Department of Pharmacy, Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine, No.1665, Kongjiang Road, Yangpu District, Shanghai, 200092, China.
Department of Pharmacology and Chemical Biology, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
Inflammation. 2019 Dec;42(6):2246-2256. doi: 10.1007/s10753-019-01089-9.
The present study investigated changes in cellular phenotype and oxidative stress during the inflammatory response in PC12 cells stimulated by lipopolysaccharide (LPS) and assessed the effects of minocycline, astragalus (AST), and baicalin on inflammation. PC12 cells were exposed to LPS with or without minocycline, AST, or baicalin. Cell viability was measured by a thiazolyl blue tetrazolium bromide (MTT) assay. Contrast and laser confocal microscopy were used to analyze changes in cellular phenotype and cytoskeleton synthesis. Western blotting tested the expression of α7nAChR and vimentin. Inhibitory ratio of superoxide dismutase (SOD) activity and leakage of lactate dehydrogenase (LDH) were detected to evaluate cellular oxidative stress. Results showed that LPS could attenuate PC12 cell viability in a time- and dose-dependent manner, which could be rescued by minocycline. In addition, minocycline could reverse PC12 cell phenotypic change and the synthesis of the mesenchymal cytoskeleton protein vimentin, both induced by LPS. During LPS-initiated inflammation, α7nAChR and vimentin expression were obviously inhibited by minocycline, AST, or baicalin. The inhibitory rate of SOD activity and LDH leakage in PC12 cells were increased by LPS and attenuated significantly when exposed to minocycline, AST, or baicalin. These findings suggest phenotype change, altered cytoskeleton protein synthesis, and oxidative stress are all involved in the inflammatory response in PC12 cells during which α7 nicotinic acetylcholine receptor (α7nAChR) is induced by LPS stimulation. Minocycline, AST, and baicalin have a protective effect against PC12 cell injury, acting as antioxidants and inhibitors of mesenchymal proteins.
本研究探讨了脂多糖（LPS）刺激 PC12 细胞炎症反应过程中细胞表型和氧化应激的变化，并评估了米诺环素、黄芪（AST）和黄芩苷对炎症的影响。将 PC12 细胞暴露于 LPS 及 LPS 加米诺环素、AST 或黄芩苷中。噻唑蓝（MTT）法检测细胞活力。对比和激光共聚焦显微镜用于分析细胞表型和细胞骨架合成的变化。Western blot 检测α7nAChR 和波形蛋白的表达。检测超氧化物歧化酶（SOD）活性抑制率和乳酸脱氢酶（LDH）漏出率，以评估细胞氧化应激。结果表明，LPS 可呈时间和剂量依赖性减弱 PC12 细胞活力，米诺环素可挽救这一现象。此外，米诺环素可逆转 LPS 诱导的 PC12 细胞表型改变和间充质细胞骨架蛋白波形蛋白的合成。在 LPS 引发的炎症中，α7nAChR 和波形蛋白的表达被米诺环素、AST 或黄芩苷明显抑制。SOD 活性抑制率和 LPS 诱导的 LDH 漏出量在 PC12 细胞中增加，当暴露于米诺环素、AST 或黄芩苷时，明显减弱。这些发现表明，表型改变、细胞骨架蛋白合成改变和氧化应激均参与 LPS 刺激的 PC12 细胞炎症反应，其中α7 烟碱型乙酰胆碱受体（α7nAChR）被 LPS 诱导。米诺环素、AST 和黄芩苷对 PC12 细胞损伤具有保护作用，作为抗氧化剂和间充质蛋白抑制剂。
