Schmidt Luke, Carrillo-Sepulveda Maria Alicia
Department of Physiology, Georgia Regents University, 1120 Fifteenth St., Augusta, GA, 30912, USA.
Pflugers Arch. 2015 Nov;467(11):2361-74. doi: 10.1007/s00424-015-1688-2. Epub 2015 Jan 21.
Increased vascular smooth muscle cell (VSMC) contraction is an early and critical contributor to the pathogenesis of vascular dysfunction in diabetes; however, knowledge regarding the underlying mechanisms is scarce. Toll-like receptor 2 (TLR2), a well-known component of the innate immunity, is expressed in VSMC and recently has been identified to be systemically activated in diabetes. Whether TLR2 is locally activated in the diabetic blood vessels and have effect on contraction is not known. In the current study, we examined the role of TLR2 in increased vascular contraction in diabetes. Utilizing rat model of type 1 diabetes (induced by streptozotocin (STZ)), we demonstrated that aortas from STZ-diabetic rats exhibit increased expression of TLR2 and its adaptor protein, myeloid differentiation primary response 88 (MyD88), as well as enhanced protein-protein interaction between TLR2 and MyD88, suggesting a TLR2 signaling activation. Blockade of TLR2 in intact aortas using anti-TLR2 antibody attenuated increased vascular contraction in STZ-diabetic rat as assessed by wire myograph. Activation of TLR2 by specific ligand in primary aortic VSMC cultures triggered activation of RhoA which was exacerbated in cells from STZ-diabetic rats than control rats. Activation of RhoA was accompanied by phosphorylation and therefore activation of its downstream targets myosin phosphatase target subunit I and myosin light chain (markers of VSMC contraction). Taken together, these results provide evidence for the role of TLR2 in increased contraction in diabetic blood vessels that involves RhoA signaling. Thus, targeting vascular TLR2 offers a promising drug target to treat vascular dysfunction in diabetes.
血管平滑肌细胞(VSMC)收缩增强是糖尿病血管功能障碍发病机制中的早期关键因素;然而,关于其潜在机制的了解却很少。Toll样受体2(TLR2)是固有免疫的一个知名组成部分,在VSMC中表达,最近已被确定在糖尿病中被全身激活。TLR2在糖尿病血管中是否局部激活以及是否对收缩有影响尚不清楚。在本研究中,我们研究了TLR2在糖尿病血管收缩增强中的作用。利用1型糖尿病大鼠模型(由链脲佐菌素(STZ)诱导),我们证明STZ糖尿病大鼠的主动脉中TLR2及其衔接蛋白髓样分化初级反应蛋白88(MyD88)的表达增加,以及TLR2与MyD88之间的蛋白质-蛋白质相互作用增强,提示TLR2信号激活。用抗TLR2抗体阻断完整主动脉中的TLR2,可减弱STZ糖尿病大鼠血管收缩的增强,这是通过线肌张力测定法评估的。在原代主动脉VSMC培养物中,特异性配体激活TLR2可触发RhoA的激活,与对照大鼠相比,STZ糖尿病大鼠细胞中的这种激活更为明显。RhoA的激活伴随着磷酸化,因此激活了其下游靶点肌球蛋白磷酸酶靶向亚基I和肌球蛋白轻链(VSMC收缩的标志物)。综上所述,这些结果为TLR2在糖尿病血管收缩增强中的作用提供了证据,该作用涉及RhoA信号传导。因此,靶向血管TLR2为治疗糖尿病血管功能障碍提供了一个有前景的药物靶点。