摘要
目的:观察电针内关穴后缺血再灌注损伤大鼠心肌能量代谢及组织形态学变化,探讨针刺对缺血再灌注损伤大鼠的心肌保护作用。方法:实验于2003-10/2004-02在湖南中医药大学、湘雅医学院完成。实验分组:将50只大鼠完全随机分为5组,每组10只。即假手术组、模型组、电针内关组、电针神门组、电针合谷组。实验干预:①假手术组:开胸、穿线、不结扎,观察实验过程120 min。②模型组:开胸、穿线20 min、结扎40 min,松扎再灌注60 min。③电针内关组:开胸、穿线,电针内关20 min后,结扎40 min,再灌注60 min,于再灌注开始时再次电针内关20 min。④电针神门组:处理同电针内关组,电针穴位为神门。⑤电针合谷组:处理同电针内关组,电针穴位为合谷。穴位定位与电针参数:穴位定位:内关穴位于大鼠腕横纹正中上5 mm处,神门穴位于大鼠腕横纹尺侧处,合谷穴位于大鼠第一、二掌骨之间中点处,针刺深度约5 mm,造模成功后分别针刺大鼠双侧上述3穴,直刺穿皮达筋间,捻转1 min后,接电子针疗仪,疏密波刺激(疏波30 Hz,密波100 Hz),以前肢出现轻微颤动为准,持续时间为20 min。实验结束后颈动脉取血5 mL,摘取心脏检测相关指标。实验评估:①应用高效液相色谱法测定三磷酸腺苷、二磷酸腺苷、一磷酸腺苷和腺苷含量。②光学显微镜观察缺血再灌注损伤心肌病理形态学的变化。结果:50只大鼠均进入结果分析。①血清三磷酸腺苷含量:假手术组最高,模型组最低。电针内关组、电针神门组高于模型组[(0.33±0.14),(0.074±0.021),(0.045±0.015)mg/L,P<0.01,P<0.05],电针合谷组与模型组比较,差异无显著性(P>0.05)。②血清中二磷酸腺苷、一磷酸腺苷、腺苷含量:假手术组最低,模型组最高,电针内关组低于模型组[(0.110±0.065),(0.20±0.10)mg/L;(0.160±0.055),(0.28±0.11)mg/L;(0.045±0.015),(0.11±0.039)mg/L;P<0.05,P<0.01],电针神门组、电针合谷组与电针内关组比较,二磷酸腺苷含量差异不显著(P>0.05),一磷酸腺苷和腺苷差异非常显著(P<0.01)。③假手术组肌纤维整齐,肌丝及肌小节结构清晰可见,线粒体丰富,正常,未见水肿及空泡变;其次为电针内关组,可见肌纤维正常走向,少量线粒体空泡变,肌丝无明显坏死、溶解。模型组的心肌修复情况最差,表现为心肌肌丝溶解、坏死,肌丝走向紊乱,细胞核浓缩,染色质靠边,线粒体水肿,肌浆网扩张。电针神门组,可见部分标本心肌水肿,间质出血,其中1只可见心肌细胞部分坏死,炎性细胞浸润。电针合谷组,部分标本可见心肌水肿,间质出血,其中2只可见心肌坏死,炎性细胞浸润。结论:电针内关穴能明显改善缺血再灌注损伤大鼠心肌的能量代谢,促进心肌组织的修复。
AIM： To observe the changes of myocardial energy metabolism and morphology in myocardial ischemia/reperfusion lesion rats by electroacupuncture （EA） on Neiguan （PC 6）, and explore the myocardial protection in ischemia/reperfusion injury by EA. METHODS： The experiment was performed at the Hunan College of Traditional Chinese Medicine and Xiangya Medical College from October 2003 to February 2004. Fifty rats were divided into 5 groups at random, with 10 rats in each.① Sham operated group： Rats underwent thoracotomy and suture but without ligation were observed for 120 minutes.② Model group： Rats underwent thoracotomy, suture.for 20 minutes, ligated for 40 minutes, and reperfusion for 60 minutes. ③EA groups： Rats were treated as above, followed by a 20-minute EA on Neiguan （PC 6） at 5 mm above transverse crease of the wrist, Shenmen （HT 7） at the ulnar end of the transverse crease of the wrist, and Hegu （LI 4） at the middle point between the first and the second metacarpal bone, respectively. EA parameters： The needling at the depth of 5 mm was conducted bilaterally on tendon through acupoints, followed by 1-minute twisting and 20-minute stimulation of disperse-dense wave （disperse wave 30 Hz, dense wave 100 Hz） using EA apparatus. The mild shivering of the rat forelimb was the indicator for the curative effect of EA. After the experiment was over, 5 mL carotid artery blood was sampled to detect the content of adenosine triphosphate （ATP）, adenosine diphosphate （ADP）, adenosine monophosphate （AMP） and adenosine （Ado） by high performance liquid chromatography method, while the change of morphology in myocardial ischemia/reperfusion lesion rats was observed by microscope.
RESULTS： All of the 50 rats were involved in the result analysis. ①The content of serum ATP was the highest in the sham operated group, and the lowest in the model group. Compared with the model group, the contents of EA Neiguan （PC 6） group and EA Shenmen （HT 7） group were increased [（0.33±0.14）, （0.074±0.021）, （0.045±0.015） mg/L, P 〈 0.01, P 〈 0.05], there was no significant difference between EA Hegu （LI 4） group and normal group （P 〉 0.05）. ②The contents of serum ADP, AMP, and Ado were the lowest in sham operated group, and the highest in model group. Compare with the model group, there was lower contents in EA Neiguan （PC 6） group [（0.110±0.065）, （0.20±0.10） mg/L; （0.160±0.055）, （0.28±0.11） mg/L; （0.045±0.015）, （0.11±0.039） mg/L; P 〈 0.05, P 〈 0.01]. There was no significant difference in the content of ADP （P 〉 0.05） but extremely significant difference in AMP and Ado among three EA groups （P 〈 0.01）.③The sham operated group： Muscle fiber was regular, myoneme and sarcomere were clear, bioblast was abundant and normal, no dropsy and vacuolus was found. EA Neiguan （PC 6） group： Muscle fiber was normal, small amount of bioblast were vacuolus, myoneme had no obvious necrosis or dissolved. Model group： Cardiac muscle repairing was the worse, myoneme was dissolved, necrosis and disordered, accompanying with caryon condensed, caryotin arranged on margin, bioblast dropsy, and sarcoplasmic reticulum expansion. EA Shenmen （HT 7） group： Parts of the cardiac muscle were dropsy, interstitial substance bled. One sample presented cadiocyte necrosis and inflammatory cell infiltration. EA Hegu （LI 4） group： The microscope result was identical with that of EA Shenmen （HT 7） group, and two samples presented cadiocyte necrosis and inflammatory cell infiltration. CONCLUSION： EA on Neiguan （PC 6） can obviously improve the myocardial energy metabolism and promote the repainng of myocardial tissue in ischemia/reperfusion lesion rats.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第47期9443-9447,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
湖南省科技厅重点资助项目(02SSY2005)~~
