Effects of Tuina on Muscle-specific microRNA and Factors Related with Proliferation and Differentiation of Muscle Satellite Cells in Denervation-induced Atrophy in Rats / 中国康复理论与实践

Objective:To investigate the effects and mechanism of Tuina on denervation-induced atrophy. Methods:A total of 42 Sprague-Dawley rats were randomly divided into sham group (n = 6), model group (n = 18) and Tuina group (n = 18). The model group and Tuina group freed and excised right tibia nerve about one centimeter, while the sham group freed the right tibia nerve only. From the second day after operation, Tuina group accepted Tuina on the injured area, while the sham group and the model group were only fixed without any intervention. Six rats were sacrificed on the 14th, 21st and 28th day after operation in the model and Tuina groups, and the sham group was sacrificed on the 28th day after operation. The gastrocnemius muscles were measured wet weight ratio. The diameter and area of muscle cells were measured under HE staining. The expression of Pax7, MyoD, MyoG, microRNA-1, microRNA-133a and microRNA-206 in the gastrocnemius muscles were detected with reverse transcription real-time quantitative polymerase chain reaction. Results:Compared with the sham group, the wet weight ratio, the area of muscle cells (except the 14-day-Tuina group) and the diameter of muscle cells decreased at each time point in the model group and Tuina group (P < 0.05); compared with the model group, the wet weight ratio, muscle cell diameter and muscle cell area increased at each time point in Tuina group (P < 0.05). Compared with the sham group, the expression of Pax7 increased in the 14-day-model group (P < 0.05) and decreased in the 28-day-model group (P < 0.05), and it increased at each time point (except 28-day) in Tuina group (P < 0.05); compared with the model group, the expression of Pax7 increased at each time point in Tuina group (P < 0.05). Compared with the sham group, the expression of MyoD and MyoG increased at each time point in the model group and Tuina group (P < 0.05); compared with the model group, the expression of MyoD and MyoG increased at each time point (except 14-day) in Tuina group (P < 0.05). Compared with the sham group, the expression of microRNA-1 and microRNA-133a decreased, and microRNA-206 increased in the model group and Tuina group at 21-day (P < 0.05); compared with the model group, the expression of microRNA-1, microRNA-133a and microRNA-206 increased in Tuina group (P < 0.05). Conclusion:Tuina may activate the Pax7/MyoD/MyoG pathway by increasing the expression of muscle-specific microRNA, to promote the proliferation and differentiation of muscle satellite cells, and delay denervation-induced atrophy.

Advance in Mechanism of microRNA Regulating Muscle Atrophy (review) / 中国康复理论与实践

Muscle loses normal function after skeletal muscle atrophy that will greatly reduce the quality of personal life. There is no effective way to treat muscle atrophy currently. microRNA (miRNA) as a small molecule of non-coding RNA brings new hope for the treatment of muscular atrophy. The mechanism of miRNA regulating muscle atrophy mainly includes: regulating abnormal muscle protein metabolism by ubiquitin-proteasome system (UPS) and mammalian target of rapamycin pathway (IGF/PI3K/Akt/mTOR), inhibiting abnormal apoptosis of muscle cells by inhibiting the expression of apoptotic factors, promoting muscle regeneration by regulating myogenic factor expression, and promoting angiogenesis by promoting the expression of angiogenic factors, and so on.

Effect of Tuina on Autophagy-related Factor Beclin-1, Vacuolar Protein Sorting and Microtubule-associated Protein Light Chain 3 in Rats with Denervated Skeletal Muscle Atrophy / 中国康复理论与实践

Objective:To explore the effect and mechanism of Tuina on denervated skeletal muscle atrophy. Methods:A total of 77 male Sprague-Dawley rats were randomly divided into sham group (n = 7), model group (n = 35) and Tuina group (n = 35). The latter two groups were established skeletal muscle atrophy model by exposing and cutting off the common tibial nerve of rats. One day after modeling, the lower limbs of the surgical side received Tuina in Tuina group. Separately, the surgical side of gastrocnemius muscle were sampled on the 0th, 7th, 14th, 21st and 28thday after modeling, and measured the wet mass ratio. The cross-sectional area and diameter of muscle fiber were measured after HE staining. The mRNA expression of autophagy-realated factor Beclin-1, vacuolar protein sorting (Vps34) and microtubule-associated protein light chain 3 (LC3) were tested with reverse transcription real-time quantitative polymerase chain reaction. Results:There was no statistical difference in the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber, and the mRNA expression of Beclin-1, Vps34 and LC3 among three groups on the 0th day (F < 1.321, P > 0.05). Compared with the sham group, the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber decreased at different time points in the model group and Tuina group (P < 0.05), the ratio of gastrocnemius wet weight was higher, and the cross-sectional area and diameter of muscle fiber were bigger, both except on the 21st day, in Tuina group than in the model group (P < 0.05). Compared with the sham group, the mRNA expression of Beclin-1, Vps34 and LC3 increased at different points in the model group than in Tuina group (P < 0.05), and all the mRNA expression was higher, except on the 14th day, in Tuina group than in the model group (P < 0.05). The ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber showed a trend of progressive decrease with time in the model group and Tuina group (P < 0.05). The mRNA expression of Beclin-1 and Vps34 increased (P < 0.05), and the mRNA expression of LC3 increased in the model group 21 days after intervention (P < 0.05). The mRNA expression of Beclin-1, Vps34 and LC3 increased first and then decreased, except the mRNA expression on the 14th day in Tuina group (P < 0.05). Conclusion:Tuina may promote the activation of autophagy by up-regulating the expression of autophagy-realated factor Beclin-1, Vps34 and LC3, remove the damaged organelles and proteins, provide certain synthetic substrate and energy for muscle fiber regeneration, thereby reduce the loss of degree of denervated skeletal muscle atrophy.

Effect of Electroacupuncture on Blood-brain Barrier in SAMP8 Mice / 中国康复理论与实践

Objective:To investigate the effects of electroacupuncture on blood-brain barrier to promote intake of drugs into brain of SAMP8 mice. Methods:The male SAMP8 mice (30-week-old) were randomly divided into model group (n = 7), drug group (n = 7), acupuncture group (n = 7) and combined group (n = 7). Other SAMR1 mice were as control group (n = 7). The acupuncture group and the combined group accepted electroacupuncture at Baihui (DU20) and Yintang (EX-HN3), the drug group and the combined group accepted Donepezil, for four weeks. They were observed hippocampus tight junction (TJ) under transmission electron microscope. The activities of acetylcholinesterase (AChE) in hippocampus were detected with immunohistochemistry. The mRNA expression of ZO-1, Claudin-5 and Occludin was detected with real-time quantitative polymerase chain reaction. Results:TJ and basement membrane in the control group, the acupuncture group and the combined group were better than those in the model group and the drug group. AChE was the most in the combined group, and then the drug group and the control group, the acupuncture group and the model group (P < 0.001). The expression of mRNA of ZO-1, Claudin-5 and Occludin was more in the acupuncture group and the combined group than in the drug group (P < 0.01). Conclusion:Electroacupuncture could ameliorate blood-brain barrier disruption and promote drug to enter the brain in SAMP8 mice, which may relate to the adjustion of ZO-1, Claudin-5 and Occludin.

Therapeutic effect of massage on denervated skeletal muscle atrophy in rats and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the therapeutic effects of massage on denervated skeletal muscle atrophy in rats and its mechanism.@*METHODS@#Forty-eight male SD rats were randomly divided into model group (n=24) and massage group (n=24). Gastrocnemius muscle atrophy model was established by transecting the right tibial nerve of rat. On the second day after operation, the gastrocnemius muscle of the rats in the massage group was given manual intervention and the model group was not intervened. Six rats were sacrificed at the four time points of 0 d, 7 d, 14 d and 21 d. The gastrocnemius of the rats were obtained and measured the wet mass ratio after weighing. Cross-sectional area and diameter of the muscle fiber were measured after HE staining. The relative expressions of miR-23a, Akt, MuRF1 and MAFbx mRNA were tested with qPCR.@*RESULTS@#Compared with 0 d, the wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle showed a progressive decline in the model group and massage group. The wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle in the massage group were higher than those in the model group on 7 d, 14 d and 21 d (P＜0.05, P＜0.01). Compared with 0 d, the expressions of MuRF1, MAFbx and Akt mRNA were increased first and then were decreased in the model group and massage group. The expression of MuRF1 mRNA in massage group was lower than that in model group on 7 d and 21 d (P＜0.05, P＜0.01). The expression of MAFbx mRNA in massage group was lower than that in model group on 7 d, 14 d and 21 d (P＜0.01, P＜0.05, P＜0.01). The expression of Akt mRNA in massage group was higher than that in model group on 7 d, 14 d and 21 d (P＜0.05, P＜0.01). Compared with 0 d, the expression of miR-23a mRNA was increased in the model group and massage group on 21 d, and the expression of miR-23a mRNA in massage group was higher than that in model group (P＜ 0.05).@*CONCLUSION@#Massage can delay the atrophy of denervated skeletal muscle. The mechanism may be related to up-regulation of the expression of miR-23a and Akt mRNA, down-regulation of the expressions of MuRF1 and MAFbx mRNA, inhibition of protein degradation rate, and reduction of skeletal muscle protein degradation.

Effects of Electroacupuncture on Autophagy-related Gene Expression in Skeletal Muscle in Denervated Rats / 中国康复理论与实践

@#Objective To explore the effects and mechanism of electroacupuncture(EA)on denervation-induced atrophy. Methods A total of 21 male Sprague-Dawley rats were divided into sham group(n=7),model group(n=7)and EA group (n=7).The latter two groups were cut off their right sciatic nerve.Since one day after modeling,EA group accept-ed electroacupuncture at right Zusanli(ST36)and Huantiao(GB30)for eight weeks.Then,the gastrocnemius of all the rats were obtained,and measured the wet mass ratio.Cross-sectional area(CSA)and fiber diameter were measured after HE staining. The expression of autophagy-related gene ULK1, Atg13, Beclin1, Atg14, Atg7, Atg12,Atg5 and Atg16L1 were tested with reverse transcription real-time quantitative polymerase chain reaction. Results Compared with the sham group,the wet mass ratio,CSA and fiber diameter of gastrocnemius were lower signifi-cantly in the model group and EA group(P<0.001),and they were more in EA group than in the model group(P<0.05).Compared with the sham group,the mRNA expression of ULK1,Atg13,Beclin1,Atg14,Atg7,Atg12,Atg5 and Atg16L1 was more significantly in the model group (P<0.001), and they decreased in EA group compared with those of the model group(P<0.05). Conclusion Electroacupuncture can inhibit the overexpression of autophagy-related gene in denervated rats,which may steady skeletal muscle cells to delay atrophy.