| 摘要: |
| 目的 利用野生型（WT）小鼠及核因子E2相关因子2（Nrf2）基因敲除（Nrf2KO）小鼠构建2型糖尿病（T2DM）模型，探究Nrf2基因对T2DM小鼠膀胱功能障碍的影响和作用机制。方法 利用雄性Nrf2基因敲除小鼠及同龄雄性WT小鼠构建T2DM小鼠模型，并随机分成Nrf2基因敲除小鼠T2DM造模组（Nrf2KO＋T2DM组）、Nrf2基因敲除小鼠对照组（Nrf2KO组）、WT小鼠T2DM造模组（T2DM组）、WT小鼠对照组（WT组），每组10只。分析小鼠生理体征及24 h饮水与排尿量。提取小鼠膀胱组织样本进行病理学检测，并采用免疫组织化学染色及蛋白质印迹法分析Nrf2信号通路相关蛋白质的表达。结果 T2DM组小鼠体重、膀胱质量、血糖、糖化血红蛋白、24 h饮水量及24 h排尿量均高于WT组（P均<0.05），提示T2DM小鼠模型建立成功。T2DM组小鼠膀胱组织中细胞核Nrf2和总Nrf2蛋白表达水平均低于WT组（P均<0.05）。Nrf2KO＋T2DM组膀胱组织凋亡相关分子caspase 3及氧化应激产物晚期糖基化终末产物、丙二醛、活性氧均高于WT组、Nrf2KO组和T2DM组（P均<0.05），抗氧化应激分子还原型辅酶/醌氧化还原酶1和血红素氧合酶1表达均低于WT组、Nrf2KO组和T2DM组（P均<0.05），神经生长因子表达低于WT组、Nrf2KO组和T2DM组（P均<0.05），而神经生长因子前体表达高于WT组、Nrf2KO组和T2DM组（P均<0.05）。结论 Nrf2通路受到抑制后T2DM小鼠可出现氧化应激调节失衡、膀胱相关神经组织病变和凋亡通路相关蛋白表达上调，可能是T2DM小鼠膀胱功能障碍的重要机制。 |
| 关键词: 2型糖尿病 糖尿病膀胱功能障碍 氧化性应激 核因子E2相关因子2 |
| DOI：10.16781/j.CN31-2187/R.20220128 |
| 投稿时间：2022-02-14修订日期：2022-07-01 |
| 基金项目: |
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| Role of nuclear factor erythroid derived 2-like 2 in pathogenesis of bladder dysfunction in type 2 diabetic mice |
| WANG Lei△,REN Guan-yu△,XU Cheng,ZHANG Xin-hui,LIU Zhi-yong* |
(Department of Urology, The First Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200433, China
△Co-first authors.
* Corresponding author) |
| Abstract: |
| Objective To construct type 2 diabetes mellitus (T2DM) models using wild-type (WT) mice and nuclear factor erythroid derived 2-like 2 (Nrf2) knockout (Nrf2KO) mice, and to explore the effect and mechanism of Nrf2 gene on bladder dysfunction in T2DM mice. Methods T2DM mouse model was constructed using male Nrf2KO mice and male WT mice in the same age, and the mice were randomly divided into Nrf2KO+T2DM group, Nrf2KO mouse control group (Nrf2KO group), T2DM modeling group (T2DM group), and WT mouse control group (WT group), with 10 mice in each group. The physiological signs and 24-h water intake and urine output of mice in each group were analyzed. The bladder tissue samples of mice were collected for histopathological analysis, and the expression of Nrf2 signaling pathway related proteins was analyzed by immunohistochemical staining and Western blotting. Results The body weight, bladder mass, blood glucose, glycosylated hemoglobin, and 24-h water intake and urine output of mice in the T2DM group were significantly higher than those in the WT group (all P<0.05), indicating that the T2DM mouse model was successfully established. The expression levels of nuclear Nrf2 and total Nrf2 proteins in the bladder tissues of mice in the T2DM group were significantly lower than those in the WT group (all P<0.05). The expression of apoptosis related molecules caspase 3 and oxidative stress-related products such as advanced glycation end products, malondialdehyde, and reactive oxygen species in bladder tissues of the Nrf2KO+T2DM group was significantly higher than that in the WT, Nrf2KO and T2DM groups (all P<0.05); the expression of antioxidant stress molecules reduced coenzyme/quinone oxidoreductase 1 and heme oxygenase 1 was significantly lower than that in the WT, Nrf2KO and T2DM groups (all P<0.05); the expression of nerve growth factor was significantly lower than that in the WT, Nrf2KO and T2DM groups (all P<0.05); however, the expression of the pro-form of nerve growth factor was significantly higher than that in the WT, Nrf2KO and T2DM groups (all P<0.05). Conclusion The inhibition of Nrf2 pathway leads to imbalance of oxidative stress regulation, bladder-related neuropathies and up-regulated expression of apoptosis pathway related proteins, and it may be an important mechanism of bladder dysfunction in T2DM mice. |
| Key words: type 2 diabetes mellitus diabetic bladder dysfunction oxidative stress nuclear factor erythroid derived 2-like 2 |
