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没食子酸单宁部位通过调控 NF-κB 和 MAPK 信号通路抑制 RAW264.7 细胞炎症反应

Modulation of NF-κB and MAPK signalling pathways by hydrolysable tannin fraction from Terminalia chebula fruits contributes to its anti-inflammatory action in RAW 264.7 cells.

机构信息

Department of Pharmaceutical Technology, University College of Engineering, BIT Campus, Anna University, Tiruchirappalli, Tamil Nadu, India.

Department of Biotechnology, University College of Engineering, BIT Campus, Anna University, Tiruchirappalli, Tamil Nadu, India.

出版信息

J Pharm Pharmacol. 2022 May 20;74(5):718-729. doi: 10.1093/jpp/rgab178.

Abstract

OBJECTIVES

Hydrolysable tannin fraction (HTF) derived from Terminalia chebula fruit pericarps was assessed for its anti-inflammatory potential in LPS-induced RAW 264.7 cells. Its molecular mechanism was also established and compared with individual tannins - chebulagic acid (CH) and corilagin (CO).

METHODS

The effect of HTF on LPS-stimulated RAW 264.7 cells was studied by estimating the release of NO, ROS, cytokines and changes in nuclear morphology by DAPI staining. Furthermore, the effect of HTF, CO and CH was compared with the expression of p65, p38 and pERK proteins by immunoblotting and the mRNA transcript level of COX-2, iNOS and TNF-α by quantitative PCR. The in-silico interactions of various hydrolysable tannins present in HTF with molecular targets of inflammation were studied using Maestro software.

KEY FINDINGS

HTF at the dose levels of 25, 50 and 100 µg/ml was able to decrease the release of NO, ROS and cytokines from LPS-induced RAW 264.7 cells without disturbing the cell nuclear morphology. Investigation of molecular mechanism revealed that inhibition of NF-κB and MAPK signalling pathways was responsible for its anti-inflammatory action. The effect of HTF was higher than the individual tannins CH and CO.

CONCLUSION

HTF can be developed as an effective anti-inflammatory agent.

摘要

目的

从诃子果皮中提取的可水解单宁(HTF),评估其在 LPS 诱导的 RAW 264.7 细胞中的抗炎潜力。并建立其分子机制,并与单体单宁 - 诃子酸(CH)和柯里拉京(CO)进行比较。

方法

通过估计 LPS 刺激的 RAW 264.7 细胞中 NO、ROS、细胞因子的释放和 DAPI 染色核形态的变化来研究 HTF 对 LPS 刺激的 RAW 264.7 细胞的影响。此外,通过免疫印迹比较 HTF、CO 和 CH 对 p65、p38 和 pERK 蛋白表达的影响,通过定量 PCR 比较 COX-2、iNOS 和 TNF-α 的 mRNA 转录水平。使用 Maestro 软件研究 HTF 中存在的各种可水解单宁与炎症分子靶点的体内相互作用。

主要发现

HTF 在 25、50 和 100 µg/ml 的剂量水平下能够降低 LPS 诱导的 RAW 264.7 细胞中 NO、ROS 和细胞因子的释放,而不会干扰核形态。对分子机制的研究表明,抑制 NF-κB 和 MAPK 信号通路是其抗炎作用的原因。HTF 的作用强于单体单宁 CH 和 CO。

结论

HTF 可开发为有效的抗炎药物。

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