Inhibition of protein kinase C alpha attenuates lipopolysaccharide-triggered acute lung injury by alleviating the hyperinflammatory response and oxidative stress.
Inhibition of protein kinase C alpha attenuates lipopolysaccharide-triggered acute lung injury by alleviating the hyperinflammatory response and oxidative stress.
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DOI：
10.21037/atm-21-6497
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发表时间：
2022-03
影响因子：
--
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Chen Y;Lin P;Nan W;Su S;Zheng H;Gao H;Zhang D;Li Y
目前可用的治疗方法在降低急性肺损伤（ALI）的死亡率或改善预后方面无效。蛋白激酶C α（PKCα）的激活最近被认为与ALI的发生有关。本研究旨在探讨PKCα抑制剂对急性肺损伤的潜在治疗效果及相关机制。检测脂多糖（LPS）处理的C57 BL/6 J小鼠（雄性）和用PKCα抑制剂预处理的巨噬细胞的肺炎症和损伤指标。收集组织以通过苏木精和伊红（H&E）染色评估肺损伤。采用二辛可宁酸（BCA）法和酶联免疫吸附法（ELISA）检测肺水肿、炎症反应和氧化应激。我们使用细胞计数试剂盒-8（CCK-8）和乳酸脱氢酶（LDH）细胞毒性测定试剂盒、流式细胞术、定量逆转录聚合酶链反应（qRT-PCR）和ELISA检测了PKCα抑制对LPS诱导的巨噬细胞增殖、细胞毒性、氧化损伤和炎性细胞因子释放的影响。采用Western blot、免疫组化和免疫荧光染色检测活化B细胞核因子κ轻链增强子（NF-κB）信号通路相关蛋白的表达。我们观察到LPS上调PKCα磷酸化，诱导高炎症反应，并引起肺损伤。而抑制PKCα可有效地减轻LPS引起的变化。此外，我们证实抑制PKCα可减弱LPS诱导的ALI下NF-κB通路的活性。提示抑制PKCα对LPS诱导的ALI高炎症反应具有保护作用，其机制可能与下调NF-κB信号通路有关。结果表明，抑制PKCα可减轻ALI，其机制可能与其抗炎、抗氧化作用密切相关。
The currently available treatment methods are ineffective in reducing mortality or improving outcomes in acute lung injury (ALI). The activation of protein kinase C alpha (PKCα) has recently been implicated in ALI development. We explored the potential therapeutic outcomes of PKCα inhibition in cases of ALI and to elucidate the related mechanisms. Indexes of lung inflammation and injury were examined in lipopolysaccharide (LPS)-treated C57BL/6J mice (male) and macrophages after pretreatment with a PKCα inhibitor. Tissues were collected to assess lung injury by hematoxylin and eosin (H&E) staining. Bronchoalveolar lavage fluid was used to measure the pulmonary edema, hyperinflammatory response, and oxidative stress by bicinchoninic acid (BCA) method and enzyme-linked immunosorbent assay (ELISA). We tested the effect of PKCα inhibition on LPS-induced proliferation, cytotoxicity, oxidative damage, and the release of inflammatory cytokines in macrophages using the Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) cytotoxicity assay kit, flow cytometry, quantitative reverse-transcription polymerase chain reaction (qRT-PCR), and ELISA. The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway related proteins were detected by Western blot, immunohistochemistry (IHC), and immunofluorescence staining. We observed that LPS upregulated PKCα phosphorylation, induced a hyperinflammatory response, and caused lung injury. However, PKCα inhibition effectively attenuated the changes caused by LPS. Moreover, we confirmed that inhibiting PKCα weakened the activity of the NF-κB pathway under LPS-induced ALI. These findings indicated that inhibition of PKCα is protective against LPS-induced hyperinflammatory response in ALI, this effect is likely to attributed to the downregulation of NF-κB signaling pathways. The results showed that PKCα inhibition could attenuate ALI which may closely related to its anti-inflammatory and anti-oxidative effects.
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影响因子：
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作者：
Hsu HT;Tseng YT;Wong WJ;Liu CM;Lo YC
通讯作者：
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影响因子：
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