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目的：探讨Rac1/JNK通路在肺气肿小鼠肺组织细胞凋亡中的潜在作用机制。方法：40只雄性C57BL/6健康小鼠，随机分为对照组、肺气肿组、肺气肿+Rac1抑制剂NSC23766干预组（肺气肿+NSC组）、肺气肿+JNK抑制剂SP600125干预组（肺气肿+SP组），每组均为10只。采用香烟熏吸结合腹腔注射香烟烟雾提取物方法构建模型，肺气肿+NSC组和肺气肿+SP组在模型建立之初每组均接受相应抑制剂进行腹腔注射，连续用药28 d。Western blot检测肺组织Rac1、JNK、p-JNK、c-Jun和Bax蛋白的表达水平。H&E染色对肺组织的病理变化进行观察，并计算肺泡的平均内衬间隔（mean linear intercept,MLI）以及肺泡的破坏指数（destruction index, DI）,qRT-PCR方法检测肺组织中Rac1 mRNA和JNK mRNA的相对表达水平，TUNEL技术评估小鼠肺组织细胞凋亡的情况，并计算凋亡指数（apoptosis index, AI）。结果：与对照组相比，肺气肿组MLI、DI、AI升高（P<0.05）,JNK、Rac1、p-JNK、c-Jun、Bax表达水平及Rac1 mRNA、JNK mRNA相对表达量升高（P<0.05）；与肺气肿组相比，用了Rac1抑制剂及JNK干预剂组的MLI、DI、AI降低（P<0.05）,JNK、Rac1、p-JNK、c-Jun、Bax表达水平及Rac1 mRNA、JNK mRNA相对表达量降低（P<0.05）。结论：抑制Rac1可抑制肺气肿小鼠肺组织中JNK的磷酸化，抑制Rac1-JNK通路可减轻肺气肿小鼠肺组织细胞凋亡，这可能与其减少促凋亡蛋白Bax的表达有关。
Abstract：Objective: To investigate the potential mechanism of Rac1/JNK pathway in apoptosis of lung tissue cells in emphysema mice. Methods: A total of 40 male C57BL/6 healthy mice were randomly divided into a control group, an emphysema group, an emphysema+Rac1 inhibitor NSC23766 intervention group(emphysema+NSC group), and an emphysema+JNK inhibitor SP600125 intervention group(emphysema+SP group), with 10 mice in each group. The models were constructed by cigarette smoke combined with intraperitoneal injection of cigarette smoke extract for the emphysema+NSC group and the emphysema+SP group. At the beginning of the establishment of the models, each group received intraperitoneal injection of the corresponding inhibitor for 28 consecutive days. Western blot was used to detect the expression levels of Rac1, JNK, p-JNK, c-Jun, and Bax proteins in lung tissues. The pathological changes of the lung tissues were observed using H&E staining, the mean lining interval(MLI) and the destruction index(DI) of alveoli were calculated, the relative expression levels of Rac1 mRNA and JNK mRNA in lung tissue were detected by qRT-PCR, the apoptosis of mouse lung tissue cells was evaluated by the TUNEL technology, and the apoptosis index(AI) was calculated. Result: Compared to the control group, MLI, DI, and AI in the emphysema group were increased(P<0.05), and the expression levels of JNK, Rac1, P-JNK, c-Jun, and Bax and the relative expression levels of Rac1 mRNA and JNK mRNA were increased(P<0.05). Compared to the emphysema group, MLI, DI, and AI in the Rac1 inhibitor and JNK intervention group were decreased(P<0.05), and the expression levels of JNK, Rac1, P-JNK, c-Jun, and Bax and the relative expression levels of Rac1 mRNA and JNK mRNA were decreased(P<0.05). Conclusion: Inhibition of Rac1 can suppress the phosphorylation of JNK in the lung tissue of mice with emphysema and inhibition of the Rac1-JNK pathway can alleviate apoptosis of lung tissue cells in mice with emphysema, which may be related to its reduction in the expression of the pro-apoptotic protein Bax.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20240912.003
中图分类号:R563.3
引用信息:
[1]金晓迪,罗兰,郑远江,等.Rac1/JNK对肺气肿小鼠肺组织细胞凋亡的影响及相关机制研究[J].海南医科大学学报,2025,31(09):659-666.DOI:10.13210/j.cnki.jhmu.20240912.003.
基金信息:
中国医学科学院中央级公益性科研院所基本科研业务费专项资金（2019PT320003）~~
2024-09-13
2024-09-13
2024-09-13