摘要
目的探讨FOXO4-DRI能否逆转放射性肺纤维化(RIPF)及其作用机制。方法将C57BL/6小鼠随机分为对照组、FOXO4-DRI组、照射组、照射+FOXO4-DRI组,照射组和照射+FOXO4-DRI组小鼠右侧全胸接受17 Gy X射线照射,FOXO4-DRI组和照射+FOXO4-DRI组小鼠照射后第16周和第20周分别腹腔注射FOXO4-DRI。照射后第24周收集小鼠右肺组织,进行HE染色和Masson三色染色,观察肺组织形态学改变和胶原沉积情况;免疫组织化学染色法检测肺组织中col1α1和α-SMA的表达;β-半乳糖苷酶(β-gal)染色观察衰老细胞;活性氧试剂盒检测肺组织活性氧水平;RT-PCR检测P21、P16^(Ink4a)和衰老相关分泌表型(SASP)mRNA的表达;Western blot检测相关蛋白表达水平。结果FOXO4-DRI减少了RIPF小鼠肺组织中胶原组织的沉积(t=6.18,P<0.05),降低了col1α1和α-SMA的表达(t=4.69、3.20,P<0.05)。FOXO4-DRI减少了RIPF小鼠肺组织中β-gal阳性细胞的数量(t=6.09,P<0.05)。FOXO4-DRI能够抑制RIPF小鼠肺组织中P21和P16^(Ink4a)基因、蛋白的表达(t=5.31、3.32和4.77、3.37,P<0.05),抑制SASP基因IL-1α、IL-1β、肿瘤坏死因子α(TNFα)和MMP2的表达(t=4.36、4.84、4.47、3.82,P<0.05)。FOXO4-DRI能够降低RIPF小鼠肺组织中活性氧水平(t=2.84,P<0.05),促进p-AKT和p-PI3K蛋白的激活(t=-7.13、-12.61,P<0.05)。结论FOXO4-DRI通过激活PI3K/AKT信号通路减少氧化应激、抑制细胞衰老,逆转RIPF。
Objective To evaluate whether FOXO4-DRI could reverse radiation-induced pulmonary fibrosis(RIPF)and to explore the underlying mechanism.Methods C57BL/6 mice were randomly divided into 4 groups:control,FOXO4-DRI,radiation,and radiation+FOXO4-DRI.Mice in radiation or radiation+FOXO4-DRI groups received 17 Gy X-ray radiation on the right side of the whole chest.Mice in FOXO4-DRI and radiation+FOXO4-DRI groups were injected with FOXO4-DRI intraperitoneally at 16 and 20 weeks after irradiation,respectively.The right lungs were collected at 24 weeks after irradiation and subjected to HE staining and Masson trichrome staining to observe the morphological changes and collagen deposition.Immunohistochemistry was used to evaluate the expressions of col1α1 andα-SMA in lung tissues.β-gal staining was used to observe senescent cells.The level of reactive oxygen species in lung tissue was detected.The expressions of P21,P16^(Ink4a)and senescenceassociated secretory phenotype(SASP)mRNA were detected by qRT-PCR,and the expression of related proteins were assessed by Western blot.Results FOXO4-DRI reduced collagen deposition(t=6.18,P<0.05),down-regulated the expression of col1α1 andα-SMA(t=4.69,3.20,P<0.05),and reduced the number ofβ-gal positive cells(t=6.09,P<0.05)in the lung tissue of RIPF mice.FOXO4-DRI also down-regulated the gene and protein expressions of P21 and P16^(Ink4a)(t=5.31,3.32 and 4.77,3.37,P<0.05)and inhibited the expressions of SASP genes IL-1α,IL-1β,TNF-αand MMP2(t=4.36,4.84,4.47,3.82,P<0.05),reduced reactive oxygen species(t=2.84,P<0.05),and promoted the activation of p-AKT and p-PI3K proteins(t=-7.13,-12.61,P<0.05)in the lung tissue of RIPF mice.Conclusions FOXO4-DRI reverses RIPF by activating the PI3K/AKT signaling pathway,reducing oxidative stress and inhibiting cellular senescence.
作者
韩晓丹
苑通
宋迪
张俊伶
石永刚
Han Xiaodan;Yuan Tong;Song Di;Zhang Junling;Shi Yonggang(Department of Radiation Oncology,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Institute of Radiation Medicine,Peking Union Medical College and Chinese Academy of Medical Science,Tianjin 300192,China;Zhengzhou University,Zhengzhou 450001,China)
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2023年第9期669-675,共7页
Chinese Journal of Radiological Medicine and Protection
基金
国家自然科学基金(81903254)。
