Changes Of Expression Levels Of MAFbx, MuRFl, Myf5mRNa And MAFbx, MuRF1, Myf5 Protein Of Skeletal Muscle In Ground Squirrels (Spermophilus Dauricus) For Different Peiords Of Hibernation And The Comparison Of Rats

Background:Studies suggest that disuse muscle atrophy in the beginning stages of protein synthesis and metabolism decreased primarily works in disuse muscle atrophy stage of development was mainly enhanced protein catabolism work. Disuse muscle atrophy occurs due to the inhibition of protein synthesis and increased protein degradation, which protein degradation of the main. Present study suggests that MAFbx and MuRF1 mRNA expression level changes may reflect the status of skeletal muscle protein degradation, increasing MAFbx and MuRF1 mRNA expression levels increased protein degradation instructions. Myogenic regulatory factors Myf-5 is to maintain growth and development of skeletal muscle cells important factor may be related to denervation of skeletal muscle atrophy. This experiment is to measure the squirrels by soleus and extensor digitorum longus MAFbx different periods of hibernation, MuRF1 and Myf-5 change, considering the variety of skeletal muscle protein metabolism, explore squirrels from protein metabolism hibernation mechanism.Objective:1. Real time PCR technique for the measurement of soleus muscle in rat extensor digitorum longus muscle atrophy, related factor MAFbx (Muscle Atrophy F-box) mRNA, MuRF1 (Muscle Ring Fingerl) and mRNA relative expression of myogenic regulatory factors Myf-5 mRNA quantity, so as to study its relationship with protein metabolism level, to further explore the mechanism and countermeasure in disuse protein metabolism level of Kamikawa waste with muscular atrophy muscle atrophy of.2. Real time PCR technique for the measurement of hibernation in different periods of Citellus dauricus soleus and extensor digitorum longus muscle atrophy associated factor MAFbx (Muscle Atrophy F-box) mRNA, MuRFl (Muscle Ring Fingerl) and mRNA relative expression of myogenic regulatory factors Myf-5 mRNA amount, in order to explore the Citellus dauricus in protein metabolism level against the disuse muscle atrophy of the possible mechanism.3. Western Blot method to measure the hibernation period of Citellus dauricus soleus muscle, extensor digitorum longus muscle atrophy associated factor MAFbx (Muscle Atrophy F-box) and MuRFl (Muscle Ring Fingerl) and myogenic regulatory factor Myf-5 protein expression, in order to explore the Citellus dauricus in protein metabolism level against the disuse muscle atrophy may mechanism.Methods:1. Real time PCR technology, real-time measurement of soleus muscle, extensor digitorum longus muscle MAFbx, MuRFl and Myf-5 mRNA;2. Expression of Western Blot in measurement of the soleus muscle, extensor digitorum longus, MAFbx MuRFl and Myf-5 protein.Results:1. Rat tail suspension after 14d, the expression quantity of MAFbx mRNA in the soleus muscle was significantly increased by 218%(P<0.01). Dose Ligustrazine synchronous rats were given after 60mg.10ml.kg-1, MAFbx mRNA expression increased 178% significantly compared with the normal group (P<0.01), and hindlimb unloaded group compared with the reduced slightly, but without statistical significance (P>0.05).Rat tail suspension after 14d, the expression quantity of MuRFl mRNA in the soleus muscle was significantly increased by 11 times (P<0.01). Dose Ligustrazine synchronous rats were given after 60mg.10ml.kg-1, MuRFl mRNA expression increased significantly compared with the normal group was 1 times (P<0.01), and hindlimb unloaded group of lower than 84.4%(P<0.01).Rat tail suspension for 14d, Myf-5 mRNA expression in the soleus muscle volume has dropped somewhat, but without statistical significance (P> 0.05).2. The rat tail suspension for 14d and synchronous after administration, changes in expression levels of both extensor digitorum longus MAFbx mRNA, MuRFl mRNA and Myf5mRNA different, but were not significant (P> 0.05).3. Before hibernation groups compared, the hibernating ground squirrel soleus muscle group the expression of MAFbx mRNA increased 11 times (P< 0.05), expression level of arousal ground squirrel soleus muscle MAFbx mRNA increased by 3 times (P< 0.05). Other changes had no statistical significance (P> 0.05).Compared with the pre hibernation group, the hibernating ground squirrel soleus muscle group the expression of MuRFl mRNA increased by 97.8%(P< 0.05), the expression amount of sleep group Citellus soleus muscle MuRFl mRNA increased about 1.3 times (P< 0.05). Other changes had no statistical significance (P> 0.05).Expression of Citellus dauricus soleus muscle Myf-5 mRNA did not change obviously, no statistical significance (P> 0.05).4. Compared the expression amount before hibernation, wake up excited group of Citellus dauricus extensor digitorum longus MAFbx mRNA increased about 8 times (P< 0.05). At the same time, the expression quantity of arousal group Citellus extensor digitorum longus MAFbx mRNA than the expression of the gene of Citellus dauricus sleep group high volume of about 7.5 times (P< 0.05). Other changes had no statistical significance (P> 0.05).Compared with the hibernation group, expression level of arousal group Citellus extensor digitorum longus MuRFl of mRNA decreased 63.7%(P< 0.05). Other changes had no statistical significance (P> 0.05).Compared with the sleep group, the expression quantity of arousal group Citellus extensor digitorum longus Myf-5 of mRNA decreased 93%(P< 0.05). Other changes had no statistical significance (P> 0.05).5. Compared with before hibernation group, hibernation group ground squirrel soleus MAFbx protein expression was increased 3 times (P< 0.01) and arousal ground squirrel soleus MAFbx protein expression increased 1.4 times (P< 0.01). Compared with the hibernation group, the arousal group decreased by 39.3%(P< 0.01), and the sleep group was decreased by 64.9%(P< 0.01). The sleep group was decreased by 42.2%(P< 0.05) compared with the excitation group. Other changes were not statistically significant (P> 0.05).The expression of Citellus dauricus soleus Myf-5 mRNA has no obvious change, no statistical significance (P> 0.05).6. The pre hibernation group, expression level of arousal group squirrel extensor digitorum longus MAFbx protein increased by about 1.15 times (P< 0.05). At the same time, wake up excited group of Citellus dauricus extensor digitorum longus MAFbx protein expression than a sleep group dauricus expression high about 1.02 times (P< 0.05). Other changes were not statistically significant (P> 0.05).Compared with the pre hibernation group, expression level of arousal group squirrel extensor digitorum longus Myf-5 protein decreased by 6.33%(P< 0.05). Other changes were not statistically significant (P> 0.05).Conclusion:1 The load of the hind legs can enhance the protein degradation and metabolism of the anti gravity muscle and the protein degradation of the muscle, but have no effect on the protein synthesis and metabolism.. The effect of Ligustrazine on the expression of the mRNA MuRFl expression of the muscular atrophy factor was increased, and the resistance to muscular dystrophy was also increased.. The muscle synthesis factor Myf-5 had no substantial changes.2 On protein metabolism in hindlimb load on non antigravity muscle extensor digitorum longus has no real impact.3 In of Daurian Ground Squirrel hibernation, the soleus muscle shrinkage factor increasing change does occur, the protein catabolism enhanced, may affect the sleep after ground squirrel soleus muscle to shrink slightly. The muscle synthesis factor Myf-5 had no substantial changes.4 In of Daurian Ground Squirrel hibernation, the extensor digitorum longus muscle shrinkage factor are subject to inter array arousal influence that array between arousal is protection of Spermophilus dauricus toe long extensor almost no shrinking method. Inter array stimulated muscle synthesis factor Myf-5 in extensor digitorum longus has certain effect.5 According to all the results, matrix between arousal is a way to fight muscle atrophy dauricus.