目的探讨不同时间给予17β-雌二醇(E2)替代治疗对apo E-/-雌鼠动脉粥样硬化(As)病变内效应性T细胞(Teff)和调节性T细胞(Treg)平衡和氧化应激的影响及其与As的关系。方法 5周龄apo E-/-鼠摘除卵巢后用高脂餐喂养,随机分为早期、延迟及持续治疗组,给予E2皮下注射(5μg/天)8周或16周,检测主动脉根部As病变大小、组成改变以及硫氧还蛋白TRX1的表达,并检测As病变内Th1细胞的细胞因子IFN-γ、Treg细胞的转录因子Foxp3和TRX1 m RNA的表达。结果 (1)和对照组相比,早期E2治疗使apo E-/-鼠脂质条纹的面积显著减少(8.82%±0.98%vs.19.3%±1.26%in controls,P<0.01);而延迟E2治疗纤维脂质斑块的面积明显增加(30.7%±0.74%vs.25.1%±0.94%in controls,P<0.05);持续E2治疗纤维脂质斑块的面积亦明显降低(22.48%±0.92%vs.25.63%±1.04%in controls,P<0.05);(2)延迟E2治疗斑块内胶原和平滑肌细胞含量明显减少,而巨噬细胞数量显著增加;持续E2治疗斑块内胶原和平滑肌细胞含量明显增加,而巨噬细胞数量明显减少;(3)早期及持续E2治疗As病变内代表促炎作用的IFN-γm RNA的表达明显降低,而延迟E2治疗斑块内IFN-γm RNA的表达明显增加;(4)早期及持续E2治疗AS病变内代表抑炎作用的Foxp3 m RNA的表达明显增加,而延迟E2治疗斑块内Foxp3 m RNA的表达明显降低;(5)早期及持续E2治疗,apo E-/-鼠血清过氧化脂质(LPO)的含量明显减少,而延迟E2治疗血清LPO含量明显增加;(6)早期及持续E2治疗As病变内TRX1 m RNA及蛋白水平的表达明显降低;而延迟E2治疗斑块内TRX1 m RNA及蛋白水平的表达却明显增加。结论 apo E-/-鼠摘除卵巢后,早期E2替代治疗可抑制As病变的形成,而缺乏雌激素一定时间后,再给予E2治疗却加速了As病变的进展并使斑块趋于不稳定,其机制与调节Teff/Treg细胞平衡以及对氧化应激的影响有关。
文摘目的:观察调脂方对动脉粥样硬化(AS)模型小鼠外周血白细胞端粒、端粒酶以及主动脉病变、血脂、炎症指标的影响,探讨其抗AS作用机制。方法: 30只8周龄雄性ApoE -/-小鼠随机分为模型组、阿托伐他汀组、中药干预组,每组10只。另取同周龄雄性C57BL/6小鼠10只做空白对照组。采用高脂饲料喂养方法建立AS模型,空白对照组、模型组小鼠用蒸馏水灌胃,中药干预组灌服中药“调脂方”,阿托伐他汀组阿托伐他汀钙灌胃给药,持续12周。运用生化仪检测脂代谢指标:总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)及炎症指标:超敏C反应蛋白(hs-CRP)、白介素-6(IL-6);采用油红“O”染色,观察动脉斑块形成及脂质堆积程度;荧光定量聚合酶链反应法检测外周血白细胞及血管细胞的端粒长度及端粒酶活性。结果:与正常空白对照组比较,模型组血清脂代谢指标和炎症指标均明显升高( P <0.01);与模型组比较,阿托伐他汀组和中药干预组均显著降低TC、LDL、HDL、hs-CRP( P <0.01),TG、IL-6( P <0.05);与阿托伐他汀组比较,中药干预组降低TC的作用较弱( P <0.05),而中药干预组在降低TG程度上优于阿托伐他汀组( P <0.05),在降低LDL、hs-CRP、IL-6及升高HDL上无明显差异。主动脉病理油红“O”染色显示模型组AS程度明显,中药干预组和阿托伐他汀组AS程度较模型组明显改善,阿托伐他汀组作用优于中药干预组。模型组外周血白细胞端粒长度明显短于空白对照组、外周血白细胞端粒酶活性低于空白对照组( P <0.05)。与空白对照组比较,中药干预组端粒酶活性、阿托伐他汀组端粒长度、端粒酶活性明显提高( P <0.01),中药干预组与阿托伐他汀组之间外周血白细胞端粒长度及端粒酶活性则无明显统计学差异。结论:调脂方具有抗小鼠AS作用,其机制可能与改善脂代谢、炎症指标及调控端粒长度及端粒酶活性相关。
基金supported by a grant from theNational Natural Science Foundation of Chinato Dr LI Dazhu (No.C03030201)
文摘Background Subtypes of T cells, called regulatory T cells （Treg cell）, play a critical role in limiting autoimmune processes and inflammatory responses, The aim of this study was to explore functional roles of FOXP3 in the manifestation of atherosclerosis in Apolipoprotein E deficient （ApoE）-/- mice. Methods Lentivirus-mediated （siRNA） was used to knock down FOXP3 and FOXP3high＋CD4＋ CD25＋ T cells adoptive transfer assays in high fat diet ApoE-/- mice were done. The resulting atherosclerotie lesions were assessed by determining FOXP3 transcript levels and investigating the expression of FOXP3 protein in different tissues. Results Animals treated with siRNA of FOXP3 showed a significant increase in atherosclerotic lesion formation and a reduction in the number of FOXP3＋CD4＋CD25＋ T cells compared with other groups. Transfer of FOXP3highCD4＋CD25＋ T cells significantly decreased atherosclerotic plaque formation and increased the number of FOXP3＋ CD4＋ CD25＋ T cells. FOXP3 protein levels and FOXP3 transcript levels were lowest in the siRNA group, and were highest in tissues from the Treg transfer group. Conclusion FOXP3 plays an important role in regulating the inflammatory response within the atherosclerotic lesion. It can inhibit significantly the progression of the atherosclerosis plaque in ApoE-/- mice.
基金supported by the grand from the Department of Education Project of Hubei Province to WANG Zhi-xiao (B20102108)
文摘Background Immune inflammatory response is throughout the entire process of atherosclerosis （AS）. It was unclear whether the mechanism of mesenchymal stem cells （MSCs） transplantation for treatment of AS is involved with inflammation regulation in the plaque area. The aim of this study was to explore the effects of MSCs in the formation of atherosclerosis plaque in hypercholesterolemic apoliprotein （apo）E-/- mice. Methods ApoE-/- mice MSCs were isolated and identified. At 8 weeks of age, 30 male ApoE-/- mice were randomly divided into negative control group （Neg, n = 10）, positive control group （Pos, n = 10） and mesenchymal stem cells group （MSCs, n = 10）. MSCs were injected through caudal vein into the body of Pos and MSCs group. The plaque area of all subjects were compared, the percentage of CD4^＋CD25^＋Tregs in different tissues were analyzed by fluorescence activated cell sorter （FACS）, proliferation response of splenocytes to MSCs was detected and cytokines in the supernatant were determined by enzyme linked immunosorbent assay （ELISA）. Results Compared with controls, MSCs resulted in a significant decrease of latherosclerotic plaques size （P 〈 0.05）, and a significant increase of CD4^＋CD25^＋ regulatory T cells in spleen （P 〈 0.05）. Specific proliferation response of CD4^＋CD25^＋ regulatory T cells in splenocytes to MCSs was significantly suppressed, the superanant level of TGF-[3 and IL-10 in MSCs group were increased while IFN-γ/ decreased significantly. Conclusion MSCs play an important role in regulating the inflammatory response and significantly inhibit the formation of the atherosclerosis plaque in ApoE-/-mice.
