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JKM > Volume 41(3); 2020 > Article
Hyun and Jeong: Effects of Jeungmiobi-tang on the Articular Cartilage Injuries Induced by Monosodium Iodoacetate in Rats

Abstract

Objectives

This study was carried out to investigate the protective effects of Jeungmiobi-tang on the articular cartilage injuries induced by monosodium iodoacetate in rats.

Methods

Twenty four rats were divided into three groups. Rats of normal group (n=8) were injected with 0.1 ml physiological saline into both knee joint cavities. In the rats of control group (n=8) and Jeungmiobi-tang group (n=8), Arthritis was induced by injecting with 0.1 ml monosodium iodoacetate (5 mg/ml) into both knee joint cavities. After the experiment, Gross and histopathological examinations on the knee joint were performed. The content of proteoglycan in articular cartilage and TNF-α and IL-1β in synovial fluid were also analyzed.

Results

Grossly, Injuries to the articular cartilage surface was observed weak in the Jeungmiobi-tang group compared to the control group. Proteoglycan content in the articular cartilage was significantly higher in the Jeungmiobi-tang group than in the control group. The chondrocyte score was significantly lower in the Jeungmiobi-tang group than in the control group.

Conclusion

According to these results, that Jeungmiobi-tang has protective effects on the articular cartilage injuries induced by monosodium iodoacetate in rats.

Fig. 1
Picture of proximal tibia of normal group. The articular cartilage surfaces of tibia are glossy and smooth.
jkm-41-3-138f1.gif
Fig. 2
Picture of proximal tibia of control group. The articular cartilage surfaces of tibia are discolored and rough(arrow head), and some areas are deeply ulcerated(arrow).
jkm-41-3-138f2.gif
Fig. 3
Picture of proximal tibia of Jeungmiobi-tang treated group. The articular cartilage surfaces of tibias are less discolored and less rough than control group.
jkm-41-3-138f3.gif
Fig. 4
Coronal section of proximal tibia of normal group. The articular cartilage(*) is thickly stained with Safranine O. EP : epiphyseal plate, Safranine O stain, bar = 300 μm.
jkm-41-3-138f4.gif
Fig. 5
Coronal section of proximal tibia of control group. The red color disappeared in a great part of articular cartilage(*). Only a portion of the periphery of cartilage and subchondral part was stained red color(arrow and arrow heads). EP : epiphyseal plate. Safranine O stain, bar = 300 μm.
jkm-41-3-138f5.gif
Fig. 6
Coronal section of proximal tibia of Jeungmiobi-tang treated group. The articular cartilage(*) is more stained with Safranine O than control group(arrow and arrow heads). EP : epiphyseal plate. Safranine O stain, bar = 300 μm.
jkm-41-3-138f6.gif
Fig. 7
Sagittal section of normal group’s knee joint. The articular cartilages(*) of synovial membrane(arrows), meniscus(M), femur(F), tibia(T), and are normal. H&E stain, bar = 300 μm.
jkm-41-3-138f7.gif
Fig. 8
Sagittal section of control group’s knee joint. A great part of articular cartilages(*) of tibia(T) and femur(F) are necrotic and degenerated. There is Cleft(arrow head) in deep part of articular cartilage. Also, reactive fibrosis(arrows) are shown. H&E stain, bar = 300 μm.
jkm-41-3-138f8.gif
Fig. 9
Higher magnification of Fig. 8. Clefts(arrow heads) and Homogenous degeneration(*) of articular cartilage are shown. Also, There are reactive fibrosis(long arrows) and activation of osteoclasts(small arrows). H&E stain, bar = 100 μm.
jkm-41-3-138f9.gif
Fig. 10
Sagittal section of Jeungmiobi-tang treated group’s knee joint. A great part of articular cartilages(*) of tibia(T) and femur(F) are degenerated. But chondrocytes (arrows) around the degenerative area are intact. Clefts in deep part of articular cartilage are not shown. H&E stain, bar = 300 μm.
jkm-41-3-138f10.gif
Table 1
Prescription of Jeungmiobi-tang per Pack
Name of herbs Pharmacognostic name Weight(g)
羌活 Ostericum Koreanum 4g
防己 Sinomenium Acutum 4g
薑黃 Curcuma Longa Rhizoma 4g
白朮 Atractylodis Rhizoma Alba 4g
海桐皮 Kalopanacis Cortex 4g
當歸 Angelicae Gigantis Radix 4g
白芍藥 Paeoniae Radix Alba 4g
甘草(炙) Glycyrrhizae Radix 3g
生薑 Zingiberis Rhizoma Crudus 30g

Total
Table 2
Criteria for Histopathological Score of Osteoarthritic Lesions
Matrix Score Chondrocyte Score
 0 Normal  0 Normal
 1 Surface irregularities  2 Reduced cells
 3 Superficial fibrillation  5 Strongly reduced cells
 6 Clefts in deep zone  8 Total loss of cartilage and cells
 8 Massive loss of cartilage

Subchondral Bone Score
 0 Normal
 3 Remodeling processes
 8 Thickening, strong sclerosis
Table 3
Effects of Jeungmiobi-tang Treatment on the Body Weight of Rats
Group Body Weight(g) Increased Ratio of Body Weight(%)
0 5 10 15 20 days
Normal (n=8) 220±6* 255±11 288±13 314±15 337±16 52.9±5.6
Control (n=8) 221±6 246±11 280±14 303±13 324±10 46.6±4.0||
Jeungmiobitang (n=8)§ 221±9 244±8 282±12 307±13 329±18 48.9±6.0

* data expressed as Mean±S.D.

† saline injected and treated with distilled water

‡ MIA injected and treated with distilled water

§ MIA injected and treated with Jeungmiobi-tang extracts

|| Statistically significant compared with normal group(p<0.05)

Table 4
Effects of Jeungmiobi-tang Treatment on the Proteoglycan Content in Tibial Articular Cartilages
Group Proteoglycan Content (%)
Normal (n=8) 100.0±8.3*
Control (n=8) 19.5±5.6||
Jeungmiobitang (n=8)§ 34.7±17.7

* data expressed as Mean±S.D.

† saline injected and treated with distilled water

‡ MIA injected and treated with distilled water

§ MIA injected and treated with Jeungmiobitang extracts

|| Statistically significant compared with normal group(p<0.001)

¶ Statistically significant compared with control group(p<0.05)

Table 5
Difference in Numbers of Total WBC, Neutrophils, Lymphocytes and Monocytes
Group Total WBC (x103/ml) Neutrophils Lymphocytes Monocytes
Normal (n=8) 4.08±0.53* 881±174 3,087±419 69±28
Control (n=8) 5.27±1.20|| 1,175±285* 3,865±1001 104±53
Jeungmiobi-tang (n=8)§ 6.35±1.17 1,330±223 4,763±892 187±100

* data expressed as Mean±S.D.

† saline injected and treated with distilled water

‡ MIA injected and treated with distilled water

§ MIA injected and treated with Jeungmiobitang extracts

|| Statistically significant compared with normal group(p<0.05)

Table 6
Effects of Jeungmiobi-tang Treatment on the Osteoarthritic Score of the Knee Joints of Rats
Group and Identification Number Matrix Score Chondrocyte Score Subchondral Bone Score Sum of Scores
Control C1 6 5 8 19
C2 3 5 3 11
C3 8 5 8 21
C4 3 5 3 11
C5 3 5 3 11
C6 6 5 8 19
C7 8 8 3 19
C8 6 5 3 14

Mean 5.4±2.0* 5.4±1.0 4.9±2.4 15.6±4.0

Jeungmiobi-tang T1 1 2 3 6
T2 3 5 3 11
T3 3 5 8 19
T4 3 2 8 13
T5 6 5 8 19
T6 1 2 0 3
T7 3 2 8 13
T8 6 5 3 14

Mean 3.3±1.8 3.5±1.5§ 5.1±3.0 12.3±5.3

* data expressed as Mean±S.D.

† MIA injected and treated with distilled water

‡ MIA injected and treated with Jeungmiobi-tang extracts

§ Statistically significant compared with control group(p<0.05)

Table 7
Effects of Jeungmiobi-tang Treatment on Tumor Necrosis Factor-α Contents in Synovial Fluids of Rats
Group Tumor Necrosis Factor-α (pg/ml)
Normal (n=8) 48.2±13.3*
Control (n=8) 101.5±16.5||
Jeungmiobi-tang (n=8)§ 84.0±18.9

* data expressed as Mean±S.D.

† saline injected and treated with distilled water

‡ MIA injected and treated with distilled water

§ MIA injected and treated with Jeungmiobi-tang extracts

|| Statistically significant compared with normal group(p<0.001)

Table 8
Effects of Jeungmiobi-tang Treatment on Interleukin-1β Contents in Synovial Fluids of Rats
Group Interleukin-1β (pg/ml)
Normal (n=8) 96.3±13.1*
Control (n=8) 114.7±13.3||
Jeungmiobi-tang (n=8)§ 102.6±8.0

* data expressed as Mean±S.D.

† saline injected and treated with distilled water

‡ MIA injected and treated with distilled water

§ MIA injected and treated with Jeungmiobi-tang extracts

|| Statistically significant compared with normal group(p<0.05)

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