秋水仙碱治疗经皮冠状动脉腔内成形术后血管再狭窄模型大鼠的效果及其机制研究

Effect of Colchicine in the Treatment of Vascular Restenosis after Percutaneous Transluminal Coronary Angioplasty Model Rats and Its Mechanism

目的分析秋水仙碱治疗经皮冠状动脉腔内成形术(PTCA)后血管再狭窄模型大鼠的效果及其机制。方法本实验时间为2021年8月—2022年8月。在45只SPF级雄性SD大鼠中选取35只并建立PTCA后血管再狭窄模型。将建模成功的30只大鼠随机分为模型组、培哚普利组、秋水仙碱组,各10只;将剩余10只未建模的大鼠设为正常组。正常组与模型组不进行干预,培哚普利组接受培哚普利0.4 mg·kg^(-1)·d^(-1)灌胃干预,秋水仙碱组接受秋水仙碱0.1 mg·kg^(-1)·d^(-1)灌胃干预,均干预30 d。干预结束后每组取3只大鼠,采用HE染色观察颈动脉病理改变情况并计算血管再狭窄面积和血管再狭窄率;干预结束后每组取3只大鼠,采用TUNEL法检测血管平滑肌细胞凋亡率;干预结束后每组取2只大鼠,采用Western blotting法检测颈动脉组织中IL-6、IL-1β、TNF-α蛋白相对表达水平;干预结束后每组取2只大鼠,采用荧光定量PCR法检测颈动脉组织中IL-6、IL-1β、TNF-αmRNA相对表达水平。结果HE染色结果显示,模型组大鼠血管内膜增厚且管腔狭窄明显,血管新生内膜排列严重紊乱;培哚普利组与秋水仙碱组大鼠管腔狭窄程度均较模型组有所减轻,其中秋水仙碱组减轻更为明显。培哚普利组、秋水仙碱组血管再狭窄面积小于模型组、血管再狭窄率低于模型组,秋水仙碱组血管再狭窄面积小于培哚普利组、血管再狭窄率低于培哚普利组(P<0.05)。模型组血管平滑肌细胞凋亡率低于正常组,培哚普利组、秋水仙碱组血管平滑肌细胞凋亡率高于正常组、模型组,秋水仙碱组血管平滑肌细胞凋亡率高于培哚普利组(P<0.05)。模型组、培哚普利组、秋水仙碱组颈动脉组织中IL-6、IL-1β、TNF-α蛋白及其mRNA相对表达水平高于正常组,培哚普利组、秋水仙碱组颈动脉组织中IL-6、IL-1β、TNF-α蛋白及其mRNA相对表达水平低于模型组,秋水仙碱组颈动脉组织中IL-6、IL-1β、TNF-α蛋白及其mRNA相对表达水平低于培哚普利组(P<0.05)。结论秋水仙碱可有效治疗PTCA后血管再狭窄模型大鼠,其机制可能与秋水仙碱可诱导血管平滑肌细胞凋亡、通过抑制炎症因子的生成来减轻炎症反应有关。

Objective To analyze the effect of colchicine in the treatment of vascular restenosis after percutaneous transluminal coronary angioplasty(PTCA)model rats and its mechanism.Methods The experiment was conducted from August 2021 to August 2022.The vascular restenosis after PTCA model was established in 35 out of 45 SPF male SD rats.The 30 successfully modeled rats were randomly divided into model group,perindopril group,and colchicine group,with 10 rats in each group.The remaining 10 rats without modeling were set as normal group.The normal group and the model group were not intervened.The perindopril group received perindopril 0.4 mg·kg^(-1)·d^(-1) gavage intervention,while the colchicine group received colchicine 0.1 mg·kg^(-1)·d^(-1) gavage intervention.Both groups were treated for 30 days.After the intervention,3 rats in each group were selected and the pathological changes of carotid artery were observed by HE staining,and the area and rate of vascular restenosis were calculated.After the intervention,3 rats in each group were selected and the apoptosis rate of vascular smooth muscle cells was detected by TUNEL method.After the intervention,2 rats in each group were selected and the relative expression levels of IL-6,IL-1βand TNF-αprotein in carotid tissues were detected by Western blotting.After the intervention,2 rats in each group were selected and the relative expression levels of IL-6,IL-1βand TNF-αmRNA in carotid tissues were detected by fluorescence quantitative PCR.Results HE staining showed that the vascular intima thickened and lumen narrowed significantly in the model group,and the arrangement of the neovascularization intima was seriously disordered;the degree of luminal stenosis of rats in the perindopril group and the colchicine group was reduced compared to the model group,especially in the colchicine group.The vascular restenosis area of perindopril group and colchicine group was samller than that of model group,the vascular restenosis rate was lower than that of model group,and the vascular restenosis area of colchicine group was samller than that of perindopril group,the vascular restenosis rate was lower than that of perindopril group(P<0.05).The apoptosis rate of vascular smooth muscle cells in model group was lower than that in normal group,the apoptosis rate of vascular smooth muscle cells in perindopril group and colchicine group was higher than that in normal group and model group,and the apoptosis rate of vascular smooth muscle cells in colchicine group was higher than that in perindopril group(P<0.05).The relative expression levels of IL-6,IL-1β,TNF-αprotein and their mRNA in carotid tissues of model group,perindopril group and colchicine group were higher than those of normal group,the relative expression levels of IL-6,IL-1β,TNF-αprotein and their mRNA in carotid tissues of perindopril group and colchicine group were lower than those of model group,the relative expression levels of IL-6,IL-1β,TNF-αprotein and their mRNA in carotid tissues of colchicine group were lower than those of perindopril group(P<0.05).Conclusion Colchicine can effectively treat vascular restenosis after PTCA model rats,and its mechanism may be related to the induction of vascular smooth muscle cell apoptosis and the reduction of inflammation by inhibiting the production of inflammatory factors.