| 赵雯月,李娜,李科君,王彦,何宁宁,杜利清,刘强.牙龈间充质干细胞抑制Ⅱ型肺泡上皮细胞衰老预防放射性肺纤维化[J].中华放射医学与防护杂志,2022,42(11):830-838 |
| 牙龈间充质干细胞抑制Ⅱ型肺泡上皮细胞衰老预防放射性肺纤维化 |
| Gingival mesenchymal stem cells inhibited senescence of type Ⅱ alveolar epithelial cells and prevented radiation-induced pulmonary fibrosis |
| 投稿时间：2022-08-23 |
| DOI：10.3760/cma.j.cn112271-20220823-00341 |
| 中文关键词: 放射性肺纤维化 细胞衰老 干细胞治疗 牙龈间充质干细胞 |
| 英文关键词:Radiation-induced pulmonary fibrosis Cellular senescence Stem cell therapy Gingival mesenchymal stem cells |
| 基金项目:国家自然科学基金(32071241,31971168);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-042) |
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| 中文摘要: |
| 目的 探讨牙龈间充质干细胞(GMSCs)移植能否抑制辐射诱导的Ⅱ型肺泡上皮细胞(AECⅡ)衰老,及其在预防放射性肺纤维化(RIPF)中的作用。方法 采用6 Gy照射小鼠Ⅱ型肺泡上皮细胞(MLE12)诱导细胞衰老,照后即刻与GMSCs共培养,通过细胞形态、β半乳糖苷酶(β-Gal)染色、衰老分泌相关表型(SASP)检测以评估细胞衰老程度;C57BL/6小鼠单侧右肺17 Gy照射构建RIPF模型,照后1 d移植GMSCs,照后180 d取材,期间记录小鼠生存率,采用肺脏器比、HE染色、Masson染色评估肺内结构及肺间质胶原沉积状况,应用β-Gal免疫组织化学法与AECⅡ免疫荧光共定位评估肺内细胞衰老程度,qRT-PCR检测肺组织SASP表达变化;采用Western blot检测P53-P21、P16通路关键蛋白表达水平,免疫荧光共定位检测AECⅡ中P21表达情况。结果 GMSCs共培养能够有效抑制辐射诱导的MLE12细胞衰老,使照射后升高的β-Gal阳染率降低11.8%(t=6.72,P<0.05),并使SASP(IL-6、IL-8、IL-1β)的表达有效降低(t=28.43、28.43、4.82,P<0.05);GMSCs移植提高了受照小鼠的生存率,改善了照射诱导的肺泡结构塌陷增厚及胶原蛋白沉积情况,照射后肺组织中衰老细胞数目降低23.9%(t=21.83,P<0.05),SASP的表达水平下降(t=8.86、20.63,P<0.05);抑制照射后MLE12及RIPF小鼠肺组织中P53-P21、P16相关蛋白的上调。结论 GMSCs通过下调衰老相关的P53-P21及P16通路蛋白,从而抑制辐射诱导的AECⅡ衰老,达到预防放射性肺纤维化发生发展的目的。 |
| 英文摘要: |
| Objective To investigate whether transplantation of gingival mesenchymal stem cells (GMSCs) can inhibit radiation-induced senescence of alveolar epithelial cells type Ⅱ (AECⅡ) and its role in the prevention of radiation-induced pulmonary fibrosis (RIPF).Methods Mouse type Ⅱ alveolar epithelial cells (MLE12) were irradiated with 6 Gy X-rays and then co-cultured with GMSCs. The extent of cellular senescence of MLE12 cells was assessed by cell morphology, β-Gal staining, and senescence secretion-associated phenotype (SASP) assay. RIPF model was constructed by unilaterally irradiating the right chest of C57BL/6 mice with 17 Gy X-rays. GMSCs were transplanted 1 d after irradiation. At 180 d after irradiation, the pulmonary organ ratio, HE staining, and Masson staining were used to assess intra-pulmonary structure and interstitial collagen deposition in the lung. β-Gal immunohistochemistry and immunofluorescence co-localization with AECⅡ were measured to assess the degree of cellular senescence in the lung. The SASP expression changes in lung tissue were detected by qRT-PCR. The protein expressions in P53-P21 and P16 pathways were detected by Western blot assay. P21 expression in AECⅡ was detected by immunofluorescence co-localization assay.Results GMSCs effectively inhibited radiation-induced senescence of MLE12 cells, reduced the ratio of radiation-elevated β-Gal positive cells by 11.8% (t=6.72, P<0.05), and decreased the expressions of SASP (IL-6, IL-8, IL-1β) (t=28.43, 28.43, 4.82, P<0.05). GMSCs transplantation improved the survival rate of irradiated mice, prevented radiation-induced alveolar structural collapse thickening and collagen deposition, reduced the number of senescent cells in the irradiated lung tissues by 23.9% (t=21.83,P<0.05), and inhibited the expressions of SASP (t=8.86, 20.63, P<0.05). GMSCs also inhibited the expression of P53-P21, P16-related proteins in MLE12 cells and lung tissues of mice after irradiation.Conclusions GMSCs inhibit senescence-related P53-P21 and P16 pathways, prevent radiation-induced AECⅡ senescence, as well as the development of RIPF. |
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