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1.
目的研究一种由鼠李糖乳杆菌DM9054和植物乳杆菌86066构成的降脂益生菌组合对非酒精性脂肪性肝病(NAFLD)小鼠胆固醇代谢的影响及其可能机制。方法 24只雄性LDLR-/-小鼠随机分为对照组、模型组和益生菌干预组。高脂饮食(HFD)15周建立小鼠NAFLD模型,造模同时干预组给予鼠李糖乳杆菌DM9054联合植物乳杆菌86066灌胃,对照组和模型组给予等量生理盐水灌胃。实验过程中监测各组小鼠体重变化。实验结束后,检测小鼠血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL)和高密度脂蛋白胆固醇(HDL)的水平差异。检测小鼠肝脏组织病理变化。使用Realtime PCR检测小鼠肠道内法尼脂受体(FXR)mRNA、顶端膜钠依赖的胆汁酸转运体(ASBT)mRNA、纤维生长因子15(FGF-15)mRNA和三磷酸腺苷结合盒转运体G5(ABCG-5)mRNA表达水平。Western blot检测小鼠肝脏胆固醇7α-羟化酶(CYP7A1)、FXR、三磷酸腺苷结合盒转运体G8(ABCG-8)、清道夫受体BI(SR-BI)、3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMGCR)、胆盐输出泵(ABCB-11)、纤维生长因子受体4(FGFR-4)和胆固醇调节元件结合蛋白-2(SREBP-2)蛋白表达水平。结果与模型组相比,降脂益生菌干预组小鼠体重减轻(P0.05);小鼠血清TC、TG、LDL水平降低,HDL水平升高(P0.05);小鼠肝脏脂肪变性和炎性细胞浸润的现象显著减少;小鼠肠道ASBT mRNA和ABCG-5mRNA表达水平明显降低(Ps0.05),FGF-15mRNA表达水平明显升高(P0.05),FXR mRNA表达水平差异无统计学意义(P0.05);小鼠肝脏FGFR-4蛋白表达水平升高(P0.05),SREBP-2和HMGCR蛋白表达水平降低(Ps0.05),FXR、CYP7A1、SR-BI、ABCG-8和ABCB-11蛋白表达水平差异无统计学意义(Ps0.05)。结论降脂益生菌可能通过激活FXR-FGF15通路调节胆汁酸代谢;通过下调SREBP-2表达水平,抑制HMGCR表达,减少胆固醇的生成,从而起到改善非酒精性脂肪肝的作用。  相似文献   

2.
目的探讨益生菌干预对高脂饮食诱导肥胖小鼠肝脏miR-33和miR-122表达的影响。方法 18只雌性C57BL/6J小鼠随机分为对照组、肥胖组和益生菌干预组,每组6只,分别给予标准饲料、高脂饲料以及高脂饲料+益生菌合剂灌胃,自由采食及饮水,连续喂养6周。每周测量3组小鼠的体质量,6周后,留取小鼠血液样本采用全自动生化仪检测小鼠血脂,安乐法处死小鼠,留取小鼠肝脏样本Hair-pin RT-PCR法检测miR-33和miR-122的含量。结果与对照组小鼠相比,肥胖组小鼠体质量明显增加(t_(2周)=3.985,t_(3周)=4.751,t_(4周)=4.380,t_(5周)=4.728,t_(6周)=4.112,均P0.01);益生菌干预组小鼠体质量较肥胖组明显降低(t_(3周)=3.694,t_(4周)=4.415,t_(5周)=3.752,t_(6周)=3.392,均P0.01);肥胖组小鼠血清总胆固醇、低密度脂蛋白含量较对照组明显升高(t=10.850,t=7.024,均P0.01),益生菌干预组小鼠较肥胖组小鼠血清总胆固醇、低密度脂蛋白含量降低(t=3.034,t=2.881,均P0.05),但与对照组仍有差异。与对照组小鼠相比,肥胖组小鼠肝脏miR-122的表达升高(t=9.170,P0.01),miR-33的表达降低(t=3.420,P0.05),益生菌干预组小鼠较肥胖组小鼠miR-122的表达降低(t=3.204,P0.05),miR-33的表达升高(t=2.070,P0.05)。结论益生菌干预能够影响高脂饮食小鼠肝脏miR-33和miR-122的表达,这可能是益生菌干预改善高脂饮食小鼠肝脏脂代谢的机制之一。  相似文献   

3.
目的探究芦笋对高脂饮食小鼠脏器指数以及血生化的影响。方法将实验动物随机分为正常组、高脂饮食组、高脂饮食低剂量芦笋组、高脂饮食中剂量芦笋组、高脂饮食高剂量芦笋组、高脂饮食阳性对照组。其中正常组喂食正常饲料,其余组喂食高脂饲料。分别灌胃蒸馏水(正常组、高脂饮食组)、芦笋1.05g/(kg·d)(高脂饮食低剂量芦笋组)、芦笋2.10g/(kg·d)(高脂饮食中剂量芦笋组)、芦笋4.20g/(kg·d)(高脂饮食高剂量芦笋组)、降脂理肝汤1.19g/(kg·d)(高脂饮食阳性对照组),每天2次,每次0.35mL。比较分析小鼠的脏器指数及血生化活性。结果脏器方面:高脂饮食中剂量芦笋组小鼠的肝脏指数最低,但与高脂饮食组相比差异无统计学意义(P0.05)。高脂饮食低剂量芦笋组小鼠的脾脏指数最高,并且高脂饮食低剂量芦笋组雌性与正常组雌性相比差异有统计学意义(t=1.486,P0.05)。胸腺指数中高脂饮食低剂量芦笋组小鼠的相对于高脂饮食中剂量芦笋组和高脂饮食高剂量芦笋组而言是最高的,但与高脂饮食组相比差异无统计学意义(P0.05)。血生化方面:高脂饮食低剂量芦笋组、高脂饮食中剂量芦笋组以及高脂饮食高剂量芦笋组小鼠的总胆固醇(TC)以及甘油三酯(TG)水平低于高脂饮食组,但与高脂饮食组相比差异无统计学意义(P0.05),其中高脂饮食低剂量芦笋组中的TC、TG水平最低,而高脂饮食高剂量芦笋组相应的数值最高。芦笋治疗组小鼠在高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)方面高于正常组与高脂饮食组,但差异无统计学意义(P0.05),其中高脂饮食低、中、高剂量芦笋三组相比,低剂量芦笋组小鼠的HDL-C数值最低,而在LDL-C方面三组差异无统计学意义。结论高脂饮食中剂量芦笋能够降低高脂饮食小鼠的肝脏指数,高脂饮食低剂量芦笋对小鼠脾脏以及胸腺等免疫器官具有一定的促进作用,而高剂量芦笋则对小鼠脾脏以及胸腺等免疫器官具有一定的抑制作用。芦笋能降低小鼠的TC、TG水平,其中低剂量的芦笋效果最好。芦笋能提高小鼠HDL-C和LDL-C的含量。  相似文献   

4.
目的探究芦笋对高脂饮食小鼠体质量的影响。方法将实验动物随机分为正常组、高脂饮食组、高脂饮食低剂量芦笋组、高脂饮食中剂量芦笋组、高脂饮食高剂量芦笋组、高脂饮食阳性对照组。其中正常组喂食正常饲料,其余组喂食高脂饲料。分别灌胃蒸馏水(正常组、高脂饮食组)、芦笋1.05g/(kg·d)(高脂饮食低剂量芦笋组)、芦笋2.10g/(kg·d)(高脂饮食中剂量芦笋组)、芦笋4.20g/(kg·d)(高脂饮食高剂量芦笋组)、降脂理肝汤1.19g/(kg·d)(高脂饮食阳性对照组),每天2次,每次0.35mL。比较分析各组小鼠的体质量变化,同时对各组之间的雌性小鼠和雄性小鼠体质量变化进行对比。结果随饲养时间的增加,服用芦笋后的三组小鼠体质量均低于高脂饮食组,但与高脂饮食组相比差异无统计学意义(P0.05)。高脂饮食低剂量芦笋组、高脂饮食中剂量芦笋组、高脂饮食高剂量芦笋组小鼠的体质量变化率低于高脂饮食组,差异无统计学意义(P0.05)。对各组之间的雌性小鼠和雄性小鼠体质量变化进行对比可以发现雄性小鼠的体质量变化大于雌性小鼠的,差异有统计学意义(P0.05)。结论芦笋能降低高脂饮食小鼠的体质量,雄性小鼠和雌性小鼠的体质量变化存在差异。  相似文献   

5.
目的初步探讨田蓟苷改善高脂饮食小鼠血脂代谢的作用机制。方法将60只C57BL/6J小鼠随机分为对照组、高脂饮食组及田蓟苷低、中、高剂量给药组(50、100、200 mg/kg)和阿托伐他汀给药组(10 mg/kg),每组10只,连续给药12周后,检测各组小鼠血清总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL C)和低密度脂蛋白胆固醇(LDL C)水平。采用Western Blot法检测各组小鼠肝脏组织中SREBP 2和LDLR蛋白表达。通过16S rDNA基因实时荧光定量PCR法检测各组小鼠肠道菌群变化。结果与对照组相比,高脂饮食组小鼠血清TC、TG、HDL C和LDL C水平明显升高(t=-3.966,P=0.001;t=-3.438,P=0.003;t=3.811,P=0.001;t=-6.591,P<0.001),肝脏中SREBP 2和LDLR蛋白表达明显下调(t=7.198,P<0.001;t=8.892,P<0.001)。与高脂饮食组相比,田蓟苷高剂量给药组和阿托伐他汀给药组小鼠血清TC、LDL C水平明显降低(TC:t=2.483,P=0.023;t=3.300,P=0.004。LDL C:t=2.535,P=0.021;t=3.836,P=0.001),肝组织中的SREBP 2和LDLR蛋白水平明显上调(SREBP 2:t=-2.188,P=0.042;t=-3.317,P=0.007。LDLR:t=-2.649,P=0.016;t=-2.249,P=0.037)。与对照组相比,高脂饮食组小鼠肠道中厚壁菌门水平明显升高(t=-2.287,P=0.047),而拟杆菌门、双歧杆菌属、乳杆菌属、嗜黏蛋白阿克曼菌和普拉梭菌水平明显降低(t=3.127,P=0.006;t=2.737,P=0.014;t=3.542,P=0.002;t=3.491,P=0.003;t=2.780,P=0.012)。与高脂饮食组相比,田蓟苷高剂量给药组小鼠肠道中拟杆菌门、乳杆菌属和嗜黏蛋白阿克曼菌水平明显升高(t=-2.613,P=0.020;t=-2.558,P=0.024;t=-2.109,P=0.049)。结论田蓟苷改善高脂饮食诱导的血脂代谢紊乱可能与肝脏中SREBP 2和LDLR蛋白表达及肠道菌群结构变化有关,但其机制仍需进一步研究。  相似文献   

6.
限制活动加剧2型糖尿病小鼠肠道菌群和糖脂代谢紊乱   总被引:1,自引:1,他引:0  
【背景】久坐行为在2型糖尿病(type 2 diabetes mellitus,T2DM)患者群体中广泛存在,对于患者的血糖控制具有严重的不良影响,但是其具体机制还缺乏较为完善的阐述。【目的】通过限制小鼠活动模拟久坐行为,从而阐明久坐导致2型糖尿病小鼠糖脂代谢紊乱加剧的具体机制,为相应的健康教育和干预提供一定的理论基础。【方法】利用C57BL/6J雄性小鼠构建糖尿病模型,小鼠随机分为3组:正常组(CON)、2型糖尿病模型组(MOD)和2型糖尿病限制活动组(SED),通过限制小鼠活动的方法使其进行8周的久坐行为;采用试剂盒和形态学观察法测定小鼠糖脂代谢紊乱情况;HE和PAS染色观察小鼠回肠组织受损情况;采用RT-qPCR法测定小鼠粪便微生物的变化;采用TBA试剂盒测定小鼠血清和肝脏中总胆汁酸含量;分别通过荧光定量PCR(RT-qPCR)和Western blotting测定小鼠回肠和肝脏组织中法尼醇X受体(farnesoid X receptor,FXR)和G蛋白偶联胆汁酸受体5(G protein coupled bile acid receptor 5,TGR5)的mRNA和蛋白表达水平。【结果】与正常组相比,模型组小鼠血糖升高、血脂增加、胰岛素抵抗和口服葡萄糖耐量紊乱,而且肠道病理学变化明显,限制活动使T2DM小鼠糖脂代谢紊乱增加(P<0.05);T2DM小鼠肠道菌群失调,在门和属水平上的有益菌减少、有害菌增加,限制活动加剧了这一情况;与正常组相比,T2DM小鼠血清和肝脏组织中总胆汁酸含量增加,限制活动组总胆汁酸进一步增加(P<0.05);模型组小鼠胆汁酸受体FXR和TGR5表达较正常组显著降低(P<0.01),限制活动后进一步抑制了这些受体的表达。同时,Spearman相关性分析也显示小鼠糖脂代谢水平与肠道菌群及肠道菌群代谢物胆汁酸存在显著相关性。【结论】限制活动致使T2DM小鼠糖脂代谢紊乱加剧,其机制可能与肠道菌群和胆汁酸代谢失调,以及胆汁酸受体的进一步下调有关。  相似文献   

7.
绞股蓝总皂苷具有显著的降低血脂的作用,然而其机制尚未明确。课题组在前期研究的基础上,将33只C57BL/6J雄性小鼠随机分为3组,对照组(ND组)、模型组(HFD组)和绞股蓝皂苷组(HFD+GP组),分别给予维持饲料和高脂饲料,从16周开始,每日分别灌胃给予绞股蓝总皂250 mg/kg和等体积的空白溶剂,至38周,收集小鼠的血清和肝脏样本。通过HE染色观察肝脏组织的病理变化,检测血清总胆固醇和低密度脂蛋白水平,利用UPLC-MS/MS技术对小鼠肝脏14种胆汁酸含量进行分析,采用实时荧光定量PCR检测Cyp7a1、Cyp8b1、Fxr、Shp、Lrh1、Hnf4α基因表达水平。与对照组比较,模型组有明显的脂肪泡结构,给予绞股蓝总皂苷可以明显减轻肝脏组织的病理改变。与模型组比较,绞股蓝总皂苷可以显著降低血清中TC和LDL-C含量,显著降低肝脏中牛磺熊去氧胆酸(TUDCA)、甘氨鹅去氧胆酸(GCDCA)和甘氨脱氧胆酸(GDCA)含量,显著升高肝脏中鹅去氧胆酸(CDCA)、脱氧胆酸(DCA)和牛磺脱氧胆酸(TDCA)含量,并在上调肝脏Cyp7a1、Cyp8b1、Fxr和Lrh1基因表达的同时下调Shp的基因表达。绞股蓝总皂苷降脂作用的潜在靶点可能与FXR介导的胆汁酸代谢通路有关,为进一步深入探讨绞股蓝总皂苷降脂作用及其机制提供了参考。  相似文献   

8.
目的:研究清血消脂片对代谢性炎症小鼠血脂和血清炎症因子IL-6、MIP-1a以及NF-κB和PPARγm RNA表达的影响。方法:60只雄性C57小鼠,随机分为正常组、模型组、立普妥组、清血消脂片组(n=15)。采用高脂饮食联合脂多糖注射造成小鼠代谢性炎症模型。造模5周后,按成人临床等效剂量换算成小鼠剂量灌胃给药,每天两次,连续灌胃5周。处死后采血和取肝脏,检测各组小鼠血清胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)水平,并采用流式细胞术检测血清炎症因子IL-6和MIP-1a水平。反转录聚合酶链式反应(RT-PCR)法测定肝脏NF-κB和PPARγm RNA的表达。结果:与模型组相比,清血消脂片组小鼠血清TC和LDL-C水平明显降低(P0.01),血清炎症因子IL-6和MIP-1a水平明显降低(P0.01),清血消脂片组小鼠肝脏组织中NF-κB m RNA的表达显著降低(P0.05),PPARγm RNA的表达显著提高(P0.05)。结论:清血消脂片可降低代谢性炎症小鼠血脂和血清炎症因子IL-6和MIP-1a水平,并可通过调控核转录因子NF-κB和PPARγ受体,抑制小鼠体内代谢性炎症,从而可能对早期动脉粥样硬化起到干预作用。  相似文献   

9.
目的探讨奥美沙坦对于高脂诱导的非酒精性脂肪肝病(NAFLD)的影响及可能机制。方法健康雄性8周龄C57BL/6小鼠24只随机分为高脂组(n=16)和正常饮食组(n=8),高脂组小鼠高脂饮食(60%的脂肪)12w后再随机分为高脂饮食对照组(n=8)、高脂饮食治疗组(n=8)。高脂饮食治疗组小鼠给予0.75mg/kg/d的奥美沙坦灌胃8w,灌胃结束后处理小鼠,留取空腹血样本检测AST和ALT。肝组织冰冻切片行油红O染色观察脂肪变;石蜡切片行HE和F4/80免疫组化染色观察肝脏炎症变化;实时荧光定量PCR检测肝脏TNF-α和IL-6mRNA的表达水平;WesternBlot检测肝组织中IκB-α、p-IκBa、NF—κB信号通路的活化。结果奥美沙坦显著抑制了高脂诱导的NAFLD脂肪变性,并明显改善肝功能。实时荧光定量PCR结果表明奥美沙坦能显著降低肝脏组织中TNF-α和IL-6mRNA表达水平(P〈0.05);Western Blot结果显示奥美沙坦显著抑制肝脏NF-κB信号通路活化。结论奥美沙坦显著抑制NAFLD小鼠肝脏炎性病变而保护肝功能,其机制与抑制NF-κB信号通路活化以及降低肝脏TNF-α和IL-6mRNA水平有关。  相似文献   

10.
目的研究美洲大蠊Periplaneta americana精粉对高脂饮食小鼠体质量、血糖和肌酐的影响。方法高脂饮食小鼠分为对照组和实验组(给予美洲大蠊精粉:低剂量100 mg·kg~(-1)、中剂量200 mg·kg~(-1)、高剂量400 mg·kg~(-1))。灌胃第42天和第49天取血后,用试剂盒测定血清血糖和肌酐的含量,实时荧光定量PCR法检测肝脏中葡萄糖-6-磷酸酶(G-6-Pase)、葡萄糖转运蛋白2(GLUT2)和肾脏中单核细胞趋化蛋白-1(MCP-1) mRNA的表达水平。结果与对照组相比,中、高剂量组体质量增长率在第42天和第49天均显著低于对照组(P 0. 05,P 0. 01)。第42天实验组小鼠的血糖、肌酐和MCP-1 mRNA水平与对照组相比有降低趋势,但差异无统计学意义(P 0. 05),中、高剂量组的G-6-Pase mRNA水平极显著低于对照组(P 0. 01),GLUT2 mRNA水平的差异无统计学意义(P 0. 05)。第49天,低、高剂量组小鼠的血糖和肌酐水平显著低于对照组(P 0. 05),且低、中、高剂量组的G-6-Pase、GLUT2和MCP-1 mRNA水平显著低于对照组(P 0. 05,P 0. 01)。结论美洲大蠊精粉可降低高脂饮食小鼠的体质量增长率以及血糖、血清肌酐水平,减少肝脏中G-6-Pase mRNA表达和调节GLUT2 mRNA表达水平可能是美洲大蠊精粉降血糖机制之一,并且可能通过降低肾脏中MCP-1 mRNA的表达减轻肾脏损伤。  相似文献   

11.
12.
《环境昆虫学报》2014,(5):790-804
综述了白蚁螱客的主要种类、共生关系及相关机制的研究进展。白蚁螱客中,已报道的动物种类达170种。在与动物的共生关系中存在偏利共生(宾主共栖和异种共栖)、互利共生和无关共生三种;在与微生物的共生关系中,存在与内生菌(原生动物、细菌、真菌和放线菌)和外生菌(蚁巢伞菌等)间的互利关系。指出了白蚁与螱客研究中存在的问题,给出了解决方案,并提出了今后可能的研究热点或方向,为白蚁的综合利用(如纤维素酶)及今后研究物种间的协同进化提供了基础资料。  相似文献   

13.
To elucidate accumulation of minerals in human iliac arteries with aging, the content of minerals was analyzed by inductively coupled plasma atomic emission spectrometry. Bilateral common, internal, and external iliac arteries of 16 men and 8 women, ranging ages from 65 to 93 yr, were examined. It was found that an extremely high accumulation of calcium and phosphorus occurred in the common iliac artery at old age, being higher than that of the internal and external iliac arteries. It should be noted that the accumulation of calcium and phosphorus is the highest in the common iliac artery among the human arteries examined to date. Regarding sexual differences, the content of calcium and phosphorus in the common and internal iliac arteries was higher in women than in men, whereas their content in the external iliac artery was lower in women than in men.  相似文献   

14.
New sulfur derivatives of phosphoramidite ligands were synthesized and the impact of the sulfur unit on the spectroscopic properties of their rhodium and iridium complexes was investigated. The new ligands Bn2NPSCH2CH2Sa(P-Sa) (Bn = benzyl, 4), Bn2NPSCHCHSa(CH2)3CaH2(P-Sa)(Ca-Sa) (6) and Bn2NP(4-XC6H4OMe)2 (X = S, 7a; X = O, 7b) were converted to the rhodium and iridium complexes trans-[Rh(CO)Cl(L)2] (L = 4, 6, 7), [RhCl(COD)(L)] (L = 4, 6, 7), [IrCl(COD)(7a)] and [IrCl2Cp∗(6)]. For comparison, some phosphoramidite complexes of these formulations also were synthesized. The new metal complexes were spectroscopically analyzed. For the carbonyl complexes, the νCO IR stretching frequencies were lower than for the corresponding phosphite and phosphoramidite ligands. The 1JPRh coupling constants for the rhodium complexes with the new ligands were also smaller than for the respective phosphoramidite and phosphite complexes. Finally, the 1JPSe coupling constants of the selenides of the new ligands were lower than those of the phosphoramidite ligands but higher than for PPh3. The spectroscopic data reveal that the new thio ligands 4, 6 and 7a are more electron donating than phosphites and phosphoramidites but less electron donating than PPh3.  相似文献   

15.
Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.  相似文献   

16.
The first and second sessions of the Workshop focussed on the basics of ultrasound and infrasound, their applications in both industry and medicine, and metrology and protection standards for ultrasound applications.  相似文献   

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18.
The ability of partially purified human and guinea-pig haematogenous cell populations, when cultured in vitro, to metabolise arachidonic acid (AA) has been studied. Supernatants from 24 hour cell culture have been subjected to analysis for products of AA metabolism by gas chromatography with electron-capture detection.The cell types studied were human peripheral blood monocytes (both glass adherent and non-adherent), neutrophils, eosinophils and leukemic leucocytes; thoracic duct lymphocytes and lung alveolar macrophages. From the guinea-pig, induced and non-induced macrophage or neutrophil enriched peritoneal exudate populations, lymph node cells, peritoneal eosinophils and peripheral blood platelets were examined. Supernatants were assayed for the presence of PGE2, PGD2, PGF, TXB2 and 6-keto-PGF. In all types studied PGE2 and TXB2 were the major products formed. The identification of PGE2 and TXB2 was confirmed by GC/MS with multiple ion monitoring.The results have been compared with other reports and their possible significance discussed in relation to the proposed role of prostaglandins as mediators and modulators in immunopathology.  相似文献   

19.
以人胎盘脐带组织为材料,提取组织总RNA,用netRTPCR方法合成人血管能抑素cDNA基因,将该cDNA克隆进pSP72载体获得重组质粒pSP72C, DNA序列分析结果与预期序列一致。用BamHⅠ和NdeⅠ双酶切,切下pSP72C上的血管能抑素cDNA,插入pET3c载体的相应位点获得重组表达质粒pETC, 转化E. coli BL21(DE3), SDSPAGE分析显示:在IPTG诱导下,血管能抑素基因获得了高效表达,表达量约占菌体总蛋白的 27.9 %,主要以包涵体形式存在。包涵体经过洗涤、裂解、蛋白复性以及Sephadex G75凝胶过滤层析等步骤后,获得了纯度达91.4 %的人血管能抑素。CAM实验证明10 μg纯化蛋白就能显著抑制鸡胚新生血管生成。  相似文献   

20.
To examine the hypothesis that foot-strike hemolysis alters vascular volumes and selected hematological properties is trained athletes, we have measured total blood volume (TBV), red cell volume (RCV) and plasma volume (PV) in cyclists (n = 21) and runners (n = 17) and compared them to those of untrained controls (n = 20). TBV (ml x kg(-1)) was calculated as the sum of RCV (ml x kg(-1)) and PV (ml x kg(-1)) obtained using 51Cr and 125I-labelled albumin, respectively. Hematological assessment was carried out using a Coulter counter. Peak aerobic power (VO2peak) was measured during progressive exercise to fatigue using both cycle and treadmill ergometry. RCV was 15% higher (P < 0.05) in male cyclists [35.4 (1.0), mean (SE); n = 12] and runners [35.3 (0.98); n = 9] compared to the controls [30.7 (0.92); n = 12]. Similar differences existed between the female cyclists [28.2 (2.1); n = 9] and runners [28.4 (1.0); n = 8] compared to the untrained controls [24.9 (1.4); n = 8]. For the male athletes, PV was between 19% (cyclists) and 28% (runners) higher (P < 0.05) in the trained athletes compared to the untrained controls. The differences in PV between the female groups were not significant. Although the males had a higher (P < 0.05) TBV, RCV and PV than the females, no differences between cyclists and runners were found for either gender. Mean cell volume was not different between the athletic groups. VO2peak (ml x kg(-1) x min(-1)) was higher (P < 0.05) in both male [68.4 (1.5)] and female [54.8 (2.1)] runners when compared to the untrained males [47.1 (1.0)] and females [40.5 (2.1)]. Although differences existed between the genders in VO2peak for both cyclists and runners, no differences were found between the athletic groups within a gender. Since the vascular volumes were not different between cyclists and runners for either the males or females, foot-strike hemolysis would not appear to have an effect on that parameter. The significant correlations (P < 0.05) found between VO2peak and RCV (r = 0.64 and 0.64) and TBV (r = 0.82 and 0.63) for the males and females, respectively, suggests a role for the vascular system in realizing a high aerobic power.  相似文献   

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