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Induced elastin regeneration by chronically activated smooth muscle cells for targeted aneurysm repair.

机译:慢性活化的平滑肌细胞诱导弹性蛋白再生,用于靶向动脉瘤修复。

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摘要

Elastin breakdown in vascular aneurysms is mediated by cytokines such as tumor necrosis factor alpha (TNF-alpha, which induces vascular smooth muscle cell (SMC) activation and regulates their deposition of matrix. We previously demonstrated that exogenous supplementation with TGF-beta1 (1 ng ml(-1)) and hyaluronan oligomers (0.786 kDa, 0.2 microg ml(-1)) cues the upregulation of elastin matrix synthesis by healthy cultured SMCs. Here, we determine whether these cues likewise enhance elastin matrix synthesis and assembly by TNF-alpha-stimulated SMCs, while restoring their healthy phenotype. Adult rat aortic SMCs were treated with TNF-alpha alone or together with TGF-beta1/hyaluronan oligomeric cues and the release of inflammatory markers were monitored during over a 21 day culture. Biochemical analysis was used to quantify cell proliferation, matrix protein synthesis and cross-linking efficiency, while immunofluorescence and electron microscopy were used to analyze the elastin matrix quality. It was observed that SMC activation with TNF-alpha (10 ng ml(-1)) induced matrix calcification and promoted production of elastolytic MMP-2 and MMP-9. However, these effects were attenuated by the addition of TGF-beta1 and HA oligomer cues to TNF-alpha-stimulated cultures, which also enhanced tropoelastin and collagen production, improved elastin matrix yield and cross-linking, promoted elastin fiber formation and suppressed elastase activity, although the release of MMP-2 and MMP-9 was not affected. Overall, the results suggest that TGF-beta1 and HA oligomers are potentially useful in suppressing SMC activation and inducing regenerative elastin repair within aneurysms.
机译:血管动脉瘤中的弹性蛋白分解由细胞因子(如肿瘤坏死因子α(TNF-alpha)介导(诱导血管平滑肌细胞(SMC)活化并调节它们对基质的沉积。我们以前表现出与TGF-BETA1的外源补充(1 ng ml(-1))和透明质酸寡聚体(0.786kda,0.2 microg ml(-1))提示通过健康培养的smc来提高Elastin基质合成的升高。在这里,我们确定这些提示是否同样增强了弹性蛋白基质合成和通过tnf- α刺激的SMCs,同时恢复健康的表型。成年大鼠主动脉SMC单独使用TNF-α处理,或与TGF-β1/透明质酸寡核苷酸寡核苷酸和炎症标志物的释放在21天培养过程中被监测。生物化学分析用于量化细胞增殖,基质蛋白质合成和交联效率,而免疫荧光和电子显微镜用于分析ELASTIN基质曲程ality。观察到具有TNF-α的SMC活化(10ng ml(-1))诱导的基质钙化并促进弹性溶解MMP-2和MMP-9的产生。然而,通过向TNF-α-刺激的培养物加入TGF-β1和HA低聚物提示来衰减这些效果,这也增强了Tropoelastin和胶原蛋白的产生,改善了弹性蛋白基质产量和交联,促进了Elastin纤维形成和抑制弹性蛋白酶活性,尽管MMP-2和MMP-9的释放不受影响。总的来说,结果表明TGF-β1和HA低聚体可能可用于抑制SMC活化和诱导动脉瘤内的再生弹性蛋白修复。

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