103pd诱导犬胆管增殖平滑肌细胞凋亡及对相关基因的影响

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目的:探讨γ射线对胱冬肽酶-3(caspase-3),Fas和bcl-2基因在犬胆管壁增殖平滑肌细胞凋亡中的活性影响及意义. 方法:犬12只随机分2组,每组6只;建立胆管损伤后狭窄模型,在犬胆管内分别植入103pd(103钯)放射性支架和普通胆管支架,采用HE,TUNEL,RT—PCR, 免疫组化检测增殖内膜中胆管平滑肌细胞增殖、凋亡及相关caspase-3,bcl-2和Fas基因表达,在JEM- 1200EX电镜下观察凋亡细胞超微结构变化,并用计算机图像检测系统检2组胆管腔面积. 结果:103pd支架组犬胆管组织中caspase-3和Fas基因表达较普通支架组明显(0.44±0.09 vs 0.16±0.02, 83.33% vs 50.00%,P<0.05),出现明显增殖平滑肌细胞凋亡(87.90±7.96 vs 5.60±0.51,P<0.05)。肝外胆管无明显狭窄;而普通支架caspase-3和Fas基因低表达,胆管未出现增殖平滑肌细胞明显凋亡,而肝外胆管有明显狭窄.103pd支架组犬胆管组织中bcl-2基因表达较普通支架组弱(16.66%vs 83.33%,P<0.05), 且bcl-2基因低表达组犬胆管出现明显增殖平滑肌细胞凋亡,而且犬肝外胆管无明显狭窄;bcl-2基因高表达组犬胆管未出现增殖平滑肌细胞明显凋亡,而肝外胆管有明显狭窄. 结论:103pd放射性支架可以激活caspase-3和Fas基因, 促进犬胆管增殖平滑肌细胞凋亡;bcl-2基因表达水平与细胞对辐射的敏感性有关,103pd放射性支架通过降低bcl-2基因表达,促进犬胆管增殖平滑肌细胞凋亡,从而抑制犬肝外胆管狭窄. Objective: To investigate the effect and significance of γ-ray on the activity of caspase-3, Fas and bcl-2 gene in the apoptosis of canine bile duct smooth muscle cells.Methods: 12 dogs were randomly divided into 2 groups, (N = 6). The model of stenosis after bile duct injury was established. 103pd (103 Pd) radioactive scaffolds and common biliary stents were implanted into the bile duct of dogs. HE, TUNEL and RT- Cell proliferation, apoptosis and the expression of caspase-3, bcl-2 and Fas genes were observed under light microscope. The ultrastructural changes of apoptotic cells were observed under JEM-1200EX electron microscope and the bile duct lumen area was detected by computer image detection system.Results: The expression of caspase-3 and Fas in the bile duct tissue of dogs was significantly higher than that of the normal scaffolds (0.44 ± 0.09 vs 0.16 ± 0.02, 83.33% vs 50.00%, P <0.05), and significantly increased the apoptosis of smooth muscle cells (87.90 ± 7.96 vs 5.60 ± 0.51, P <0.05). The extrahepatic bile ducts had no obvious stenosis. However, the expression of caspase-3 and Fas genes in common scaffolds was low, but there was no significant apoptosis in the bile duct, while the extrahepatic bile duct had obvious stenosis. The common bracketed group was weaker (16.66% vs 83.33%, P <0.05), and the bcl-2 low expression group showed significant proliferation of smooth muscle cells and no obvious stenosis of canine extrahepatic bile duct. The bcl-2 gene overexpression group The bile duct did not appear obvious apoptosis in proliferating smooth muscle cells, while the extrahepatic bile duct had obvious stenosis.Conclusion: The 103pd radioactive scaffold can activate caspase-3 and Fas gene and promote the apoptosis of canine biliary duct proliferation smooth muscle cells. The expression of bcl- The sensitivity of radiation, 103pd radioactive stent through reducing bcl-2 gene expression, promote canine bile duct smooth muscle cell apoptosis, thereby inhibiting canine bile duct stricture.
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