Top

BMC Complementary Medicine and Therapies

Published in:

Open Access 01-12-2018 | Research article

Anti-inflammatory activities of Guang-Pheretima extract in lipopolysaccharide-stimulated RAW 264.7 murine macrophages

Authors: Chuanqi Huang, Wei Li, Qiufeng Zhang, Lihong Chen, Weiming Chen, Hongchao Zhang, Yuxin Ni

Published in: BMC Complementary Medicine and Therapies | Issue 1/2018

Abstract

Background

Guang-Pheretima, which is originated from Pheretima aspergillum, has been documented in academic Chinese herbal studies for nearly 2000 years for its prominent treating effects of various inflammatory diseases such as asthma, cough and fever. However, the anti-inflammatory activity and mechanism of Guang-Pheretima has been rarely reported. Hence, we investigated the inhibitory effect and the underlying mechanism of Guang-Pheretima aqueous extracts on inflammatory response in RAW 264.7 cells.

Method

RAW 264.7 macrophages were pretreated with various concentrations of Guang-Pheretima decoction (GPD) or protein-free Guang-Pheretima decoction (PF-GPD) and subsequently stimulated with lipopolysaccharide (LPS) to trigger the inflammatory response. Productions of nitric oxide (NO) were determined by Griess reaction, and prostaglandin E₂ (PGE₂), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 were measured by enzyme-linked immunosorbent assays (ELISA). The protein expressions and messenger ribonucleic acid (mRNA) amounts of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were analyzed by Western Blot and Real-Time polymerase chain reaction (PCR), respectively. Finally, the translocation of nuclear factor (NF)-κB was observed by Western Blot.

Results

GPD of the experimental concentrations showed no anti-inflammatory activity. In contrast, PF-GPD at concentrations of 40–320 μg/mL significantly inhibited NF-κB activation and reduced the production of inflammatory mediators, such as NO, PGE₂, TNF-α, as well as the related key synthases including iNOS and COX-2. Moreover, PF-GPD markedly suppressed the release of inflammatory cytokines, such as IL-1β and IL-6.

Conclusion

These results demonstrate the excellent anti-inflammatory properties of PF-GPD, and suggest that Guang-Pheretima may be used to treat and prevent certain inflammatory diseases.

McGettrick AF, O'Neill LA. How metabolism generates signals during innate immunity and inflammation. J Biol Chem. 2013;288(32):22893–8.CrossRefPubMedPubMedCentral

Kaufmann S, Ilg K, Mashaghi A, Textor M, Priem B, Aeb M, Reimhult E. Supported lipopolysaccharide bilayers. Langmuir. 2012;28(33):12199–208.CrossRefPubMed

Wang L, Trebicka E, Fu Y, Waggoner L, Akira S, Fitzgerald K, Kagan J, Cherayil B. Regulation of lipopolysaccharide-induced translation of tumor necrosis factor-alpha by the toll-like receptor 4 adaptor protein TRAM. J Innate Immun. 2011;3(5):437–46.CrossRefPubMedPubMedCentral

Moreno B, Jukes J, Vergarairigaray N, Errea O, Villoslada P, Perry V, Newman T. Systemic inflammation induces axon injury during brain inflammation. Ann Neurol. 2011;70(6):932–42.CrossRefPubMed

Kroncke KD, Fehsel K, Kolb-Bachofen V. Inducible nitric oxide synthase in human diseases. Clin Exp Immunol. 1998;113:147–56.CrossRefPubMedPubMedCentral

Satoh H, Amagase K, Ebara S, Akiba Y, Takeuchi K. Cyclooxygenase (COX)-1 and COX-2 both play an important role in the protection of the duodenal mucosa in cats. J Pharmacol Exp Ther. 2013;344(1):189–95.CrossRefPubMed

Chen J, Song Y, Yang J, Zhang Y, Zhao P, Zhu X, Su H. The contribution of TNF-α in the amygdala to anxiety in mice with persistent inflammatory pain. Neurosci Lett. 2013;541:275–80.CrossRefPubMed

Ogawa H, Mukai K, Kawano Y, Minegishi Y, Karasuyama H. Th2-inducing cytokines IL-4 and IL-33 synergistically elicit the expression of transmembrane TNF-α on macrophages through the autocrine action of IL-6. Biochem Bioph Res Co. 2012;420(1):114–8.CrossRef

Duque J, Diaz-Munoz MD, Fresno M, Iniguez MA. Up-regulation of cyclooxygenase-2 by interleukin-1β in colon carcinoma cell. Cell Signal. 2006;18(8):1262–9.CrossRefPubMed

10.

Lee J, Nakagiri T, Oto T, Harada M, Morii E, Shintani Y, Inoue M, Iwakura Y, Miyoshi S, Okumura M, Hirano T, Murakami M. IL-6 amplifier, NF-κB-triggered positive feedback for IL-6 signaling, in grafts is involved in allogeneic rejection responses. J Immunol. 2012;189(4):1928–36.CrossRefPubMed

11.

Kolyada AY, Savikovsky N, Madias NE. Transcriptional regulation of the human iNOS gene in vascular-smooth-muscle cells and macrophages: evidence for tissue specificity. Biochem Bioph Res Co. 1996;220(3):600–5.CrossRef

12.

Schmidt N, Pautz A, Art J, Rauschkolb P, Jung M, Erkel G, Goldring M, Kleinert H. Transcriptional and post-transcriptional regulation of iNOS expression in human chondrocytes. Biochem Pharmacol. 2010;79(5):722–32.CrossRefPubMed

13.

Bagga D, Wang L, Farias-Eisnert R, Glaspy JA, Reddy ST. Differential effects of prostaglandin derived from ω-6 and ω-3 polyunsaturated fatty acids on COX-2 expression and IL-6 secretion. P Natl Acad Sci USA. 2003;100(4):1751–6.CrossRef

14.

Madonna R, Caterina R. Relevance of new drug discovery to reduce NF-κB activation in cardiovascular disease. Vasc Pharmacol. 2012;57(1):41–7.CrossRef

15.

Abdel-Latif MM, Kelleher D, Reynolds JV. Potential role of NF-κB in esophageal adenocarcinoma: as an emerging molecular target. J Surg Res. 2009;153(1):172–80.CrossRefPubMed

16.

Nguyen DP, Li J, Yadav SS, Tewari AK. Recent insights into NF-κB signalling pathways and the link between inflammation and prostate cancer. BJU Int. 2014;114(2):168–76.CrossRefPubMed

17.

DiDonato JA, Mercurio F, Karin M. NF-κB and the link between inflammation and cancer. Immunol Rev. 2012;246(1):379–400.CrossRefPubMed

18.

Poligone B, Baldwin AS. Positive and negative regulation of NF-κB by COX-2: roles of different prostaglandins. J Bio Chem. 2001;276(42):38658–64.CrossRef

19.

Hayden MS, Ghosh S. NF-κB in immunobiology. Cell Res. 2011;21(2):223–44.CrossRefPubMedPubMedCentral

20.

Ma J. The edition and annotation of Sheng Nong's herbal classic. Beijing: People’s Medical Publishing House; 2013. p. 313–4.

21.

Chinese-Pharmacopoeia-Commission. Pharmacopoeia of People's Republic of China, in part 1. Beijing: China Medical Science Press; 2015. p. 113–4.

22.

Cooper EL, Balamurugan M, Huang CY, Tsao CR, Heredia JJB, Tommaseoponzetta M, Paoletti MG. Earthworms dilong: ancient, inexpensive, noncontroversial models may help clarify approaches to integrated medicine emphasizing neuroimmune systems. Evid Based Compl Alt. 2012;2012:164152.CrossRef

23.

Li PC, Tien YC, Day CH, Pai P, Kuo W, Chen T, Kuo C, Tsai C, Ju D, Huang C. Impact of LPS-induced cardiomyoblast cell apoptosis inhibited by earthworm extracts. Cardiovasc Toxicol. 2015;15(2):172–9.CrossRefPubMed

24.

Fu YT, Chen KY, Chen YS, Yao CH. Earthworm (Pheretima aspergillum) extract stimulates osteoblast activity and inhibits osteoclast differentiation. BMC Complem Altern Med. 2014;14:440–9.CrossRef

25.

Wang KY, Tull L, Cooper E, Wang N, Liu D. Recombinant protein production of earthworm lumbrokinase for potential antithrombotic application. Evid Based Compl Alt. 2013;2013:783971.

26.

Gong Q. Chinese medicine processing. Beijing: Chinese Medicine Press; 2012.

27.

Huang C, Li W, Zhang Q, Chen L, Chen W, Zhang H, Ni Y. Pheretima aspergillum decoction suppresses inflammation and relieves asthma in a mouse model of bronchial asthma by NF-κB inhibition. J Ethnopharmacol. 2016;189(2):22–30.CrossRefPubMed

28.

Oh BK, Mun J, Seo HW, Ryu SY, Kim YS, Lee BH, Oh K. Euonymus alatus extract attenuates LPS-induced NF-κB activation via IKKβ inhibition in RAW 264.7 cells. J Ethnopharmacol. 2011;134(2):288–93.CrossRefPubMed

29.

Chen X, Zhang S, Xuan Z, Ge D, Chen X, Zhang J, Wang Q, Wu Y, Liu B. The phenolic fraction of Mentha Haplocalyx and its constituent linarin ameliorate inflammatory response through inactivation of NF-κB and MAPKs in lipopolysaccharide-induced RAW264.7 cells. Molecules. 2017;22:811.CrossRef

30.

Liu GJ, Geng M, Xu R, Hui R, Zhao J, Xu Q, Xu H, Li J. Synthesis of a novel series of diphenolic chromone derivatives as inhibitors of NO production in LPS-activated RAW264.7 macrophages. Bioorg Med Chem Lett. 2010;18(8):2864–71.CrossRef

31.

Yang J, Li S, Xie C, Ye H, Tang H, Chen L, Peng A. Anti-inflammatory activity of ethyl acetate fraction of the seeds of Brucea Javanica. J Ethnopharmacol. 2013;147(2):442–6.CrossRefPubMed

32.

Udit AK, Bellistonbittner W, Glazer EC, Nguyen YHL, Gillan JM, Hill MG, Marletta MA, Goodin DB, Gray HB. Redox couples of inducible nitric oxide synthase. J Am Chem Soc. 2005;127(32):11212–3.CrossRefPubMedPubMedCentral

33.

Chen EP, Smyth EM. COX-2 and PGE2-dependent immunomodulation in breast cancer. Prostag Oth Lipid M. 2011;96(1–4):14–20.CrossRef

34.

Wang G, Hu D, Chen J, Zou Y, Chen P. Analysis of chemical components from Pheretima. J Chin Med Mater. 1998;21:133–5.

35.

Korns D, Frasch SC, Fernandez-Boyanapalli R, Henson PM, Bratton DL. Modulation of macrophage efferocytosis in inflammation. Front Immunol. 2011;2:1–10.CrossRef

36.

Guha M, Mackman N. LPS induction of gene expression in human monocytes. Cell Signal. 2011;2:1–10.

37.

Bessueille L, Magne D. Inflammation: a culprit for vascular calcification in atherosclerosis and diabetes. Cell Mol Life Sci. 2015;72(13):2475–89.CrossRefPubMed

38.

Kleinert H, Pautz A, Linker K, Schwarz PM. Regulation of the expression of inducible nitric oxide synthase. Eur J Pharmacol. 2004;500(1–3):255–66.CrossRefPubMed

39.

Yeoh A, Gibson R, Yeoh EK, Bowen JM, Stringer AM, Giam K, Logan RM, Keefe DMK. Radiation therapy-induced mucositis: relationships between fractionated radiation, NF-κB, COX-1, and COX-2. Cancer Treat Rev. 2006;32(8):645–51.CrossRefPubMed

40.

Lu T, Stark GR. NF-κB: regulation by methylation. Cancer Res. 2015;75(18):3692–5.CrossRefPubMedPubMedCentral

Title: Anti-inflammatory activities of Guang-Pheretima extract in lipopolysaccharide-stimulated RAW 264.7 murine macrophages
Authors: Chuanqi Huang
Wei Li
Qiufeng Zhang
Lihong Chen
Weiming Chen
Hongchao Zhang
Yuxin Ni
Publication date: 01-12-2018
Publisher: BioMed Central
Published in: BMC Complementary Medicine and Therapies / Issue 1/2018
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-018-2086-z

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Anti-inflammatory activities of Guang-Pheretima extract in lipopolysaccharide-stimulated RAW 264.7 murine macrophages

Abstract

Background

Method

Results

Conclusion

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Method

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2018

Maoberry (Antidesma bunius) ameliorates oxidative stress and inflammation in cardiac tissues of rats fed a high-fat diet

Use of biologically-based complementary medicine in breast and gynecological cancer patients during systemic therapy

Feifukang ameliorates pulmonary fibrosis by inhibiting JAK-STAT signaling pathway

Astragalus membranaceus (Fisch.) Bunge repairs intestinal mucosal injury induced by LPS in mice

Associations between the use of herbal medicines and adverse pregnancy outcomes in rural Malawi: a secondary analysis of randomised controlled trial data

Five traditional Nigerian Polyherbal remedies protect against high fructose fed, Streptozotocin-induced type 2 diabetes in male Wistar rats