The Effects of Haedoksamul-tang on Oxidative Stress and Hyperlipidemia in LPS-induced ICR Mouse

Gyu-ho Choi; Yu-sun Jung; Hyeon-cheol Shin; Choi, Gyu-ho; Jung, Yu-sun; Shin, Hyeon-cheol

doi:10.13048/jkm.16008

JKM > Volume 37(1); 2016 > Article

Choi, Jung, and Shin: The Effects of Haedoksamul-tang on Oxidative Stress and Hyperlipidemia in LPS-induced ICR Mouse

Original Article

The Journal of Korean Oriental Medicine 2016; 37(1): 77-89.

Published online: March 31, 2016

DOI: https://doi.org/10.13048/jkm.16008

The Effects of Haedoksamul-tang on Oxidative Stress and Hyperlipidemia in LPS-induced ICR Mouse

Gyu-ho Choi, Yu-sun Jung, Hyeon-cheol Shin

Department of Internal Medicine of Korean Medicine, College of Korean Medicine, Dae-gu Haany University

Correspondence to: 신현철, 대구한의대학교 한의과대학 내과학교실, Tel: +82-54-281-0055, Fax: +82-54-281-7464, E-mail: ungaeshin@naver.com

Received February 15, 2016 Revised March 21, 2016 Accepted March 28, 2016

Abstract

Objectives:

The present study was conducted to examine whether Haedoksamul-tang (HS), a traditional oriental herbal medicine, have beneficail effects on anti-inflammation and dyslipidemia in lipopolysaccharide (LPS)-induced ICR mouse.

Methods:

Twenty four 8-week old male ICR mouse were divided into four groups: normal untreated; LPS treatment only; HS 10 mg/kg plus LPS treatment; and HS 30 mg/kg plus LPS treatment. HS was orally administered per day for 2days. Twenty-four hours after LPS injection (10 mg/kg/day, i.p.), all the mice were sacrificed, and serological changes were evaluated. The levels of nuclear factor-κB (NF-κB), sterol regulatory element-binding transcription protein 1 (SREBP-1) activity and cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor a (TNF-a), monocyte chemotactic protein 1 (MCP-1), acetyl-CoA carboxylase a (ACCa) expression were analyzed in Western blot analysis.

Results:

HS inhibited oxidative stress in the liver of LPS-induced ICR mice. The LPS-induced ICR mice exhibited the increase of NF-κB activity and COX-2, iNOS, TNF-a, MCP-1 expressions in the liver, while HS treatment significantly inhibited them. Moreover, The administration of HS significantly decreased the elevated serum triglyceride and down-regulated the levels of SREBP-1, ACCa in the liver of LPS-induced ICR mice.

Conclusions:

In conclusion, HS could have hepato-protective effects against the oxidative stress-related inflammation and abnormal lipid metabolism.

Keywords: Haedoksamul-tang, inflammation, dyslipidemia, NF-κB, SREBP-1

Fig. 1.

Inhibition effects of HS on serum and hepatic oxidative stress in LPS-induced ICR mice. Serum ROS (A), hepatic ROS (B), hepatic TBARS (C) levels. N: normal group, Veh: vehicle-treated mice, HS10: HS 10 mg/kg treated mice, HS30: HS 30 mg/kg treated mice. Bars represent means ± SD. ^** P < 0.01 versus vehicle-treated mice values.

Fig. 2.

Effects of HS on NF-κBp65 activity in LPS-induced ICR mice liver. N: normal group, Veh: vehicle-treated mice, HS10: HS 10 mg/kg treated mice, HS30: HS 30 mg/kg treated mice. Histone was used for loading control. Bars represent means ± SD. ^* P < 0.05, ^*** P < 0.001 versus vehicle-treated mice values.

Fig. 3.

Effects of HS on COX-2 and iNOS expressions in LPS-induced ICR mice liver. COX-2 (A) and iNOS (B) protein expressions in liver. N: normal group, Veh: vehicle -treated mice, HS10: HS 10 mg/kg treated mice, HS30: HS 30 mg/kg treated mice. ß-actin was used for loading control. Bars represent means ± SD. ^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 versus vehicle-treated mice values.

Fig. 4.

Effects of HS on TNF-a and MCP-1 expressions in LPS-induced ICR mice liver. TNF-a (A) and MCP-1 (B) protein expressions in liver. N: normal group, Veh: vehicle-treated mice, HS10: HS 10 mg/kg treated mice, HS30: HS 30 mg/kg treated mice. ß-actin was used for loading control. Bars represent means ± SD. ^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 versus vehicle-treated mice values.

Fig. 5.

Effects of HS on serum and hepatic triglyceride levels in LPS-induced ICR mice. Serum triglyceride (A), hepatic triglyceride (B) levels. N: normal group, Veh: vehicle-treated mice, HS10: HS 10 mg/kg treated mice, HS30: HS 30 mg/kg treated mice. Bars represent means ± SD. ^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 versus vehicle-treated mice values.

Fig. 6.

Effects of HS on SREBP-1 activity and ACCa expression in LPS-induced ICR mice liver. SREBP-1 (A) activity and ACCa (B) protein expression in liver. N: normal group, Veh: vehicle-treated mice, HS10: HS 10 mg/kg treated mice, HS30: HS 30 mg/kg treated mice. Histone or ß-actin was used for loading control. Bars represent means ± SD. ^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 versus vehicle-treated mice values.

Table 1.

Composition of Haedoksamul-tang

Herb name	Scientific name	Amounts (g)
黃芩	Scutellaria baicalensis George	4
黃連	Coptis chinensis Franch.	4
黃柏	Phellodendron amurense Ruprecht	4
梔子	Gardenia jasminoides Ellis	4
當歸	Angelica acutiloba (S. et Z.) Kitagawa	4
川芎	Cnidium officinale Makino	4
白芍藥	Paeonia latiflora Pall.	4
生乾地黃	Rehmannia glutinosa (Gaetner) Libosch.	4

	Total	32

참고문헌

1.. Johan, F. Immunity, atherosclerosis and cardiovascular disease. BMC med, (2013). 11, 117-29.

2.. Feng, X, Zhang, Y, Xu, R, Xie, X, Tao, L, & Gao, H, et al. Lipopolysaccharide up-regulates the expression of Fcalpha/mu receptor and promotes the binding of oxidized low-density lipoprotein and its IgM antibody comples to activated human macrophages. Atherosclerosis, (2010). 208, 396-405.

3.. Wiesner, P, Choi, SH, Almazan, F, Benner, C, Huang, W, & Diehl, CJ, et al. Low doses of lipopolysaccharide and minimally oxidized low-density lipoprotein cooperatively activate macrophages via nuclear factor kappa B and activator protein-1: possible mechanism for acceleration of atherosclerosis by subclinical endotoxemia. Circ Res, (2010). 107, 56-65.

4.. Maitra, U, & Li, L. Molecular mechanisms responsible for the ruduced expression of cholesterol transporters from macrophages by low-dose endotoxin. Arterioscler Thromb Vasc Biol, (2013). 33, 24-33.

5.. Vikatmaa, P, Lajunen, T, Ikonen, TS, Pussinen, PJ, Leinonen, M, & Saikku, P, et al. Chlamydial lipopolysaccharide (cLPS) is present in atherosclerotic and aneurysmal arterial wall--cLPS levels depend on disease manifestation. Cardiovasc Pathol, (2010). 19(1), 48-54.

6.. Jean, L, Bret, D, & Chistophe, C. Systemic capsaicin pretreatment fails to bolck the decrease in food-motivated behavior induced by lipopolysaccharide and interleukin-1ß. Brain Research Bul, (1997). 42, 443-9.

7.. Sharifov, OF, Nawar, G, Ternovov, W, Mishra, VK, Litovsky, SH, & Palqunachare, MN, et al. Cationic peptide mR18L with lipid lowering properties inhibits LPS-induced systemic and liver inflammation in rats. Biochem Biophys Res Commun, (2013). 436(4), 705-10.

8.. Kim, ID, Kang, KS, Kwon, RH, Yang, JO, Lee, JS, & Ha, BJ. The effect of Rubus coreanum Miquel against lipopolysaccharide-induced oxidative stress and lipid metaobolism. J. Fd Hyg. Safety, (2007). 22(3), 213-7.

9.. Ju JH. Dan-gye-sim-beop-bu-yeo. Seoul. Daesung Publishing Co. Ltd;(1982). p. 714

10.. Lee SI. Cheon-jin-cheo-bang-hea-sheol. Seoul. Sheong-bo-sa;(1995). p. 289-91.

11.. Kim, ES. A experimental study of Hwangryeonheadock-Tang and Onchung-Eum on hyperlipidemia & hypertension. J. of Korean Medicine, (1999). 20(1), 185-96.

12.. Kim, YH, & Jo, HB. Anti-inflammatory effects of Haedoksamul-tang in RAW 264.7 cells. J. of Oriental Obstetrics & Gynecology, (2008). 21(2), 166-83.

13.. Beom, HB. Effects of Onchungeum and Gamionchungeum on the antiallergic response and blood coagulation. J. of Kyung Hee University, (1990). 6(4), 490-9.

14.. Jeon, YG. Effects of Haedoksamul-tang on Trimelliticanhydride-induced contact hypersensitivity in a mouse model, (2010). 1-41.

15.. Seo, M, Jeon, BH, Woo, WH, & Jeong, WY. Effect of Onchengyeum on the damaged liver cell by carbon tetrachloride in rats. Korean J. Oriental Physiology & Pathology, (1989). 11(2), 27-36.

16.. Im, HJ, Hwang, CY, Gang, HC, Kim, NG, & Gwon, IH. Inhibitory effects of Onchumgeum on cytokine production from phytohaemagglutin-stimulated peripheral blood mononuclear cells of behcets patients. Korean J. Oriental Physiology & Pathology, (2002). 16(4), 768-73.

17.. Zhu, GF, Guo, HJ, Huang, Y, Wu, CT, & Zhang, XF. Eriodictyol, a plant flavonoid, attenuates LPS-induced acute lung injury through its antioxidative and anti-inflammatory activity. Exp Ther Med, (2015). 10(6), 2259-66.

18.. Heo J. Dong-ui-bo-gam. Seoul. Bubin Publishing Co. Ltd;(2007). 416:p. 455

19.. Rush, GF, Gorski, JR, Ripple, MG, Sowinski, J, & Bugelski, P. Organic hydroperoxide -induced lipid peroxidation and cell death in isolated hepatocytes. Toxicol Appl Pharmacol, (1985). 78, 473-83.

20.. Izeboud, CA, Hoebe, KH, Grootendorst, AF, Nijmeijer, SM, van Miert, AS, & Witkamp, RF, et al. Endotoxin-induced liver damage in rats is minimized by beta 2-adrenoceptor stimulation. Inflamm Res, (2004). 53, 93-9.

21.. Munford, RS. Severe sepsis and septic sock: the role of gram-negative bacteremia. Annu Rev Pathol, (2006). 1, 467-96.

22.. Kleemann, R, Verschuren, L, van Erk, M, Nikolsky, Y, Verheij, ER, & Smilde, AK, et al. Atherosclerosis and liver inflammation induced by increased dietary cholesterol intake: a combined transcriptomics and metabolomics analysis. Genome Biol, (2007). 8, R200.

23.. Xu, X, Hu, J, McGrath, BC, & Cavener, DR. GCN2 in the brain programs PPARγ2 and triglyceride storage in the liver during perinatal development in response to maternal dietary fat. PLos One, (2013). 8(10), e75917.

24.. Liu, RH, & Hotchkiss, JH. Potential genotoxicity of chronically elevated nitirc oxide: a review. Mutat. Res, (1996). 339, 73-89.

25.. Collins, T, & Cybulsky, MI. NF-kappaB: pivotal mediator or innocent bystander in atherogenesis. J Clin Invest, (2001). 107, 255-64.

26.. Thuberg, BL, & Collins, T. The nuclear factor-B/inhibitor of B autoregulatory system and atherosclerosis. Curr Opin Lipidol, (1998). 9, 387-96.

27.. Wang, QS, Xiang, Y, Cui, YL, Lin, KM, & Zhang, XF. Dietary blue pigments derived from Genipin, attenuate inflammation by inhibiting LPS-Induced iNOS and COX-2 expression via the NF-κB inactivation. PLos One, (2012). 7(3), 1-11.

28.. Kwon, HJ, Sung, BK, Kim, JW, Lee, JH, Kim, ND, & Yoo, MA, et al. The effect of lipopolysaccharide on enhanced inflammatory process with age: modulation of NF-κB. J. Amer. Aging Assoc, (2001). 24, 163-72.

29.. Napoli, C, de Nigris, F, Williams-Ignarro, S, Pignalosa, O, Sica, V, & Ignarro, LJ. Nitric oxide and atherosclerosis: an update. Nitric Oxide, (2006). 15, 265-79.

30.. Baldwin, AS. The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol, (1996). 14, 649-83.

31.. DeGraba, TJ. Expression of inflammatory mediators and adhesion molecules in human atherosclerotic plaque. Neurology, (1997). 49(Supple 4), S15-9.

32.. Fassbender, K, Rossol, S, Kammer, T, Daffertshofer, M, Wirth, S, & Dollman, M, et al. Proinflammatory cytokines in serum of patients with acute cerebral ischemia: kinetics of secretion and relation to the extent of brain damage and outcome of disease. J Neurol Sci, (1994). 122(2), 135-9.

33.. Adams, DH, & Lioyd, AF. Chemokines: leukocyte recruitment and activation cytokines. Lancet, (1997). 349, 490-5.

34.. Brown, MS, & Goldstein, JL. The SREBP pathway: regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor. Cell, (1997). 89, 331-40.

35.. Weber, LW, Boll, M, & Stampfl, A. Maintaining cholesterol homeostasis: sterol regulatory element-binding proteins. World J Gastroenterol, (2004). 10(21), 3081-7.

36.. Horton, JD, Goldstein, JL, & Brown, MS. SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. J Clin Invest, (2002). 109, 1125-31.

37.. Foufelle, F, & Ferre, P. New perspectives in the regulation of hepatic glycolytic and lipogenic genes by insulin and glucose: a role for the transcription factor sterol regulatory element binding protein-1c. Biochem J, (2002). 366, 377-91.

38.. Ferre, P, Foretz, M, Azzout-Marniche, D, Becard, D, & Foufelle, F. Sterol-regulatory-element-binding protein 1c mediates insulin action on hepatic gene expression. Biochemical Society Transactions, (2001). 29(4), 547-52.

39.. Horton, JD. Sterol regulatory element-binding proteins: transcriptional activators of lipid synthesis. Biochemical Society Transactions, (2001). 30(6), 1091-5.

40.. Tong, L. Acetyl-coenzyme A carboxylase: crucial metabolic enzyme and attractive target for drug discovery. Cell Mol Life Sci, (2005). 62(16), 1784-803.

41.. Munday, MR. Regulation of mammalian acetyl-CoA carboxylase. Biochem Soc Trans, (2002). 30(6), 1059-64.