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J Acupunct Res > Volume 32(3); 2015 > Article
Lee, Kim, Kim, Park, Yang, Choi, Na, and Wei: Effects of Sweet Bee Venom Pharmacopuncture and Low Level Laser Acupuncture on a Rat Model with Complete Freund’s Adjuvant-induced Arthritis
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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
acupunct-32-3-1f1.gif
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.
acupunct-32-3-1f2.gif
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.

VI.
VI.

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