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Objective: To investigate the therapeutic effects of Tuina (Chinese therapeutic massage) in a knee osteoarthritis (KOA) rat model and its influence on proteins associated with the Wnt/β-catenin signaling pathway. Methods: A total of 32 specific-pathogen-free grade Sprague-Dawley rats were used. Eight rats were randomly selected as the control group (CG). The remaining 24 rats underwent intra-articular injections with 0.2 mL of 4％ papain to prepare the KOA rat models. After the model was established, the 24 rats were randomly and equally assigned to 3 groups, including a model group (MG), a Tuina group (TG), and a positive medicine group (PMG), with 8 rats in each group. The Lequesne score was applied to evaluate the success of model development. After the model was successfully established, the CG did not receive any intervention, and the TG was treated with local, clockwise annular Rou-Kneading around the knee joint with the thumbs. The pressure in the longitudinal direction was 3 N, and the frequency was designed to be 120-140 times/min for 15 min, followed by flexing the joint 10 times. The PMG was intragastrically administered with celecoxib [24 mg/(kg·bw)] every day. These interventions were performed once a day, 6 d per week, for a total of 4 weeks. After treatment, the Lequesne score was applied again to assess the severity of the KOA in the rats; hematoxylin-eosin (HE) staining and a mixture of equal volumes of aqueous solutions of safranin O-fast green were used to stain and observe the cartilage morphology and structure; the modified Mankin score was applied to evaluate the pathology; enzyme-linked immunosorbent assay method was used to quantify the C-telopeptide fragments of type Ⅱ collagen (CTX-Ⅱ) and cartilage oligomeric matrix protein (COMP); Western blotting was then applied to quantify Wnt4, β-catenin, matrix metalloproteinase 13 (MMP-13), and bone morphogenetic protein 2 (BMP-2) protein expression; immunohistochemistry was conducted to determine the percentage of collagen type X (ColX)-positive cells. Results: The Lequesne score of the TG and PMG was both lower than that of the MG (P<0.01); the HE staining, safranin O-fast green stained morphology and structure, and modified Mankin scores of the TG and the PMG were also better than those in the MG (P<0.01). Compared with the CG, the amounts of CTX-Ⅱ and COMP in the serum were significantly increased (P<0.01); the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins in the cartilage tissue was significantly increased (P<0.01), and the percentage of ColX-positive chondrocytes was significantly increased (P<0.01) in the MG. In comparison with those in the MG, the amounts of CTX-Ⅱ and COMP were significantly decreased (P<0.01), the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins was significantly decreased (P<0.01), and the percentage of ColX-positive chondrocytes was significantly decreased (P<0.01) in the TG and PMG. Compared with the PMG, the contents of CTX-Ⅱ and COMP and the expression of Wnt4, β-catenin, MMP-13, and BMP-2 proteins were decreased (P<0.05 or P<0.01); the percentage of ColX-positive chondrocytes was significantly decreased (P<0.01) in the TG. Conclusion: Tuina can relieve the degeneration of KOA, and the mechanism may be related to the down-regulation of the Wnt/β-catenin signaling pathway, the decrease in MMP-13 and BMP-2 protein expression, the reduction in chondrocyte extracellular matrix degradation, and slowing down the terminal cell differentiation.
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Objective:To investigate the effects of tuina, treadmill running or both on the expression of factors related to gastrocnemius muscle proteins after acute muscle injury and to explore the mechanisms involved.Methods:Thirty adult male Sprague-Dawley rats were randomly divided into a control group, a natural recovery group, a tuina group, a treadmill running group and a combined treatment group, each of 6. An impactor was used to induce an acute skeletal muscle injury in the right hind legs of all of the rats except those of the control group. One day after the successful modelling, the tuina, treadmill running and combined groups were given interventions as their name implied, 5 times a week for 3 weeks. The gait of rats in each group was analyzed and the number of times the rats fell into and striken by the electrical grid was counted. The injured muscles′ cross-sectional areas (CSAs) and the diameters of muscle fibers were observed using hematoxylin and eosin staining. The expression of mTOR, p-mTOR, p70S6K, p-p70S6K and smad2/3 protein were tested using western blotting. The relative expression of myostatin (MSTN) mRNA was measured using real-time quantitative polymerase chain reactions.Results:Compared with the natural recovery group, all the other groups fell into the electrical grid significantly less often. The average CSA and wet weight of the affected gastrocnemius had increased significantly in the tuina, treadmill running and combined treatment groups, with the average CSA increases in the treadmill running and combined treatment groups significantly greater compared with the tuina group. The average relative expression of mTOR, p-mTOR, p70S6K, and p-p70S6K in the other four groups increased significantly compared with the control group, while that of Smad2/3 and MSTN decreased significantly. Compared with the natural recovery group, the average increases in the other groups were significantly greater. Compared with the tuina group, the treadmill running and combined treatment groups showed significantly better improvements, on average.Conclusions:Tuina, treadmill running and their combination all can improve recovery from skeletal muscle trauma, at least in rats. However, treadmill running and combined treatment are more effect than tuina alone.
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Objective To observe the effects of massage on inflammation,oxidative stress and autophagy during the repair of acute contusion of skeletal muscles so as to explore its biological mechanisms.Methods Forty-two adult Sprague-Dawley rats were randomly divided into a control group (n =6),a model group (n =18),and a treatment group (n =18).Acute contusion of the gastrocnemius muscles of the rats in the model and treatment groups was inflicted using a home-made impactor.Beginning forty-eight hours later,15 minutes of massage was administered daily for two weeks.After one,7 and 14 days of the massage treatment,the injured gastrocnemius was resected from 6 rats of both the model and treatment groups.Morphological changes were observed using haematoxylin and eosin (HE) staining.The serum content of tumor necrosis factor alpha (TNF-α),interleukin1β (IL-1 β),C reactive protein (CRP) and prostaglandin E2 (PGE2) were detected using enzyme-linked immunosorbent assay (ELISA).The serum content of superoxide (SOD) and malondialdehyde (MDA) were detected using spectrophotometry.The expression of microtubule-associated protein 1 light chain 3 (LC3),Bcl-2 homeodomain protein Beclin1 and ubiquitin binding protein P62 were detected using Western blotting.Results The HE staining showed more significant collapse and swelling of cells in the model group than in the control group at each time point.New muscle cells were observed at days 7 and 14 in the model group.At each time point,significantly better recovery was observed in the treatment group compared to the model group,with more new muscle cells and better cell morphology.According to the ELISA results,a significant increase in serum pro-inflammatory factors occurred in the model group compared to the control group and compared to the treatment group after one day and 7 days of treatment.The average serum content of SOD and MDA in the model group was significantly higher than in the control group,while the average serum content of SOD in the treatment group was significantly higher than in the model group and that of MDA was significantly lower.Western blotting showed a significant decrease in LC3 (Ⅱ/Ⅰ) and Beclin1,as well as a significant increase in P62 in the model group at each time point compared with the treatment group and the controls.Conclusion Inflammation and oxidative stress increase significantly in a skeletal muscle after injury,but autophagy decreases significantly.Massage can effectively reduce the inflammatory response and oxidative stress and promote autophagy,which leads to quicker repair of skeletal muscles.
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@# Irrespective of physiological or pathological skeletal muscle atrophy, the endocrine and motor functions of skeletal muscle are impaired. The mechanism of acupuncture to prevent and treat skeletal muscle atrophy is not only related to classical protein synthesis and decomposition, but also involves apoptosis, autophagy, muscle satellite cell proliferation and differentiation, muscle fiber type conversion, neuromuscular junction conduction, and cell energy metabolism conversion.
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@#Objective To explore the effects and mechanism of electroacupuncture (EA) on denervation-induced atrophy in rats. Methods A total of 18 male Sprague-Dawley rats were divided into sham group (n=6), model group (n=6) and EA group (n=6). The latter two groups were clamped right sciatic nerve to establish atrophy model of skeletal muscle. On the second day after modeling, EA group accepted electroacupuncture on right Zusanli (ST36) and Huantiao (GB30) for two weeks. Their gastrocnemius muscles were obtained after intervention, and the wet weight ratio of the gastrocnemius muscles was calculated. The cross-sectional area (CSA) and diameter of muscle fibers were measured after HE staining. The protein expression of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), 70-KD ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6k (p-p70S6k) was tested with Western blotting. The gene expression of mTOR and p70S6K was detected with real-time quantitative polymerase chain reaction. Results Compared with the sham group, the wet weight ratio of the gastrocnemius muscle, CSA and diameter of the muscle fibers decreased in the model group and EA group (P<0.001), which were more in EA group than in the model group (P<0.01); the protein expression of mTOR, p-mTOR, p70S6K and p-p70S6K increased in the model group (P<0.01), and increased more in EA group (P<0.05); the gene expression of mTOR and p70S6K increased in the model group (P<0.05) , and increased more in EA group (P<0.05).Conclusion Electroacupuncture delays the atrophy of denervated skeletal muscles, which may relate to activation of mTOR/p70S6K signal pathway to impact synthesis of skeletal muscle proteins.
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<p><b>OBJECTIVE</b>To observe the effects of acupuncture at opposite acupoints on expression of hepatocyte growth factor (HGF) in rats with skeletal muscle contusion, and to explore the mechanism of acupuncture at opposite acupoints on skeletal muscle contusion.</p><p><b>METHODS</b>Fifty-four Sprague Dawley (SD) rats were randomly divided into a blank group (6 rats), a model group (24 rats) and an opposing needling group (24 rats). The model group and opposing needling group were further divided into 1-day subgroup, 3-day subgroup, 5-day subgroup and 7-day subgroup, 6 rats in each one. No intervention was given in the blank group, while the model of skeletal muscle contusion was established in the model group and opposing needling group by self-made contusion device. 24 hours after contusion, electroacupuncture (EA) was applied at "Zusanli" (ST 36) and the corresponding points ofpoints at health side for 15 min, once a day. The subgroups of opposing needling group were treated for 1 day, 3 days, 5 days and 7 days, respectively. No treatment was given in the model group. Samples were collected in the subgroups 1 day, 3 days, 5 days and 7 days after treatment. The morphological change of injured gastrocnemius muscle was observed by using microscope after HE staining. The positive cell rate of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. The expression levels of HGF protein and PCNA protein were observed by Western blot.</p><p><b>RESULTS</b>① The results of HE staining showed that, 1 day after contusion, the inflammatory cells of gastrocnemius muscle in the opposing needling group were less than those in the model group; 3 days and 5 days after contusion, myoblasts and myotubes in the opposing needling group were more than those in the model group; 7 days after contusion, the neonatal muscle cells in the opposing needling group were more than those in the model group. ② The results of immunohistochemistry showed that, 1 day, 3 days and 5 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly higher than that in the model group (all<0.001); 7 days after contusion, the positive cell rate of PCNA in the opposing needling group was significantly less than that in the model group (<0.001). ③ The results of Western blot showed that, 1 day, 3 days and 5 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly higher than that in the model group (all<0.05); 7 days after contusion, the expression of HGF protein and PCNA protein in the opposing needling group was significantly lower than that in the model group (all<0.05).</p><p><b>CONCLUSION</b>Acupuncture at opposite acupoints could regulate the expression of HGF and promote the activation, proliferation, migration and differentiation of muscle satellite cells in rats with skeletal muscle contusion, which could speed up the process of skeletal muscle injury repair.</p>
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@#Objective To observe effects and mechanism of electroacupuncture (EA) on denervated skeletal muscle atrophy. Methods Forty-nine male Sprague-Dawley rats were randomly divided into normal group (group A, n=7), natural recovery group (group B, n=21) and EA group (group C, n=21). The groups B and C, established the model of denervated skeletal muscle atrophy by transecting the sciatic nerve of rats, were divided into subgroups of 7 days, 14 days, 21 days postoperation, seven in each subgroup. Electroacupuncture was given to the group C at Zusanli (ST36) and Chengshan (BL57) once a day since 24 hours after modeling. The muscle wet weight ratio of the affected gastrocnemius was determined. Cross-sectional area and fiber diameter of the gastrocnemius were measured with HE staining. The expression of insulin-like growth factor-1 (IGF-1), Myostatin and proliferating cell nuclear antigen (PCNA) protein and gene in the gastrocnemius were detected with Western blotting and RT-PCR. Results The wet weight ratio, cross-sectional area and fiber diameter were less in the groups B and C than in the group A (P<0.001), and they were more in the group C than in the group B (P<0.001). Compared with the group B, the protein and gene of IGF-1, PCNA increased in the group C (P<0.05), while the Myostatin decreased (P<0.05). Conclusion Electroacupuncture can increase the expression of IGF-1 and decrease the expression of Myostatin, to promote the proliferation of satellite cell, which may relate with the prevention of denervated skeletal muscle atrophy.
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Objective To observe the effect of the electroacupuncture ( EA) on the expression of cyclin-de-pendent kinase 5 ( Cdk5 ) in rats with muscle contusion and to explore its mechanism. Methods Thirty-two Sprague-Dawley rats were randomly divided into a normal group of 4, a model group of 4, a natural recovery group ( NR) of 12 and an EA group of 12. All except those in the normal group had acute skeletal muscle contusion induced through a heavy blow. The EA group was treated with 15 minutes of EA daily beginning 48 h after the injury while the other rats received no EA. The model group was sacrificed 24 h after modeling, and rats from the NR and EA groups were sacrificed on the 7th, 14th and 21st day after the modeling to collect tissues. Hematoxylin eosin ( HE) staining, Western blotting and quantitative real-time fluorescence PCR were used to observe any histological changes, as well as Cdk5 protein and mRNA expression. Results The HE staining showed that the other 3 groups displayed larger a-mounts of muscle fiber fracture, dissolution and inflammatory cell invasion than was observed in the normal group. Compared with the NR group, quicker recovery was seen in the EA group as evidenced by faster muscle satellite cell proliferation and more new muscle fiber generation. The average Cdk5 protein expression in both the NR and EA groups was higher than in the normal group, and that of the EA group was significantly lower than that of the NR group. Conclusions Muscle contusion can increase Cdk5 expression in skeletal muscles, at least in rats. EA can promote the restoration of skeletal muscle function, probably by inhibiting CDK5 protein and mRNA expression.
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<p><b>OBJECTIVE</b>To investigate the effect of electro-acupuncture on Toll-like receptor 4 (TLR4) nuclear factor κB (NF-κB) expressions in the liver of rats with non-alcoholic fatty liver disease (NAFLD).</p><p><b>METHODS</b>SD rat models of NAFLD induced by feeding high-fat diet for 5 weeks were subjected to early or late intervention with electro-acupuncture at the acupoints Zusanli, Fenglong and Sanyinjiao for 20 min once daily for 21 consecutive days. The changes in serum ALT, AST, TC, and TG levels were determined in the rats and hepatic expressions of TLR4 and NF-κB detected with quantitative fluorescence PCR and Western blotting after treatments.</p><p><b>RESULTS</b>Serum ALT, AST, TC, and TG levels and TLR4 and NF-κB levels increased significantly after NAFLD onset (P<0.05), and were markedly reduced by interventions with electro-acupuncture (P<0.05).</p><p><b>CONCLUSION</b>Electro-acupuncture at the acupoints Zusanli, Fenglong and Sanyinjiao can modify blood lipid and hepatic function to produce positive effect on NAFLD possibly by modifying lipid metabolism, lowering expressions of TLR4 and NF-κB in the liver, and reducing the impact of inflammation on NAFLD.</p>
Subject(s)
Animals , Rats , Diet, High-Fat , Disease Models, Animal , Electroacupuncture , Inflammation , NF-kappa B , Metabolism , Non-alcoholic Fatty Liver Disease , Metabolism , Therapeutics , Rats, Sprague-Dawley , Toll-Like Receptor 4 , MetabolismABSTRACT
Traditional Chinese Medicine as a rare heritage from our progenitor is becoming more and more important in the world. A lot of foreign students have come to China for learning TCM , and the number of foreign students has increased with time. A lot of issues occur during the teaching process for the foreign students. The problems including the source of students' quality, language, teaching material and clinic practice during the teaching process for the foreign students were ana-lyzed in the article, and the resolutions according to the problems were put forward to attract more and more foreign students to learn TCM and promote TCM in the world.
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In order to explore the possible mechanism of EA (Electroacupuncture) in improving cognitive impairment, the present study was to research the effect of EA on the behavior and expression of the hippocampal neuronal nitric oxide synthase (nNOS) protein of the ovariectomized rats. Forty female SD rats were randomly and equally divided into four groups: sham group (ovarian fat around removal), model group (ovariectomy), sham EA group (sham EA stimulation after ovariectomy) and EA group (EA stimulation after ovariectomy). Two weeks after the ovariectomy, EA and sham EA were applied continuously for 3 months. In the experiment, Morris water maze was used to test the ability of spatial learning and memory, while enzyme-linked immuno sorbent assay (ELISA) and Western blot (WB) were used to detect the concentration of serum estradiol (E2) and relative expression of nNOS protein. The results showed that the escape latency of the EA and sham EA groups was shortened and the number of platform-crossing was significantly increased. The concentration of serum E2 and the expression of nNOS protein in both EA group and sham EA group were significantly elevated compared to those in the model group, while increment in the EA group was more significant (P<0.01). It could be concluded that from above experimental results, EA could be capable of improving learning and memory in ovariectomized rats by promoting the expression of nNOS protein through increasing the concentration of estrogen.
Subject(s)
Animals , Female , Rats , Behavior, Animal , Cognition Disorders , Electroacupuncture , Estradiol , Blood , Hippocampus , Learning , Memory , Neurons , Nitric Oxide Synthase Type I , Metabolism , Ovariectomy , Rats, Sprague-DawleyABSTRACT
Objective To investigate the effect of massage on quadriceps femoris repair after injury by external force and the expression of transforming growth factor β1 (TGF-β1) and collagen-Ⅰ (COL-Ⅰ) mRNA.To explore the molecular mechanisms inhibiting scar tissue formation and promoting muscle repair.Methods Forty New Zealand white rabbits weighing (2.0 ±0.5) kg were randomly divided into a normal control group (A) (n =4),a selfrepair group (B) (n =20,further divided into the 3rd,7th,11th,15th and 19th day time points),and a massage group (C) (n =16,further divided as in group B).In group A the rabbits were not treated,as normal controls.In groups B and C rabbit models of quadriceps femoris injury were prepared using a self-made beater.In group B no massage therapy was given as a natural recovery control; in group C,massage therapy was given after 5 days.Realtime quantitative PCRs were used to detect TGF-β1 and COL-Ⅰ mRNA expression.Resnlts There was no significant difference between groups B and C in the expression of TGF-β1 or COL-Ⅰ mRNA on day 7.At the later time points,expression of both mRNAs in group C was significantly less than in group B.Conclusion Massage can effectively reduce the expression of TGF-β1 and COL-Ⅰ mRNA,inhibit excessive scar formation and promote repair of injured tissue.
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Some traditional Chinese medicine technologies,with strong safety, quick and significant effect, standerdized manipulation, simple operation and good prospect for promotion and application are selected in the research project for application and demonstration of rural health appropriate technology. Through the application of the project, we have new experience in medical teaching.