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| Qinggan Dongyin inhibits LPS-induced inflammation of RAW264.7 macrophages by regulating NF-κB/iNOS/NO signaling pathway |
| Hits 1285 Download times 712 Received:August 20, 2022 |
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| DOI
10.11656/j.issn.1673-9043.2022.06.13 |
| Key Words
Qinggan Dongyin;RAW264.7 macrophages;lipopolysaccharides;anti-inflammation;NF-κB/iNOS/NO signaling pathway |
| Author Name | Affiliation | E-mail | | SU Rui | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | LU Jia | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | XI Zhinan | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | WANG Jiabao | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | SONG Xinbo | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin Modern Innovation Traditional Chinese Medicine Technology Co. Ltd., Tianjin 300392, China | | | ZHANG Han | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | MIAO Lin | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | mmmlin@tjutcm.cn |
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| Abstract
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| [Objective] To investigate the anti-inflammatory effects and mechanism of Qinggan Dongyin(QGDY) by establishing lipopolysaccharide (LPS)-induced inflammation model in RAW264.7 macrophages. [Methods] CCK-8 assay and Cytotoxicity LDH Assay were used to determine the effects of different concentrations of QGDY on cell viability and LDH leakage in HEK293T cells and RAW264.7 macrophages. Antioxidant reaction element (ARE) and nuclear factor-κB (NF-κB) dual luciferase reporter systems were established to examine the effects of QGDY on the expression of ARE and NF-κB in HEK293T cells,respectively. LPS-induced RAW264.7 macrophages inflammatory model was established in vitro. RAW264.7 cells were randomly divided into control group,model group,and QGDY low-dose group,medium-dosegroup,high-dose group (1,10,and 100 μg/mL). The morphological differences of cells in each group were observed by inverted microscope. The release of nitric oxide(NO),tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in supernatant were determined by Griess assay and ELISA kits. The mRNA expression levels of TNF-α,IL-6 and IL-1β were detected by qRT-PCR assay. The nuclear translocation of p65 was observed by immunofluorescence. The protein expressions of p65 and iNOS were detected in each group by western blot. [Results] The 0.01-10 μg/mL QGDY had no significant effect on cell viability of HEK293T cells, 1-10 μg/mL QGDY obviously inhibited the activity of NF-κB promoter induced by TNF-α,and QGDY had no significant effect on the activity of ARE promoter,suggesting that QGDY had anti-inflammatory effect but no obvious antioxidant effect. QGDY(0.01-100 μg/mL) had no cytotoxic effect on RAW264.7 macrophages after 24 h of intervention. Compared with the model group,the release of NO and inflammatory factors TNF-α,IL-6 and IL-1β were significantly inhibited by 1-100 μg/mL QGDY in LPS-induced RAW264.7 macrophages. Compared with model group,10-100 μg/mL QGDY groups not only decresed the mRNA expression levels of TNF-α,IL-6 and IL-1β,but also inhibited the nuclear translocation of p65 in RAW264.7 macrophages,significantly. In addition,100 μg/mL QGDY significantly inhibited the expression of iNOS and nuclear protein p65,and significantly promoted the expression of plasma protein p65. [Conclusion] QGDY could inhibit the inflammatory response of RAW264.7 macrophages induced by LPS,which might be related to the regulation of NF-κB/iNOS/NO signaling pathway and the inhibition of the release of pro-inflammatory factors. |
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