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1.
目的 分离和培养人腹膜微血管内皮细胞.方法 取非炎症且非转移性肿瘤患者施行腹部外科手术的大网膜组织,经Ⅰ型胶原酶消化,两次筛网过滤、离心等步骤获取人微血管段,接种24 h待内皮细胞从微血管段爬出贴壁后,去除微血管段获得内皮细胞进行传代培养.组织形态学观察及免疫组化法鉴定所培养的细胞.结果 运用此法进行的原代培养,血细胞在换液和传代过程中被去除,成纤维细胞污染被减至最少;组织形态学观察显示内皮细胞呈铺路石样生长,免疫组化鉴定表明Ⅷ因子阳性;证实所培养的细胞为人腹膜微血管内皮细胞,其纯度较高,生长状态良好.结论 实验成功分离和培养了人腹膜微血管内皮细胞,为体外研究腹膜透析的病理生理学改变,提供了细胞来源. 相似文献
2.
长期腹膜透析会引起腹膜血管生成,对腹膜功能有重大的影响。本文就尿毒症腹膜透析患者腹膜血管生成的原因,血管形成的病理生理过程及腹膜血管新生对腹膜功能的影响做一综述。 相似文献
3.
Objective To investigate the association between angiopoietin-2 (Angpt-2) and peritoneal angiogenesis in a uremic peritoneal dialysis (PD) rat model. Methods Uremic (subtotal nephrectomy) rats were established and divided into non-PD, 10 d-PD, 28 d-PD and 56 d-PD groups. Standard PD solution was applied in the study. Rats undergone sham operation without PD were used as control group. Vessel density of the peritoneum was detected and quantified with anti-CD31 immunohistochemical staining. Expressive levels of Angpt-2 and vascular endothelial growth factor (VEGF) were examined in the peritoneum by real-time PCR and Western blotting. Results The non-PD group was characterized by increased vessel density in the peritoneum compared with that of the control group [(5±3)/HP vs (1±1)/HP]. Progressive angiogenesis was found in 10 d-PD, 28 d-PD and 56 d-PD groups [(10±5)/HP, (17±5)/HP, (19±4)/HP]. Furthermore, expressive levels of Angpt-2 and VEGF increased significantly in the non-PD group compared with the control (P<0.01), and such expressions were significantly higher in the PD groups as compared to non-PD group (P<0.01), but no difference was found among the PD groups. Both VEGF and Angpt-2 levels were positively correlated with vessel density(r=0.7756, P<0.01; r=0.5223, P<0.05). Conclusions Uremia and PD promote peritoneal angiogenesis in rats. Increased expression level of Angpt-2 in peritoneum is positively correlated with peritoneal angiogenesis. Angpt-2 may be a new therapeutic target of peritoneal angiogenesis. 相似文献
4.
目的血管内皮细胞功能紊乱是腹膜透析患者心血管疾病(cardiovascular disease,CVD)发生的重要原因。本研究模拟腹膜透析液的高糖环境,观察高糖作用下脂肪细胞对微血管内皮细胞细胞间粘附分子-1(intercellular adhesion molecule,ICAM-1)表达的影响,探讨腹膜透析患者内皮功能紊乱导致CVD发生的可能机制。方法体外诱导3T3-L1分化成为脂肪细胞。将实验所用内皮细胞分成6组,分别是:对照组,高糖组,高渗组,对照脂肪细胞组(普通培养基培养脂肪细胞所得上清干预内皮细胞),高糖脂肪细胞组(高糖培养脂肪细胞所得上清干预内皮细胞)及高渗脂肪细胞组(高渗培养脂肪细胞所得上清作干预内皮细胞)。细胞增殖检测试剂盒检测b End.3增殖能力。酶联免疫吸附试验检测脂肪细胞高糖、高渗作用后上清中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达量的改变。Western blot法检测脂肪细胞条件培养基对内皮细胞ICAM-1表达量的影响,及加用丝裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路阻断剂SB203580作用于内皮细胞后ICAM-1表达量的变化。结果与对照组相比,高糖培养液直接干预内皮细胞或培养脂肪细胞后干预内皮细胞均可以促进内皮细胞增殖(高糖组/对照组:1.600±0.104比1.000±0.000,t=9.954,P=0.010;高糖脂肪细胞组/对照脂肪细胞组:1.563±0.181比1.213±0.097,t=2.945,P=0.042)。高糖可显著促进脂肪细胞TNF-α的表达(2515.313±277.434比788.683±167.267,t=9.232,P=0.001)。高糖培养脂肪细胞的上清显著促进内皮细胞ICAM-1表达(1.533±0.058比1.133±0.153,t=4.243,P=0.013),该作用可被p38 MAPK信号通路阻断剂SB203580部分抑制(0.850±0.111比1.124±0.108,t=-3.053,P=0.038)。结论高糖培养脂肪细胞后,可通过TNF-α-p38 MAPK信号通路促进内皮细胞ICAM-1表达,可能和腹膜透析患者内皮细胞功能紊乱及CVD发生有关。 相似文献
5.
目的观察高浓度葡萄糖对体外培养的人脐静脉内皮细胞(HUVECs)血管生成素2(Ang2)和酪氨酸激酶2受体(Tie2)表达的影响,探讨腹膜透析患者腹膜血管新生的可能机制。方法分别以4.25%葡萄糖(高糖组)、4.25%甘露醇(甘露醇组)和800μmol/L葡萄糖降解产物甲基乙二醛(MGO)(MGO组)对体外培养的HUVECs进行干预,设立空白对照组。以MTT比色法测定的D(490 nm)比较各组HUVECs干预后12、24、48 h时点的细胞增殖情况;分别采用Real-Time PCR技术、ELISA法和Western blotting法检测各组HUVECs中Ang2和Tie2 mRNA表达、细胞培养上清液中Ang2分泌量及HUVECs中Tie2蛋白的表达。结果高糖组、甘露醇组和MGO组HUVECs干预后12 h时点的D(490 nm)均明显高于空白对照组(P<0.05),干预后24 h时点的D(490 nm)则显著低于空白对照组(P<0.05)。与空白对照组比较,高糖组、甘露醇组和MGO组HUVECs中Ang2mRNA表达均明显上调,高糖组和甘露醇组细胞培养上清液中Ang2分泌量均显著增加(P<0.05),... 相似文献
6.
Objective To investigate the association between angiopoietin-2 (Angpt-2) and peritoneal angiogenesis in a uremic peritoneal dialysis (PD) rat model. Methods Uremic (subtotal nephrectomy) rats were established and divided into non-PD, 10 d-PD, 28 d-PD and 56 d-PD groups. Standard PD solution was applied in the study. Rats undergone sham operation without PD were used as control group. Vessel density of the peritoneum was detected and quantified with anti-CD31 immunohistochemical staining. Expressive levels of Angpt-2 and vascular endothelial growth factor (VEGF) were examined in the peritoneum by real-time PCR and Western blotting. Results The non-PD group was characterized by increased vessel density in the peritoneum compared with that of the control group [(5±3)/HP vs (1±1)/HP]. Progressive angiogenesis was found in 10 d-PD, 28 d-PD and 56 d-PD groups [(10±5)/HP, (17±5)/HP, (19±4)/HP]. Furthermore, expressive levels of Angpt-2 and VEGF increased significantly in the non-PD group compared with the control (P<0.01), and such expressions were significantly higher in the PD groups as compared to non-PD group (P<0.01), but no difference was found among the PD groups. Both VEGF and Angpt-2 levels were positively correlated with vessel density(r=0.7756, P<0.01; r=0.5223, P<0.05). Conclusions Uremia and PD promote peritoneal angiogenesis in rats. Increased expression level of Angpt-2 in peritoneum is positively correlated with peritoneal angiogenesis. Angpt-2 may be a new therapeutic target of peritoneal angiogenesis. 相似文献
7.
Objective To investigate the association between angiopoietin-2 (Angpt-2) and peritoneal angiogenesis in a uremic peritoneal dialysis (PD) rat model. Methods Uremic (subtotal nephrectomy) rats were established and divided into non-PD, 10 d-PD, 28 d-PD and 56 d-PD groups. Standard PD solution was applied in the study. Rats undergone sham operation without PD were used as control group. Vessel density of the peritoneum was detected and quantified with anti-CD31 immunohistochemical staining. Expressive levels of Angpt-2 and vascular endothelial growth factor (VEGF) were examined in the peritoneum by real-time PCR and Western blotting. Results The non-PD group was characterized by increased vessel density in the peritoneum compared with that of the control group [(5±3)/HP vs (1±1)/HP]. Progressive angiogenesis was found in 10 d-PD, 28 d-PD and 56 d-PD groups [(10±5)/HP, (17±5)/HP, (19±4)/HP]. Furthermore, expressive levels of Angpt-2 and VEGF increased significantly in the non-PD group compared with the control (P<0.01), and such expressions were significantly higher in the PD groups as compared to non-PD group (P<0.01), but no difference was found among the PD groups. Both VEGF and Angpt-2 levels were positively correlated with vessel density(r=0.7756, P<0.01; r=0.5223, P<0.05). Conclusions Uremia and PD promote peritoneal angiogenesis in rats. Increased expression level of Angpt-2 in peritoneum is positively correlated with peritoneal angiogenesis. Angpt-2 may be a new therapeutic target of peritoneal angiogenesis. 相似文献
8.
目的观察尿毒症腹透对大鼠腹膜血管新生和转运、超滤功能的影响。方法35只雄性Wistar大鼠,随机分为对照组(假手术组,n=6)、尿毒症组(5/6肾切除,n=6)和尿毒症腹透组(n=18);尿毒症腹透组大鼠根据腹透时间不同分成10 d、4周和8周组(n=6)。各组大鼠处死前行腹膜平衡试验,检测腹膜转运功能,并取大网膜进行组织形态学检查。结果大鼠大网膜上每一高倍视野下的血管数,对照组、尿毒症组和尿毒症腹透各时间组分别为1±1、5±3、10±5、17±5和19±4,尿毒症组和尿毒症腹透组显著多于对照组,而尿毒症腹透组又较尿毒症组明显增多(均P<0.05)。腹膜平衡试验结果显示,尿毒症组和尿毒症腹透组大鼠2 h腹透液和血浆肌酐比值(D/PCr)较对照组升高,净超滤量(Net UF)较对照组减少(P<0.05)。血管数变化与2 h D/PCr呈显著正相关(r=0.9038,P<0.001),与Net UF呈显著负相关(r=-0.9168,P<0.05)。结论尿毒症及非生物相容性腹透液均会引起腹膜血管新生,从而导致超滤减少,最终使患者退出腹透。 相似文献
9.
可溶性酪氨酸激酶2融合蛋白对尿毒症腹膜透析大鼠腹膜形态和功能的影响 总被引:3,自引:0,他引:3
目的 研究可溶性酪氨酸激酶2融合蛋白(sTie-2-Fc)对尿毒症腹膜透析大鼠腹膜血管新生、溶质转运和超滤功能的影响。 方法 32只雄性Wistar大鼠按数字随机法分为假手术组、尿毒症组、尿毒症腹透组和sTie-2-Fc干预组(均n=8)。尿毒症腹透组和sTie-2-Fc干预组大鼠经腹透管每天2次腹腔灌注4.25%葡萄糖透析液(3 ml/100 g体质量)共4周,sTie-2-Fc干预组大鼠每次灌注时在透析液中加入1 μg sTie-2-Fc。各组大鼠处死前行腹膜平衡试验,检测腹膜转运和超滤功能,取大网膜标本行抗CD31免疫组化染色并计血管数。 结果 与假手术组大鼠相比,尿毒症组大鼠的2 h腹透液和血肌酐比值(D/Pcr)增高(0.78±0.05比0.70±0.09,P = 0.028),腹透液2 h与0 h葡萄糖比值(D/D0)降低(0.69±0.05比0.76±0.07,P = 0.033),腹膜超滤量(UF,ml)减少(2.29±0.50比4.58±1.64,P = 0.005),腹膜血管数量增加[(5.8±3.0)/HP比(1.6±0.5)/HP,P < 0.01]。尿毒症腹透组大鼠的溶质转运较尿毒症组大鼠进一步增高(D/Pcr: 0.89±0.05比0.78±0.05,P < 0.01;D/D0:0.47±0.09 比0.69±0.05, P < 0.01),UF(ml)减少(0.40±0.59比2.29±0.50,P = 0.005),腹膜血管数量增多[(16.7±1.2)/HP比(5.8±3.0)/HP,P < 0.01]。干预组大鼠使用sTie-2-Fc后,UF(ml)较尿毒症腹透组大鼠显著增加(1.56±0.48比0.40±0.59,P = 0.014),腹膜血管数量显著减少[(9.2±1.2)/HP比(16.7±1.2)/HP,P < 0.01],但两组大鼠的D/Pcr和D/D0差异均无统计学意义。 结论 sTie-2-Fc使尿毒症腹透大鼠腹膜血管新生减少,超滤增加,有利于保护腹膜结构和功能,可能是防治腹透后腹膜结构和功能改变的另一靶点。 相似文献
10.
人腹膜微血管内皮细胞的原代培养 总被引:1,自引:0,他引:1
目的分离和培养人腹膜微血管内皮细胞。方法取非炎症且非转移性肿瘤患者施行腹部外科手术的大网膜组织,经Ⅰ型胶原酶消化,两次筛网过滤、离心等步骤获取人微血管段,接种24 h待内皮细胞从微血管段爬出贴壁后,去除微血管段获得内皮细胞进行传代培养。组织形态学观察及免疫组化法鉴定所培养的细胞。结果运用此法进行的原代培养,血细胞在换液和传代过程中被去除,成纤维细胞污染被减至最少;组织形态学观察显示内皮细胞呈铺路石样生长,免疫组化鉴定表明Ⅷ因子阳性;证实所培养的细胞为人腹膜微血管内皮细胞,其纯度较高,生长状态良好。结论实验成功分离和培养了人腹膜微血管内皮细胞,为体外研究腹膜透析的病理生理学改变,提供了细胞来源。 相似文献