文章摘要
王芳,王改兰,朱莹,等.姜黄素介导的光动力疗法通过PI3K/AKT信号通路对PDGF-BB诱导的血管平滑肌细胞增殖的影响[J].中华物理医学与康复杂志,2024,46(5):393-400
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姜黄素介导的光动力疗法通过PI3K/AKT信号通路对PDGF-BB诱导的血管平滑肌细胞增殖的影响
  
DOI:10.3760/cma.j.issn.0254-1424.2024.05.002
中文关键词: 姜黄素  光动力  血小板衍生生长因子  血管平滑肌细胞  增殖
英文关键词: Curcumin  Photodynamic therapy  Platelet-derived growth factor BB  Vascular smooth muscle cells  Cell proliferation
基金项目:国家自然科学基金资助项目(81871853);重庆市自然科学基金资助项目(cstc2020jcyj-msxmX0484)
作者单位
王芳 重庆医科大学附属第一医院重庆医科大学附属第一医院康复医学科重庆 400016 
王改兰 重庆医科大学附属第一医院重庆医科大学附属第一医院康复医学科重庆 400016 
朱莹 重庆医科大学附属第一医院重庆医科大学附属第一医院康复医学科重庆 400016 
白定群 重庆医科大学附属第一医院重庆医科大学附属第一医院康复医学科重庆 400016 
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中文摘要:
      目的 观察姜黄素(CUR)介导的光动力疗法(PDT)通过磷酸酰肌醇3-激酶(PI3k)/丝苏氨酸蛋白激酶(AKT)信号通路对血小板衍生生长因子-BB(PDGF-BB)诱导的血管平滑肌细胞(VSMCs)增殖的影响及其可能的作用机制。 方法 分别选取大鼠胸主动脉平滑肌细胞系(A7R5)和小鼠主动脉平滑肌细胞系(MOVAS)进行平行实验,选择20 μg/L的PDGF-BB诱导VSMCs增殖。将培养好的VSMCs分为对照组、PDGF-BB组、姜黄素组、光动力组,其中光动力组再根据照射剂量的不同,又分为低剂量光动力组(照射60 s)、中剂量光动力组(照射120 s)、高剂量光动力组(照射180 s),共3个亚组。对照组不予任何刺激,PDGF-BB组饥饿过夜后加入含20 μg/L的PDGF-BB的新鲜培养基刺激24 h,姜黄素组先给予与PDGF-BB组相同的干预,然后换液加入含20 μmol/L CUR的新鲜培养基刺激6 h,光动力组先给予与姜黄素组相同的干预,再换液后采用425 nm激光对光动力组各亚组进行对应时长的激光照射。采用细胞活力检测法(CCK8)检测光动力组中不同激光剂量照射后,VSMCs的存活率;然后采用5-乙炔基-2′-脱氧尿苷(EDU)阳性标记率法检测4组细胞的增殖活性,采用流式细胞术对4组细胞进行细胞周期检测,采用Western blot法检测PI3K/AKT通路相关磷酸化蛋白和细胞增殖核蛋白(PCNA)的表达水平。 结果 经CCK8法检测,本研究选择120 s作为激光照射的最佳剂量。干预后,中、高剂量光动力组的细胞存活率与PDGF-BB组比较,差异均有统计学意义(P<0.01)。光动力组增殖的阳性细胞比例显著低于PDGF-BB组,差异有统计学意义(P<0.05)。光动力组细胞在G0/G1期的比例明显低于对照组(P<0.01),而在G2/M期的比例则明显高于对照组和PDGF-BB组(P<0.01)。光动力组增殖细胞核抗原(PCNA)、PI3K/AKT通路磷酸化蛋白的表达较PDGF-BB组显著下降,差异均有统计学意义(P<0.01)。 结论 CUR介导的PDT可抑制PDGF-BB诱导下的血管平滑肌细胞的增殖,其机制可能与CUR介导的PDT可下调PI3K/AKT通路相关磷酸化蛋白表达水平有关。
英文摘要:
      Objective To observe any effect of photodynamic therapy (PDT) mediated by curcumin (CUR) on the proliferation of vascular smooth muscle cells (VSMCs) induced by platelet-derived growth factor-BB (PDGF-BB) through the phosphatidylinositol 3-kinase (PI3k)/serine threonine protein kinase (AKT) signaling pathway and explore its possible mechanism. Methods Twenty μg/L of PDGF-BB was used to induce proliferation of VSMCs in A7R5 rat aortic smooth muscle cells and mouse aortic smooth muscle cells. Well-cultured VSMCs were randomly divided into a normal group, a PDGF-BB group, a curcumin group, and a low-dose PDT group (irradiated for 60s), a medium-dose PDT group (irradiated for 120s), and a high-dose PDT group (irradiated for 180s). The normal group was not given any intervention, and the PDGF-BB group was starved overnight and then stimulated for 24h by adding fresh medium containing 20μg/L of PDGF-BB. The curcumin group was first given the same intervention as the PDGF-BB group, and then placed in a medium containing 20μmol/L of curcumin for 6h. The PDT groups were first given the same intervention as the curcumin group, and then irradiated using a 425nm laser after removal of the medium. A CCK8 cell counting kit was used to detect the survival rate of VSMCs in the PDT groups after irradiation with the different laser doses. VSMC proliferation was detected using 5-ethynyl-2′-deoxyuridine (EDU) positive labeling. The cell cycle was observed using flow cytometry, and the expression levels of related phosphorylated proteins of the PI3K/AKT pathway and proliferating cell nuclear antigen were detected using western blotting. Results The CCK-8 counts suggested that 120s was the optimum irradiation dose. The cell survival rates in the medium-dose and high-dose PDT groups were significantly different from the PDGF-BB group′s average. The proportion of proliferating cells in the PDT groups was significantly lower than in the PDGF-BB group. The proportion of cells in the G0 or G1 phase was significantly lower in the PDT group than in the normal group, while that in the G2 or M phase was significantly higher than in the normal and PDGF-BB groups. The expression of proliferating cell nuclear antigen and the phosphorylated proteins of the PI3K/AKT pathway had decreased significantly in the PDT group compared with the PDGF-BB group. Conclusion CUR-mediated PDT inhibits PDGF-BB-induced proliferation of vascular smooth muscle cells, probably by down-regulating the expression of phosphorylated proteins related to the PI3K/AKT pathway.
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