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Sonic Hedgehog信号通路在重症急性胰腺炎大鼠模型肠黏膜屏障损伤中的作用探讨

文琳 曾家月 马凤雨 陈霞

引用本文:
Citation:

Sonic Hedgehog信号通路在重症急性胰腺炎大鼠模型肠黏膜屏障损伤中的作用探讨

DOI: 10.3969/j.issn.1001-5256.2023.05.020
基金项目: 

国家自然科学青年基金 (81600420)

泸州市人民政府-西南医科大学科技战略合作项目 (2019LZXNYDJ48)

伦理学声明:本研究方案于2022年3月9日经由西南医科大学动物保护与福利委员会批准,批号:swmu20220019,符合实验室动物管理与使用准则。
利益冲突声明:本文不存在任何利益冲突。
作者贡献声明:文琳负责课题设计,资料分析,撰写论文;曾家月、马风雨参与收集数据,修改论文;陈霞负责拟定写作思路,指导撰写文章并最后定稿。
详细信息
    通信作者:

    陈霞,970217858@qq.com (ORCID: 0000-0003-3675-5336)

Role of the Sonic Hedgehog signaling pathway in intestinal mucosal barrier injury in rats with severe acute pancreatitis

Research funding: 

National Natural Science Foundation of China (81600420);

Luzhou Municipal People's Government-Southwest Medical University Science and Technology Strategic Cooperation Project (2019LZXNYDJ48)

More Information
  • 摘要:   目的  探讨Sonic Hedgehog(Shh)信号通路在重症急性胰腺炎(SAP)大鼠肠道黏膜屏障损伤中的表达及作用。  方法  48只SD大鼠按随机数字表法分为:假手术组(Sham组)、SAP模型组(SAP组)、SAP模型组+Shh信号通路特异性激动剂嘌呤胺组(PUR+SAP组)、SAP模型组+Shh信号通路特异性抑制剂环巴胺组(CYC+SAP组),各组又分为12 h、24 h两个亚组,每个亚组6只大鼠。SAP造模采用5%牛磺胆酸钠胰胆管逆行注射,干预组分别在造模前腹腔注射0.69 mg/kg嘌呤胺及0.69 mg/kg环巴胺。在造模后12 h和24 h取材。HE染色观察大鼠胰腺及回肠病理学变化;ELISA法检测大鼠血清淀粉酶、脂肪酶、DAO和EndoCAb的表达水平;TUNEL法检测肠上皮细胞凋亡情况;Western Blot检测回肠组织Shh、Ptch1和Gli1的表达。计量资料符合正态分布时多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验法;计量资料不符合正态分布时多组间比较及进一步两两比较均采用Kruskal-Wallis H检验法。  结果  与Sham组相比,SAP组胰腺和回肠组织病理学评分均明显增加,血清脂肪酶、淀粉酶、DAO和EndoCAb水平明显升高,肠上皮细胞凋亡增加,回肠组织Shh、Ptch1和Gli1蛋白表达升高(P值均<0.05)。与SAP组对比,PUR+SAP组胰腺和肠道病理损伤及功能障碍减轻,肠道上皮细胞凋亡显著减少,回肠组织中的Shh、Ptch1和Gli1蛋白表达明显升高(P值均<0.05)。与SAP组对比,CYC+SAP组胰腺和肠道病理损伤及功能障碍加重,肠道上皮细胞凋亡显著增加,回肠组织中的Shh、Ptch1和Gli1蛋白表达明显降低(P值均<0.05)。  结论  Shh信号通路可能参与了SAP肠黏膜屏障损伤并且发挥保护作用。

     

  • 图  1  胰腺组织病理学改变(HE染色,×200)

    Figure  1.  Histological analyses of pancreatic tissues(HE, ×200)

    图  2  回肠组织病理学改变(HE染色,×200)

    Figure  2.  Histological analyses of ileum tissues(HE, ×200)

    图  3  血清中淀粉酶、脂肪酶、DAO和EndoCAb水平

    Figure  3.  Serum levels of amylase, lipase, DAO and EndoCAb

    图  4  TUNEL法检测回肠黏膜细胞凋亡水平

    注:光镜观察,原始放大倍数×200;凋亡细胞的特征是细胞核呈绿色。

    Figure  4.  Apoptosis of ileal mucosal cells detected by TdT-mediated dUTP nick end labeling assay

    图  5  细胞凋亡的积分光密度值

    Figure  5.  Integral optical density values of apoptosis

    图  6  大鼠回肠组织中Shh、Ptch1和Gli1的蛋白表达水平

    注:a,Western Blot检测Shh、Ptch1、Gli1通路相关蛋白表达,GAPDH作为对照; b、c,Shh、Ptch1和Gli1蛋白相对表达量。

    Figure  6.  The protein expression levels of Shh、Ptch1and Gli1in ileum

    表  1  胰腺和回肠组织的病理学评分

    Table  1.   The pancreas and ileum tissue pathology score

    组别 动物数(只) 胰腺组织12 h 胰腺组织24 h 回肠组织12 h 回肠组织24 h
    Sham组 6 0.50(0.50~0.63) 0.50(0.50~0.63) 0.50(0.38~0.50) 0.75(0.50~1.00)
    SAP组 6 7.76(7.36~8.50)1) 8.75(8.38~9.50)1) 3.62(3.53~3.73)1) 4.25(4.07~4.73)1)
    PUR+SAP组 6 2.25(1.88~2.50)1)2) 2.75(2.38~3.13)1)2) 2.16(2.08~2.29)1)2) 1.20(1.08~1.60)1)2)
    CYC+SAP组 6 13.25(12.38~13.63)1)2) 15.50(15.00~15.63)1)2) 4.75(4.50~5.00)1)2) 5.00(5.00~5.00)1)2)
    H 21.84 21.88 21.78 21.73
    P <0.05 <0.05 <0.05 <0.05
    注: 与Sham组比较,1)P<0.05;与SAP组比较,2)P<0.05。
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  • 收稿日期:  2022-10-27
  • 录用日期:  2022-12-17
  • 出版日期:  2023-05-20
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