Abstract
ECa 233 has been reported for the treatment of inflammatory-related disorders. Therefore, the objective of this study is to examine the effects of ECa 233 and their mechanisms on RAW264.7 macrophage-conditioned medium and nutrient stress-induced adipocytes under condition. Preadipocytes were differentiated into mature adipocyte and apoptotic cell death was observed by Hoechst 33,342 staining. The inflammatory cytokine levels were analyzed using ELISA. The protein expression of B-cell lymphoma 2 (Bcl-2), Bcl2 associated x (Bax), pro-caspase 3, cleaved caspase 3, p38, extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor-κB (NF-κB) were performed by western blot. Our results showed that macrophage-conditioned medium and nutrient stress significantly increased (P < 0.05) the number of apoptotic cells, and released the inflammatory cytokines including tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) via the activation of ERK1/2, p38 MAPK and NF-κB involving the development of inflammation in 3T3-L1 adipocytes. ECa 233 at 10–100 μg/mL significantly inhibited the number of apoptotic cell death. At mechanistic level, the exposure of ECa 233 to 3T3-L1 adipocytes showed a reduction of Bax/Bcl2 ratio, and subsequently inhibited the proteolytic activity of caspase-3. The production of TNF-α and IL-1β were significantly decreased after treatment with ECa 233 (1–100 μg/mL). Additionally, ECa 233 inhibited adipocytokine levels by downregulated ERK1/2, p38 MAPK, and NF-κB protein expression. Taken together, the present findings indicate that ECa 233 possesses anti-inflammatory effect in adipocytes though suppressing NF-κB and MAPK pathway. This study provides the information regarding the protective effect of ECa 233 against dysregulated adipocytes related to diseases.
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Data availability
The data generated and analyzed during the current study are available from the corresponding authors on reasonable request by permission of institute and department chairman’s.
Abbreviations
- AAA:
-
Abdominal aortic aneurysm
- Akt:
-
Protein kinase B
- Bcl-2:
-
B-cell lymphoma 2
- Bax:
-
Bcl2 associated x
- ELISA:
-
Enzyme-linked immunosorbent assay
- ERK:
-
Extracellular signal-regulated kinase
- FBS:
-
Fetal bovine serum
- IL-1β:
-
Interleukin-1β
- LPS:
-
Lipopolysaccharide
- MAPKs:
-
Mitogen-activated protein kinases
- MTT:
-
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- NF-κB:
-
Nuclear factor-κB
- TNF-α:
-
Tumor necrosis factor alpha
References
Anukunwithaya T, Tantisira MH, Tantisira B, Khemawoot P (2017) Pharmacokinetics of a standardized extract of Centella asiatica ECa 233 in rats. Planta Med 83:710–717. https://doi.org/10.1055/s-0042-122344
Belwal T, Andola HC, Atanassova MS, Joshi B, Suyal R, Thakur S, Bisht A, Jantwal A, Bhatt ID, Rawal RS (2019) Gotu Kola (Centella asiatica). In: Nabavi SM, Silva AS (eds) Nonvitamin and nonmineral nutritional supplements. Academic Press, pp 265–275
Cargnello M, Roux PP (2011) Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev 75:50–83. https://doi.org/10.1128/MMBR.00031-10
Catrysse L, Loo GV (2017) Inflammation and the metabolic syndrome: the tissue specific functions of NF-kB. Trends Cell Biol 27:417–429. https://doi.org/10.1016/j.tcb.2017.01.006
Doderer SA, Gäbel G, Kokje VBC, Northoff BH, Holdt LM, Hamming JF, Lindeman JHN (2017) Adventitial adipogenic degeneration is an unidentified contributor to aortic wall weakening in the abdominal aortic aneurysm. J Vasc Surg 67:1891–1900. https://doi.org/10.1016/j.jvs.2017.05.088
Dou HX, Wang T, Su HX, Gao DD, Xu YC, Li YX, Wang HY (2020) Exogenous FABP4 interferes with differentiation, promotes lipolysis and inflammation in adipocytes. Endocrine 67:587–596. https://doi.org/10.1007/s12020-019-02157-8
He B, Wang X, Jin X, Xue Z, Ni Y, Zhu J, Wang C, Jin Y, Fu Z (2020) β-cypermethrin alleviated the inhibitory effect of medium from RAW 264.7 cells on 3T3-L1 cell maturation into adipocytes. Lipids 55:251–260. https://doi.org/10.1002/lipd.12234
Heilbronn L, Smith SR, Ravussin E (2004) Failure of fat cell proliferation, mitochondrial function and fat oxidation results in ectopic fat storage, insulin resistance and type 2 diabetes mellitus. Int J Obes 28:S12–S21. https://doi.org/10.1038/sj.ijo.0802853
Hou Q, Li M, Lu YH, Liu DH, Li CC (2016) Burn wound healing properties of asiaticoside and madecassoside. Exp Ther Med 12:1269–1274. https://doi.org/10.3892/etm.2016.3459
Hsieh CC, Wang CH (2018) Aspirin disrupts the crosstalk of angiogenic and inflammatory cytokines between 4T1 breast cancer cells and macrophages. Mediators Inflamm 2018:6380643. https://doi.org/10.1155/2018/6380643
Klöting N, Blüher M (2014) Adipocyte dysfunction, inflammation and metabolic syndrome. Rev Endocr Metab Disord 15:277–287. https://doi.org/10.1007/s11154-014-9301-0
Kugo H, Zaima N, Tanaka H, Mouri Y, Yanagimoto K, Hayamizu K, Hashimoto K, Sasaki T, Sano M, Yata T, Uran T, Setou M, Unno N, Moriyama T (2016) Adipocyte in vascular wall can induce the rupture of abdominal aortic aneurysm. Sci Rep 6:31268. https://doi.org/10.1038/srep31268
Kugo H, Tanaka H, Moriyama T, Zaima N (2018) Pathological implication of adipocytes in AAA development and the rupture. Ann Vasc Dis 11:159–168. https://doi.org/10.3400/avd.ra.17-00130
Kugo H, Sukketsiri W, Iwamoto K, Suihara S, Moriyama T, Zaima N (2021) Low glucose and serum levels cause an increased inflammatory factor in 3T3-L1 cell through Akt, MAPKs and NF-кB activation. Adipocyte 10:232–241. https://doi.org/10.1080/21623945.2021.1914420
Kwak DH, Seo YN, Lee JH, Park SJ, Cho YH, Kim JS, Kim SU, Choo YK (2018) GM1 induced the inflammatory response related to the Raf-1/MEK1/2/ERK1/2 pathway in coculture of pig mesenchymal stem cells with RAW264.7. Anim Cells Syst (seoul) 22:157–164. https://doi.org/10.1080/19768354.2018.1453546
Lu Y, Ma J, Zhao J, Song Z, Zhou C, Liu X, Teng W, Wang W, Zhang Q, Yan W, Jiao P (2020) The role of MKP-5 in adipocyte-macrophage interactions during obesity. Obes Facts 13:86–101. https://doi.org/10.1159/000505343
Moolsap F, Tanasawet S, Tantisira M, Hutamekalin P, Tipmanee V, Sukketsiri W (2020) Standardized extract of Centella asiatica ECa 233 inhibits lipopolysaccharide-induced cytokine release in skin keratinocytes by suppressing ERK1/2 pathways. Asian Pac J Trop Biomed 10:273–280. https://doi.org/10.4103/2221-1691.283941
Niestrawska JA, Regitnig P, Viertler C, Cohnert TU, Babu AR, Holzapfel GA (2019) The role of tissue remodeling in mechanics and pathogenesis of abdominal aortic aneurysms. Acta Biomater 88:149–161. https://doi.org/10.1016/j.actbio.2019.01.070
Park JH, Choi JY, Son DJ, Park EK, Song MJ, Hellström M, Hong JT (2017) Anti-inflammatory effect of titrated extract of Centella asiatica in phthalic anhydride induced allergic dermatitis animal model. Int J Mol Sci 18:738. https://doi.org/10.3390/ijms18040738
Park HJ, Jo SM, Seo SH, Lee M, Lee Y, Kang I (2020) Anti-inflammatory potential of cultured ginseng roots extract in lipopolysaccharide-stimulated mouse macrophages and adipocytes. Int J Environ Res Public Health 17:4716. https://doi.org/10.3390/ijerph17134716
Ramella M, Boccafoschi F, Bellofatto K, Follenzi A, Fusaro L, Boldorini R, Casella F, Porta C, Settembrini P, Cannas M (2017) Endothelial MMP-9 drives the inflammatory response in abdominal aortic aneurysm (AAA). Am J Transl Res 9:5485–5495
Ramesh BN, Girish TK, Raghavendra RH, Naidu KA, Rao UJ, Rao KS (2014) Comparative study on anti-oxidant and anti-inflammatory activities of Caesalpinia crista and Centella asiatica leaf extracts. J Pharm Bioallied Sci 6:86–91. https://doi.org/10.4103/0975-7406.129172
Saifah E, Suttisri R, Patarapanich C, Laungchonlatan S, Tantisira MH, Tantisira B (2009) Preparation methods of colorless mixture between madecassoside and asiaticoside from Centella asiatica. Department of Intellectual Property MoC ed. C07H 1/00 ed. Chulalongkorn University, Bangkok, Thailand.
Singkhorn S, Tantisira MH, Tanasawet S, Hutamekalin P, Wongtawatchai T, Sukketsiri W (2018) Induction of keratinocyte migration by ECa 233 is mediated through FAK/Akt, ERK, and p38 MAPK signaling. Phytother Res 32:1397–1403. https://doi.org/10.1002/ptr.6075
Sukketsiri W, Tanasawet S, Moolsap F, Tantisira MH, Hutamekalin P, Tipmanee V (2019) ECa 233 suppresses LPS-induced proinflammatory responses in macrophages via suppressing ERK1/2, p38 MAPK and Akt pathways. Biol Pharm Bull 42:1358–1365. https://doi.org/10.1248/bpb.b19-00248
Sultana A, Cochran BJ, Tabet F, Patel M, Torres LC, Barter PJ, Rye KA (2016) Inhibition of inflammatory signaling pathways in 3T3-L1 adipocytes by apolipoprotein A-I. FASEB J 30:2324–2335. https://doi.org/10.1096/fj.201500026R
Sun K, Kusminski CM, Scherer PE (2011) Adipose tissue remodeling and obesity. J Clin Investig 121:2094–2101. https://doi.org/10.1172/JCI45887
Sun W, von Meyenn F, Peleg-Raibstein D, Wolfrum C (2019) Environmental and nutritional effects regulating adipose tissue function and metabolism across generations. Adv Sci 6:1900275. https://doi.org/10.1002/advs.201900275
Tanaka H, Zaima N, Sasaki T, Sano M, Yamamoto N, Saito T, Inuzuka K, Hayasaka T, Goto-Inoue N, Sugiura Y, Sato K, Kugo H, Moriyama T, Konno H, Setou M, Unno N (2015) Hypoperfusion of the adventitial vasa vasorum develops an abdominal aortic aneurysm. PLoS One 10:e0134386. https://doi.org/10.1371/journal.pone.0134386
Taylor EB (2021) The complex role of adipokines in obesity, inflammation, and autoimmunity. Clin Sci (lond) 135:731–752. https://doi.org/10.1042/CS20200895
Teerapattarakan N, Benya-aphikul H, Tansawat R, Wanakhachornkrai O, Tantisira MH, Rodsiri R (2018) Neuroprotective effect of a standardized extract of Centella asiatica ECa233 in rotenone-induced parkinsonism rats. Phytomedicine 44:65–73. https://doi.org/10.1016/j.phymed.2018.04.028
Tsai SH, Wang JC, Liao WI, Hsu YJ, Lin CY, Liao MT, Huang PH, Lin SJ (2018) Fucoidan attenuates angiotensin II-induced abdominal aortic aneurysms through the inhibition of c-Jun N-terminal kinase and nuclear factor kappaB activation. J Vasc Surg 68:72S-81S. https://doi.org/10.1016/j.jvs.2017.09.042
Wanasuntronwong A, Tantisira MH, Tantisira B, Watanabe H (2012) Anxiolytic effects of standardized extract of Centella asiatica (ECa 233) after chronic immobilization stress in mice. J Ethnopharmacol 143:579–585. https://doi.org/10.1016/j.jep.2012.07.010
Wang W, Wu L, Li Q, Zhang Z, Xu L, Lin C, Gao L, Zhao K, Liang F, Zhang Q, Zhou M, Jiang W (2018) Madecassoside prevents acute liver failure in LPS/D-GalN-induced mice by inhibiting p38/NF-κB and activating Nrf2/HO-1 signaling. Biomed Pharmacother 103:1137–1145. https://doi.org/10.1016/j.biopha.2018.04.162
Xing Y, Ji Q, Li X, Ming J, Zhang N, Zha D, Lin Y (2017) Asiaticoside protects cochlear hair cells from high glucose-induced oxidative stress via suppressing AGEs/RAGE/NF-κB pathway. Biomed Pharmacother 86:531–536. https://doi.org/10.1016/j.biopha.2016.12.025
Zhang C, Chen S, Zhang Z, Xu H, Zhang W, Xu D, Lin B, Mei Y (2020) Asiaticoside alleviates cerebral ischemia-reperfusion injury via NOD2/mitogen-activated protein kinase (MAPK)/nuclear factor kappa B (NF-κB) signaling pathway. Med Sci Monit 26:e920325 https://doi.org/10.12659/MSM.920325
Zhong L, He X, Si X, Wang H, Li B, Hu Y, Li M, Chen X, Liao W, Liao Y, Bin J (2019) SM22α (Smooth Muscle 22α) prevents aortic aneurysm formation by inhibiting smooth muscle cell phenotypic switching through suppressing reactive oxygen species/NF-kappaB. Arterioscler Thromb Vasc Biol 39:e10–e25. https://doi.org/10.1161/ATVBAHA.118.311917
Acknowledgements
The authors are grateful to the Faculty of Science Research Fund, Prince of Songkla University, Contract no. 1-2562-02-016. We thank Publication clinic, Prince of Songkla University for the assistance in manuscript language editing.
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Nareenath Muneerungsee, Wanida Sukketsiri and Furoida Moolsap performed the experimental study; Nobuhiro Zaima and Supita Tanasawet conceived and drafted, edited, and approved the paper; Wandee Udomuksorna and Mayuree Tantisira edited the paper. Wanida Sukketsiri designed the study, funding acquisition, writing- review, editing and approved the paper.
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Muneerungsee, N., Tanasawet, S., Moolsap, F. et al. The standardized Centella asiatica extract suppressed the inflammation and apoptosis in macrophage-conditioned medium and nutrient stress-induced adipocytes. Biologia 77, 3545–3554 (2022). https://doi.org/10.1007/s11756-022-01194-5
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DOI: https://doi.org/10.1007/s11756-022-01194-5