Siegesbeckia pubescens Makino inhibits Pam3CSK4-induced inflammation in RAW 264.7 macrophages through suppressing TLR1/TLR2-mediated NF-κB activation

被引：37

作者：

Sang, Wei ^{[1
]}

Zhong, Zhangfeng ^{[1
,4
]}

Linghu, Kegang ^{[1
]}

Xiong, Wei ^{[1
]}

Tse, Anfernee Kai Wing ^{[5
]}

Cheang, Wai San ^{[1
]}

Yu, Hua ^{[1
,2
,3
,6
]}

Wang, Yitao ^{[1
,7
]}

机构：

[1] Univ Macau, Inst Chinese Med Sci, State Key Lab Qual Res Chinese Med, Macau, Peoples R China

[2] HKBU Shenzhen Res Ctr, Shenzhen, Guangdong, Peoples R China

[3] Hong Kong Baptist Univ, Sch Chinese Med, Kowloon Tong, Hong Kong, Peoples R China

[4] Guangdong Med Univ, Guangdong Key Lab Res & Dev Nat Drugs, Zhanjiang, Peoples R China

[5] Southern Univ Sci & Technol, Acad Adv Interdisciplinary Studies, Shenzhen, Guangdong, Peoples R China

[6] Univ Macau, Inst Chinese Med Sci, Room 8008,Bldg N22,Ave Univ, Taipa, Macao, Peoples R China

[7] Univ Macau, Inst Chinese Med Sci, Room 1050,Bldg N22,Ave Univ, Taipa, Macao, Peoples R China

来源：

CHINESE MEDICINE | 2018年 / 13卷

基金：

中国国家自然科学基金; 中国博士后科学基金;

关键词：

Siegesbeckia pubescens Makino; Pam(3)CSK(4); Inflammation; Toll-like receptor 1/2; NF-kappa B; TOLL-LIKE RECEPTORS; THERAPEUTIC TARGETS; DISEASES; CELLS; DITERPENOIDS; CYTOKINES; RECOGNITION; EXTRACTION; APOPTOSIS;

D O I：

10.1186/s13020-018-0193-x

中图分类号：

R [医药、卫生];

学科分类号：

10 ;

摘要：

Background: Siegesbeckia pubescens Makino (SP) is one of the important plant origins for the anti-inflammatory Chinese herbal medicine of Siegesbeckiae Herba. The current investigations indicated that the anti-inflammatory effects of SP were associated with the toll-like receptors (TLRs)-mediated nuclear factor-kB (NF-kB) and the mitogen-activated protein kinase (MAPK) signaling pathways. Methods: Raw 264.7 macrophages were pretreated with the 50% ethanol extract of SP (SPE, 50-200 mu g/mL) and then co-treated with Pam(3)CSK(4) (200 ng/mL) for another 12 h. The inhibitory effect of SPE on Pam(3)CSK(4)-stimulated NO release and post-inflammatory cytokines secretions were determined using Griess reagent and Elisa kits, respectively. The influence of SPE on NF-kB and MAPKs signaling relevant proteins was measured by Western blotting analysis, while the intracellular nitric oxide (NO) generation and NF-kB/p65 nuclear translocation were determined using Leica TCS SP8 laser scanning confocal microscope. Moreover, the effect of SPE on luciferase reporter gene in NF-kB-Iuc DNA transfected raw 264.7 cells was determined using the Dual-Glo luciferase assay system kit. Results: SPE dose-dependently (50-200 mu g/mL) attenuated Pam(3)CSK(4)-induced NO release, post-inflammatory cytokines (IL-6,TNF-alpha and MCP-1) secretions and intracellular NO generation in raw 264.7 cells. Biologically, SPE suppressed Pam(3)CSK(4)-induced expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), phosphorylation ofNF-kB/p65 and IkB alpha, but did not significantly show effect on the proteins involved in MAPKs signaling (p38, ERK and JNK). The results were further confirmed by NF-kB-luc reporter gene assay and p65 nuclear translocation assay. Conclusions: In conclusion, SPE ameliorated Pam(3)CSK(4)-induced inflammation in raw 264.7 cells through suppressing TLR 1/2-mediated NF-kB activation.

引用

页数：10