论文部分内容阅读
目的:观察生胃酮对大鼠颈动脉损伤后新生内膜过度增殖的影响。方法:采用组织贴块法培养大鼠主动脉平滑肌细胞,Western blot检测生胃酮对大鼠主动脉平滑肌细胞缝隙连接蛋白43表达的影响。采用球囊损伤建立大鼠颈动脉损伤模型,对照组给予生理盐水2 mL、生胃酮组给予生胃酮3 mg/kg腹腔注射,1次/d,HE染色观察新生内膜形成情况,免疫荧光染色检测新生内膜中缝隙连接蛋白43的表达,伊文思蓝-DAPI双染色以评价新生内膜形成情况。结果:所培养的细胞抗SMα-actin免疫荧光染色阳性。50μmol/L生胃酮处理平滑肌细胞24 h后对缝隙连接蛋白43的表达无影响(0.85±0.06vs0.83±0.03,n=3,P>0.05)。球囊损伤大鼠颈动脉14 d后可见血管管腔狭窄、新生内膜增生明显。免疫荧光染色显示形成的新生内膜中缝隙连接蛋白43表达丰富。经生胃酮干预后新生内膜增生明显减轻,新生内膜细胞计数显著低于对照组(89±28.40vs236±15.04,n=5,P<0.01)。结论:缝隙连接在血管损伤后新生内膜形成过程中扮演重要角色,而缝隙连接阻断剂生胃酮能抑制新生内膜过度增生。
Objective: To observe the effect of carbenoxolone on neointimal hyperproliferation after carotid artery injury in rats. Methods: Rat aorta smooth muscle cells (VSMCs) were cultured by tissue patch method and the effect of carbenoxolone on the expression of connexin43 in rat aortic smooth muscle cells was detected by Western blot. The model of carotid artery injury was established by balloon injury in rats. The control group was given 2 mL of normal saline. The rats in group B were given intragastric administration of ketamine 3 mg / kg intraperitoneally once a day for HE staining to observe the formation of neointima. Fluorescent staining was used to detect the expression of connexin 43 in neointima, Evans blue-DAPI double staining to evaluate neointimal formation. Results: The cultured cells were positive for anti-SMα-actin immunofluorescence staining. Smooth muscle cells were treated with 50 micromol / L carbenoxolone for 24 h and had no effect on the expression of connexin43 (0.85 ± 0.06 vs 0.83 ± 0.03, n = 3, P> 0.05). Balloon injury of the carotid artery in rats after 14 days showed stenosis of blood vessels, neointimal hyperplasia. Immunofluorescence staining revealed that neointimal formation of connexin 43 was abundantly expressed. Neonatal endometrial hyperplasia was significantly reduced after intervention of ketamine, and neointimal cell count was significantly lower than that of the control group (89 ± 28.40 vs 236 ± 15.04, n = 5, P <0.01). CONCLUSIONS: Gap junctions play an important role in the formation of neointima after vascular injury, while the gap junction blocker carbenoxolone inhibits neointimal hyperplasia.