低蛋白饮食加α-酮酸对减轻腺嘌呤慢性肾功能衰竭大鼠模型血管钙化的作用

目的:观察低蛋白饮食加α-酮酸(LPD+α-KA)这一终末期肾功能衰竭(ESRD)病人营养治疗方法对减轻血管钙化的作用.方法:将18只大鼠随机分为正常对照组、正常蛋白-慢性肾功能衰竭(NPD-CRF)组和LPD+α-KA-CRF组.两组CRF大鼠饲料中含0.75%腺嘌呤.于喂养6周后留取血标本和腹主动脉,测定血钙、磷等...     

舒洛地特抑制单侧输尿管梗阻小鼠肾间质纤维化

目的 探讨舒洛地特对单侧输尿管梗阻(UUO)小鼠肾脏间质纤维化的作用,及其对肾小管上皮-间充质转化(EMT)和肾组织Jagged-1、Noteh1表达的影响.方法 雄性CD1小鼠60只随机分为:假手术组(A组),仅行腹膜切开后缝合;对照组(B组),行单侧输尿管结扎,术后予生理盐水0.1ml/d皮下注射;低剂量治疗组(C...     

Inhibitory effect of vascular endothelial growth factor on transforming growth factor β1-induced epithelial-mesenchymal transition of HK2 cells and its relationship with connective tissue growth factor and PI3K-Akt pathway

Objective To examine the relationship of the inhibitory effect of vascular endothelial growth factor(VEGF) on epithelial-mesenchymal transition (EMT) induced by TGF-β1 in HK2 cells with the expression of connective tissue growth factor (CTGF) and PI3K-Akt pathway. Methods The cultured HK2 cells were divided into the following groups: normal control group, TGF-β1 (5 μg/L) group, VEGF (100 μg/L) group, TGF-β1 plus VEGF group. LY294002 (25 μmol/L), the blocker of PI3K-Akt pathway, was added to each of above-mentioned groups for the second part of the study, α-smooth muscle actin (α-SMA) and E-cadherin expressions of HK2 cells were assessed with double-stain immunocytochemistry method. The mRNA and protein expressions of α-SMA and CTGF of cells were assessed with RT-PCR and Western blot. The expressions of fibronectin (FN) and collagen Ⅰ (Col Ⅰ) in medium were assessed with ELISA. Results The expressions of α-SMA and CTGF significantly increased in HK2 cells treated with TGF-β1 compared with those in normal control (P<0.05), while significantly decreased in cells co-treated with TGF-β1 and VEGF compared with those treated with TGF-β1 alone (P<0.05, respectively). The expression of E-cadherin was exactly opposite to that of α-SMA. When LY294002 was added to TGF-β1 and VEGF co-treated cells, the expressions of α-SMA, CTGF, FN and Col Ⅰ were markedly up-regulated, when compared with those without LY294002 treatment (P<0.05). Conclusion Inhibitory effect of VEGF on TGF-β1-induced EMT of HK2 ceils in vitro may be related to down-regulation of CTGF expression and reduction of FN and Col Ⅰ, which may be partly dependent on PI3K-Akt pathway.     

水盐限制联合利尿治疗CAPD患者容量超负荷的对照研究

目的观察水盐限制联合利尿治疗对CAPD患者容量超负荷的疗效。方法第四军医大学西京医院血液净化中心CAPD52例符合条件的患者随机分为A、B两组,A组单纯水盐限制,B组在水盐限制的基础上,加服速尿120～240mg/日,分2次口服,检测治疗前及治疗1月后体重、平均动脉压、尿量、超滤量、浮肿程度、心胸比例等变化。结果①较治疗前相比,A组的体重、平均动脉压、心胸比例均有所前下降,但无统计学意义,平均尿量和平均超滤量无变化。B组的体重、平均动脉压、平均尿量、心胸比例均较治疗前明显下降(P<0.05),其中体重和平均尿量在两组间差异具有显著性(P<0.05);②两组的水肿程度均有不同程度的下降,B组更为明显。③A组总有效率为53.8%,B组总有效率为84.7%。两组相比总有效率差异有统计学意义(P<0.05)。结论单纯限制水盐摄入或联用利尿治疗均能改善CAPD患者容量超负荷状况,但后者的效果更明显,水盐限制基础上加用利尿是治疗CAPD患者容量超负荷的安全有效的方法。     

连续性血液净化对心脏术后多脏器功能障碍综合征患者炎性介质和氧化应激的影响

目的 前瞻性观察连续性血液净化(CBP)对心脏术后多脏器功能障碍综合征(MODS)存活者与死亡者炎性介质和氧化应激指标的影响.方法 16例心脏术后MODS伴ARF的患者使用HF700型聚砜膜血液滤过器行CBP治疗,8例存活(A组),8例死亡(B组);分别在治疗前(0h)和治疗后2、6、12、24和48h取血检测两组患者血清TNF-α、IL-6、IL-8、IL-4、丙二醛(MDA)、超氧化物歧化酶(SOD)和总抗氧化能力(TAC)等指标的变化.结果 CBP治疗2h后A组促炎因子TNF-α、IL-6迅速下降,24h后IL-8明显下降,而死亡组治疗过程中上述炎性介质水平较治疗前无明显变化;两组抗炎因子IL-4在CBP治疗过程中较治疗前有所下降,但无明显统计意义.A组TAC在CBP治疗6h后明显下降,MDA在12h后明显下降,SOD在24h后明显下降;B组SOD、TAC在治疗2h后即开始明显下降,MDA在48h的治疗过程中持续保持高水平,较治疗前无明显下降.结论 CBP有助于改善MODS患者促炎因子与抗炎因子的平衡,减轻炎性损伤.CBP能改善MODS患者的氧化应激,减轻氧化损伤.在CBP治疗过程中,TNF-α、IL-6、IL-8、MDA浓度持续高水平及SOD早期下降者预后差;动态监测上述指标可能有助于判断心脏术后MODS患者的预后.     

Inhibitory effect of vascular endothelial growth factor on transforming growth factor β1-induced epithelial-mesenchymal transition of HK2 cells and its relationship with connective tissue growth factor and PI3K-Akt pathway

Objective To examine the relationship of the inhibitory effect of vascular endothelial growth factor(VEGF) on epithelial-mesenchymal transition (EMT) induced by TGF-β1 in HK2 cells with the expression of connective tissue growth factor (CTGF) and PI3K-Akt pathway. Methods The cultured HK2 cells were divided into the following groups: normal control group, TGF-β1 (5 μg/L) group, VEGF (100 μg/L) group, TGF-β1 plus VEGF group. LY294002 (25 μmol/L), the blocker of PI3K-Akt pathway, was added to each of above-mentioned groups for the second part of the study, α-smooth muscle actin (α-SMA) and E-cadherin expressions of HK2 cells were assessed with double-stain immunocytochemistry method. The mRNA and protein expressions of α-SMA and CTGF of cells were assessed with RT-PCR and Western blot. The expressions of fibronectin (FN) and collagen Ⅰ (Col Ⅰ) in medium were assessed with ELISA. Results The expressions of α-SMA and CTGF significantly increased in HK2 cells treated with TGF-β1 compared with those in normal control (P<0.05), while significantly decreased in cells co-treated with TGF-β1 and VEGF compared with those treated with TGF-β1 alone (P<0.05, respectively). The expression of E-cadherin was exactly opposite to that of α-SMA. When LY294002 was added to TGF-β1 and VEGF co-treated cells, the expressions of α-SMA, CTGF, FN and Col Ⅰ were markedly up-regulated, when compared with those without LY294002 treatment (P<0.05). Conclusion Inhibitory effect of VEGF on TGF-β1-induced EMT of HK2 ceils in vitro may be related to down-regulation of CTGF expression and reduction of FN and Col Ⅰ, which may be partly dependent on PI3K-Akt pathway.     

氧化应激与急性肾衰及血液净化

本文简要叙述了氧化应激与急性肾衰竭、血液净化之间的相互影响与作用.     

心脏术后发生MODS伴ARF时机体氧化应激状态的初步研究

目的初步观察经体外循环心脏术后多脏器功能障碍综合征(MODS)伴急性肾功能衰竭(ARF)患者的氧化应激水平,为临床治疗提供理论基础。方法测定16例经体外循环心脏术后MODS伴ARF患者及16例健康人的血清丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(TAC),同时对患者进行MODS评分,并将评分与氧化应激参数进行相关性分析。结果116例患者,8例存活,8例死亡(病死率50%);2与健康对照组相比,MODS伴ARF组在性别、年龄方面无显著性差异,TAC,SOD代偿性升高,但GSH-Px明显下降,MDA显著升高。3MDA同MODS评分呈正相关;GSH-Px同MODS评分呈负相关。SOD、TAC同评分无明显相关性。结论1经体外循环心脏手术后MODS伴ARF的患者发生了氧化应激。2监测MDA和GSH-Px有助于判断病情。3在MODS伴ARF的治疗中,有必要采用各种治疗措施恢复机体的氧化抗氧化。     

血管内皮生长因子对肾小管上皮-间充质细胞转化的作用及其与骨形成蛋白-7、分化抑制因子表达的关系

目的探讨血管内皮生长因子(VEGF)对肾小管上皮-间充质细胞转化(EMT)的作用及其与骨形成蛋白-7(BMP-7)、分化抑制因子(Id)2、Id3表达的关系。方法体外培养的人肾小管上皮细胞(HK-2)经转化生长因子-β1(TGF-β1,5ng/ml)与不同浓度VEGF165(0.1、1、10、100ng/ml)共同作用,或TGF-β1(5ng/ml)与血管内皮生长因子受体-1(VEGFR1)抗体(10μg/ml)共同作用48h后,免疫组织化学双染方法检测细胞α-平滑肌肌动蛋白(α-SMA)、E-钙黏素表达,实时荧光定量PCR法、Westernblot法检测细胞α-SMA、BMP-7、Id2和Id3的表达。TGF-β1(5ng/ml)、VEGF165(100ng/ml)与激活素受体样激酶-6/Fc嵌合体(Alk6/FcChimera)(2μg/ml,中和内源性BMP-7)共同作用48h后,Westernblot法检测细胞α-SMA、Id2的表达情况。结果TGF-β1作用后,α-SMA表达比正常对照显著增强(P<0.05),E-钙黏素、BMP-7、Id2及Id3的mRNA及蛋白质表达均显著减弱(P<0.05)。VEGF165以浓度依赖方式增强BMP-7及Id2蛋白表达,而显著降低TGF-β1上调α-SMA表达的作用(P<0.05)。加入VEGFR1抗体(10μg/ml)后,TGF-β1上调α-SMA表达的作用显著增强(P<0.05),而E-钙黏素、BMP-7、Id2表达显著减弱(P<0.05)。TGF-β1 VEGF165 Alk6/FcChimera共同作用后α-SMA蛋白表达比TGF-β1 VEGF165共同作用后显著增强(P<0.05),而Id2表达差异无显著性。结论VEGF165可抑制TGF-β1诱导HK-2细胞发生EMT;其机制可能与BMP-7和Id2表达上调有关,而与Id3表达变化无关。Id2表达增强可能与VEGF165的直接作用有关,而与BMP-7无关。     

四种气管上皮细胞分离培养方法的比较研究

目的：比较四种不同的方法在培养气管上皮细胞中的异同，改进其培养技术，为组织工程气管提供种子细胞。方法：用两步酶消化法（使用0.1％ Dispase 4℃消化18h后，用0．25％的胰酶37℃消化5min）、Dispase冷消化法（用0．1％Dispase 4℃消化18h）、胰酶温消化法（0．25％的胰酶37℃消化10min）和机械刮刷法四种方法分离、培养兔气管上皮细胞，测定分离细胞的数量和纯度。结果：两步酶消化法、Dispase冷消化法较其他两种方法得到的气管上皮细胞纯度高，细胞状态好；Dispase冷消化法得到细胞数量较其他方法少，其余三种方法得到的细胞数统计学上不存在差异。结论：两步酶消化法较其他方法所得细胞数量多、纯度高、细胞成活率高，是一种较好的气管上皮细胞体外分离培养方法。