Yuan-Yuan Zhang1, Xi-Hai Li2, Ming-Xia Wu3. 1. College of Acupuncture and Moxibustion, Fujian University of TCM, Fuzhou 350122, China. 2. Institute of Integrated Chinese and Western Medicine, Fujian University of TCM. 3. Department of Acupuncture-Moxibustion and Tuina, Second People's Hospital Affiliated to Fujian University of TCM, Fuzhou 350003.
Abstract
OBJECTIVE: To explore the action mechanism of electroacupuncture (EA) for knee osteoarthritis (KOA) based on Wnt/beta-catenin (Wnt/β-catenin) signaling pathway. METHODS: Ten rats were randomly selected into a sham-operation group among 50 male 2-month-old SD rats, and the KOA model was established in the remaining 40 rats by modified Hulth method. Four weeks after the model establishment, the rats were randomly divided into a model group, an experimental A group, an experimental B group and an experimental C group, 10 rats in each group. The rats in the sham-operation group and model group did not receive any intervention. The rats in the experimental A group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 15 min. The rats in the experimental B group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 30 min. The rats in the experimental C group were treated with EA at non-acupoint for 15 min. EA intervention was given once a day, five times a week, and totally 12-week treatment was given. After 12 weeks, the knee cartilage tissues were stained and the morphological changes were observed under light microscopy; the severity of cartilage degeneration was evaluated by modified Mankin's score; the content of interleukin-1β (IL-1β) in synovium tissues was detected by ELISA method; the content of Wnt-4, β-catenin and matrix metalloprotein-13 (MMP-13) in cartilage tissues was detected by Western blot method. RESULTS: Compared with the sham-operation group, in the model group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and tidal line was incomplete; the Mankin's score was significantly increased (P<0.01), the content of IL-1β in synovium tissues were significantly increased (P<0.01), and the expressions of Wnt-4, β-catenin and MMP-13 at protein level were significantly increased (P<0.01). Compared with the model group, in the experimental A group and experimental B group the morphology and structure of cartilage were more orderly, the number of cells was increased, the matrix staining was deepened and the tidal line was more complete; Mankin's scores were decreased significantly (P<0.01); the contents of IL-1β in synovium tissues were decreased significantly (P<0.01), and the expressions of Wnt-4, β-catenin and MMP-13 at protein level were decreased significantly (P<0.01). Compared with the model group, no improvement was observed in the experimental C group. Compared with the experimental A group, in the experimental C group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and the tidal line was incomplete; Mankin's score was significantly increased (P<0.01), the content of IL-1β in synovium tissues was significantly increased (P<0.01), and the expressions of Wnt-4, β-catenin and MMP-13 at protein level were significantly increased (P<0.01). The morphological structure of cartilage in the experimental B group was similar to that in the experimental A group, and there was no significant difference in Mankin's score, IL-1β content in synovium tissues and the expressions of Wnt-4, β- catenin and MMP-13 at protein level between the two groups (P>0.05). CONCLUSION: EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) may reduce the expression of MMP-13 and the production of inflammatory factor IL-1β through Wnt/β-catenin signaling pathway, thus inhibit the degradation of cartilage matrix and the apoptosis of chondrocyte, and improve the morphology and structure of cartilage.
OBJECTIVE: To explore the action mechanism of electroacupuncture (EA) for knee osteoarthritis (KOA) based on Wnt/beta-catenin (Wnt/β-catenin) signaling pathway. METHODS: Ten rats were randomly selected into a sham-operation group among 50 male 2-month-old SD rats, and the KOA model was established in the remaining 40 rats by modified Hulth method. Four weeks after the model establishment, the rats were randomly divided into a model group, an experimental A group, an experimental B group and an experimental C group, 10 rats in each group. The rats in the sham-operation group and model group did not receive any intervention. The rats in the experimental A group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 15 min. The rats in the experimental B group were treated with EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) for 30 min. The rats in the experimental C group were treated with EA at non-acupoint for 15 min. EA intervention was given once a day, five times a week, and totally 12-week treatment was given. After 12 weeks, the knee cartilage tissues were stained and the morphological changes were observed under light microscopy; the severity of cartilage degeneration was evaluated by modified Mankin's score; the content of interleukin-1β (IL-1β) in synovium tissues was detected by ELISA method; the content of Wnt-4, β-catenin and matrix metalloprotein-13 (MMP-13) in cartilage tissues was detected by Western blot method. RESULTS: Compared with the sham-operation group, in the model group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and tidal line was incomplete; the Mankin's score was significantly increased (P<0.01), the content of IL-1β in synovium tissues were significantly increased (P<0.01), and the expressions of Wnt-4, β-catenin and MMP-13 at protein level were significantly increased (P<0.01). Compared with the model group, in the experimental A group and experimental B group the morphology and structure of cartilage were more orderly, the number of cells was increased, the matrix staining was deepened and the tidal line was more complete; Mankin's scores were decreased significantly (P<0.01); the contents of IL-1β in synovium tissues were decreased significantly (P<0.01), and the expressions of Wnt-4, β-catenin and MMP-13 at protein level were decreased significantly (P<0.01). Compared with the model group, no improvement was observed in the experimental C group. Compared with the experimental A group, in the experimental C group the morphology and structure of cartilage were disordered, the number of cells was significantly reduced, the matrix was decontaminated and the tidal line was incomplete; Mankin's score was significantly increased (P<0.01), the content of IL-1β in synovium tissues was significantly increased (P<0.01), and the expressions of Wnt-4, β-catenin and MMP-13 at protein level were significantly increased (P<0.01). The morphological structure of cartilage in the experimental B group was similar to that in the experimental A group, and there was no significant difference in Mankin's score, IL-1β content in synovium tissues and the expressions of Wnt-4, β- catenin and MMP-13 at protein level between the two groups (P>0.05). CONCLUSION: EA at "Neixiyan" (EX-LE 4) and "Dubi"(ST 35) may reduce the expression of MMP-13 and the production of inflammatory factor IL-1β through Wnt/β-catenin signaling pathway, thus inhibit the degradation of cartilage matrix and the apoptosis of chondrocyte, and improve the morphology and structure of cartilage.
Authors: Mina Wang; Lu Liu; Claire Shuiqing Zhang; Zehuan Liao; Xianghong Jing; Marc Fishers; Luopeng Zhao; Xiaobai Xu; Bin Li Journal: J Pain Res Date: 2020-06-15 Impact factor: 3.133