Department of Internal Medicine and Institute of Hypertension, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, People's Republic of China.
Hum Gene Ther. 2012 Jul;23(7):688-99. doi: 10.1089/hum.2011.135. Epub 2012 Mar 29.
The cytochrome P450 epoxygenase, CYP2J2, converts arachidonic acid to four regioisomeric epoxyeicosatrienoic acids (EETs), which are highly abundant in the kidney and considered renoprotective. Accumulating evidence suggests that EETs are important in regulating renal and cardiovascular function. Further, EETs have been confirmed to exert diverse biological activities including potent vasodilation; fibrinolytic properties; and antiinflammatory, antiapoptotic, and mitogenic effects. In the current study, we investigated the effects of overexpression of CYP2J2 via recombinant adeno-associated virus (rAAV) in protection against renal damage in a rat 5/6 nephrectomy (5/6-Nx) model of chronic renal failure. The rAAV-CYP2J2 gene delivery in vivo increased EET generation; attenuated the rise in blood pressure; and reduced the levels of proteinuria, serum creatinine, and blood urea nitrogen. Morphological analysis indicated that rAAV-CYP2J2 gene delivery reduced 5/6 nephrectomy-induced glomerular sclerosis, tubular dilatation, luminal protein cast formation, and tubulointerstitial fibrosis. rAAV-CYP2J2 gene delivery also significantly lowered collagen I and IV deposition, as well as renal cell apoptosis detected by TUNEL staining, caspase-3 activity, and the loss of mitochondrial membrane potential (ΔΨ(m)). Furthermore, rAAV-CYP2J2 gene delivery regulated the level of protein expression including transforming growth factor (TGF)-β(1)/SMADs; matrix metalloproteinases (MMPs); mitogen-activated protein kinases (MAPKs); and apoptosis-related proteins Bax, Bcl-2, and Bcl-x(L). Together, these findings demonstrated that rAAV-CYP2J2 gene delivery can protect remnant kidney against renal injury in 5/6-Nx rats by inhibiting apoptosis and fibrosis via regulation of protein expression including TGF-β(1)/SMADs, MMPs, MAPKs, and apoptosis-related proteins.
细胞色素 P450 环氧合酶，CYP2J2，将花生四烯酸转化为四个区域异构的环氧二十碳三烯酸（EETs），这些物质在肾脏中含量丰富，被认为具有肾保护作用。越来越多的证据表明，EETs 在调节肾脏和心血管功能方面起着重要作用。此外，EETs 已被证实具有多种生物学活性，包括强效血管舒张作用、纤维蛋白溶解特性以及抗炎、抗凋亡和有丝分裂作用。在本研究中，我们通过重组腺相关病毒（rAAV）过表达 CYP2J2 研究了其在慢性肾衰竭大鼠 5/6 肾切除（5/6-Nx）模型中保护肾脏免受损伤的作用。体内 rAAV-CYP2J2 基因传递增加了 EET 的产生；减轻了血压升高；并降低了蛋白尿、血清肌酐和血尿素氮的水平。形态学分析表明，rAAV-CYP2J2 基因传递减少了 5/6 肾切除诱导的肾小球硬化、肾小管扩张、管腔蛋白铸型形成和肾小管间质纤维化。rAAV-CYP2J2 基因传递还显著降低了胶原 I 和 IV 的沉积以及通过 TUNEL 染色、caspase-3 活性和线粒体膜电位（ΔΨ(m)）丧失检测到的肾细胞凋亡。此外，rAAV-CYP2J2 基因传递调节了蛋白表达水平，包括转化生长因子（TGF）-β(1)/SMADs；基质金属蛋白酶（MMPs）；丝裂原活化蛋白激酶（MAPKs）；和凋亡相关蛋白 Bax、Bcl-2 和 Bcl-x(L)。综上所述，这些发现表明 rAAV-CYP2J2 基因传递可以通过调节 TGF-β(1)/SMADs、MMPs、MAPKs 和凋亡相关蛋白等蛋白表达来抑制凋亡和纤维化，从而保护 5/6-Nx 大鼠残肾免受肾损伤。
