Hwangryunhaedoktang exerts anti-inflammation on LPS-induced NO production by suppressing MAPK and NF-κB activation in RAW264.7 macrophages

doi:10.1016/S2095-4964(17)60350-9

Abstract

Objective

This study aimed to evaluate whether Hwangryunhaedoktang (HHT), a herbal compound, has an inhibitory effect on lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophages.

Methods

The effects of HHT were evaluated by confirming nitric oxide (NO) production and expression of inducible NO synthase (iNOS) and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW264.7 macrophages via the Griess assay, Western blotting, and real-time reverse transcription quantitative polymerase chain reaction. Western blot analyses and luciferase assays were used to evaluate whether HHT has an effect on the phosphorylation and translocation of nuclear factor-κB (NF-κB). The secretion and expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were determined via enzyme-linked immunosorbent assay and Western blot analyses.

Results

HHT suppressed LPS-induced NO production and expression of iNOS in a dose-dependent manner. Additionally, MAPKs activation was also attenuated via inhibition of phosphorylation of extracellular signal-regulated kinases 1/2, c-Jun N-terminal kinase and p38 which were related to inflammatory pathway. Furthermore, HHT also effectively attenuated NF-κB activation and its translocation to the nucleus, a process that is closely linked to inflammation. LPS normally induced the expression of inflammatory cytokines such as TNF-α and IL-6, but the secretion and expression of TNF-α and IL-6 were significantly attenuated by pretreating the cells with HHT.

Conclusion

HHT suppressed LPS-induced NO production by blocking the activation of NF-κB and MAPK signaling pathways in RAW264.7 macrophages. Furthermore, HHT may have an anti-inflammatory effect by suppressing the LPS-induced secretion of TNF-α and IL-6. Therefore, the traditional herbal formula HHT might be a useful potential therapeutic agent for inflammation.

Key words: Medicine, Chinese traditional, Lipopolysaccharide, Nitric oxide, Nuclear factor-κB;, Mitogen-activated protein kinases, Tumor necrosis factor-α, Inflammation

Byung Hyuk Han, Yun Jung Lee, Jung Joo Yoon, Eun Sik Choi, Seung Namgung, Xian Jun Jin, Da Hye Jeong, Dae Gill Kang, Ho Sub Lee. Hwangryunhaedoktang exerts anti-inflammation on LPS-induced NO production by suppressing MAPK and NF-κB activation in RAW264.7 macrophages. Journal of Integrative Medicine, 2017, 15(4): 326-336.

/ / Recommend

Add to citation manager EndNote| Reference Manager|ProCite|BibTeX|RefWorks

URL: http://www.jcimjournal.com/EN/10.1016/S2095-4964(17)60350-9

http://www.jcimjournal.com/EN/Y2017/V15/I4/326

TrendMD

References

1 Sharma JN, Al-Omran A, Pavathy SS.Role of nitric oxide in inflammatory diseases.Inflammopharmacology. 2007; 15(6): 252-259.
2 Yun HY, Dawson VL, Dawson TM.Neurobiology of nitric oxide.Crit Rev Neurobiol. 1996; 10(3-4): 291-316.
3 Posadas I, Terencio MC, Guillén I, Ferrándiz ML, Coloma J, Payá M, Alcaraz MJ.Co-regulation between cyclooxygenase- 2 and inducible nitric oxide synthase expression in the time-course of murine inflammation.Naunyn Schmiedebergs Arch Pharmacol. 2000; 361(1): 98-106.
4 Isomäki P, Punnonen J.Pro- and anti-inflammatory cytokines in rheumatoid arthritis.Ann Med. 1997; 29(6): 499-507.
5 Chen JW, Chen YH, Lin FY, Chen YL, Lin SJ.Ginkgo biloba extract inhibits tumor necrosis factor-α-induced reactive oxygen species generation, transcription factor activation, and cell adhesion molecule expression in human aortic endothelial cells. Arterioscler Thromb Vasc Biol. 2003; 23(9): 1559-1566.
6 Gloire G, Legrand-Poels S, Piette J.NF-κB activation by reactive oxygen species: fifteen years later.Biochem Pharmacol. 2006; 72(11): 1493-1505.
7 Lawrence T, Gilroy DW, Colville-Nash PR, Willoughby DA.Possible new role for NF-κB in the resolution of inflammation.Nat Med. 2001; 7(12): 1291-1297.
8 Hawiger J, Veach RA, Liu XY, Timmons S, Ballard DW.IκB kinase complex is an intracellular target for endotoxic lipopolysaccharide in human monocytic cells.Blood. 1999; 94(5): 1711-1716.
9 Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K, Cobb MH.Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions.Endocr Rev. 2001; 22(2): 153-183.
10 Christman JW, Lancaster LH, Blackwell TS.Nuclear factor κB: a pivotal role in the systemic inflammatory response syndrome and new target for therapy.Intensive Care Med. 1998; 24(11): 1131-1138.
11 Guha M, Mackman N.LPS induction of gene expression in human monocytes.Cell Signal. 2001; 13(2): 85-94.
12 Angel P, Karin M.The role of Jun, Fos and the AP-1 complex in cell proliferation and transformation.Biochim Biophys Acta. 1991; 1072(2-3): 129-157.
13 Cobb MH.MAP kinase pathways.Prog Biophys Mol Biol. 1999; 71(3-4): 479-500.
14 Davis RJ.Signal transduction by the c-Jun N-terminal kinase.Biochem Soc Symp. 1999; 64: 1-12.
15 Potapova O, Basu S, Mercola D, Holbrook NJ.Protective role for c-Jun in the cellular response to DNA damage.J Biol Chem. 2001; 276(30): 28546-28553.
16 Delgado M.Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide inhibit the MEKK1/MEK4/JNK signaling pathway in endotoxinactivated microglia.Biochem Biophys Res Commun. 2002; 293(2): 771-776.
17 Wallach D, Varfolomeev EE, Malinin NL, Glitsev YV, Kovalenko AV, Boldin MP.Tumor necrosis factor receptor and Fas signaling mechanisms.Ann Rev Immunol. 1999; 17: 331-367.
18 Ettinger R, Mebius R, Browning JL, Michie SA, van Tuijl S, Kraal G, van Ewijk W, McDevitt HO. Effects of tumor necrosis factor and lymphotoxin on peripheral lymphoid tissue development.Int Immunol. 1998; 10(6): 727-741.
19 Yoon HJ, Moon ME, Park HS, IM SY, Kim YH.Chitosan oligosaccharide (COS) inhibits LPS-induced inflammatory effects in RAW264.7 macrophage cells.Biochem Biophys Res Commun. 2007; 358(3): 954-959.
20 Hirano T, Matsuda T, Nakajima K.Signal transduction through gp130 that is shared among the receptors for the interleukin 6 related cytokine subfamily.Stem Cells. 1994; 12(3): 262-277.
21 Kim MC, Lee CH, Yook TH.Effects of anti-inflammatory andRehmanniae radix pharmacopuncture on atopic dermatitis in NC/Nga mice. J Acupunct Meridian Stud. 2013; 6(2): 98-109.
22 Zeng H, Dou S, Zhao J, Fan S, Yuan X, Zhu S, Li L, Zhang W, Liu R.The inhibitory activities of the components of Huang-Lian-Jie-Du-Tang (HLJDT) on eicosanoid generation via lipoxygenase pathway.J Ethnopharmacol. 2011; 135(2): 561-568.
23 Kim NK, Lee DH, Seo HS, Sun SH, Oh YL, Kim JE, Yoon IH, Seo ES, Shim GS, Zaslawski C.Hwangryunhaedoktang in adult patients with atopic dermatitis: a randomised, double-blind, placebo-controlled, two-centre trial—study protocol.BMC Complement Altern Med. 2011; 11: 68.
24 Seo CS, Kim OS, Kim JH, Shin HK.Simultaneous quantification and antiatherosclerosis effect of the traditional Korean medicine, Hwangryunhaedok-tang.BMC Complement Altern Med. 2015; 15: 108.
25 Gilroy DW, Colville-Nash PR.New insights into the role of COX 2 in inflammation.J Mol Med (Berl). 2000; 78(3): 121-129.
26 Lin MW, Tsao LT, Chan LC, Chen YL, Huang LJ, Kuo SC, Tzeng CC, Lee MR, Wang JP.Inhibition of lipopolysaccharide-stimulated NO production by a novel synthetic compound CYL-4d in RAW264.7 macrophages involving the blockade of MEK4/JNK/AP-1 pathway.Biochem Pharmacol. 2007; 73(11): 1796-1806.
27 Ahmad N, Chen LC, Gordon MA, Laskin JD, Laskin DL.Regulation of cyclooxygenase-2 by nitric oxide in activated hepatic macrophages during acute endotoxemia.J Leukoc Biol. 2002; 71(6): 1005-1011.
28 Seibert K, Zhang Y, Leahy K, Hauser S, Masferrer J, Perkins W, Lee L, Isakson P.Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain.Proc Natl Acad Sci U S A. 1994; 91(25): 12013-12017.
29 Xie QW, Kashiwabara Y, Nathan C.Role of transcription factor NF-κB/Rel in induction of nitric oxide synthase.J Biol Chem. 1994; 269(7): 4705-4708.
30 Brasier AR.The nuclear factor-κB-interleukin-6 signalling pathway mediating vascular inflammation.Cardiovasc Res. 2010; 86(2): 211-218.
31 Surh YJ, Chun KS, Cha HH, Han SS, Keum YS, Park KK, Lee SS. Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: downregulation of COX-2 and iNOS through suppression of NF- κB activation. Mutat Res.2001; 480-481: 243-268.
32 Tsao LT, Tsai PS, Lin RH, Huang LJ, Kuo SC, Wang JP.Inhibition of lipopolysaccharide-induced expression of inducible nitric oxide synthase by phenolic (3E)-4-(2- hydroxyphenyl)but-3-en-2-one in RAW264.7 macrophages.Biochem Pharmacol. 2005; 70(4): 618-626.
33 Zhang JM, An JX.Cytokines, inflammation and pain.Int Anesthesiol Clin. 2009; 45(2): 27-37.
34 Aggarwal BB, Natarajan K.Tumor necrosis factor: developments during the last decade.Eur Cytokine Netw. 1996; 7(2): 93-124.

Related Articles 15

[1]	Keat Lam Ho, Phaik Har Yong, Chee Woon Wang, Umah Rani Kuppusamy, Chek Tung Ngo, Festo Massawe, Zhi Xiang Ng. Peperomia pellucida (L.) Kunth and eye diseases: A review on phytochemistry, pharmacology and toxicology. Journal of Integrative Medicine, 2022, 20(4): 292-304.
[2]	Jia-yi Zhao, Jin Pu, Jian Fan, Xin-yu Feng, Jian-wen Xu, Rong Zhang, Yan Shang. Tanshinone IIA prevents acute lung injury by regulating macrophage polarization. Journal of Integrative Medicine, 2022, 20(3): 274-280.
[3]	Peter Bablis, Henry Pollard, Anthony L. Rosner. Stress reduction via neuro-emotional technique to achieve the simultaneous resolution of chronic low back pain with multiple inflammatory and biobehavioural indicators: A randomized, double-blinded, placebo-controlled trial. Journal of Integrative Medicine, 2022, 20(2): 135-144.
[4]	Victor Abou Nehmi Filho, Gilson Masahiro Murata, Ruan Carlos Macêdo de Moraes, Gabriely Cristina Alves Lima, Danielle Araujo de Miranda, Katrin Radloff, Raquel Galvão Figuerêdo Costa, Joyce de Cassia Rosa de Jesus, Jéssica Alves De Freitas, Nayara Izabel Viana, Ruan Pimenta, Katia Ramos Moreira Leite, José Pinhata Otoch, Ana Flávia Marçal Pessoa. A novel supplement with yeast β-glucan, prebiotic, minerals and Silybum marianum synergistically modulates metabolic and inflammatory pathways and improves steatosis in obese mice. Journal of Integrative Medicine, 2021, 19(5): 439-450.
[5]	Na Zhi, Qian Mo, Shuo Yang, Yuan-xing Qin, Hao Chen, Zeng-guang Wu, Cai-hong Lan, Jun Zhang, Yin-long Li. Treatment of pulmonary fibrosis in one convalescent patient with corona virus disease 2019 by oral traditional Chinese medicine decoction: A case report. Journal of Integrative Medicine, 2021, 19(2): 185-190.
[6]	Swatantra Kumar, Vimal K Maurya, Debadatta Nayak, Anil Khurana, Raj K Manchanda, Srinivasulu Gadugu, Madan LB Bhatt, Shailendra K Saxena. Calcarea carbonica treatment rescues lipopolysaccharide-induced inflammatory response in human mononuclear cells via downregulation of inducible cyclooxygenase pathway. Journal of Integrative Medicine, 2020, 18(5): 441-449.
[7]	The Su Moe, Htet Htet Win, Thin Thin Hlaing, War War Lwin, Zaw Min Htet, Khin Mar Mya . Evaluation of in vitro antioxidant, antiglycation and antimicrobial potential of indigenous Myanmar medicinal plants. Journal of Integrative Medicine, 2018, 16(5): 358-366.
[8]	Uday P. Pratap, Hannah P. Priyanka, Karthik R. Ramanathan, Vishak Raman, Lalgi Hima, Srinivasan Thyagarajan. Noni (Morinda citrifolia L.) fruit juice delays immunosenescence in the lymphocytes in lymph nodes of old F344 rats. Journal of Integrative Medicine, 2018, 16(3): 199-207.
[9]	Omowumi Titilola Kayode, Musa Toyin Yakubu. Parquetina nigrescens leaves: chemical profile and influence on the physical and biochemical indices of sexual activity of male Wistar rats. Journal of Integrative Medicine, 2017, 15(1): 64-76.
[10]	Anirudha A. Lande, Shirishkumar D. Ambavade, Uma S. Swami, Prafulla P. Adkara Prashant D. Ambavade, Arun B. Waghamare. Saponins isolated from roots of Chlorophytum borivilianum reduce acute and chronic inflammation and histone deacetylase. Journal of Integrative Medicine, 2015, 13(1): 25-33.
[11]	Benjamin Perry, Junzeng Zhang, Tarek Saleh, Yanwen Wang. Liuwei Dihuang, a traditional Chinese herbal formula, suppresses chronic inflammation and oxidative stress in obese rats. Journal of Integrative Medicine, 2014, 12(5): 447-454.
[12]	Xin-fang Zhang, Ji Zhu, Wen-ye Geng, Shu-jun Zhao, Chuan-wei Jiang, Sheng-rong Cai, Miao Cheng, Chuan-yun Zhou, Zi-bing Liu. Electroacupuncture at Feishu (BL13) and Zusanli (ST36) down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease. Journal of Integrative Medicine, 2014, 12(5): 417-424.
[13]	Rajendra Karki, Cheol-Ho Park, Dong-Wook Kim. Extract of buckwheat sprouts scavenges oxidation and inhibits pro-inflammatory mediators in lipopolysaccharide-stimulated macrophages (RAW264.7). Journal of Chinese Integrative Medicine, 2013, 11(4): 246-252.
[14]	Jian-sheng Li, Ya Li, Su-yun Li, Yuan-yuan Wang, Li Deng, Yan-ge Tian, Su-li Jiang , Ying Wang. Long-term effects of Tiaobu Feishen therapies on systemic and local inflammation responses in rats with stable chronic obstructive pulmonary disease. Journal of Chinese Integrative Medicine, 2012, 10(9): 1039-1048.
[15]	Ju-yong Wang, Jing Guo, Chang-sheng Dong, Zhan Zheng, Qing Wang. Effects of Feiyanning Decoction, a compound traditional Chinese medicine, on iNOS and COX-2 expressions induced by tumor necrosis factor-α in lung adenocarcinoma cell line. Journal of Chinese Integrative Medicine, 2012, 10(6): 695-700.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 10

[1]	Wei-xiong Liang. Problems-solving strategies in clinical treatment guideline for traditional Chinese medicine and integrative medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 1-4
[2]	Zhao-guo Li. Discussion on English translation of commonly used sentences in traditional Chinese medicine: part one. Journal of Chinese Integrative Medicine, 2008, 6(1): 107-110
[3]	Jun Hu, Jian-ping Liu. Non-invasive physical treatments for chronic/recurrent headache. Journal of Chinese Integrative Medicine, 2008, 6(1): 31
[4]	Xue-mei Liu, Qi-fu Huang, Yun-ling Zhang, Jin-li Lou, Hong-sheng Liu, Hong Zheng. Effects of Tribulus terrestris L. saponion on apoptosis of cortical neurons induced by hypoxia-reoxygenation in rats. Journal of Chinese Integrative Medicine, 2008, 6(1): 45-50
[5]	. Uniform requirements for manuscripts submitted to biomedical journals: Writing and editing for biomedical publication (Chinese version, part two). Journal of Chinese Integrative Medicine, 2010, 8(11): 1001-1005
[6]	Daniel Weber, Janelle M Wheat, Geoffrey M Currie. Inflammation and cancer: Tumor initiation, progression and metastasis,and Chinese botanical medicines. Journal of Chinese Integrative Medicine, 2010, 8(11): 1006-1013
[7]	Hong Liu , Guo-liang Zhang, Li Shen , Zhen Zeng, Bao-luo Zhou, Cheng-hai Liu, Guang Nie . Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5Nl avian influenza virus. Journal of Chinese Integrative Medicine, 2010, 8(11): 1036-1040
[8]	Zhao-guo Li . A discussion of English translation of 1995 and 1997 Chinese National Standards of Traditional Chinese Medical Terminologies for Clinical Diagnosis and Treatment. Journal of Chinese Integrative Medicine, 2010, 8(11): 1090-1096
[9]	Rui Jin, Bing Zhang. A complexity analysis of Chinese herbal property theory: the multiple formations of herbal property (Part 1). Journal of Chinese Integrative Medicine, 2012, 10(11): 1198-1205
[10]	Hui-min Liu, Xian-bo Wang, Yu-juan Chang, Li-li Gu. Systematic review and meta-analysis of randomized controlled trials of integrative medicine therapy for treatment of chronic severe hepatitis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1211-1228