| BackgroundIn the process of acupuncture clinical treatment of diseases,the selection of acupuncture points,Operation method techniques,acupuncture depth,and duration of treatment are the basic factors for acupuncture prescription.The ancients decided on the timing of acupuncture according to the seasons change,the alternation of the sun and the moon,and the monthly surplus and monthly loss.The modern acupuncture clinical doctors choose the best acupuncture period based on the characters of the disease,the etiology,the pathogenesis,and the course of the disease.The timing of acupuncture intervention is a key influencing factor for the difference in acupuncture efficacy,and is often neglecteed during the treatment of acupuncture.Paying attention to grasping the correlation between the acupuncture time and the acupuncture effect of can not only improve the clinical efficacy of acupuncture,but also provide new ideas for acupuncture treatment of diseases.Multifidus muscle has been widely concerned in the field of acupuncture treatment of low back pain,rehabilitation,sports medicine,etc.It is deep in the back of the human body,originating from the ankle,and is widely associated with each vertebra.It can stabilize the spine and prevent the spine from being dislocated、writhing and other pathological changes;Once the Multifidus muscle is damaged,or muscle atrophy,dysfunction,etc.,the human spine will be unstable,and there will be low back pain,lumbar disc herniation and other symptoms;Myod proved to be an indicator of muscle regeneration,Foxol,Myostatin are the upstream of Myod,and can directly act on Myod,inhibit its expression,and directly act on myoblasts,which is negatively correlated with the early differentiation of myoblasts;In the early stage of the research,it has been confirmed from the research of muscle cell growth related factors,protein,myocyte autophagy,apoptosis and inflammation that electroacupuncture at Weizhong and Shenshu points can effectively promote the repairation of injured multifidus muscle in model rats.On the basis of the previous research,combined with the timing of acupuncture intervention,the treatment of lumbar multifidus muscle injury with electroacupuncture at the Weizhong and Shenshu points will be further explored.ObjectiveThe expressions of Foxo1,Myostatin and Myod in the rat model of multifidus muscle will be detected by immunohistochemistry and Western blotting.The effects of acupuncture intervention time on the treatment of multifidus muscle injury with acupuncture will be also explored.Good acupuncture intervention time provides a accordance with clinical acupuncture treatment of skeletal muscle injuryMethods152 adult male Sprague-Dawley rats were randomly divided into blank group(n=8),electroacupuncture group(n=72)and model group(n=72).The electroacupuncture group and the model group were divided into 9 subgroups according to the intervention immediately after injury,24h intervention,and 48h intervention,and 1d,3d,7d three different material time points;electroacupuncture group and model group rats were selected to inject bupivacaine at four points on both sides of L4 and L5 levels of bilateral spine to establish Rat model of lumbar multifidus muscle injury,the blank group did not do any treatment,feeding normaly;electroacupuncture group according to different acupuncture intervention time points to select bilateral side of the Weizhong point and bilateral Shenshu points for electroacupuncture intervention,connect Korean Electroacupuncture instrument HANS-200A,2/100Hz sparse wave,current intensity 1mA,lasts 20 minutes,1 time/day,until the day before the material is taken,the corresponding model group is synchronously bundled and fixed;The expressions of Foxo1,Myostatin and Myod in the multifidus muscle tissue of the rats were detected by immunohistochemistry and Western blotting.Results1.Changes of Foxol expression in the lumbar spine muscleThe results of Western blotting showed that the expression of Foxol protein was increased in the 24h model group and the 48h model group compared with the blank group(P<0.05).Compared with the model group which is corresponding,the protein expression of Foxo1 was decreased in the 24h electroacupuncture group(P<0.05).<0.05),there was no statistical difference in the expression of Foxo1 in the immediate and 48h acupuncture group(P>0.05).Compared with the immediate and 48h electroacupuncture group,the Foxol protein content in the 24h electroacupuncture group was lower(P<0.05).Compared with the blank group,the expression of Foxol protein in the model group was increased at 24h and 48h(P<0.05).There was no significant difference in the expression of Foxo1 protein in the immediate model group(P>0.05);compared with the corresponding model group.The protein expression of Foxol was decreased in the 24h electroacupuncture group(P<0.01).There was no significant difference in the expression of Foxo1 protein between the 48 and electroacupuncture groups(P>0.05).Compared with the immediate electroacupuncture group,the 24h electroacupuncture group The content of Foxol protein was increased(P<0.05).Compared with the blank group,the expression of Foxol protein in the model group was increased at 24h and 48h(P<0.01).There was no significant difference in the expression of Foxol protein in the immediate model group(P>0.05);compared with the corresponding model group.The protein expression of Foxo1 was decreased in 24h electroacupuncture group(P<0.01).There was no statistical difference in the expression of Foxo1 protein in the 48h electroacupuncture group(P>0.05).Compared with the 48h electroacupuncture group,24h electroacupuncture histone expression Decrease(P<0.05).The results of immunohistochemistry showed that the expression of Foxol protein was significantly increased in the model group at 1 h,compared with the blank group(P<0.01).Compared with the model group which is corresponding,the electroacupuncture group was immediately,24h,48h.The expression of Foxo1 protein was decreased(P<0.05).Compared with the immediate electroacupuncture group,the histone content of 24h electroacupuncture was significantly increased(P<0.01);the expression of histone protein was significantly increased at 48h(P<0.01).Compared with the blank group,the expression of Foxo1 protein in the model group was significantly increased immediately(P<0.01).Compared with the corresponding model group,the protein expression of Foxol was significantly decreased in the 24h electroacupuncture group(P<0.01).There was no statistical difference in the expression of Foxol protein between the immediate and 48h electroacupuncture groups(P>0.05).Compared with the immediate and 48h electroacupuncture group,the expression of Foxo1 protein was decreased in the 24h electroacupuncture group(P<0.01).Compared with the blank group,the expression of Foxo1 protein was significantly increased in the model group at 24h and 48h(P<0.01).Compared with the corresponding model group,the protein expression of Foxol was significantly decreased in the 48 h electroacupuncture group(P<0.01).There was no statistical difference in the expression of Foxo1 protein between the immediate and 24h electroacupuncture groups(P>0.05).There was no statistical difference in the expression of Foxo1 protein between the two groups(P>0.05).2.Changes of the expression of Myostatin in the lumbar muscleThe results of Western blot showed that the expression of Myostatin protein in the model group was decreased at 24h and 48h compared with the blank group(P<0.05).There was no difference in the expression of Myostatin protein in the immediate model group(P>0.05).Compared with the corresponding model group,the expression of Myostatin protein was decreased in the model group.The protein expression of Myostatin was increased in the electroacupuncture group(P<0.05).There was no difference in the expression of Myostatin in the electroacupuncture group at 24h and 48h(P>0.05).Compared with the immediate electroacupuncture group,the electroacupuncture group Myostatin at 24h and 48h Protein expression was reduced(P<0.05).Compar ed with the blank group,the expression of Myostatin protein in the model group was decreased immediately(P<0.05),and the expression of Myostatin protein in the model group was significantly increased(P<0.01).Compared with the corresponding model group,the 24h electroacupuncture group was compared with the corresponding model group.The protein expression of Myostatin was significantly decreased(P<0.01).There was no significant difference in the expression of Myostatin between the immediate and 48h electroacupuncture groups(P>0.05).Compared with the 48h electroacupuncture group,the expression of Myostatin protein was decreased in the 24h electroacupuncture group(P<0.05).<0.05).Compared with the blank group,the expression of Myostatin protein in the model group was decreased immediately(P<0.05).There was no significant difference in the expression of Myostatin protein in the 24h model group(P>0.05).Compared with the corresponding model group,24h.The protein expression of Myostatin was significantly decreased in the electroacupuncture group(P<0.01).There was no statistical difference in the protein expression of Myostatin between the immediate and 48h electroacupuncture groups(P>0.05).Compared with the immediate 48h electroacupuncture group,24h electroacupuncture histone protein The content was reduced(P<0.05).The results of immunohistochemistry showed that the expression of Myostatin protein was significantly increased in the model group at 1 h,compared with the blank group(P<0.01).Compared with the corresponding model group,the protein expression of Myostatin in the electroacupuncture group at 24 h was compared.There was no significant difference in the protein expression of Myostatin between the immediate and 48h electroacupuncture groups(P>0.05).Compared with the 48h electroacupuncture group,the expression of Myostatin protein in the electroacupuncture group was significantly lower(P<0.01).Compared with the blank group,the expression of Myostatin protein in the model group was significantly higher than that in the blank group(P<0.01).Compared with the model group which is corresponding,the protein expression of Myostatin was decreased in the 24h electroacupuncture group(P<0.05).The protein expression of Myostatin was significantly increased in the 48h electroacupuncture group(P<0.01).There was no statistical difference in the protein expression of Myostatin in the electroacupuncture group(P>0.05).Compared with the 48h electroacupuncture group,the 24h electroacupuncture had significant protein content.Decrease(P<0.01).Compared with the blank group,the expression of Myostatin protein in the model group was increased at 24h and 48h(P<0.05,P<0.01).Compared with the model group which is corresponding,the protein expression of Myostatin was significantly decreased in the electroacupuncture group at 24h and 48h.(P<0.01),there was no statistical difference in the protein expression of Myostatin in the electroacupuncture group(P>0.05).There was no statistical difference in the protein expression of Myostatin between the three groups in the immediate,24h,and 48h electroacupuncture groups(P>0.05).3.Changes of the expression of Myod in the lumbar spine muscleThe results of Western blotting showed:Compared with the blank group,the expression of Myod protein in the model group was significantly higher than that in the blank group(P<0.01,P<0.05).Compared with the model group,the protein expression of Myod was increased immediately in the 24h electroacupuncture group.High(P<0.05,P<0.01);compared with the 48h electroacupuncture group,Myod protein expression was significantly increased in the 24h electroacupuncture group(P<0.01).Compared with the blank group,the expression of Myod protein in the model group was increased immediately(P<0.05,P<0.01).Compared with the model group,the Myod protein in the electroacupuncture group was immediately,24h,48h.The expression was increased(P<0.05).Compared with the immed-iate and 48h electroacupuncture group,the expression of Myod protein was significantly increased in the 24h electroacupuncture group(P<0.01).Compared with the blank group,the expression of Myod protein in the model group was increased immediately(P<0.05).Compared with the corresponding model group,the protein expression of Myod was signifi-cantly increased in the 24h electroacupuncture group(P<0.01).There was no statistical difference in the expression of Myod protein between the 24h and 48h electroacupuncture groups(P>0.05).Compared with the immediate and 48h electroacupuncture group,the 24h electroacupuncture histone protein content was significantly increased(P<0.01).The results of immunohistochemistry showed that the expression of Myod protein was increased in the model group at 1 d,3d and 7d,and immediately after 24h and 48h(P<0.05);compared with the corresponding model group,immediately,24h There was no difference in the protein expression of Myod in the 48h electroacupuncture group(P>0.05).There was no difference in the expression of Myod protein between the three groups in the electroacupuncture group at 24h and 48h(P>0.05).ConclusionsBupivacaine can effectively establish a model of rat lumbar multifidus muscle injury;Electroacupuncture can effectively promote the repair of multifidus muscle in model rats by down-regulating the expression of Foxo1 and Myostatin and up-regulating the expression of Myod;different acupuncture intervention times can be a factor on the effect of acupuncture,and the best time for electroacupuncture intervention is 24h after injury. |
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