Anti-inflammatory Effects of Ojeok-san in LPS-induced Inflammatory Rat Model

Joo-Hee Kim; Dong Hyuk Lee; Ji Hye Kim; Sung Eun Jung; Seong Ho Ham; Woong Mo Yang; Bo-in Kwon; Kim, Joo-Hee; Lee, Dong Hyuk; Kim, Ji Hye; Jung, Sung Eun; Ham, Seong Ho; Yang, Woong Mo; Kwon, Bo-in

doi:10.13048/jkm.21012

JKM > Volume 42(2); 2021 > Article

Kim, Lee, Kim, Jung, Ham, Yang, and Kwon: Anti-inflammatory Effects of Ojeok-san in LPS-induced Inflammatory Rat Model

Original Article

The Journal of Korean Medicine 2021; 42(2): 21-30.

Published online: June 01, 2021

DOI: https://doi.org/10.13048/jkm.21012

Anti-inflammatory Effects of Ojeok-san in LPS-induced Inflammatory Rat Model

Joo-Hee Kim¹^{, 6}^{, #}

, Dong Hyuk Lee²^{, 6}^{, #}

, Ji Hye Kim³, Sung Eun Jung³, Seong Ho Ham³, Woong Mo Yang⁴, Bo-in Kwon⁵^{, 6}^{, *}

¹Department of Acupuncture and Moxibustion Medicine, College of Korean Medicine, Sangji University

²College of Korean Medicine, Sangji University

³National Development Institute of Korean Medicine

⁴Department of Convergence Korean Medical Science, College of Korean Medicine, Kyung Hee University

⁵Department of Pathology, College of Korean Medicine, Sangji University

⁶Research Institute of Korean Medicine, Sangji Univeristy

Correspondence to: Bo-In Kwon, Department of Pathology, College of Korean Medicine, Sangji University, 83 Sangjidae-gil, Wonju-si, Gangwon-do 26339, Republic of Korea, Tel: +82-33-730-0662, Fax: +82-33-730-0654, E-mail: kbi34812@sangji.ac.kr

Co-First: Joo-Hee Kim, Department of Acupuncture and Moxibustion Medicine, College of Korean Medicine, Sangji University, 83 Sangjidae-gil, Wonju-si, Gangwon-do 26339, Republic of Korea. E-mail: jhkim712@sangji.ac.kr

Co-First: Dong Hyuk Lee, College of Korean Medicine, Sangji University, 83 Sangjidae-gil, Wonju-si, Gangwon-do 26339, Republic of Korea. E-mail: neurohani_ldh@sangji.ac.kr

Received January 29, 2021 Revised April 9, 2021 Accepted April 13, 2021

Abstract

Objectives

The aim of this study is to investigate the anti-inflammatory effects of Ojeok-san and compare the therapeutic effects according to its formation.

Methods

We evaluated the anti-inflammatory effects of Ojeok-san using lipopolysaccharide (LPS) induced inflammatory animal model. Male SD rats were administered intra-orally with two different formulation types of Ojeok-san according to prescribed dosage. One hour later, to induce inflammatory responses, subsequent intra-peritoneal injection of LPS was conducted. After 5 hours later, serum TNF-α, IL-1β, IL-6 and PGE2 levels were measured by ELISA to assess the alteration of pro-inflammatory markers.

Results

In our experiment, regardless of its formation, administration of Ojeok-san decreased TNF-α, IL-1β, IL-6 and PGE2 level in serum. Furthermore, LPS-induced toxicity of liver and kidney was not detected by Ojeok-san administration.

Conclusions

The anti-inflammatory effect of Ojeok-san was shown in LPS-induced inflammatory model by decreasing pro-inflammatory markers, and there would be no significant difference in therapeutic effect between two formulation types of Ojeok-san.

Keywords: Ojeok-san, lipopolysaccharide(LPS)-induced model, anti-inflammatory effect, pro-inflammatory markers

Acknowledgement

This work was supported by a grant from Pharmacological Standardization Project of Traditional Korean Medicine by Ministry of Health and welfare (MOHW) and JeollaNamdo, Republic of Korea.

Fig. 1

Principles of prescription composition in Ojeok-san

Fig. 2

Anti-inflammatory effects between Ojeok-san Extract Powder group and Soft Extract groups in LPS induced inflammatory models (n=5, mean±SD, p: p value of ANOVA test vs. LPS + Vehicle group; *: p<0.05, **: p<0.01)

Fig. 3

Liver function test results after the injection of LPS according to the groups (n=6, mean±SD)

Fig. 4

Kidney function test results after the injection of LPS according to the groups

(n=6, mean±SD; p: p value of ANOVA test vs. LPS + Vehicle group ; ^*: p<0.05)

Table 1

Comparison of anti-inflammatory effects between Ojeok-san Extract Powder group and Soft Extract groups in LPS induced inflammatory models

(n=5)

Group	Dosage (mg/kg)	LPS (mg/kg, ip)	Blood concentration

			TNF-α (pg/ml)	IL-1β (pg/ml)	IL-6 (pg/ml)	PGE2 (pg/ml)
No stimulation	D.W.	-	41.01 ± 17.01	22.54 ± 2.07	22.38 ± 3.57	97.60 ± 19.29
LPS+Vehicle	D.W.	5	147.98 ± 19.84	156.59± 32.84	67.10 ± 7.32	401.12 ± 18.88
LPS+Powder	435	5	65.91 ± 19.35^**	74.38 ± 19.7^**	36.55 ± 10.71^**	327.96 ± 15.23^**
	1,305	5	57.26 ± 7.92^**	70.79 ± 20.99^**	27.12 ± 7.73^**	259.69 ± 35.32^**
	6,525	5	50.09 ± 11.9^**	64.50 ± 15.78^**	27.96 ± 5.80^**	267.60 ± 30.98^**
LPS+Soft Extract	1,983	5	74.21 ± 4.16^**	63.27 ± 17.69^**	42.33 ± 6.84^**	332.71 ± 35.28^**
	5,949	5	61.05 ± 12.61^**	49.06 ± 13.59^**	28.23 ± 3.62^**	280.30 ± 9.74^**
	32,625	5	48.83 ± 15.51^**	50.59 ± 17.12^**	24.49 ± 5.85^**	242.50 ± 23.14^**

The values indicate mean±SD, p: p value of ANOVA test vs. the LPS + vehicle group;

* p<0.05,

** p<0.01

Table 2

Regulation of Pro-inflammatory cytokines in Ojeok-san Extract powder and Soft Extract groups

Formula (mg/kg)		TNF-α	p^*	IL-1β	p^*	IL-6	p^*	PGE2	p^*
LPS+ Powder	435	76.71	0.69^†	61.32	0.71^†	68.31	0.76^†	25.26	0.43^†
	1,305	84.80	0.88^‡	64.00	0.43^‡	89.38	0.91^‡	44.74	0.62^‡
	6,525	91.50	0.95^§	68.69	0.52^§	87.51	0.73^§	42.64	0.58^§
LPS+ Soft Extract	1,983	68.96	-	69.61	-	55.37	-	22.78	-
	5,949	81.26	-	80.21	-	86.90	--	39.91	-
	32,625	92.69	-	79.07	-	95.26		50.89	-

* p: p value of Student’s t-test,

†, ‡, § p value of corresponding dosage between Ojeok-san Extract powder and Soft Extract group.

Table 3

Liver function test values after the injection of LPS according to the groups

(n=6)

Formula (mg/kg)		LPS (mg/kg)	AST (U/L)	ALT (U/L)	GGT (U/L)	Total protein (g/dL)
No stimulation			92.00 ± 14.55	46.80 ± 16.17	1.00 ± 0.00	5.84 ± 0.23

LPS+Vehicle		5	111.83 ± 10.43	50.29 ± 8.91	1.43 ± 0.05	5.27 ± 0.39

LPS+ Powder	435	5	90.50 ± 22.41	56.80 ± 11.29	1.00 ± 0.00	6.73 ± 0.37
	1,305	5	98.67 ± 12.51	54.33 ± 11.70	1.50 ± 0.50	6.48 ± 0.64
	6,525	5	98.67 ± 27.15	53.67 ± 9.88	1.20 ± 0.40	5.82 ± 0.62

LPS+Soft Extract	1,983	5	106.20 ± 12.89	54.17 ± 18.63	1.00 ± 0.00	5.87 ± 0.27
	5,949	5	92.17 ± 10.48	45.50 ± 6.65	1.00 ± 0.00	5.88 ± 0.39
	32,625	5	100.50 ± 16.09	51.00 ± 9.73	1.00 ± 0.40	5.95 ± 0.60

The values indicate mean±SD. p: p value of ANOVA test vs. LPS + Vehicle group

Table 4

Kidney function test values after the injection of LPS according to the groups

(n=6)

Formula (mg/kg)		LPS (mg/kg)	Creatinine (mg/dL)	BUN (mg/dL)
No stimulation			0.32 ± 0.12	15.10 ± 1.34
LPS +Vehicle		5	0.49 ± 0.06	18.00 ± 4.54
LPS+ Powder	435	5	0.33 ± 0.07	12.22 ± 2.30
	1,305	5	0.35 ± 0.10	12.02 ± 3.11
	6,525	5	0.40 ± 0.10	12.92 ± 2.92
LPS+Soft Extract	1,983	5	0.33 ± 0.09	12.24 ± 4.65
	5,949	5	0.35 ± 0.08	11.37 ± 1.48^*
	32,625	5	0.35 ± 0.05	12.87 ± 3.69

The values indicate mean±SD. p: p value of ANOVA test vs. LPS + Vehicle group

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