壳聚糖抑制家兔动静脉内瘘内膜增生的实验研究  

Experimental study of chitosan inhibiting vascular intimal hyperplasia of rabbit arteriovenous fistula

作　　者：郑婕[1] 鄢艳[2] 陈钦开[2] 苏晓霞[2] 张莉[2] 杨柳[2] 吕金雷[2] 

机构地区：[1]赣南医学院第一附属医院肾内科 [2]南昌大学第一附属医院肾内科,330006

出　　处：《中华肾脏病杂志》2015年第5期367-371,共5页Chinese Journal of Nephrology

基　　金：江西省教育厅课题（GJJ13144）

摘　　要：目的 观察壳聚糖对家兔颈总动脉-颈内静脉内瘘术后血管内膜增生的影响,探讨Toll样受体4/核因子κB（TLR4/NF-κB）通路对动静脉瘘（AVF）术后血管内膜增生的调控作用.方法28只新西兰大耳白兔被随机分为对照组（n=4）、模型组（n=12）及壳聚糖组（n=12）.模型组和壳聚糖组家兔建立家兔颈总动脉-颈内静脉内瘘模型,壳聚糖组动静脉瘘术后静脉端血管及吻合口处涂抹壳聚糖,术后4周、6周及8周分别取AVF静脉端血管组织.观察各组家兔AVF静脉端血管组织的病理改变;免疫组化法检测各组血管组织α平滑肌肌动蛋白（α-SMA）表达;RT-PCR法检测增殖细胞核抗原（PCNA）和TLR4 mRNA的表达;Western印迹法检测血管组织PCNA、TLR4、髓样分化因子88（MyD88）和NF-κB蛋白的表达.用双因素方差分析法比较各组实验数据的差异.结果 （1）与对照组相比,模型组家兔AVF术后4周静脉端血管内膜增厚,在增生的血管内膜中α-SMA染色阳性,血管平滑肌细胞（VSMCs）增殖明显.随时间延长,内膜增生改变更为显著变窄,α-SMA表达增加（均P＜0.05）.与模型组相比,壳聚糖组内膜增生程度明显减轻,α-SMA表达下降,VSMCs增殖改变减轻（均P＜0.05）.（2）与对照组比较,模型组家兔血管组织PCNA、TLR4、MyD88和NF-κB表达量随着AVF时间的延长而增加（均P＜ 0.05）;壳聚糖组各指标表达量高于对照组,但明显低于模型组（P＜0.05）.结论 壳聚糖可抑制家兔血管平滑肌细胞（VSMCs）增殖,其机制可能与下调TLR4介导的信号途径减少家兔AVF静脉端血管内膜增生有关.Objective To investigate the effete of chitosan on rabbit carotid artery internal jugular vein fistula intimal hyperplasia and its regulation on TLR4/NF-κB signaling.Methods A total of 28 New Zealand white rabbits were randomly divided into the control group（n=4）,the model group（n=12） and the chitosan group（n=12）.Model group and chitosan group rabbits were established respectively carotid artery internal jugular vein fistula models.After AVF surgery,chitosan was smeared on venous blood vessels and anastomosis.After 4,6 and 8 weeks,the rabbits were separately sacrificed and the AVF venous vascular tissues were taken.The pathological changes of AVF venous vascular tissue in each group were observed.The changes of α-SMA were detected by immunohistochemistry method.The mRNA expressions of PCNA and TLR4 in the tissues were measured by Real-time PCR.At the same time,the protein expressions of PCNA,TLR4,MyD88 and NF-κB were detected by Western blotting.The experimental data were processed by two-factor analysis of variance in statistics.Results （1） After 4 weeks,vascular intimal was thicked in mdel group.In intimal hyperplasia,α-SMA was staining,and then proliferation of vascular smooth muscle cell was significant.As time increasing,more intimal hyperplasia shown obviously,the expression of α-SMA significantly increased.Compared with model group,chitosan group significantly reduced the degree of intimal hyperplasia,the level of α-SMA was significantly decreased,vascular smooth muscle cell proliferation was also extraordinarily decreased.（2） Compared with control group,the expression levels of PCNA,TLR4,MyD88 and NF-κB increased with time.The indices of Chitosan group were markedly higher than control group,but significantly lower than model groups.Conclusion Chitosan can inhibit the proliferation of rabbit VSMCs.The mechanism may be concerned in down regulating TLR4-mediated signaling pathway,reducing the possibility of intimal hyperplasia of rabbit AVF venous blood vessels.

关 键 词：动静脉瘘 壳聚糖 血管内膜 Toll样受体4 血管平滑肌细胞 TOLL-LIKE RECEPTOR 4 

分 类 号：R459.5[医药卫生—治疗学]