College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400010.
China Beibei Traditional Chinese Medicine Hospital, Chongqing 400700, China.
J Tradit Chin Med. 2022 Apr;42(2):194-199. doi: 10.19852/j.cnki.jtcm.20220311.005.
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.
Fifty Specific pathogen Free Sprague-Dawley rats were randomly divided into five groups ( = 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•d 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.
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 ( 0.01), while the expression of ROCK, LIMK1, Phospho-LIMK1, Cofilin, and Phospho-Cofilin in cartilage were all higher ( 0.01). Also, the serum levels of IL-1β and TNF-α in each treatment group were lower (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 (< 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 ( > 0.05).
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.
观察手法治疗对膝骨关节炎大鼠的作用及对 Rho 相关蛋白激酶(ROCK)/ LIM 激酶 1(LIMK1)/ 丝切蛋白(Cofilin)信号通路的影响。
将 50 只 SPF 级 SD 大鼠随机分为 5 组(每组 8 只):空白组、模型组、手法组、塞来昔布组、手法联合塞来昔布组(MC 组)。采用向关节腔内注射 0.2 mL 4%木瓜蛋白酶的方法制备骨关节炎模型。造模成功后,手法组采用一指禅推法依次作用于内膝眼(EX-LE4)、外膝眼(EX-LE5)、血海(SP10)、梁丘(ST34)、足三里(ST36)穴位,每次 10 min;塞来昔布组灌胃给予 24 mg•kg•d 塞来昔布;MC 组采用上述两种方法联合治疗。4 周后,取右侧股骨软骨进行苏木精-伊红染色,观察软骨组织病理形态变化,采用 Mankin 评分法进行评估;采用酶联免疫吸附法检测血清中白细胞介素 1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的表达;采用 Western blot 法检测软骨组织中 ROCK、LIMK1、磷酸化 LIM 激酶 1(Phospho-LIMK1)、丝切蛋白(Cofilin)和磷酸化丝切蛋白(Phospho-Cofilin)的表达。
与模型组比较,手法组、塞来昔布组和 MC 组的软骨病理形态学变化均有不同程度改善,Mankin 评分降低( 0.01);与空白组比较,模型组血清 IL-1β和 TNF-α水平均升高( 0.01),软骨组织中 ROCK、LIMK1、Phospho-LIMK1、Cofilin 和 Phospho-Cofilin 表达均升高( 0.01);与模型组比较,各治疗组血清 IL-1β和 TNF-α水平均降低( 0.01),软骨组织中 ROCK、LIMK1、Phospho-LIMK1、Cofilin 和 Phospho-Cofilin 表达均降低( 0.01)。与模型组比较,塞来昔布组软骨组织中 ROCK、LIMK1、Phospho-LIMK1、Cofilin 和 Phospho-Cofilin 表达差异无统计学意义( > 0.05)。
手法治疗膝骨关节炎的疗效优于塞来昔布,其通过调节细胞骨架信号通路相关蛋白的表达,抑制软骨细胞骨架损伤的发展,减缓关节退变。