Impaired endothelium-mediated vasodilation and/or increased sensitivity to vasoconstrictors lead to vascular smooth muscle cell (VSMC) dysfunction in individuals with diabetes. Diabetic nephropathy is associated with a considerably higher risk of cardiovascular disease and death than their nondiabetic counterparts. We studied the activity of Cullin 3 RING ubiquitin ligase (CRL3) and its substrates in mice using an intraperitoneal injection of streptozotocin (STZ) and db/db mice. The levels of CRL3 adaptors, including Kelch-like 2/3 (KLHL2/3) and Rho-related BTB domain-containing protein 1, were significantly decreased in the aortic tissues and heart of the STZ group, whereas the levels of Cullin 3 (CUL3) and its neddylated derivatives were substantially increased. Decreased KLHL3 expression and significantly increased expression of NEDD8 conjugates were observed in the kidneys of db/db mice. The neddylation inhibitor MLN4924 decreased the degradation of KLHL2/KLHL3 under high-glucose conditions with/without insulin, and transfection with KLHL2 promoted the degradation of its substrates with-no-lysine (WNK) kinases. Increased abundance of WNK3, RhoA/ROCK activity and phosphodiesterase 5 enhanced the sensibility to vasoconstrictors and impaired vasodilation. Moreover, WNK3 localized in VSMCs undergoing cell division, and high-glucose medium increased WNK3 signaling in VSMCs undergoing mitosis, which might explain the increased thickness of aortic tissues in subjects with diabetes. Increases in WNK4 abundance resulted in increased sodium reabsorption in the distal renal tubules. Thus, KLHL2/RhoBTB1/KLHL3 inactivation in the aortic tissues and kidney is a result of excessive activation of neddylation in hyperglycemia and hyperinsulinemia, which affects vascular tone and sodium reabsorption.

受损的内皮介导的血管舒张和/或对血管收缩剂的敏感性增加导致糖尿病患者的血管平滑肌细胞 (VSMC) 功能障碍。与非糖尿病肾病相比，糖尿病肾病与心血管疾病和死亡风险显着升高相关。我们使用链脲佐菌素 (STZ) 和 db/db 小鼠腹腔注射，研究了 Cullin 3 RING 泛素连接酶 (CRL3) 及其底物在小鼠中的活性。在STZ组的主动脉组织和心脏中，CRL3接头的水平，包括Kelch样2/3（KLHL2/3）和Rho相关的BTB结构域蛋白1，显着降低，而Cullin 3的水平（ CUL3) 及其 neddylated 衍生物显着增加。在 db/db 小鼠的肾脏中观察到 KLHL3 表达降低和 NEDD8 缀合物表达显着增加。neddylation 抑制剂 MLN4924 在有/无胰岛素的高糖条件下减少了 KLHL2/KLHL3 的降解，并且用 KLHL2 转染促进了其底物的降解，其中有无赖氨酸 (WNK) 激酶。WNK3、RhoA/ROCK 活性和磷酸二酯酶 5 丰度的增加增强了对血管收缩剂的敏感性和受损的血管舒张。此外，WNK3 定位于进行细胞分裂的 VSMC 中，高糖培养基增加了进行有丝分裂的 VSMC 中的 WNK3 信号传导，这可能解释了糖尿病患者主动脉组织厚度增加的原因。WNK4 丰度的增加导致远端肾小管的钠重吸收增加。因此，