Effect of manipulation on cartilage in rats with knee osteoarthritis based on the Rho-associated protein kinase/LIM kinase 1/Cofilin signaling pathways

doi:10.19852/j.cnki.jtcm.20220311.005

Abstract

Abstract:

OBJECTIVE: To investigate the effect of manipulation treatment on knee osteoarthritis rats and the effect on Rho-associated protein kinase (ROCK)/LIM-kinase1 (LIMK1)/Cofilin signaling pathway.

METHOD: Fifty Specific pathogen Free Sprague-Dawley rats were randomly divided into five groups (n = 8 each): blank group, model group, manipulation group, celecoxib group, and manipulation combined with celecoxib group (MC group). The osteoarthritis model was established by injecting 0.2 mL 4% papain into the articular disc of the rats. After successfully establishing the model, we treated the manipulation group with pushing manipulation using one-finger-meditation to the Neixiyan (EX-LE4), Waixiyan (EX-LE5), Xuehai (SP10), Liangqiu (ST34), and Zusanli (ST36) acupoints for 10 min each time. Also, the celecoxib group was gavaged with 24 mg•kg^-1•d^-1 celecoxib, while the MC group was treated using both of these two methods. After four weeks, the cartilage of the right femur was removed for hematoxylin-eosin staining of the cartilage tissue. The expressions of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in serum were observed using the enzyme-linked immunosorbent assay. Besides, we detected the expressions of ROCK, LIMK1, Phospho-LIM-kinase1 (Phospho-LIMK1), Cofilin, and Phospho-Cofilin by Western blot.

RESULTS: Compared to the model group, the manipulation group, celecoxib group, and MC group all exhibited superior results concerning pathological morphologic changes of cartilage, as observed by hematoxylin-eosin staining and calculated using the Mankin score. Besides, in contrast to the blank group, the model group exhibited elevated serum levels of IL-1β and TNF-α (P < 0.01), while the expression of ROCK, LIMK1, Phospho-LIMK1, Cofilin, and Phospho-Cofilin in cartilage were all higher (P < 0.01). Also, the serum levels of IL-1β and TNF-α in each treatment group were lower (P < 0.01) than in the model group. Moreover, there were lower expressions of ROCK, LIMK1, Phospho-LIMK1, Cofilin, and Phospho-Cofilin in cartilage in the manipulation group and the MC group (P < 0.01). Compared with the model group, the expression of ROCK, LIMK1, Phospho-LIMK1, Cofilin, and Phospho-Cofilin in cartilage in the celecoxib group were not statistically different (P > 0.05).

CONCLUSION: In this study, we established that manipulation has a better curative effect than celecoxib. Manipulation inhibits the development of cytoskeleton damage in cartilage and slows articular degeneration by regulating the expression of related proteins in the cytoskeletal signaling pathway.

Key words: osteoarthritis, knee, manipulation, cytoskeletal proteins, Rho-associated kinases, LIM kinases, actin depoly-merizing factors, signal transduction

GUO Xiao, YANG Yunhao, LIAO Dongmei, PANG Fang, YANG Zhixue, ZHU Zhengwei, LUO Ao, TANG Chenglin. Effect of manipulation on cartilage in rats with knee osteoarthritis based on the Rho-associated protein kinase/LIM kinase 1/Cofilin signaling pathways[J]. Journal of Traditional Chinese Medicine, 2022, 42(2): 194-199.

Figures/Tables 3

Figure 1 Morphological changes in rat knee cartilage (hematoxylin-eosin staining, × 200) A: the blank group did not undergo any treatment; B: the model group were injected with 4% papain 0.2 mL into the knee joint on days 1, 4 and 7; C: the manipulation group were treat with manipulation once daily for six consecutive days, then stopped for one day; D: the celecoxib group was given 24 mg·kg–1·d–1 celecoxib by gavageonce daily for six consecutive days, then stopped for one day; E: the manipulation combined with Celecoxib group (MC group), the above 2 treatments were given once daily for six consecutive days, then stopped for one day. Each group was treated for 4 consecutive weeks.

Table 1 Comparison of the level of serum IL-1β, TNF-α (pg/mL, $\bar{x}±s$)

Group	n	IL-1β	TNF-α
Blank	8	42.6±1.1	116.5±1.9
Model	8	73.1±1.6^a	300.7±1.8^a
Manipulation	8	53.8±0.8^b	201.2±2.9^b
Celecoxib	8	60.7±1.9^bc	250.6±3.4^bc
MC	8	48.9±1.4^b	172.2±1.9^b
P value		<0.01	<0.01

Figure 2 ROCK, LIMK1, Phospho-LIMK1, Cofilin, and Phospho-Cofilin protein expression in the cartilage tissue of rats in each group detected by Western blot A, B: Western blot analysis. The Blank group did not undergo any treatment; the Model group were injected with 4% papain 0.2 mL into the knee joint on days 1, 4 and 7; the Manipulation group were treat with manipulation once daily for six consecutive days, then stopped for one day ; the celecoxib group was given 24 mg·kg–1·d–1 celecoxib by gavageonce daily for six consecutive days, then stopped for one day; the Manipulation combined with Celecoxib group (MC group), the above 2 treatments were given once daily for six consecutive days, then stopped for one day. ROCK: Rho-associated protein kinase; LIMK1: LIM-kinase 1; P-LIMK1: phospho-LIM-kinase 1; P-Cofilin: phospho-Cofilin. aP < 0.01, compared with the blank group; bP < 0.01, cP > 0.05, compared with the model group.

References 21.

1.	Lü SM, Zhang RL. Progress of epidemiological study on osteoarthritis of the knee in middle-aged and elderly people. Zhong Guo Lao Nian Xue Za Zhi 2016; 6: 4133-35.
2.	Tang X, Wang S, Zhan S, et al. The prevalence of symptomatic knee osteoarthritis in China: results from the China health and retirement longitudinal study. Arthritis Rheumatology 2016; 68: 648-53. DOI URL
3.	Zhang SY, Guan XF. Latest development of external treatment with Traditional Chinese Medicine for knee osteoarthritis. Zhong Guo Gu Zhi Su Song Za Zhi 2016; 22: 907-11.
4.	Zheng XB, Lin WR, Lin YW. Clinical efficiency of manipulations combined with warm-needle acupuncture on improving muscle balance of knee osteoarthritis patients. Guangzhou Zhong Yi Yao Da Xue Xue Bao 2016; 33: 822-27.
5.	Blain EJ. Involvement of the cytoskeletal elements in articular cartilage homeostasis and pathology. Int J Exp Pathol 2010; 90: 1-15. DOI URL
6.	Li ZH, Zhu LG. Cofilin gene and protein expression in rabbit chondrocytes under pressure loading conditions. Zhong Guo Lao Nian Xue Za Zhi 2014; 34: 5172-73.
7.	He Q, Yin H, Dai FL, et al. Establishment and verification of rat models of early knee osteoarthritis. Zhong Guo Zu Zhi Gong Cheng Yan Jiu Za Zhi 2019; 23: 4338-43.
8.	Guo Y. Experimental acupuncture and moxibustion experimental guidance. 4th ed. Beijing: China Traditional Chinese Medicine Publishing House, 2016: 78-80.
9.	Wei W, Wu XM, Li YJ. Experimental methodology in pharmacology. 4th ed. Beijing: China People's Medical Publishing House, 2010: 1698.
10.	Liang J, Feng J, Wu W, et al. Leptin-mediated cytoskeletal remodeling in chondrocytes occurs via the RhoA/ROCK pathway. J Orthop Res 2011; 29: 369-74. DOI URL
11.	Romarowski A, Battistone MA, Spina F, et al. PKA-dependent phosphorylation of LIMK1 and cofilin is essential for mouse sperm acrosomal exocytosis. Dev Biol 2015, 405: 237-49. DOI PMID
12.	Duan X, Zhang Y, Chen KL, et al. The small GTPase RhoA regulates the LIMK1/2-cofilin pathway to modulate cytoskeletal dynamics in oocyte meiosis. J Cell Physiol 2018; 233: 6088-97. DOI URL
13.	Bergstrom B, Carlsten H, Ekwall AKH. Methotrexate inhibits effects of platelet-derived growth factor and interleukin-1β on rheumatoid arthritis fibroblast-like synoviocytes. J Arthritis Res Ther 2018; 20: 49.
14.	Fan XL, Liu ZH, Yue T, He DY. Changes in serum tumor necrosis factor-α, interleukin-1β, interleukin-6, and interleukin-17 levels and their predictive value in clinical responses to adalimumab treatment in patients with rheumatoid arthritis. Zhong Guo Yi Ke Da Xue Xue Bao 2018; 47: 556-61.
15.	Salmeri FM, Laganà AS, Sofo V, et al. Behavior of tumor necrosis factor-α and tumor necrosis factor receptor 1/tumor necrosis factor receptor 2 system in mononuclear cells recovered from peritoneal fluid of women with endometriosis at different stages. J Reprod Sci 2015; 22: 165-72.
16.	Liu J, Wan L, Huang CB. Discussion on Bi disease caused by spleen deficiency. Zhong Hua Zhong Yi Yao Za Zhi 2017; 32: 2440-44.
17.	Wu B, Li YP. Excavating osteoarthritis syndrome characteristics and prescription rules by ROST-CM. Shi Zhen Guo Yi Guo Yao Za Zhi 2017; 4: 253-55.
18.	Li ZG. Clinical application and adverse reactions of non-steroidal anti-inflammatory drugs. Shu Li Yi Yao Xue Za Zhi 2017; 30: 1837-38.
19.	Zhang SY, Guan XF. Latest development of external treatment with Traditional Chinese Medicine for knee osteoarthritis. Zhong Guo Gu Zhi Su Song Za Zhi 2016; 22: 907-11.
20.	Li PZ, He GY. Clinical application of a Finger Zen massage technique. Zhong Yi Yao Lin Chuang Za Zhi 2018; 30: 64-7.
21.	Hu BL, Gong L, Li JH, et al. A clinical study of one-finger Tuina on knee osteoarthritis. Zhong Yi Yao Lin Chuang Dao Bao 2017; 2: 72-5.

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