First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, China.
Apoptosis. 2023 Apr;28(3-4):639-652. doi: 10.1007/s10495-022-01800-6. Epub 2023 Jan 31.
Chronic obstructive pulmonary disease is the 3rd leading cause of death worldwide, and the available treatments are unsatisfactory, resulting in a major economic burden. As cellular therapy is commonly used for lung disease, we investigated a treatment with CXCR4-overexpressing BMSCs in a COPD model. We extracted and purified Bone marrow mesenchymal stem cells (BMSCs) from SD rats. COPD apoptosis model was established by cigarette smoke exposure. BMSCs (1 × 10 cells per injection)were transplanted in vivo twice a month during model establishment, and alveolar rupture in the lung was assessed. Lung cell apoptosis was assessed by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) analysis, and the concentrations of apoptotic proteins in the lungs were detected by Western blotting. We successfully isolated BMSCs and established CXCR4-overexpressing BMSCs. qRT‒PCR and Western blotting detection both reveal that CXCR4 mRNA level and protein both significantly higher expression in CXCR4-BMSCs than the pBABE-BMSCs. Continuous cigarette smoke exposure caused alveolar septal rupture: In the model group, the alveolar mean linear intercept in the first month was significantly lower than that in the third month (p < 0.05). In the third month, the alveolar mean linear intercept values of the control and CXCR4-BMSC groups were lower than those of the model group (control group p < 0.01, CXCR4-BMSC group p < 0.05), and TUNEL staining revealed that the apoptosis rates of the control and CXCR4-BMSC groups were significantly lower than those of the model group (p < 0.01). Furthermore, the levels of the apoptotic proteins cleaved caspase-8, cleaved caspase-3 and cleaved PARP-1 were higher in the model group than in the control group (p < 0.05) and significantly lower in the CXCR4-BMSC group than in the model group (p < 0.05). The transplantation of CXCR4-overexpressing BMSCs during COPD model generation significantly inhibited apoptosis via the extrinsic apoptosis pathway. CXCR4 enhances the inhibitory effects of bone mesenchymal stem cells on lung cell apoptosis in a rat model of smoking-induced COPD.
慢性阻塞性肺疾病是全球第 3 大死因，现有的治疗方法并不令人满意，导致了巨大的经济负担。由于细胞疗法常用于肺部疾病，我们研究了在 COPD 模型中使用 CXCR4 过表达骨髓间充质干细胞（BMSCs）的治疗方法。我们从 SD 大鼠中提取并纯化了骨髓间充质干细胞（BMSCs）。通过香烟烟雾暴露建立 COPD 细胞凋亡模型。在模型建立过程中，每月两次进行体内移植，每次 BMSCs（1×10 个细胞/注射），评估肺部肺泡破裂情况。通过末端脱氧核苷酸转移酶生物素-dUTP 缺口末端标记（TUNEL）分析评估肺细胞凋亡，通过 Western blot 检测肺组织中凋亡蛋白的浓度。我们成功分离了 BMSCs 并建立了 CXCR4 过表达 BMSCs。qRT-PCR 和 Western blot 检测均显示 CXCR4-BMSCs 中的 CXCR4 mRNA 水平和蛋白表达均明显高于 pBABE-BMSCs。连续香烟烟雾暴露导致肺泡隔破裂：在模型组中，第一个月的肺泡平均线性截距明显低于第三个月（p<0.05）。在第三个月，对照组和 CXCR4-BMSC 组的肺泡平均线性截距值均低于模型组（对照组 p<0.01，CXCR4-BMSC 组 p<0.05），TUNEL 染色显示对照组和 CXCR4-BMSC 组的细胞凋亡率明显低于模型组（p<0.01）。此外，模型组中裂解的 caspase-8、裂解的 caspase-3 和裂解的 PARP-1 凋亡蛋白水平高于对照组（p<0.05），而 CXCR4-BMSC 组显著低于模型组（p<0.05）。在 COPD 模型生成期间移植 CXCR4 过表达 BMSCs 可通过外在凋亡途径显著抑制细胞凋亡。CXCR4 增强了骨髓间充质干细胞对吸烟诱导的 COPD 大鼠肺细胞凋亡的抑制作用。
