江津, 张松灵, 赵舒雅, 冯国兴, 樊赛军. 海藻糖对小鼠放射性肠损伤的防护作用及机制研究[J]. 国际放射医学核医学杂志. DOI: 10.3760/cma.j.cn121381-202403011-00414
引用本文: 江津, 张松灵, 赵舒雅, 冯国兴, 樊赛军. 海藻糖对小鼠放射性肠损伤的防护作用及机制研究[J]. 国际放射医学核医学杂志. DOI: 10.3760/cma.j.cn121381-202403011-00414
Jin Jiang, Songling Zhang, Shuya Zhao, Guoxing Feng, Saijun Fan. The protective effect and mechanism of trehalose on ionizing radiation-induced intestinal damage in mice[J]. Int J Radiat Med Nucl Med. DOI: 10.3760/cma.j.cn121381-202403011-00414
Citation: Jin Jiang, Songling Zhang, Shuya Zhao, Guoxing Feng, Saijun Fan. The protective effect and mechanism of trehalose on ionizing radiation-induced intestinal damage in mice[J]. Int J Radiat Med Nucl Med. DOI: 10.3760/cma.j.cn121381-202403011-00414

海藻糖对小鼠放射性肠损伤的防护作用及机制研究

The protective effect and mechanism of trehalose on ionizing radiation-induced intestinal damage in mice

  • 摘要:
    目的 探讨海藻糖对小鼠放射性肠损伤的防护作用及分子机制。
    方法 采用简单随机抽样法将8周龄的雄性C57 BL/6 J 小鼠分为4组:对照组(不给予任何处理)、海藻糖组(饮用2%海藻糖水)、照射组(以13 Gy 剂量对小鼠腹部照射)、海藻糖+照射组(饮用2%海藻糖水+以13 Gy 剂量对小鼠腹部照射),每组6只。经13 Gy照射和2%(g/ml)海藻糖饮水处理后每天记录小鼠体重,14 d后安乐处死小鼠并采集小鼠的血液、小肠和结肠,并测量结肠长度。采用苏木素-伊红染色法(HE)和过碘酸雪夫染色法(PAS)评估小鼠肠道损伤程度;酶联免疫吸附实验检测血清和小肠组织中炎症因子肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)的水平;细胞增值实验、细胞克隆形成实验、细胞凋亡检测实验、活性氧测定实验和DNA损伤检测实验分别检测细胞活力、克隆形成能力、凋亡、活性氧水平和DNA双链断裂数量;实时定量聚合酶链式反应(qRT-PCR)和Western blot 法分别检测自噬相关基因mRNA和蛋白水平的表达情况。组间比较采用t检验。
    结果 动物实验结果显示,海藻糖显著缓解电离辐照后小鼠体重下降,并且促进其恢复(t=4.064,P<0.05)。与照射组比较,海藻糖+照射组小鼠结肠缩短程度较轻,差异有统计学意义(t=4.044,P<0.05)。HE和PAS评估结果显示,海藻糖可缓解电离辐照后小鼠小肠黏膜损伤,与照射组比较,海藻糖+照射组小鼠绒毛较长、隐窝较深、杯状细胞较多,2组间的差异有统计学意义(t=8.414,4.373,6.515,均P<0.05)。ELISA检测结果显示,海藻糖显著降低电离辐照后小鼠血清及小肠组织中TNF-α和IL-6的水平,差异有统计学意义(t=8.686,4.475,6.007,3.993,均P<0.05)。细胞增殖和克隆实验结果显示,海藻糖促进电离辐照后MODE-K细胞活性和增殖能力(0.885±0.127)对(0.6441±0.151)、(97.330±5.937)对(64.000±7.324),差异有统计学意义(t=4.431、4.411,均P<0.05);细胞凋亡检测实验结果显示,海藻糖抑制电离辐照后小鼠小肠MODE-K细胞凋亡(15.27±0.647)%对(9.334±0.854)%,差异有统计学意义(t=8.315,P<0.05);活性氧测定实验和DNA损伤检测实验结果显示,海藻糖降低辐照后细胞内ROS水平和DNA双链断裂数量(t=8.884、4.802,均P<0.05)。qRT-PCR和Western blot检测结果显示,与照射组比较,海藻糖+照射组小鼠小肠组织和MODE-K细胞中自噬相关基因TFEB、MAP1LC3B表达显著增加(t=8.875,8.461,均P<0.05);与对照组比较,海藻糖处理组小肠细胞中TFEB表达水平显著增加(t=8.877,均P<0.05)。
    结论 海藻糖通过调控TFEB介导的自噬反应激活缓解小鼠放射性肠损伤。

     

    Abstract:
    Objective To investigate the protective effect and molecular mechanism of trehalose on ionizing radiation (IR)-induced small intestinal damage.
    Methods 8-week-old male C57 BL/6 J mice were randomly divided into 4 groups and each group contains six mice: control, trehalose-treated group, whole abdominal irradiation group treated with 13 Gy γ-ray and total abdominal irradiation treated with13 Gy γ-ray + trehalose group. The body weight was monitored daily after irradiation. After 14 days, the mice were euthanized, and samples from blood, small intestine, and colon were collected. Intestinal injury was assessed using hematoxylin-eosin (H&E) and periodic acid-Schiff (PAS) staining. The levels of TNF-α and IL-6 in serum and small intestine tissues were quantified by ELISA. Cell viability, proliferation, apoptosis, ROS levels, and DNA double-strand breaks (DSBs) were evaluated using CCK-8, cell clone formation, Annexin V/PI staining, DCFH-DA staining, and γ-H2AX staining, respectively. The expression levels of autophagy-related genes and proteins were determined by qRT-PCR and Western blot.
    Results The animal study results demonstrated that trehalose significantly alleviated the weight loss and facilitated the recovery of irradiated mice. Additionally, trehalose mitigated IR-induced colon shortening in irradiated mice. Immunohistochemical analysis revealed that trehalose mitigated mucosal damage in the small intestines of irradiated mice. ELISA assays demonstrated that trehalose substantially decreased TNF-α and IL-6 levels in both mouse serum and small intestinal tissues after IR. In vitro, trehalose enhanced MODE-K cell activity and proliferation after irradiation, and decreased apoptosis as demonstrated by Annexin V/PI staining. Furthermore, trehalose reduced intracellular ROS and DSBs in irradiated cells, as shown by DCFH-DA and γ-H2AX foci assays. qRT-PCR and Western blot analysis revealed that an upregulation of autophagy-related genes TFEB and MAP1LC3B were in response to co-treatment with IR and trehalose (P<0.05). Compared with the control group, the expression level of TFEB was significantly increased in the trehalose-treated group (P<0.05).
    Conclusion Trehalose alleviates IR-induced intestinal damage in mice by regulating TFEB mediated autophagy activation.

     

/

返回文章
返回