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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

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.

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
jkm-42-2-21f1.gif
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)
jkm-42-2-21f2.gif
Fig. 3
Liver function test results after the injection of LPS according to the groups (n=6, mean±SD)
jkm-42-2-21f3.gif
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)
jkm-42-2-21f4.gif
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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