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目的观察晚期糖基化终产物(advanced glycation end products,AGEs)对足细胞内肾素-血管紧张素系统(renin-angiotensin system,RAS)的影响及作用机制。方法不同浓度的AGEs干预小鼠足细胞24 h,分别检测肾素(renin)、血管紧张素原(renin-angiotensin system,AGT)、血管紧张素Ⅱ1型、2型受体(AT1R、AT2R)的表达,血管紧张素转换酶(angiotensin-converting enzyme,ACE)的活性和血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)的浓度,观察蛋白激酶B(Akt)的磷酸化,然后分别加入磷酸肌醇3激酶抑制剂LY294002、losartan、captopril和chymastatin,观察足细胞粘附性的变化。结果与对照组相比,AGEs(80μg/mL)明显上调AGT和AT1R的表达[(183.0±19.0)%vs 100%,(179.0±17.0)%vs100%,P<0.05],裂解液中ACE活性明显增加[(142.8±10.3)U/μg vs(85.0±9.2)U/μg,P<0.05],细胞上清中AngⅡ的浓度明显增加[(11.2±0.8)pg/mL vs(7.0±0.7)pg/mL,P<0.05];Akt的磷酸化上调100%(P<0.05),而LY294002可减轻AGEs介导的足细胞内RAS的激活;与AGEs组相比,LY294002可改善AGEs介导的足细胞粘附性的下降[(82±13)%vs(40±12)%;(78±14)%vs(42±13)%,P<0.05]。结论 AGEs可能通过磷酸肌醇3激酶途径激活足细胞内的RAS,降低足细胞粘附性。
Objective To investigate the effect and mechanism of advanced glycation end products (AGEs) on renin-angiotensin system (RAS) in podocytes. Methods Different concentrations of AGEs were applied to the podocytes of mice for 24 hours to detect the expression of renin, renin-angiotensin system (AGT), angiotensin Ⅱ type 1 and type 2 receptor (AT1R, AT2R) , The activity of angiotensin-converting enzyme (ACE) and the concentration of angiotensinⅡ (AngⅡ) were observed. The phosphorylation of protein kinase B (Akt) was observed, and then phosphoinositide 3 kinase inhibition The agents LY294002, losartan, captopril and chymastatin were used to observe the changes of podocyte adhesion. Results Compared with the control group, AGEs (80μg / mL) significantly upregulated the expression of AGT and AT1R [(183.0 ± 19.0)% vs 100%, (179.0 ± 17.0)% vs100%, P <0.05] (142.8 ± 10.3) U / μg vs (85.0 ± 9.2) U / μg, P <0.05]. The concentration of AngⅡ in the supernatant of the cells increased significantly [(11.2 ± 0.8) pg / mL vs pg / mL, P <0.05]. The phosphorylation of Akt was up-regulated by 100% (P <0.05), while LY294002 reduced the AGEs-mediated activation of RAS in podocytes. Compared with AGEs, LY294002 improved AGEs-mediated Podocyte adhesion decreased [(82 ± 13)% vs (40 ± 12)%; (78 ± 14)% vs (42 ± 13)%, P <0.05]. Conclusion AGEs may activate RAS in podocytes via phosphoinositide 3 kinase pathway and decrease podocyte adhesion.