2025 年第 6 期 第 20 卷

人胚胎干细胞外泌体通过缓解肺血管外膜纤维化治疗小鼠高原肺动脉高压的实验研究

Experimental study of human embryonic stem cell exosomes in the treatment of high-altitude pulmonary hypertension by alleviating pulmonary vascular adventitial fibrosis in mice

作者：孙家树1赵启旭1邓玉松1肖飞2吕力知1张晗1王晟2王强1李刚1

英文作者：Sun Jiashu1 Zhao Qixu1 Deng Yusong1 Xiao Fei2 Lyu Lizhi1 Zhang Han1 Wang Sheng2 Wang Qiang1 Li Gang1

单位：1首都医科大学附属北京安贞医院小儿心脏中心北京市心肺血管疾病研究所，北京100029；2首都医科大学附属北京安贞医院麻醉手术中心北京市心肺血管疾病研究所，北京100029

英文单位：1Pediatric Cardiac Center Beijing Anzhen Hospital Capital Medical University Beijing Institute of Heart Lung and Blood Vessel Diseases Beijing 100029 China; 2Anesthesia Surgery Center Beijing Anzhen Hospital Capital Medical University Beijing Institute of Heart Lung and Blood Vessel Diseases Beijing 100029 China

关键词：肺动脉高压；低压低氧；人胚胎干细胞外泌体；血管外膜纤维化

英文关键词：Pulmonaryhypertension;Hypobarichypoxia;Humanembryonicstemcellexosomes;Vascularadventitialfibrosis

• 摘要：

• 目的 探讨高原肺动脉高压（HAPH）的形成机制及人胚胎干细胞外泌体（hESC-exo）对HAPH的干预效果。方法 将30只8周龄C57BL/6雄性小鼠完全随机分为3组：对照组、低压低氧组（HH组）、低压低氧hESC-exo干预组（HH-exo组），每组10只。对照组在常压常氧环境中饲养35 d；HH组和HH-exo组在低压低氧仓中饲养28 d，接着转至常压常氧环境下继续饲养7 d；实验第29、30、31天，HH-exo组小鼠尾静脉注射20 μg hESC-exo，对照组和低压低氧组小鼠注射等体积的磷酸盐缓冲液。第35天评估小鼠右心室收缩压（RVSP）、右心室肥厚指数（RVHI）、肺小动脉血管壁厚度（PA thickness）；实时荧光定量聚合酶链反应和蛋白质印迹法检测α-平滑肌肌动蛋白（α-SMA）、α-1型胶原蛋白（Col1-a1）及纤维连接蛋白（FN）的mRNA及蛋白表达水平。结果 建模成功后HH组小鼠RVSP、RVHI和PA thickness显著高于/大于对照组，差异均有统计学意义(均P＜0.05)。末次外泌体注射后4 d，重新评估各组小鼠的肺血管阻力及肺血管形态。结果显示，HH-exo组小鼠的RVSP、RVHI以及PA thickness均显著低于/小于HH组［（21.0±3.8）mmHg（1 mmHg=0.133 kPa）比（26.8±3.6）mmHg、（0.256±0.016）比（0.361±0.018）、（0.54±0.11）比（0.79±0.13）］（均P<0.05）。HH-exo组α-SMA、Col1-a1、FN的mRNA和蛋白表达水平均显著低于HH组(均P＜0．05)。结论 肺血管外膜纤维化是低压低氧相关的小鼠肺动脉高压的主要组织病理学特征。hESC-exo可通过减轻肺血管外膜纤维化，有效缓解低压低氧所致的小鼠HAPH。

• Objective To investigate the mechanism of high-altitude pulmonary hypertension (HAPH) and the intervention effect of human embryonic stem cell exosomes (hESC-exo) on HAPH. Methods Thirty 8-week-old C57BL/6 male mice were randomly divided into three groups: control group, hypobaric hypoxia group (HH group), hypobaric hypoxia hESC-exo intervention group (HH-exo group), with 10 mice in each group. The mice in the control group were fed in the normobaric and normoxic environment for 35 d. The mice in HH and HH-exo groups were exposed to hypobaric hypoxia for 28 d and then exposed to normobaric and normoxic environment for another 7 d. On day 29, 30 and 31 of the experiment, the HH-exo group was injected with 20 μg hESC-exo via the tail vein, and the control group and the hypobaric hypoxia group were injected with the same volume of phosphate buffer solution. Right ventricular systolic pressure (RVSP), right ventricular hypertrophy index (RVHI) and pulmonary arterial wall thickness (PA thickness) were evaluated on day 35. The mRNA and protein expression levels of α-smooth muscle actin (α-SMA), α-1 collagen (Col1-a1) and fibronectin (FN) were detected by real-time fluorescent quantitative polymerase chain reaction and Western blotting. Results After successful modeling, RVSP, RVHI and PA thickness in HH group were significantly higher/larger than those in control group (all P<0.05). Four days after the last exosome injection, the pulmonary vascular resistance and pulmonary vascular morphology of mice in each group were re-evaluated. The results  showed that the RVSP, RVHI and PA thickness of the HH-exo group were significantly lower/smaller than those of the HH group ［(21.0±3.8)mmHg vs (26.8±3.6)mmHg, (0.256±0.016) vs (0.361±0.018), (0.54±0.11) vs (0.79±0.13)］(all P<0.05). The mRNA and protein expression levels of α-SMA, Col1-a1, and FN in the HH-exo group were significantly lower than those in the HH group (all P<0.05). Conclusions Pulmonary adventitial fibrosis is the main histopathological feature of hypoxia-induced pulmonary hypertension in mice. hESC-exo can effectively alleviate HAPH induced by hypobaric hypoxia in mice by reducing pulmonary adventitial fibrosis.