消肿止痛膏对腓肠肌损伤大鼠MyoD mRNA与蛋白表达的影响
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摘要
目的基于微小RNA表达机制,探讨消肿止痛膏对大鼠骨骼肌损伤后重塑和修复的影响。方法通过钝挫伤模型的方法建立大鼠腓肠肌损伤模型,观察消肿止痛膏对大鼠损伤后4、7、14、21天定量PCR检测腓肠肌MyoD mRNA与蛋白表达水平,探讨消肿止痛膏对大鼠腓肠肌钝挫伤模型的肌肉重塑与修复的作用机制。结果治疗组MyoD mRNA表达与蛋白表达水平在造模后14天、21天均显著高于空白组与模型组(P<0.05),进一步证实消肿止痛膏在骨骼肌重塑及修复过程中所起的重要作用。MyoD在损伤后7天上升,其后表达一直维持在较高水平,至损伤后21天仍可以表达出较高水平。与模型组相比较,其表达的高峰期在14天左右,其后则基本回归到一般状态。结论大鼠腓肠肌损伤后,在自然修复过程中,生肌调节因子的表达水平呈现上升-回落的趋势,其中损伤后7～14天,为其表达的高峰期。MyoD的表达水平呈现上升趋势,即使至损伤后21天,其表达仍未见明显下降趋势,可见消肿止痛软膏可以促进MyoD的表达。消肿止痛软膏可以促进MyoD的表达,可以调节骨骼肌的再生修复,提示消肿止痛膏可以通过基因调控,发挥对骨骼肌损伤后的再生修复作用。
Objective To investigate the effects of Xiaozhong Zhitong Ointment on remodeling and repair after skeletal muscle injury based on mRNA expression mechanism.Methods After rat model of injured gastrocnemius muscle with the method of contusion model were established,Xiaozhong Zhitong Ointment was used on rats of 4 d,7 d,14 d,21 d,quantitative PCR was used to detect MyoD mRNA and expression level of protein after the injury to explore the mechanism of muscle remodeling and repair for gastrocnemius muscle contusion model rat.Results The expression of MyoD mRNA and expression level of protein in the treatment group were significantly higher than that of the control group and model group(P<0.05),which further confirmed the important role of Xiaozhong Zhitong Ointment in remodeling and repair process.MyoD was rise at 7 d after the injury,and it expression had maintained at a high level until 21 d after the injury.Compared with the model group,the expression peak was around 14 d,and then it returned to a general state.Conclusion After the rat gastrocnemius muscle get injured,in the natural repair process,the expression level of myogenic regulatory factors shows a trend of rising and dropping,the period of 7 to 14 days after the injury is in the peak of the expression.The expression level of MyoD shows a rising trend,and the decline trend of expression is still not obvious even at 21 d after the injury,so it is can be seen that Xiaozhong Zhitong Ointment can promote the expression of MyoD.Xiaozhong Zhitong Ointment can promote the expression of MyoD,and adjust the regeneration and repair of skeleton muscle,which suggest that Xiaozhong Zhitong Ointment can play a role of regenerating and repairing skeletal muscle after the injury through gene regulation.
Objective To investigate the effects of Xiaozhong Zhitong Ointment on remodeling and repair after skeletal muscle injury based on mRNA expression mechanism.Methods After rat model of injured gastrocnemius muscle with the method of contusion model were established,Xiaozhong Zhitong Ointment was used on rats of 4 d,7 d,14 d,21 d,quantitative PCR was used to detect MyoD mRNA and expression level of protein after the injury to explore the mechanism of muscle remodeling and repair for gastrocnemius muscle contusion model rat.Results The expression of MyoD mRNA and expression level of protein in the treatment group were significantly higher than that of the control group and model group(P<0.05),which further confirmed the important role of Xiaozhong Zhitong Ointment in remodeling and repair process.MyoD was rise at 7 d after the injury,and it expression had maintained at a high level until 21 d after the injury.Compared with the model group,the expression peak was around 14 d,and then it returned to a general state.Conclusion After the rat gastrocnemius muscle get injured,in the natural repair process,the expression level of myogenic regulatory factors shows a trend of rising and dropping,the period of 7 to 14 days after the injury is in the peak of the expression.The expression level of MyoD shows a rising trend,and the decline trend of expression is still not obvious even at 21 d after the injury,so it is can be seen that Xiaozhong Zhitong Ointment can promote the expression of MyoD.Xiaozhong Zhitong Ointment can promote the expression of MyoD,and adjust the regeneration and repair of skeleton muscle,which suggest that Xiaozhong Zhitong Ointment can play a role of regenerating and repairing skeletal muscle after the injury through gene regulation.
引文
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[4] Tilghman SL,Rhodes LV,Bratton MR,et al.Phytoalexins,miRNAs and breast cancer:a review of phytochemical-mediated miRNA regulationg in breast cancer[J].J Health Care Poor Underserved,2013,24(1 Supple):36-46.
[5] Lodewijk L,Prins AM,Kist JW,et al.The value of miRNA in diagnosing thyroid cancer:a systematicreview[J].CancerBiomark,2012,11(6):229-238.
[6] Abouheif MM,Nakasa T,Shibuya H et al.Silencing microRNA-34a inhibits chondrocyte apoptosis in a rat osteoarthritis model in vitro[J].Rheumatology(Oxford),2010,49(11):2054-2060.
[7] 程文.微小RNA与肿瘤相关性研究[J].医学研究生学报,2011,24(2):203-207.
[8] 赵勇,王钢.MicroRNA基因调控在膝骨关节炎发生发展作用中的研究进展[J].中国骨伤,2016,29(4):96-98.
[9] 张海平,宋洁锐.急性骨骼肌损伤动物实验模型构建及应用[J].中国组织工程研究与临床康复,2007,11(49):9984-9988.
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