组织蛋白酶S对犬颈动脉支架置入后再狭窄的影响及法舒地尔对其干预的研究

The analysis of Cathepsin S on the restenosis of carotid stent and the effect of Farshudil on it

目的观察法舒地尔能否通过抑制比格犬支架置入后颈动脉中组织蛋白酶S(Cat S)表达从而抑制再狭窄进程。方法雄性比格犬12只,分为A组(假手术组),B组(裸支架组),C组(裸支架+fasudil组)。将8枚金属裸支架分别置入B组和C组的颈总动脉最狭窄处。支架置入2 h后对C组静脉输注法舒地尔进行干预,2 m后行DSA和颈部血管彩超检查,观察颈动脉再狭窄情况;取手术侧血管,进行观察及检测。结果 (1)彩超和DSA结果显示:支架置入后7 d和2 m后比格犬管腔内无血栓形成;支架置入2 m后A组管腔光滑,无狭窄形成;B组狭窄率显著大于C组(P<0.05)。(2)HE染色及弹力纤维染色结果显示:A组颈动脉无内膜增生,中膜由环形排列的弹力纤维和平滑肌细胞组成;B、C组颈动脉中膜平滑肌细胞层排列稍紊乱,内膜增生明显;B组新生内膜厚度、面积、狭窄率均显著大于C组(P<0.05)。(3)Cat S mRNA RT-PCR结果显示:A组比格犬颈动脉有少量Cat S mRNA表达;与A组比较,B、C组Cat S mRNA表达显著升高(P<0.05);B组Cat S mRNA表达显著高于C组(P<0.05)。(4)α-SMA免疫组化结果显示:A、B、C组中膜上均有大量α-SMA阳性表达。A组内弹力膜外无α-SMA阳性表达,B、C组新生内膜上见α-SMA阳性细胞。(5)PCNA免疫组化结果显示:A组PCNA免疫组化阳性染色较少,B、C组在支架周围及新生内膜上PCNA阳性表达增多,B组新生内膜上PCNA平均光密度值显著高于C组(P<0.05)。结论法舒地尔能通过抑制比格犬支架置入后颈动脉中组织蛋白酶S的表达,从而抑制再狭窄的进展。

Objective To observe the process of in-stent restenosis whether by Fasudil can effect the expression of cathepsin S after Carotid artery stenting.Methods 12 healthy beagles in male were randomly divided into 3 groups,group A(sham operation group,)and group B(group of bare stent)and group C(group of bare metal stent+fasudil).8 bare metal stents were placed in the common carotid artery stenosis of group B and group C.Fasudil was given to group C two hours after intervention.Two months later,DSA and cervical vascular ultrasonography were performed to observe the restenosis of the carotid artery.The side blood vessels were taken for observation and detection.Results (1)Vascular ultrasound showed no thrombosis in the lumens two months later after stenting.Results showed the lumens of group A were smooth without stenosis;whose differences has statistically significances(P<0.05);group B is higher than group C in stenosis rate(P<0.05).(2)The results of HE staining and elastic fiber staining showed that there was no intimal hyperplasia in the carotid artery in group A.The middle membrane was composed of elastic fibers and smooth muscle cells arranged in a circle;the carotid medium smooth muscle cell layer was slightly disordered and the intimal hyperplasia was evident in groups B and C.The thickness,area and stenosis rate of neointima in group B were significantly higher than those in group C(P<0.05).(3)The expression of Cat S mRNA by RT-PCR suggested Cat S mRNA appeared low expression in group A,but increased in group B and C,the differences has statistically significances(P<0.05).And the level of expression in group B is higher than group C(P<0.05).(4)Immunohistochemical staining of a-SMA showed a large number of a-SMA expressed in media of all three groups.Group A appeared on expression of a-SMA-positive outside of the elastic intima,but opposite in the new intima of groups B and C.(5)The results of PCNA immunohistochemistry showed that the positive staining of PCNA immunohistochemistry was less in group A,and positive more around the stents and in neointimal of group B and C;The value of average optical density in neointimal of group B was higher than C(P<0.05).Conclusion Fasudil can inhibit the process of restenosis by inhibiting the expression of cathepsin S in beagle dogs after beagle dogs implantation.