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Objective:To study the effect of treadmill exercise and massage shortly after acute injury on expression of key growth factor in muscle satellite cells activation, insulin-like growth factor-1 (IGF-1), and mitogen-activated protein kinase (MAPK) / mitogen-activated protein kinase kinase (MEK) / extracellular regulated protein kinases (ERK)1/2 signaling pathways in muscle satellite cell in rats. Methods:A total of 40 SPF male adult Sprague-Dawley rats were randomly divided into groups A (n = 8), B (n = 8), C (n = 8), D (n = 8) and E (n = 8). Group A did not receive any treatment, while the other rats were contused the gastrocnemius muscle with self-made impactor. Group B received no intervention, groups C and D received massage and treadmill exercise shortly after injury, respectively, while group E received both treadmill exercise and massage shortly after injury. As the model was established, samples of gastrocnemius were obtained from all the rats 24 hours after injury, and observed under HE staining, detected the expression of MyoD1 and MyoG mRNA with reverse transcription polymerase chain reaction (RT-PCR), and expression of IGF-1, p-MAPK, p-MEK and p-ERKI/2 protein with Western blotting. Results:The expression of MyoD1 mRNA was more in groups C, D and E than in group B, which was the most in group C; less expression of MyoG mRNA, which was the most in group E (P < 0.05). The expression of p-MAPK, p-MEK and p-EPK1/2 was more in groups C, D and E than in group B, which was the most in group D (P < 0.05). The expression of IGF-1 increased in group C compared with that in group B (P < 0.05), and it decreased in group D (P < 0.05). Conclusion:Early intervention of treadmill exercise and massage may promote the activation of muscle satellite cells in different ways.
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Objective:To observe the effect of electroacupuncture at Baihui (DU20) and Shenshu (BL23) acupoints on learning-memory ability and expression of the relative protein of P35/P25-cyclin-dependent kinase 5 (CDK5)-Tau phosphorylation signaling pathway in the prefrontal cortex (PFC) in rats with Alzheimer's disease (AD), so as to reveal its potential mechanisms in treating AD. Methods:Male adult Sprague-Dawley rats were randomly divided into normal control group, sham group, model group and treatment group with six rats in each group. The AD model was constructed by bilateral hippocampal injection of Aβ25-35 in latter two groups. Equal amount of normal saline was injected into the sham group. The treatment group was acupunctured at Baihui and Shenshu once a day for ten days. All the rats were tested with Morris Water Maze. Immunohistochemistry staining and Western blotting were used to detect the related protein of P35/P25-CDK5-Tau protein phosphorylation in the PFC. Results:Compared with the normal control group and the sham group, the escape latency and escape length increased (P < 0.05) and the times crossing the platform reduced (P < 0.05) in the model group; compared with the model group, the escape latency and escape length reduced (P < 0.05), and the times crossing the platform increased (P < 0.05) in the treatment group. The optical density of P35/P25 and CDK5 were significantly higher in the model group than in the normal control group and the sham group (P < 0.01), and they were lower in the treatment group than in the model group (P < 0.001). The relative expression of P35/P25, CDK5, Tau[pS199] and Tau[pS202] were higher in the model group than in the normal control group and the sham group (P < 0.05), and the expression of the above proteins was lower in the treatment group than in the model group (P < 0.05). Conclusion:Electroacupuncture could improve the learning-memory and spatial exploration ability, which associate with inhabiting the P35/P25-CDK5-Tau protein phosphorylation signaling pathway in the PFC to delay the development of AD.
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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.
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OBJECTIVE@#To observe the effect of electroacupuncture (EA) on the expression of insulin phosphatidylinositol-3 kinase/glycogen synthetase kinase-3α (PI3K/GSK3α) signal pathway related proteins in the hippocampus in mice with Alzheimer's disease (AD), and to explore the regulatory mechanism of EA on improving the pathological characteristics of AD.@*METHODS@#Twelve male APP/PS1 double transgenic mice were randomly divided a model group and a treatment group, 6 mice in each group; another 6 wild-type male mice were taken as the control group. The mice in the treatment group were treated with EA (continuous wave, 2 Hz of frequency) at "Baihui" (GV 20) and bilateral "Shenshu" (BL 23), once a day; 7-day treatment was taken as a course of treatment, and 2 courses of treatment were given. The immunohistochemistry method and Western blot method were used to detect the distribution and expression level of hippocampal PI3K/GSK3α signal pathway related proteins P85α, P110α, GSK3α and pSGSK3α, and the number of hippocampal senile plaques (SP) was observed.@*RESULTS@#The proteins of P85α, P110α, GSK3α and pSGSK3α were mainly distributed in the cytoplasm of hippocampal neurons, and the GSK3α was also distributed in the axons of neurons in the model group and the treatment group. The immunohistochemistry results showed that the distribution level of GSK3α in the hippocampus in the model group was significantly higher than that in the control group (<0.001), and the distribution level of pSGSK3α, P85α and P110α was significantly decreased (<0.01, <0.001); compared with the model group, the distribution level of GSK3α in the hippocampus in the treatment group was significantly decreased (<0.001), and the distribution level of pSGSK3α, P85α and P110α in hippocampus was significantly increased (<0.05, <0.001). The Western blot results showed compared with the control group, the expression of pSGSK3α, P85α and P110α as well as the ratio of pSGSK3α/GSK3α in the hippocampus in the model group were significantly decreased (<0.001), and the expression of GSK3α was increased (<0.05); compared with the model group, the expression of pSGSK3α, P85α, P110α and the ratio of pSGSK3α/GSK3α in the hippocampus in the treatment group were significantly increased (<0.01, <0.001), and the expression of GSK3α was decreased (<0.05). Compared with the control group, the number of hippocampal SP in the model group was significantly increased (<0.001); compared with the model group, the number of hippocampal SP in the treatment group was significantly decreased (<0.01).@*CONCLUSION@#EA could effectively regulate the expression of PI3K/GSK3α signal pathway related proteins in the hippocampus in mice with AD, so as to reduce the formation and deposition of SP.
Subject(s)
Animals , Male , Mice , Alzheimer Disease , Therapeutics , Electroacupuncture , Hippocampus , Physiology , Insulin , Physiology , Mice, Transgenic , Random Allocation , Signal TransductionABSTRACT
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.
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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.
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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.
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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.
Subject(s)
Animals , Male , Rats , Massage , MicroRNAs , Metabolism , Muscle Fibers, Skeletal , Muscle Proteins , Metabolism , Muscle, Skeletal , Muscular Atrophy , Therapeutics , Proto-Oncogene Proteins c-akt , Metabolism , Rats, Sprague-Dawley , SKP Cullin F-Box Protein Ligases , Metabolism , Tripartite Motif Proteins , Metabolism , Ubiquitin-Protein Ligases , MetabolismABSTRACT
@#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.
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<p><b>OBJECTIVES</b>To explore the possible biological mechanism of skeletal muscle contusion repair through researching the changes in expression of autophagy-related genes and proteins in SD rats with acute skeletal muscle contusion.</p><p><b>METHODS</b>Six rats were randomly selected as the control group from 30 male SD rats, acute skeletal muscle contusion model were established in the remaining 24 rats with self-made hitter, then the model rats were randomly divided into 4 groups (3 d, 5 d, 7 d, 14 d groups, =6). On the 3, 5, 7 and 14 day after injury, injured gastrocnemius of each group was harvested. The morphological and the ultra-microstructure changes of gastrocnemius after injury were observed by HE staining and transmission electron microscope (TEM) respectively. The relative protein levels of (LC3-Ⅱ) and P62 of each group were observed by Western blot. The relative mRNA levels of atg7, atg10, atg12, atg16L1 of each group were observed by RTPCR.</p><p><b>RESULTS</b>The results of HE staining showed that compared with the control group, the inflammation reached its peak on the 5 day after injury, new muscle fibers were clearly observed in 7 d group. The results of TEM showed that, compared with the control group, oncotic mitochondria could be clearly seen in the 3 d, 5 d, 7 d groups. Also, the Z line changed from disappearing to drift thickening, sarcoplasmic reticulum dilatation gradually improved, there was no evident difference between the 14 d group and the control group, suggesting that the damage has preliminarily healed. The results of Western blot showed that the expressions of LC3-Ⅱand P62 were increased at first and then decreased. The expression of LC3-Ⅱwas markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (<0.01). Similarly, compared with the control group, the expression of P62 reached its peak on the 3 day after injury (<0. 01), and returned to normal level on the 14 day. The results of RT-PCR showed that the expression of atg10 mRNA in the natural recovery group of 3 d, 5 d, 7 d, 14 d was firstly decreased and then increased, the atg10 mRNA was markedly down-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (<0. 01). The expression of atg7, atg12, atg16L1 mRNA was generally increased at first and then decreased, it was markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (<0.01, <0.05, <0.01).</p><p><b>CONCLUSIONS</b>The above results indicate that the autophagy is involved in repair of skeletal muscle injury by its autophagyrelated factors,regularly changes after contusion, and the rate of damage repair may be related to the level of autophagy.</p>
Subject(s)
Animals , Male , Rats , Autophagy , Contusions , Muscle, Skeletal , Wounds and Injuries , RNA, Messenger , Rats, Sprague-DawleyABSTRACT
The present study was aimed to investigate the effect of electroacupuncture (EA) on learning-memory of rats with low estrogen-induced cognitive impairment and the possible mechanism. The rat model was established by ovariectomy, which resulted in low estrogen-induced cognitive impairment. EA was applied continuously for 3 months 2 weeks after ovariectomy. Morris water maze was used to test the ability of spatial learning and memory. Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative RT-PCR were used to detect the concentration of serum estradiol (E2) and relative expression of choline acetyltransferase (ChAT) mRNA in hippocampus, respectively. The result showed that, compared with the sham group, the ovariectomy model group exhibited longer escape latency, reduced number of platform-crossing, lower concentration of serum E2, and decreased expression of ChAT mRNA in hippocampus. EA shortened the escape latency and increased the number of platform-crossing in the ovariectomy model group. Moreover, the concentration of serum E2 and the hippocampal expression of ChAT mRNA in the ovariectomy model group were significantly elevated by EA treatment. These results suggest EA is capable of improving learning and memory in ovariectomized rats, and the mechanism involves the up-regulation of the expression of ChAT mRNA in hippocampus induced by the increase of the serum concentration of estrogen.