电针“委中”对布比卡因致大鼠腰多裂肌损伤后形态学及CK、IL-17表达的影响
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摘要
目的:观察电针"委中"穴对布比卡因(bupivacaine,BPVC)致大鼠腰多裂肌损伤后组织形态学及磷酸肌酸激酶(CK)、白介素17(IL-17)表达水平的影响。方法:将32只雄性SD大鼠随机分为对照组、模型组、电针委中组和电针肾俞组,每组8只。模型组、电针委中组和电针肾俞组采用0.5%BPVC肌内注射制备多裂肌损伤模型;对照组采用同样的方法注射0.9%Na Cl溶液。电针委中组、电针肾俞穴分别电针"委中"与"肾俞"穴,选用疏密波,频率2 Hz/10 Hz,电流强度1~2 m A,持续20 min;对照组与模型组不进行针刺干预。电针干预14 d后,通过苏木精-伊红(HE)染色和马松三色染色法(Masson)观察多裂肌炎细胞计数、瘢痕面积和肌纤维横截面积的变化。通过酶联免疫吸附法(ELISA)检测血清CK活性和IL-17的含量,并用免疫组化法检测多裂肌损伤部位的IL-17表达。结果:干预后,模型组、电针委中组和电针肾俞组的炎细胞计数、瘢痕面积明显高于对照组(均P<0.01),肌纤维横截面积明显减少(均P<0.01);电针委中组和电针肾俞组炎细胞计数、瘢痕面积均少于模型组(均P<0.01),肌纤维横截面积大于模型组(P<0.01,P<0.05)。干预后,模型组、电针委中组和电针肾俞组多裂肌损伤局部的IL-17表达、血清IL-17含量及CK活性均明显高于对照组(均P<0.01);电针委中组和电针肾俞组多裂肌中IL-17的表达、血清IL-17含量及CK活性均低于模型组(P<0.01,P<0.05);与电针肾俞组比较,电针委中组下调IL-17的趋势更明显(P<0.01)。结论:电针"委中"穴可通过下调血清CK和白介素-17的过度表达,减轻炎性反应,促进多裂肌的良性修复。
Objective To observe effects of electroacupuncture(EA) at "Weizhong"(BL 40) on morphology and expression of creatine kinase(CK) and interleukin-17(IL-17) in rats with bupivacaine-induced multifidus muscle injury.Methods A total of 32 male SD rats were randomly divided into a control group, a model group, a Weizhong group and a Shenshu group, 8 rats in each one. The rats in the model group, Weizhong group and Shenshu group were treated with intramuscular injection of 0.5% bupivacaine to establish the model of multifidus muscle injury; the rats in the control group were injected with 0.9% sodium chloride solution. The rats in the Weizhong group and Shenshu group were treated with EA(2 Hz/10 Hz in frequency, 1~2 m A in intensity) at "Weizhong"(BL 40) and "Shenshu"(BL 23), 20 min per treatment. No treatment was given in the control group and model group. After 14-day treatment of EA, the inflammatory cell count, scar tissues area and muscle fiber cross sectional area of multifidus muscle were observed with HE and Masson staining method. The activity of CK and serum content of IL-17 were test with enzyme-linked immunosorbent assay(ELISA) method; the expression of IL-17 in multifidus muscle was measured with immunohistochcmical method. Results After intervention, the inflammatory cell count and scar tissues area in the model group, Weizhong group and Shenshu group were higher than those in the control group(all P<0.01),but the muscle fiber cross sectional area was significantly reduced(all P<0.01); the inflammatory cell count and scar tissues area in the Weizhong group and Shenshu group were lower than those in the model group(all P<0.01), and the muscle fiber cross sectional area was significantly increased(P<0.01, P<0.05). After intervention, the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the model group, Weizhong group and Shenshu group were higher than those in the control group(all P<0.01); the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the Weizhong group and Shenshu group were lower than those in the model group(P<0.01, P<0.05); compared with the Shenshu group, the down-regulation of IL-17 was more obvisous in the Weizhong group(P<0.01). Conclusion EA at "Weizhong"(BL 40) can down-regulate the overexpression of serum CK and IL-17, alleviate inflammation reaction and improve the repair of multifidus muscle.
Objective To observe effects of electroacupuncture(EA) at "Weizhong"(BL 40) on morphology and expression of creatine kinase(CK) and interleukin-17(IL-17) in rats with bupivacaine-induced multifidus muscle injury.Methods A total of 32 male SD rats were randomly divided into a control group, a model group, a Weizhong group and a Shenshu group, 8 rats in each one. The rats in the model group, Weizhong group and Shenshu group were treated with intramuscular injection of 0.5% bupivacaine to establish the model of multifidus muscle injury; the rats in the control group were injected with 0.9% sodium chloride solution. The rats in the Weizhong group and Shenshu group were treated with EA(2 Hz/10 Hz in frequency, 1~2 m A in intensity) at "Weizhong"(BL 40) and "Shenshu"(BL 23), 20 min per treatment. No treatment was given in the control group and model group. After 14-day treatment of EA, the inflammatory cell count, scar tissues area and muscle fiber cross sectional area of multifidus muscle were observed with HE and Masson staining method. The activity of CK and serum content of IL-17 were test with enzyme-linked immunosorbent assay(ELISA) method; the expression of IL-17 in multifidus muscle was measured with immunohistochcmical method. Results After intervention, the inflammatory cell count and scar tissues area in the model group, Weizhong group and Shenshu group were higher than those in the control group(all P<0.01),but the muscle fiber cross sectional area was significantly reduced(all P<0.01); the inflammatory cell count and scar tissues area in the Weizhong group and Shenshu group were lower than those in the model group(all P<0.01), and the muscle fiber cross sectional area was significantly increased(P<0.01, P<0.05). After intervention, the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the model group, Weizhong group and Shenshu group were higher than those in the control group(all P<0.01); the expression of IL-17 in multifidus muscle, serum content of IL-7 and activity of CK in the Weizhong group and Shenshu group were lower than those in the model group(P<0.01, P<0.05); compared with the Shenshu group, the down-regulation of IL-17 was more obvisous in the Weizhong group(P<0.01). Conclusion EA at "Weizhong"(BL 40) can down-regulate the overexpression of serum CK and IL-17, alleviate inflammation reaction and improve the repair of multifidus muscle.
引文
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[11]张泽高,肖琳,詹欣宇,等.维药小茴香抗肝纤维化作用及对TGF-β/smad信号转导通路的影响[J].中国肝脏病杂志(电子版),2014,6(1):32-37.
[12]管海涛,赵金丽,汤瑞,等.白细胞介素17促进肝纤维化发生发展的机制[J].临床肝胆病杂志,2013,29(5):370-374.
[2]陈玉佩,刘通,邹德辉,等.局部注射布比卡因建立大鼠骨骼肌损伤模型的组织形态学评价[J].中国组织工程研究,2016,20(18):2615-2621.
[3]刘通,于佳妮,邹德辉,等.电针血清对多裂肌卫星细胞增殖及Pax-7、成肌分化抗原、磷酸化蛋白激酶B表达的影响[J].针刺研究,2016,41(5):402-409.
[4]陈欢,张莉,崔强,等.电针阿是穴对骨骼肌损伤大鼠肌肉再生及碱性成纤维细胞生长因子表达的影响[J].中国康复理论与实践,2013,19(4):334-340.
[5]彭博,张莉,陈欢,等.电针委中穴对腰肌损伤家兔CK、VEGF和MVD的影响[J].中国康复,2014,29(1):3-7.
[6]马忆南,杨华元,冯麟,等.针刺诱导胶原信号变化对骨骼肌损伤的修复作用[J].中国组织工程研究,2012,16(42):7888-7892.
[7]Beringer A,Noack M,Miossec P.IL-17 in chronic inflammation:from discovery to targeting[J].Trends Mol Med,2016,22(3):230-241.
[8]赵永军,卢健.骨骼肌自噬的调控机制与Sarcopenia的关联及运动的影响[J].中国运动医学杂志,2013,32(10):929-937.
[9]苏兆亮,王映梅,刘嫣方,等.IL-17对小鼠心肌成纤维细胞Ⅰ/Ⅲ胶原表达的作用研究[J].细胞与分子免疫学杂志,2012,28(9):897-898.
[10]夏鑫钰,耿莹,叶宇,等.白细胞介素-17基因多态性与侵袭性牙周炎的相关性研究[J].口腔医学,2015(6):417-420.
[11]张泽高,肖琳,詹欣宇,等.维药小茴香抗肝纤维化作用及对TGF-β/smad信号转导通路的影响[J].中国肝脏病杂志(电子版),2014,6(1):32-37.
[12]管海涛,赵金丽,汤瑞,等.白细胞介素17促进肝纤维化发生发展的机制[J].临床肝胆病杂志,2013,29(5):370-374.