Effects of <i>Jeungmiobi-tang</i> on the Articular Cartilage Injuries Induced by Monosodium Iodoacetate in Rats

Jae-Cheol Hyun; Su-Hyeon Jeong; Hyun, Jae-Cheol; Jeong, Su-Hyeon

doi:10.13048/jkm.20030

JKM > Volume 41(3); 2020 > Article

Hyun and Jeong: Effects of Jeungmiobi-tang on the Articular Cartilage Injuries Induced by Monosodium Iodoacetate in Rats

Original Article

The Journal of Korean Medicine 2020; 41(3): 138-150.

Published online: September 01, 2020

DOI: https://doi.org/10.13048/jkm.20030

Effects of Jeungmiobi-tang on the Articular Cartilage Injuries Induced by Monosodium Iodoacetate in Rats

Jae-Cheol Hyun

, Su-Hyeon Jeong

Department of Rehabilitation medicine of Korean Medicine, College of Korean Medicine, Se-Myung University

Correspondence to: Su-Hyeon Jeong Department of Rehabilitation Medicine of Korean Medicine, College of Korean Medicine, Se-Myung University, 63, Sangbang 4-gil, Chungju-si, Chungcheongbuk-do, Republic of Korea Tel: +82-43-841-1737, Fax: +82-43-856-1731, E-mail: js365a@hanmail.net

Received July 31, 2020 Revised August 13, 2020 Accepted August 19, 2020

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.

Keywords: Jeungmiobi-tang, Monosodium iodoacetate, Articular cartilage injury, Proteoglycan. Osteoarthritis

Fig. 1

Picture of proximal tibia of normal group. The articular cartilage surfaces of tibia are glossy and smooth.

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).

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.

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.

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.

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.

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.

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.

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.

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.

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(%)
Group	0	5	10	15	20 days	Increased Ratio of Body Weight(%)
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)

참고문헌

1. The Korean Orthopaedic Association. Orthopaedics. The 7th edition Second. Seoul: Choisin publisher;2013. p. 317–22.

2. Rehabilitation Medicine of Korean Medicine. Korean Rehabilitation Medicine. 4th edition. Seoul: Kunja Publishing INC;2015. p. 102–16.

3. Felson DT, Lawrence RC, Dieppe PA, et al. Osteoarthritis: new insights: part 1: the disease and its risk factors. Ann Intern Med. 2000; 133:635–646.

4. Spector TD, Cicuttini F, Baker J, Loughlin J, Hart D. Genetic influences on osteoarthritis in women: a twin study. BMJ. 1996; 312:940–3.

5. Greene MA, Loeser RF. Aging-related inflammation in osteoarthritis. Osteoarthritis Cartilage. 2015; 23:11. 1966–71.

6. Won YK. The Effect of Population Aging on the Economic Growth. Pusan national university;2020. p. 1

7. Heo J. Donguibogam. Bupin publishes;2009. p. 1048–50.

8. Park HJ, Bae GS, Kim DY, Seo SW, Park KB, Kim BJ, et al. Inhibitory Effect of Extract from Ostericum koreanum on LPS-induced Proinflammatory Cytokines Production in RAW264.7 Cells. Kor J Herbology. 2008; 23:3. 127–34.

9. Kim YJ. Effects of Curcuma longa Rhizoma on MIA-induced Osteoarthritis in Rat Model. J Korean Med. 2019; 40:3. 35–58.

10. Bin CH, Song CH. Ameliorating Effects of Atractylodes macrocephala Koidzumi on TNF-α-induced 3T3-L1 Adipocyte Dysfunction. Korean Journal of Acupuncture. 2015; 32:3. 116–23.

11. Son HC, Keum DH, Lee MJ. Effects of ELV(Erythrina variegata L inne Var. orientalis(L.) Merr.) on Type II Collagen-Induced Arthritis. J Oriental Rehab Med. 2003; 13:1. 67–76.

12. Han HS. Anti-inflammatory Effect of Angelicae Gigantis Radix Water Extract on LPS-stimulated Mouse Macrophages. Kor J Herbology. 2013; 28:5. 113–9.

13. Herbology editorial committee of Korean medicine schools. Herbology. Seoul: Younglimsa;2007. p. 634–5.

14. Jung HW, Yoon CH, Park KM, Han HS, Park YK. Hexane fraction of Zingiberis Rhizoma Crudus extract inhibits the production of nitric oxide and proinflammatory cytokines in LPS-stimulated BV2 microglial cells via the NF-kappaB pathway. Food and Chemical Toxicology. 2009; 47:6. 1190–7.

15. Yang DH, Woo CH, An HD. Effects of Jinmu-tang on the Osteoarthritis by MIA in Rats. Journal of Korean Medicine Rehabilitation. 2018; 28:1. 19–31.

16. Seo IB, Jeong SH, Park DS. Inhibitory Effects of Daeyeoungjeon on the Injury of Articular Cartilage Induced by Monosodium Iodoacetate in Rats. Journal of Korean Medicine Rehabilitation. 2017; 27:2. 9–17.

17. Jung HC, Jeong SH, Seo IB. Protective Effects of Banggi-eum (FangchiYin) on the Articular Cartilage Injuries Induced by Monosodium Iodoacetate in Rats. Journal of Korean Medicine Rehabilitation. 2014; 24:3. 39–50.

18. Ahn HB, Kim SJ, Seo IB, Jeong SH. Effects of Imyo-san Treatment on the Monosodium Iodoacetate-induced Osteoarthritis in Rats. Journal of Korean Medicine Rehabilitation. 2010; 20:3. 13–26.

19. Janusz MJ, Hookfin EB, Heitmeyer SA, Woessner JF, Freemont AJ, Hoyland JA, et al. Moderation of iodoacetate-induced experimental osteoarthritis in rats by matrix metalloproteinase inhibitors. Osteoarthritis and Cartilage. 2001; 9:751–60.

20. Rudolphi K, Gerwin N, Verziji N, van der Kraan P, van den Berg W. Pralnacasan an inhibitor of interleukin-1β converting enzyme, reduces joint damage in two murine models of osteoarthritis. Osteoarthritis and Cartilage. 2003; 11:10. 738–46.

21. Goldring MB, Otero M, Plumb DA, et al. Roles of inflammatory and anabolic cytokines in cartilage metabolism: signals and multiple effectors converge upon MMP-13 regulation in osteoarthritis. Eur Cell Mater. 2011; 21:202–20.

22. Kim CS, Yoo KS, Hong SH. The Effects of Foot Reflexology on Arthralgia, Ankylosis, Depression, and Sleep in Community dwelling Elderly Women with Osteoarthritis. Journal of Korean Academy of Community Health Nursing. 2014; 25:3. 207–16.

23. Kim HJ. Influence of Depression on Physical Activity, Symptoms of Chronic Pain And Sleep Disorders in the Female Elderly with Knee Osteoarthritis. Exercise Science. 2017; 28:2. 191–7.

24. Grazio S. Osteoarthritis-epidemiology. economics and quality of life. Reumatizam. 2005; 52:2. 21–9.

25. Dijkgraaf LC, de Bont LG, Boering G, Liem RS. Normal cartilage structure, biochemistry, and metabolism: a review of the literature. J Oral Maxillofac Surg. 1995; 53:8. 924–9.

26. Mary KC, Shawn OF, Kwok HS, Kim JW, Kim HG, Jung HS, Tae GS. translation. Biochemistry. 5th ed. Seoul: Life Science Publisher;2007. p. 451

27. Frank H, Netter MD. The CIBA collection of medical illustrations. Seoul: Jeongdam;2000. p. 178

28. Ferrero-Miliani L, Nielsen OH, Andersen PS, Girardin SE. Chronic inflammation: importance of NOD2 and NALP3 in interleukin-1β generation. Clin Exp Immunol. 2007; 147:2. 227–35.

29. Park SK. An interpretation on abnormal finding of CBC. The Korean Journal of Medicine. 2010; 78:5. 531–9.

30. Smolen JS, Aletaha D, Koeller M, Weisman MH, Emery P. New therapies for treatment on rheumatoid arthritis. Lancet. 2007; 370:9602. 1861–74.

31. Iwanami K, Matsumoto I, Tanaka-Watanabe Y, Inoue A, Mihara M, Ohsugi Y, et al. Crucial Role of the Interleukin-6/interleukin-17 Cytokine Axis in the Induction of Arthritis by Glucose-6-phosphate Isomerase. Arthritis Rheum. 2008; 58:3. 754–63.

32. Hulejova H, Baresova V, Klezl Z, Polanska M, Adam M, Senolt L. Increased level of cytokines and matrix metalloproteinases in osteoarthritic subchondral bone. Cytokine. 2007; 38:3. 151–6.

33. Legrand A, Fermor B, Fink C, Pisetsky DS, Weinberg JB, Vail TP, Guilak F. Interleukin-1, tumor necrosis factor alpha, and interleukin-17 synergistically upregulate nitric oxide and prostaglandin E2 production in explants of human osteoarthritic knee menisci. Arthritis Rheum. 2001; 44:9. 2078–83.