Effects of Sweet Bee Venom Pharmacopuncture and Low Level Laser Acupuncture on a Rat Model with Complete Freund’s Adjuvant-induced Arthritis

Article information

Acupunct. 2015;32(3):1-13
1Department of Acupuncture & Moxibustion Medicine, College of Oriental Medicine, Dongshin University
2Department of Oriental Neuropsychiatry, College of Oriental Medicine, Dongshin University
3College of Oriental Medicine, Dongshin University
4Department of Oriental Opthalmology, Otolaryngology & Dermatology, College of Oriental Medicine, Dongshin University
5Department of Oriental Gynecology, College of Oriental Medicine, Dongshin University
6Department of Oriental Internal Medicine, College of Oriental Medicine, Dongshin University
7Department of Acupoint & Meridian, College of Oriental Medicine, Dongshin University
*Corresponding author: Department of Acupuncture & Moxibustion Medicine, Suncheon Hospital fo Traditional Korean Medicine, Dongshin University, 331, Isu-ro, Suncheon-si, Jeollanam-do, 57949, Republic of Korea, Tel: +82-61-729-7133, E-mail: tiger-tung@hanmail.net
Received 2015 May 08; Revised 2015 July 03; Accepted 2015 September 02.

Abstract

Purpose:

This research was conducted to investigate the effect of sweet bee venom pharmacopuncture and low level laser acupuncture on paw edema, pain index, anti-inflammatory factor, AST, ALT and complete blood cell count of a rat model with Complete Freund’s Adjuvant-induced arthritis.

Methods:

Five experimental groups were formed with each consisting of six rats: normal group, control group, sweet bee venom pharmacopuncture group, lower level laser acupuncture group, and sweet bee venom pharmacopuncture, lower level laser acupuncture group. The experimental model of arthritis was induced by two injections of Freund’s adjuvant into the left knee joint of Sprague Dawley(SD) rats. The second injection of Freund’s adjuvant was given ten days after the first one. Ten days later, sweet bee venom pharmacopuncture and low level laser acupuncture were administered separately or together by assigned groups at GB34 and GB39 of rats twice a week for a total of six times. Thereafter, edema rate, pain index, tumor necrosis factor-α, interleukin-6, aspartate aminortansferase, alanine aminotransferase and complete blood cell count were measured.

Results:

We noticed synergic effects of sweet bee venom pharmacopuncture and low level laser acupuncture according to the results of the paw edema and Von Frey pain index. The sweet bee venom pharmacopuncture(BVA) and sweet bee venom pharmacopuncture+ low level laser acupuncture(BVA+LLA) groups experienced a more significant effect when compared with the control group.

Conclusions:

These results suggest that Sweet Bee Venom Pharmacopuncture and low level laser acupuncture at GB34 and GB39 have a significant anti-inflammatory effect on Freund’s adjuvant arthritis in rats.

Fig. 1

Laser acupuncture system

Fig. 2

Changes on the gross lesion of hind paws after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

A: normal group. B: CFA control. C: BVA treated group. D: LLA treated group. E: BVA+LLA treated group.

The Specifications of Laser Acupuncture System

Group Classification and Treatment

Changes on the Paw Edema after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats

Changes on the Reaction Time Contents after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats (sec)

Changes on the Force Contents after VA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats (g)

Changes on the Aspartate Aminotransferase Contents after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats

Changes on the Alanine Aminotransferase Contents after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats

Changes on the TNF-α contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Changes on the Interleukin-6 contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Changes on the blood WBC, Neutrophils, Lymphocytes and Monocytes contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Changes on the blood RBC, HGB, HCT, MCV, MCH, MCHC and PLT contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

References

1. James B Wyngeeden Cecil textbook of medicine Philadelphia: WA Sauders Company; 1992. p. 1508–15.
2. Korean Acupuncture & Moxibustion Society Textbook Compilation Committee. The Acupuncture and Moxibustion. Gyeonggi. Jipmoondang 2008;127
3. Ahn YG. A series of meridian point theory Seoul: Sungbosa; 2000. p. 584–5. p. 592–3.
4. Korean Pharmacopuncture Institute. Pharmacopuncture Seoul: Elsevier Korea LLC; 2008. p. 167–9.
5. Kim CW, Lee YH, Lee GH. The case report of an anaphylaxis occurred when using sweet bee venom and common bee venom at the same time. Journal of Pharmacopuncture 2011;14(4):59–61.
6. Jang IS, Cho GH, Kim YS, et al. Study on clinical application of low level laser therapy in oriental medicine by reviewing recent research. Journal of Korean Medicine 2001;22(3):11–20.
7. Yook TH. Clinical observation of effects of He-Ne laser acupuncture on pain. The Acupuncture 1997;14(1):1–8.
8. Kim GD, Kim GS, Ahn CB, et al. Study on the effect of acupuncture, moxibustion and laser beam acupuncture upon the allviation of pain in mice. The Acupuncture 1987;4(1):39–48.
9. Wei TS, Yoon DH, Yoon YC, et al. Comparative study of acupuncture and invasive laser acupuncture therapy at SI3 BL40 on the tibial, sural nerve injury and L5 spinal nerve ligation model in rats. J Kor AM-M&P Soc 2005;22(2):9–24.
10. Nah SY. Effects of low level laser on pain and cartilage regeneration for a rat model with degenerative arthritis Naju: Dongshin University; 2012. Korean.
11. Kim YS, Lee JH, Lee SH, et al. Effects of electro acupuncture and low level laser theraphy at GB39 and GB34 on neuropathic pain in rats induced by tibial and sural nerves ligation. The Acupuncture 2013;30(3):125–34.
12. Han TR, Bang MS. Rehabilitation nedication Seoul: Goonja Publisher; 2008. 908
13. Bae SC. Mechanics and causes of rheumarthritis. Journal of the Korean Medical Association 2010;53(10):843–52.
14. Koh EM. Medical treatment of rheumatoid arthritis (I): Nonsteroidal anti-inflammatory drugs, disease modifying antirheumatic drugs and glucocorticoids. Journal of the Korean Medical Association 2010;53(1):871–9.
15. Hong WS. Junggyo Hwangjaenagyong Somoon Seoul: Oriental Medicine Research Center; 1981. p. 39–41.
16. Jang JK. Jingguiyaolue Taiwan: Daeryungulpung Company; 1973. p. 32–3.
17. Jang GA. Gyongakjunseo Seoul: Jungdam Publisher; 1999. 217
18. Kim DH, Jung SH, Seo IB, et al. Inhibiting effects of Hwallakdan on the adjuvant-induced arthritis in Lewis rats. JORM 2011;21(1)
19. Ryu MS, Yoon YC, Kim JH. The effect of Angelica gigas NAKAI pharmacopuncture at ST36 and BL23 on Freund’s adjuvant arthritis in rats. The Acupuncture 2010;27(5):25–34.
20. Doh WS, Kim GS. The effect of aqua-acupuncture(Ulmi Pumilae, Ramulus Cinnamomum, Radix Achyranthis, Apitoxin, OK) on LPS-induced arthritis in mice. The Journal of The Korea Institute of Oriental Medicine Informatics 1998;4(1):81–93.
21. Ahn HJ, Doh WS, Jang JH, et al. Effects of acupuncture on rats with LPS-induced arthritis: focusing on macrophage, NK-cell, ICAM. The Journal of the Korea Institute of Oriental Medicine Informatics 1998;4(1):35–48.
22. Song US, Ahn BC, Park DS. Studies of Ga-mi-so-pung-whal tang aqua-acupuncture treatment on the adjuvant arthritis induced by Freundś adjuvant complete. 1990. The Acupuncture 1990;7(1):19–37.
23. Nah CS, Kim JS, Gwon OS, et al. Comparison of effects of acupuncture with bear gall bladder, cow bezoar and bee venom. The Acupuncture 1999;16(1):533–47.
24. Lee HS, Lee JD, Goh HG. The review on the study of bee venom in the domestic papers. The Acupuncture 2003;20(3):154–65.
25. Yoon HM, Ahn CB, Cho EJ. An investigation of directions of research on bee venom in the sphere of oriental medicine in Korea in last decade. Dong-eui-han-eui-yeon 2001;5:23–42.
26. Lee SH, Lee HJ, Paek YH, et al. Effects of bee venom on the pain, edema, and acute inflammatory reactant of rheumatoid arthritis patients. The Acupuncture 2003;20(2):77–84.
27. Hwang YJ, Lee GM, Hwang WJ, et al. Clinical research of bee venom Acupuncture effects on rheumatoid arthritis. The Acupuncture 2001;18(5):33–42.
28. Kim TW, Choi DY, Lee JD. The effect of bee venom on type 2 collagen-induced arthritis. The Acupuncture 2002;19(2):92–104.
29. Yoo DS, Yeom SR, Lee SG, et al. The effect of bee venom pharmacopuncture therapy with concentrated treatment condition in rheumatoid arthritis rat model. JORM 2011;21(2):101–23.
30. Lee SW, Kim YJ, Kim EJ, et al. The effect of varying concentrations of bee venom pharmacoupuncture treatments on type ii collagen induced arthritis in mice. The Acupuncture 2012;29(1):75–87.
31. Tunér Jan, Hode Lars. Laser treatment Seoul: Jungdam; 2006. p. 8–112.
32. Kim JH, Huh W. Medical application of laser for ophthalmology. Journal of the Korean Medical Association 1984;27(2):103–12.
33. Shir Y, Seltzer Z. Effects of sympathectomy in sciatic nerve injury in rats. Pain 1991;45(3):309–20.
34. Lee GN. Clinical pathology file Seoul: Eui-Hak-Moon-Hwa-Sa; 1990. p. 82–7. p. 740–1.
35. Lee GN. Lab test 2000 directory Seoul: Goryeo Medicine; 1999. p. 71–4.
36. Scott D, Wolfe F, Huizinga TW. Rheumatoid arthritis. Lacet 2010;376(9746):1094–108.
37. Park MH, Noh MH, Lee HW, et al. The effects of swimming and low power laser on the concentration of interleukin-6 for rheumatoid arthritic rats. Physical Therapy Korea 2008;15(1):69–76.
38. Ueuji Murakawa. Understanding of Internal Medicine with Pathophysiology Seoul: Jungdam; 2008. p. 71–89.
39. Kwon Yl, Gang SC, Gang JS, et al. Clinical pathology for experimental animals Seoul: Academia; 2009. 202p. 207–10.

Article information Continued

Fig. 1

Laser acupuncture system

Fig. 2

Changes on the gross lesion of hind paws after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

A: normal group. B: CFA control. C: BVA treated group. D: LLA treated group. E: BVA+LLA treated group.

Table 1

The Specifications of Laser Acupuncture System

Specifications LD
Optical properties CW output power 60 mW
Wavelength 904±4 nm
Beam mode Multimode
Mode quality < 1.5
Power stability < 10 %
Electrical properties Operating voltage(V0) < 3 V
Threshold current(Ith) 60±5 mA
Operating current(I0) 100±10 mA
Maximum current(Imax) 150 mA
Accessory properties Optical fiber type Multimode
Core/cladding diameter 50/125 μm
Numerical aperture 0.22
Transmission loss 0.3±0.05 dB/Km
Connector type FC/PC

Table 2

Group Classification and Treatment

Group Classification Treatment
NOR No arthritis inducement and investigational therapies
CON Arthritis inducement with the CFA but no investigational therapies
BVA Arthritis inducement with the CFA and the sweet bee venom(10 %, 100 ul) injection on both GB34 and GB39 Arthritis inducement with the CFA
LLA and administration of 904nm laser acupuncture on GB34 and GB39
BVA+LLA Arthritis inducement with the CFA and concomitant administration of sweet bee venom and 904nm laser acupuncture on GB34 and GB39

Table 3

Changes on the Paw Edema after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats

CFA Treatment

Before After 1st 2nd 3rd 4th 5th 6th
NOR 1.44±0.04 1.57±0.02 1.64±0.05 1.73±0.06 1.80±0.04 1.89±0.04 1.93±0.02 1.92±0.05
CON 1.42±0.03 4.10±0.29## 4.0 3±0.31## 3.79±0.25## 3.84±0.30## 3.88±0.37## 3.84±0.34## 3.97±0.33##
BVA 1.40±0.04 3.63±0.19 3.27±0.14* 2.9 6±0.13* 3.01±0.12 3.06±0.13 2.83±0.09* 2.51±0.10**
LLA 1.40±0.04 4.08±0.42 3.52±0.39 3.11±0.29 3.39±0.19 3.69±0.16 3.01±0.07* 2.85±0.12*
BVA+LLA 1.42±0.09 3.45±0.31 3.16±0.34 2.84±0.26* 3.04±0.17* 3.23±0.10 2.60±0.14 2.54±0.17**

Values are expressed mean±SE. CFA: Complete Freund’s Adjuvant.

##

p<0.01 compared with normal.

*

p<0.05,

**

p<0.01 compared with CFA control.

Table 4

Changes on the Reaction Time Contents after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats (sec)

CFA Treatment

Before After 1st 2nd 3rd 4th 5th 6th
NOR 7.2±0.1 7.0±0.2 7.1±0.5 6.8±0.2 6.6±0.5 6.7±0.3 6.9±0.5 6.2±0.3
CON 7.1±0.1 3.2±0.3## 3.1±0.2## 2.9±0.4## 2.8±0.4## 2.8±0.5## 2.7±0.4## 2.7±0.3##
BVA 7.1±0.2 3.4±0.3 3.0±0.3 3.4±0.2 3.8±0.3 4.1±0.2* 5.1±0.3** 5.5±0.4**
LLA 7.3±0.2 3.1±0.4 3.7±0.5 4.0±0.4 4.4±0.4 4.8±0.4** 5.4±0.4** 5.1±0.4**
BVA+LLA 7.2±0.2 3.2±0.5 3.2±0.3 3.3±0.3 4.8±0.2** 5.7±0.4** 5.5±0.6** 5.5±0.3**

Values are expressed mean±SE. CFA: Complete Freund’s Adjuvant.

##

p<0.01 compared with normal.

*

p<0.05,

**

p<0.01 compared with CFA control.

Table 5

Changes on the Force Contents after VA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats (g)

CFA Treatment

Before After 1st 2nd 3rd 4th 5th 6th
NOR 35.6±0.3 34.9±1.0 35.4±2.3 34.2±1.2 33.1±2.3 33.7±1.4 34.5±2.3 31.1±1.3
CON 35.3±0.4 16.1±1.4## 15.6±1.2## 14.8±2.2## 14.3±2.2## 14.3±2.5## 14.0±2.0## 13.8±1.5##
BVA 35.4±0.8 17.1±1.5 15.2±1.4 17.5±0.8 19.3±1.6 20.7±1.2* 25.6±1.7** 28.2±1.9**
LLA 35.9±0.7 15.6±2.2 18.8±2.4 20.4±1.8 22.0±2.1* 24.3±2.0** 27.1±1.9** 25.9±2.0**
BVA+LLA 36.0±0.9 16.0±2.3 16.3±1.5 16.6±1.6 24.2±1.2** 28.8±1.8** 27.8±2.8** 27.4±1.6**

Values are expressed mean±SE. CFA: Complete Freund’s Adjuvant.

##

p<0.01 compared with normal.

*

p<0.05,

**

p<0.01 compared with CFA control.

Table 6

Changes on the Aspartate Aminotransferase Contents after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats

Group Aspartate aminotransferase(U/l)
NOR 65.7±3.0
CON 101.5±3.9##
BVA 98.6±4.5
LLA 87.0±3.5*
BVA+LLA 98.6±6.2

Values are expressed mean±SE.

CFA: Complete Freund’s Adjuvant.

##

p<0.01 compared with normal.

*

p<0.05 compared with CFA control.

Table 7

Changes on the Alanine Aminotransferase Contents after BVA, LLA and BVA+LLA Treatments in CFA Induced Arthritis Rats

Group Alanine aminotransterase(U/l)
NOR 32.5±2.2
CON 42.5±4.4
BVA 34.0±1.1
LLA 31.3±2.6*
BVA+LLA 35.8±3.2

Values are expressed mean±SE.

CFA: Complete Freund’s Adjuvant.

*

p<0.05 compared with CFA control.

Table 8

Changes on the TNF-α contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Group Tumor necrosis factor-α(pg/ml)
NOR 1.35±0.18
CON 3.23±0.09#
BVA 1.12±0.09*
LLA 1.04±0.08**
BVA+LLA 1.82±0.32

Values are expressed mean±SE. CFA: Complete Freund’s Adjuvant.

#

p<0.05 compared with normal.

*

p<0.05,

**

p<0.01 compared with CFA control.

Table 9

Changes on the Interleukin-6 contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Group Interleukin-6(pg/ml)
NOR 7.43±0.12
CON 7.89±0.09#
BVA 7.35±0.08**
LLA 7.13±0.07**
BVA+LLA 7.01±0.05**

Values are expressed mean±SE.

CFA: Complete Freund’s Adjuvant.

#

p<0.05 compared with normal.

**

p<0.01 compared with CFA control.

Table 10

Changes on the blood WBC, Neutrophils, Lymphocytes and Monocytes contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Group\Parameters WBC (K/uL) Neutrophils (K/uL) Lymphocytes (K/uL) Monocytes (K/uL) Eosinophils (K/uL) Basophils (K/uL)
NOR 4.4±0.2 1.03±0.13 3.1±0.179 0.17±0.04 0.03±0.02 0.003±0.002
CON 4.8±0.4 1.32±0.17 3.22±0.34 0.21±0.03 0.02±0.00 0.002±0.002
BVA 4.4±0.4 1.21±0.16 2.93±0.32 0.18±0.02 0.03±0.01 0.005±0.005
LLA 4.7±0.4 1.45±0.23 3.01±0.22 0.18±0.02 0.02±0.00 0.001±0.000
BVA+LLA 4.1±0.2 1.33±0.13 2.62±0.13 0.12±0.02* 0.04±0.02 0.017±0.012

Values are expressed mean±SE.

CFA: Complete Freund’s Adjuvant.

*

p<0.05 compared with CFA control.

Table 11

Changes on the blood RBC, HGB, HCT, MCV, MCH, MCHC and PLT contents after BVA, LLA and BVA+LLA treatments in CFA induced arthritis rats

Group\Parameters RBC (K/uL) HGB (g/dL) HCT (%) MCV (fL) MCH (pg) MCHC (g/dL) PLT (K/uL)
NOR 7.8±0.2 13.9±0.1 52.1±0.7 67.1±1.2 17.9±0.4 26.7±0.3 675.8±104.8
CON 8.1±0.1 13.9±0.2 51.0±0.5 63.2±0.8 17.2±0.3 27.2±0.3 864.5±33.6
BVA 7.7±0.2 13.7±0.3 48.8±1.2 63.3±1.1 17.7±0.3 28.0±0.3 797.2±41.6
LLA 7.9±0.1 14.1±0.3 51.2±0.8 64.5±0.8 17.8±0.3 27.6±0.3 814.5±15.4
BVA+LLA 7.4±0.2* 13.6±0.3 48.5±1.3 65.5±1.0 18.4±0.2* 28.0±0.5 785.7±87.7

Values are expressed mean±SE.

CFA: Complete Freund’s Adjuvant.

*

p<0.05 compared with CFA control.