匹他伐他汀促进小鼠血管新生作用及其机制的研究

Pitavastatin enhances angiogenesis and perfusion in a murine model of limb ischemia

目的探讨匹他伐他汀对小鼠血管新生的促进作用及其可能的作用机制。方法建立野生型C3H/He小鼠下肢缺血模型并分为2组,缺血对照组,匹他伐他汀组。使用激光多普勒血流测定仪测定实验小鼠投药前、下肢缺血手术后双下肢血流。免疫荧光组化SP法计数缺血肢毛细血管数。免疫酶组化直接法计数缺血肢磷酸化蛋白激酶Akt(p-Akt)阳性细胞数。蛋白印迹杂交方法检测缺血肢血管内皮生长因子蛋白表达。镉还原Griess法测定实验结束后血清一氧化氮代谢产物含量。结果匹他伐他汀使实验小鼠术后缺血肢血流恢复明显,缺血肢与非缺血肢血流面积比明显增加;缺血肢毛细血管密度明显增加、p-Akt活性增加(p-Akt阳性细胞数明显增加);血中一氧化氮代谢产物含量明显增高;缺血肢血管内皮生长因子蛋白表达增强。结论匹他伐他汀有促进小鼠血管新生的作用。

Objective We investigated the effects of pitavastatin on angiogenesis and perfusion in C3 H/He mice with unilateral hind limb ischemia. Methods C3 H/He mice treated with saline （n = 15） or pitavastatin （1mg · kg^-1 · d^-1, n = 15）per garage for 1 week underwent unilateral hind limb ischemia surgery and were treated for another 5 weeks. Hind- limb blood flow was measured by Laser Doppler perfusion imager （LDPI, ischemic/nonischemic limb, % ） at baseline, immediately after ischemia and weekly thereafter for 5 weeks. Endpoints included local vessel counts by immunofluorescence, phospho-Akt positive cell counts by immunoenzyme histochemical technique, vascular endothelial growth factors （VEGFs） expression in ischemic limbs by Western blot and serum nitric oxide metabolite （NOx） by chrome deoxide Griess method. Restdts Lower extremity perfusion was significantly improved in pitavastatin treated mice vs. controls as measured by LDPI% at 1 week post ischemia and thereafter （P 〈 0. 05 ）. Pitavastatin treatment was associated with significantly increased capillary count E （47 ± 11 ） vs. （26 ± 14 ）/per highpower field （ × 200）, P 〈 0. 05 ）and greater percentage of phospho-Akt positive cells E （6 ± 1 ） vs. （2 ± 0）/per high-power field （ ×200）, P 〈0. 051 in ischemic limbs. Serum NOx [ （77.3±21.8） vs. （52. 1 ± 11.2 ） moL/L, P 〈 0. 05 ） and VEGF protein expression in ischemic limbs were also significantly increased in pitavastatin group than those in control group. Conclusions Pitavastatin enhances angiogenesis and perfusion in CsH/He mice with limb ischemia.