Mitochondrial catalase overexpressed transgenic mice are protected against lung fibrosis in part via preventing alveolar epithelial cell mitochondrial DNA damage.
Mitochondrial catalase overexpressed transgenic mice are protected against lung fibrosis in part via preventing alveolar epithelial cell mitochondrial DNA damage.
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DOI：
10.1016/j.freeradbiomed.2016.11.007
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发表时间：
2016-12
影响因子：
7.4
通讯作者：
Kamp DW
中科院分区：
文献类型：
--
作者：
Kim SJ;Cheresh P;Jablonski RP;Morales-Nebreda L;Cheng Y;Hogan E;Yeldandi A;Chi M;Piseaux R;Ridge K;Michael Hart C;Chandel N;Scott Budinger GR;Kamp DW
肺泡上皮细胞（AEC）损伤和线粒体功能障碍在肺纤维化的发展中很重要。我们的研究小组已经证明，在石棉暴露的肺中，AEC 中线粒体活性氧 (ROS) 的产生介导了肺纤维化所必需的线粒体 DNA (mtDNA) 损伤和细胞凋亡。这些数据表明，针对线粒体的抗氧化剂应该可以改善石棉引起的肺损伤。确定表达线粒体靶向过氧化氢酶 (MCAT) 的转基因小鼠在暴露于石棉或博来霉素后是否减少了肺纤维化，如果是这样，这是否与减少 AEC mtDNA 损伤和细胞凋亡有关。将青石棉 (100 μg/50 μL)、TiO2（阴性对照）、博来霉素 (0.025 单位/50 μL) 或 PBS 经气管内滴注到 8-10 周龄野生型 (WT - C57Bl/6 J) 或 MCAT 小鼠中。 21天时收获肺。肺纤维化通过胶原水平（Sircol）和肺纤维化评分进行量化。通过裂解 caspase-3 (CC-3)/表面活性蛋白 C (SFTPC) 免疫组织化学 (IHC) 和半定量分析评估 AEC 凋亡。 AEC（来自 WT 和 MCAT 小鼠的原代 AT2 细胞以及 MLE-12 细胞）通过基于定量 PCR 的测定评估 mtDNA 损伤，通过 DNA 片段化评估细胞凋亡，并通过 Mito-Sox 测定评估 ROS 产生。与 WT 相比，通过肺胶原水平和肺纤维化评分测量，青石棉暴露的 MCAT 小鼠表现出肺纤维化减少。对 MCAT 小鼠的保护作用伴随着 AEC mtDNA 损伤和细胞凋亡的减少。接触博莱霉素后也发现了类似的结果。 Euk-134 是一种线粒体 SOD/过氧化氢酶模拟物，可减轻 MLE-12 细胞 DNA 损伤和细胞凋亡。最后，与 WT 相比，石棉诱导的 MCAT AT2 细胞 ROS 产生减少。我们发现，MCAT 小鼠在暴露于石棉或博来霉素后，肺纤维化、AEC mtDNA 损伤和细胞凋亡减少，这表明 AEC 线粒体 H2O2 诱导的 mtDNA 损伤在促进肺纤维化中发挥着重要作用。我们推断，旨在限制线粒体 H2O2 产生过多导致的 AEC mtDNA 损伤的策略可能是减轻肺纤维化的新治疗靶点。
Alveolar epithelial cell (AEC) injury and mitochondrial dysfunction are important in the development of lung fibrosis. Our group has shown that in the asbestos exposed lung, the generation of mitochondrial reactive oxygen species (ROS) in AEC mediate mitochondrial DNA (mtDNA) damage and apoptosis which are necessary for lung fibrosis. These data suggest that mitochondrial-targeted antioxidants should ameliorate asbestos-induced lung. To determine whether transgenic mice that express mitochondrial-targeted catalase (MCAT) have reduced lung fibrosis following exposure to asbestos or bleomycin and, if so, whether this occurs in association with reduced AEC mtDNA damage and apoptosis. Crocidolite asbestos (100 μg/50 μL), TiO2 (negative control), bleomycin (0.025 units/50 μL), or PBS was instilled intratracheally in 8–10 week-old wild-type (WT - C57Bl/6 J) or MCAT mice. The lungs were harvested at 21 d. Lung fibrosis was quantified by collagen levels (Sircol) and lung fibrosis scores. AEC apoptosis was assessed by cleaved caspase-3 (CC-3)/Surfactant protein C (SFTPC) immunohistochemistry (IHC) and semi-quantitative analysis. AEC (primary AT2 cells from WT and MCAT mice and MLE-12 cells) mtDNA damage was assessed by a quantitative PCR-based assay, apoptosis was assessed by DNA fragmentation, and ROS production was assessed by a Mito-Sox assay. Compared to WT, crocidolite-exposed MCAT mice exhibit reduced pulmonary fibrosis as measured by lung collagen levels and lung fibrosis score. The protective effects in MCAT mice were accompanied by reduced AEC mtDNA damage and apoptosis. Similar findings were noted following bleomycin exposure. Euk-134, a mitochondrial SOD/catalase mimetic, attenuated MLE-12 cell DNA damage and apoptosis. Finally, compared to WT, asbestos-induced MCAT AT2 cell ROS production was reduced. Our finding that MCAT mice have reduced pulmonary fibrosis, AEC mtDNA damage and apoptosis following exposure to asbestos or bleomycin suggests an important role for AEC mitochondrial H2O2-induced mtDNA damage in promoting lung fibrosis. We reason that strategies aimed at limiting AEC mtDNA damage arising from excess mitochondrial H2O2 production may be a novel therapeutic target for mitigating pulmonary fibrosis.
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影响因子：
82.9
作者：
Cloonan SM;Glass K;Laucho-Contreras ME;Bhashyam AR;Cervo M;Pabón MA;Konrad C;Polverino F;Siempos II;Perez E;Mizumura K;Ghosh MC;Parameswaran H;Williams NC;Rooney KT;Chen ZH;Goldklang MP;Yuan GC;Moore SC;Demeo DL;Rouault TA;D'Armiento JM;Schon EA;Manfredi G;Quackenbush J;Mahmood A;Silverman EK;Owen CA;Choi AM
通讯作者：
Choi AM
影响因子：
10.8
作者：
Dai, Dao-Fu;Hsieh, Edward J.;Rabinovitch, Peter S.
通讯作者：
Rabinovitch, Peter S.
影响因子：
4.8
作者：
Jain, Manu;Rivera, Stephanie;Chandel, Navdeep S.
通讯作者：
Chandel, Navdeep S.
影响因子：
4.8
作者：
Handy, Diane E.;Lubos, Edith;Loscalzo, Joseph
通讯作者：
Loscalzo, Joseph
影响因子：
4.8
作者：
He, Chao;Ryan, Alan J.;Carter, A. Brent
通讯作者：
Carter, A. Brent