医源性输尿管损伤后梗阻形成机制及其防治的研究
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摘要
目的：构建输尿管损伤动物模型,筛选输尿管损伤后纤维化进程中敏感的组织标志物并探索其波动情况,研究卡托普利对阻断输尿管损伤后纤维化的意义。
材料与方法：以成年雄性新西兰兔为实验对象,构建输尿管热损伤后狭窄动物模型。通过RT-PCR技术,于损伤后2周、3周及4周动态测定EGF、TGF-β、FN、Col Ia1、Col Ia2及Col Ⅲ在受累输尿管中的表达水平,与对侧正常输尿管组织对比,寻找较为敏感的纤维化指标,并探索不同指标表达高峰时间。设置卡托普利组(实验组)与对照组,干预组予复方卡托普利10mg/Kg/d口服,对照组予安慰剂。给药2周取患侧输尿管狭窄段及对侧2cm正常输尿管。通过RT-PCR技术分别测定实验组与对照组患侧输尿管与对侧正常输尿管EGF、TGF-β、Col Ia1、 Col Ia2及FN水平,对比干预组与实验组各指标上升幅度。
结果：动物模型构建成功。在动物模型构建过程中,测得EGF、TGF-β、FN、 Col Ia1及Col Ia2较对侧正常输尿管组织显著升高(p<0.01)。其中,EGF、TGF-β、 Col Ial高表达期均为输尿管损伤后2周,而FN与Col Ia2高表达期分别为损伤后3周与损伤后4周。在卡托普利干预研究中,实验组EGF、Fn及Col Ia2表达量较对照组显著降低(p<0.05)；而TGF-β与Col Ial表达量较对照组无显著差异(p=0.18,p=0.58)。
结论：在众多细胞因子中,EGF、TGF-β、Col Ia1、Col Ia2及FN在输尿管瘢痕形成中起重要作用。其表达高峰主要为损伤后2周,而2周之后仍有不同的细胞因子参与纤维瘢痕的终末期构建。卡托普利可部分阻断输尿管损伤后的纤维化进程,降低EGF、Col Ia2及FN表达,而对TGF-β与Col la的表达抑制作用不明显,因此未来还需进一步研究阐明更完整的输尿管纤维化调控通路,从而为进一步临床干预提供线索。
Purpose:To establish a reliable animal mode of the ureteral injury and to screen the sensitive markers of the fibrotic process after the ureteral injury. Make clear the fluctuation characteristic of the different sensitive makers during the tissue scarring and evaluate the anti-fibrotic role of Captopril.
Materials and methods:The New Zealand rabbit was taken to establish the animal model of ureteral injury. The EGF, TGF-β, FN, Col Ial, Col Ia2and Col III of the impaired ureter and the contralateral normal ureter were quantified by RT-PCR technique to find out the sensitive biomarkers of the ureteral fibrosis. The fluctuation of the markers during2to4weeks was also noted. A Captopril group and a control group were set to evaluate the anti-fibrotic efficacy of this medication in ureteral fibrosis. The Captopril10mg/Kg/d was given in experimental group while the placebo was given in control group. The specimens were harvest2weeks after the intervention and the RT-PCR was carried out.
Result:The ureteral injury animal mode was established successfully. By RT-PCR screening, EGF, TGF-β, FN, Col Ial and Col Ia2were found to be significantly elevated than the other markers.(p<0.01) The peak level of EGF, TGF-β and Col Ial appeared at2weeks after the injury while for Fn and Col Ia2the peak level appeared at3weeks and4weeks after the surgery, respectively. For the Captopril experimentation, the expression of EGF, Fn and Col Ia2in Captopril group was significantly lower than control group. However, there was no statistic significance for TGF-B and Col Ial between the two groups.
Conclusion:EGF, TGF-β, Col Ial, Col Ia2and FN seemed to have important role in the ureteral fibrosis and scarring after the ureteral lesion. The peak of the fibrosis was at2weeks after the injury. The Captopril might partially inhibit the fibrotic process by blocking the EGF, Col Ia2and FN pathway. Further research work should be carried out to clarify the ureteral fibrotic network, which may offer new clinical methods to prevent the ureteral scarring.
材料与方法：以成年雄性新西兰兔为实验对象,构建输尿管热损伤后狭窄动物模型。通过RT-PCR技术,于损伤后2周、3周及4周动态测定EGF、TGF-β、FN、Col Ia1、Col Ia2及Col Ⅲ在受累输尿管中的表达水平,与对侧正常输尿管组织对比,寻找较为敏感的纤维化指标,并探索不同指标表达高峰时间。设置卡托普利组(实验组)与对照组,干预组予复方卡托普利10mg/Kg/d口服,对照组予安慰剂。给药2周取患侧输尿管狭窄段及对侧2cm正常输尿管。通过RT-PCR技术分别测定实验组与对照组患侧输尿管与对侧正常输尿管EGF、TGF-β、Col Ia1、 Col Ia2及FN水平,对比干预组与实验组各指标上升幅度。
结果：动物模型构建成功。在动物模型构建过程中,测得EGF、TGF-β、FN、 Col Ia1及Col Ia2较对侧正常输尿管组织显著升高(p<0.01)。其中,EGF、TGF-β、 Col Ial高表达期均为输尿管损伤后2周,而FN与Col Ia2高表达期分别为损伤后3周与损伤后4周。在卡托普利干预研究中,实验组EGF、Fn及Col Ia2表达量较对照组显著降低(p<0.05)；而TGF-β与Col Ial表达量较对照组无显著差异(p=0.18,p=0.58)。
结论：在众多细胞因子中,EGF、TGF-β、Col Ia1、Col Ia2及FN在输尿管瘢痕形成中起重要作用。其表达高峰主要为损伤后2周,而2周之后仍有不同的细胞因子参与纤维瘢痕的终末期构建。卡托普利可部分阻断输尿管损伤后的纤维化进程,降低EGF、Col Ia2及FN表达,而对TGF-β与Col la的表达抑制作用不明显,因此未来还需进一步研究阐明更完整的输尿管纤维化调控通路,从而为进一步临床干预提供线索。
Purpose:To establish a reliable animal mode of the ureteral injury and to screen the sensitive markers of the fibrotic process after the ureteral injury. Make clear the fluctuation characteristic of the different sensitive makers during the tissue scarring and evaluate the anti-fibrotic role of Captopril.
Materials and methods:The New Zealand rabbit was taken to establish the animal model of ureteral injury. The EGF, TGF-β, FN, Col Ial, Col Ia2and Col III of the impaired ureter and the contralateral normal ureter were quantified by RT-PCR technique to find out the sensitive biomarkers of the ureteral fibrosis. The fluctuation of the markers during2to4weeks was also noted. A Captopril group and a control group were set to evaluate the anti-fibrotic efficacy of this medication in ureteral fibrosis. The Captopril10mg/Kg/d was given in experimental group while the placebo was given in control group. The specimens were harvest2weeks after the intervention and the RT-PCR was carried out.
Result:The ureteral injury animal mode was established successfully. By RT-PCR screening, EGF, TGF-β, FN, Col Ial and Col Ia2were found to be significantly elevated than the other markers.(p<0.01) The peak level of EGF, TGF-β and Col Ial appeared at2weeks after the injury while for Fn and Col Ia2the peak level appeared at3weeks and4weeks after the surgery, respectively. For the Captopril experimentation, the expression of EGF, Fn and Col Ia2in Captopril group was significantly lower than control group. However, there was no statistic significance for TGF-B and Col Ial between the two groups.
Conclusion:EGF, TGF-β, Col Ial, Col Ia2and FN seemed to have important role in the ureteral fibrosis and scarring after the ureteral lesion. The peak of the fibrosis was at2weeks after the injury. The Captopril might partially inhibit the fibrotic process by blocking the EGF, Col Ia2and FN pathway. Further research work should be carried out to clarify the ureteral fibrotic network, which may offer new clinical methods to prevent the ureteral scarring.
引文
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