Klotho蛋白对血管平滑肌细胞钙化的影响及其机制研究

发布时间：2018-05-02 13:15

  本文选题：Klotho蛋白 + 血管平滑肌细胞　； 参考：《南京医科大学》2015年硕士论文

【摘要】：目的:探讨Klotho蛋白对高磷诱导的大鼠血管平滑肌细胞成骨样转化、钙化作用的影响及可能的机制。方法:体外培养大鼠的血管平滑肌细胞,用10mmol/L的β甘油磷酸诱导其钙化,在培养基中加入不同浓度的重组小鼠Klotho蛋白(rm Klotho),并加或不加Wnt/β-catenin通路激动剂Li Cl。干预12天后,茜素红染色观察钙盐沉积,分光光度计测定细胞外基质钙含量。Western印记法分别检测骨形态形成蛋白2(BMP2)、Runt相关转录因子2(Runx2)、β连环蛋白(β-catenin)的表达,免疫荧光法检测α-SMA的表达。结果:β甘油磷酸诱导大鼠血管平滑肌细胞12天后,茜素红染色钙盐沉积明显,细胞外基质钙含量增加,细胞BMP2,Runx2,β-catenin表达较正常对照组上调,α-SMA表达较正常对照下调。而在加rm Klotho蛋白干预后,细胞钙化减轻,BMP2,Runx2,β-catenin表达均下调,α-SMA表达上调。但是,Klotho干预的同时加Li Cl,则BMP2、Runx2及β-catenin表达较Klotho干预组上调。结论:Klotho蛋白能减轻高磷诱导的血管平滑肌细胞的成骨细胞转化、钙化作用,且这一保护作用可能与Wnt/β-catenin通路的抑制有关。目的:探讨容量超负荷(FO)水平在评价危重病患者预后中的意义。方法:对2013年3月至2013年10月在南京医科大学第一附属医院综合ICU住院的100例危重患者,评估其病程中的FO水平,分析FO与APACHEⅡ评分及危重症患者的住院病死率、ICU治疗时间、机械通气时间等主要预后指标的关系。结果:100例患者存活80例,死亡20例,病死率为20.0%。其中死亡组APACHEⅡ评分值和FO水平均高于存活组(P0.01)。APACHEⅡ评分、ICU治疗时间、机械通气时间及病死率随着FO水平升高而升高。结论:FO和APACHEⅡ评分的升高对危重症患者的存活、机械通气时间、ICU治疗时间等主要预后指标的预测有重要意义。
[Abstract]:Aim: to investigate the effect of Klotho protein on osteoblast-like transformation and calcification of rat vascular smooth muscle cells induced by high phosphorus and its possible mechanism. Methods: rat vascular smooth muscle cells were cultured in vitro. Calcification was induced by 尾 -glycerophosphoric acid (尾 -glycerophosphoric acid) of 10mmol/L. Different concentrations of recombinant mouse Klotho protein rm Klothotrol were added to the medium and Wnt/ 尾 -catenin pathway agonist LiCl was added or not. After 12 days of intervention, calcium deposition was observed by alizarin red staining. The expression of bone morphogenetic protein 2BMP2U Runt, 尾 -catenin and 伪 -SMA were detected by spectrophotometer and immunofluorescence respectively. Results: after 12 days of 尾 -glycerophosphoric acid induced vascular smooth muscle cells, alizarin red staining calcium salt deposition was obvious, extracellular matrix calcium content increased, BMP2 + Runx2, 尾 -catenin expression was higher than normal control group, 伪 -SMA expression was down-regulated. However, after the addition of RM Klotho protein, calcification decreased the expression of BMP2nRunx2 and 尾 -catenin, and up-regulated the expression of 伪 -SMA. However, the expression of BMP2 + Runx2 and 尾 -catenin in Klotho treated group was higher than that in Klotho group. Conclusion Klotho protein can attenuate the osteoblast transformation and calcification of vascular smooth muscle cells induced by high phosphorus, and this protective effect may be related to the inhibition of Wnt/ 尾 -catenin pathway. Objective: to evaluate the prognostic value of volume overload FOO level in critically ill patients. Methods: from March 2013 to October 2013, 100 critically ill patients with ICU in the first affiliated Hospital of Nanjing Medical University were enrolled. The FO levels in the course of the disease were evaluated, and the scores of FO and APACHE 鈪，

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