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J Acupunct Res > Volume 33(2); 2016 > Article
Kim, Jeon, and Kim: A Study on the Effect of Erycibae Caulis and Corydalis Tuber Pharmacopuncture on a Mouse Model with Collagen Induced Rheumatoid Arthritis
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Abstract

Objectives:

The aim of the present study is to examine the effect and mechanism of Erycibae Caulis and Corydalis Tuber Pharmacopuncture (ECP) on a mouse model with collagen induced rheumatoid arthritis (CIA).

Methods:

We evaluated the Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Creatinine, and the Blood urea nitrogen (BUN) of serum to examine the safety of this study. In vivo, we compared the results of the non-treated group, the normal saline pharmacopuncture treated control group, the indomethacin treated group and the ECP group. We evaluated rheumatoid arthritis manifestation and the Rheumatoid Arthritis Index (AI). Also, immune cells in blood affected by ECP were evaluated by calculating the level of white blood cells (WBC), neutrophil, lympocytes and monocytes. Next, the level of Immunoglobulin M (IgM), Immunoglobulin G (IgG), Interleukin (IL)-1β, IL-6, IL-17, Tumor Necrosis Factor (TNF)-α and Granulocyte-macrophage Stimulating Factor (GM-CSF)in serum were measured. We examined the imaging of cartilage degeneration using micro CT-arthrography of the hind paw. Additionally, we examined the effects of reducing bone volume (BV) ratio and bone surface/bone volume (BS/BV) ratio with 3D Micro-CT. Finally, we did a histopathologic examination analysis.

Results:

The absence of liver and kidney toxicity was evident.
In vivo, edema of the joints of the ECP group decreased greatly in macroscopic observation. AI measurement of the ECP group also decreased significantly compared to the control group. The level of WBC, neutrophil, lympocytes, and monocytes in the blood decreased but there was no statistical significance of this data. IgM of the ECP group decreased significantly compared to the control group. IL-1β, IL-6, TNF-α, and GM-CSF production of the ECP group decreased significantly compared to the control group. As a result of examining joint condition with 3D micro CT, deformation and destruction of the joint was shown to have decreased. Bone density of ECP group increased at a statistically significant level compared to the control group. Degree of joint inflammation of ECP group decreased significantly compared to the control group. After H&E and M-T staining, infiltration of immune cells, subsidence of the cartilage, damage to the synovial cells and joint erosion decreased.

Conclusion:

This study showed that ECP hindered the process of rheumatoid arthritis and protected joints and cartilage.

Fig. 1
Comparison of Rheumatoid Arthritis manifestation in DBA/1 mice between control and experimental group
*: W: Weeks
Control: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
Indo: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
ECP: CIA-induced Rheumatoid Arthritis group treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f1.gif
Fig. 2
Effects of ECP on Rheumatoid Arthritis index score in CIA-induced DBA/1 mice
Rheumatoid Arthritis index was defined as the sum of the individual scores graded as 0 (no arthritis), 1 (small degree of arthritis), 2 (light swelling), 3 (medium swelling), 4 (severe swelling). The results represent the mean ± standard deviation. Statistically significant value was calculated by compared with Control group by student’s t-test (Significance of results, *: p<0.001, †: p<0.01).
Control: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
Indo: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
ECP: CIA-induced Rheumatoid Arthritis group treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f2.jpg
Fig. 3
Effect of ECP on the level of Immunoglobulins in the serum of CIA-induced Rheumatoid Arthritis model of DBA/1 mice
The results were represent the mean ± standard deviation. Statistically significant value was calculated by compared with Control group by student’s t-test (Significance of results, *: p<0.01, †: p<0.05).
(A) Immunoglobulin M (B) Immunoglobulin G.
Normal: Non treated DBA/1 mice.
Control: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
Indo: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
ECP: CIA-induced Rheumatoid Arthritis group treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f3.gif
Fig. 4
Effect of ECP on the level of Cytokines and Chemokines in the serum of CIA-induced Rheumatoid Arthritis model of DBA/1 mice
The results were represent the mean ± standard deviation. Statistically significant value was calculated by compared with Control group by student’s t-test (Significance of results, *: p<0.001, †: p<0.01, ‡: p<0.05).
(A) Interleukin 1β (B) Interleukin 6 (C) Interleukin 17.
(D) Tumor Necrosis Factor-α (E) Granulocyte-macrophage Stimulating Factor.
Normal: Non treated DBA/1 mice
Control: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
Indo: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
ECP: CIA-induced Rheumatoid Arthritis group treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f4.gif
Fig. 5
Effects of ECP on imaging of cartilage degeneration using micro CT-arthrography in paw hind of CIA-induced Rheumatoid Arthritis model of DBA/1 mice
A: Non treated DBA/1 mice.
B: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
C: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
D: CIA-induced Rheumatoid Arthritis group treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f5.gif
Fig. 6
Effects of ECP on bone using 3D Micro-CT of CIA-induced Rheumatoid Arthritis model of DBA/1 mice
The results were expressed as mean ± standard deviation. Statistically significant value was calculated by compared with Control group by student’s t-test (Significance of results, *: p<0.01, †: p<0.05).
(A) Bone volume ratio (B) Bone surface / bone volume ratio.
A: Non treated DBA/1 mice.
B: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
C: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
D: CIA-induced Rheumatoid Arthritis group treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f6.gif
Fig. 7
Effects of ECP histopathologic examination analysis of CIA-induced Rheumatoid Arthritis model of DBA/1 mice
A: Non treated DBA/1 mice.
B: CIA-induced Rheumatoid Arthritis group treated with Normal Saline pharmacopuncture (every two days).
C: CIA-induced Rheumatoid Arthritis group treated orally with indomethacin (200 mg/kg/every two days).
D: CIA-induced Rheumatoid Arthritis group were treated with Erycibae Caulis and Corydalis Tuber pharmacopuncture (200 mg/kg/every two days).
acup-33-2-21f7.gif
Table 1
The Prescription of Erycibae Caulis and Corydalis Tuber
Pharmacognostic name Weight(g)
Erycibae Caulis 30
Corydalis Tuber 30

Total amount 60
Table 2
Standard of Rheumatoid Arthritis Index Score
Rheumatoid Arthritis index Inflammation-induced degree
0 no arthritis
1 small degree of arthritis
2 light swelling
3 medium swelling
4 severe swelling

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