摘要
目的:应用钙离子敏感微电极技术,分析肌肉损伤与细胞内Ca2+浓度的关系及针刺对骨骼肌细胞内Ca2+活度的影响。方法:实验于2000-09/2001-05在河北医科大学中医学院实验针灸实验室完成。选用同种健康青紫蓝家兔70只,随机数字表法进行分组,分1次针刺处理组和4次针刺处理组,1次针刺处理组30只,分为3个亚组,即针刺组10只,对照组11只,空白组9只。4次针刺处理组40只,分为3个亚组,即针刺组15只,对照组13只,空白组12只。针刺组和对照组通过固定在兔左后肢的表面电极使股四头肌反复被动强直收缩运动,空白组不予电刺激。制造超负荷运动状态的疲劳模型。同时针刺组选取家兔股外侧肌局部(相当于伏兔和梁丘穴位置)于第4天刺激运动结束后即刻进行第1次针刺,4次针刺处理组还将于其后的第1天上,下午,第2天上午各针刺1次,对照组和空白组除不针刺外,亦以同样方式固定于实验架上。标本经处理后通过自制并测定的钙离子敏感微电极进行细胞膜电位的测定,同时测定细胞内外Ca2+有效浓度的电位差,计算细胞内钙离子活度。组间比较采用t检验。结果:70只实验动物最终进入结果分析51只。包括1次针刺处理组中针刺组7只,对照组8只,空白组7只。4次针刺处理中针刺组12只,对照组7只,空白组10只。①家兔过度运动后,针刺组及对照组与空白组相比,骨骼肌膜电位的绝对值均显著降低(64.3±1.38),(64.1±1.13),(80.7±3.09)-mV,P<0.01,细胞内钙离子活度显著升高(1.39±0.029),(1.38±0.031),(0.14±0.018)mmol/L,P<0.01。针刺组与对照组相比,膜电位或细胞内钙离子活度的对比无显著差别(P>0.05)。②运动结束后46h,针刺组膜电位绝对值与对照组比较,显著升高(79.0±2.59),(67.6±1.58)-mV,P<0.01,针刺组膜电位的显著升高,与空白组膜电位基本接近(81.3±2.69)-mV,P>0.05。细胞内钙离子活度值针刺组均细胞内钙离子活度值比对照组显著降低,但仍高于空白组,差别显著(P<0.01)。结论:超负荷运动导致肌细胞超微结构的改变;细胞内钙离子活度升高;骨骼肌膜电位的绝对值均显著降低,可能是肌肉的损伤所致。针刺能够改善细胞内钙离子的代谢,从而促进超微结构损伤的修复。
AIM： To analyze the association between muscle injury and intraeellular calcium ion （Ca^2＋） concentration, and the influence of acupuncture on Ca^2＋ activity coefficient in the skeletal muscle cells by using the technique of Ca^2＋ sensitive microeleetrode. METHODS： The experiment was carried out in the acupuncture laboratory of School of Traditional Chinese Medicine, Hebei Medical University between September 2000 and May 2001. Seventy healthy rabbits were randomly divided into once acupuncture group （n=30） and four-time acupuncture group （n=40）. The once acupuncture group was subdivided into 3 groups： acupuncture group （n=10）, eontrol group （n=11） and blank group （n =9）. The 4-time acupuncture group was subdivided into 3 groups： acupuncture group （n=15）, control group （n=13） and blank group （n=12）. The quadriceps of the rabhits in the acupuncture group and the control group repetitively passively made rigid contractive movements under the stimulation of the surface electrode, no electrosimulation was given in the blank group. Fatigue models under overloaded exercise were made. The rabbits in the acupuncture .groups were treated with acupuncture for once and 4 times respectively. The cell membrane potential was detected with the Ca^2＋ sensitive microelectrode, the potential difference of effective Ca^2＋ concentration in and out of cells was determined, the intracellular Ca^2＋ activity coefficient was calculated. The t test was used for the intergroup comparison. RESULTS： Of the 70 rabbits, 51 were involved in the analysis of results, including in the once acupuncture group： acupuncture group （n=7）, control group （n=8） and hlank group （n=7）; in the 4-time acupuncture group： acupuncture group （n=12）, control group （n=7） and blank group （n=10）. ① After overloaded exercise, the absolute values of skeletal muscle membrane poteutial in the acupuncture group and control group was significantly decreased as compared with that in the blank group [（64.3±1.38）, （64.1±1.13）, （80.7±3.09） -mV, P 〈 0.01], and the intracellular Ca^2＋ activity coefficients was significantly increased [（1.39±0.029）, （1.38±0.031 ）, （0.14±0.018） mmol/L, P 〈 0.01]. There were insignificant differences between the acupuncture group and control group （P 〉 0.05）. ② At 46 hours after exercises, the absolute value of membrane potential was significantly increased in the acupuncture group as compared with that in the control group [（79.0±2.59）, （67.6±1.58） -mV, P 〈 0.01], the membrane potential in the acupuncture group was significantly increased, and close to that in the blank group [（81.3±2.69）-mV, P 〉 0.05]. The intracellular Ca^2＋ activity coefficient in the acupuncture group was significantly decreased as compared with that in the control group, but still higher than that in the blank group （P 〈 0.01）. CONCLUSION： Overloaded exercise leads to the change in uhrastrueture of muscle cells, increase of Ca^2＋ activity coefficient, remarkable decrease of the absolute value of skeletal muscle membrane potential, and it is presumed that it is caused by muscle injury. Acupuncture can ameliorate the Ca^2＋ metabolism in cells and speed up the recovery of the ultrastructural injury.
出处
《中国临床康复》
CSCD
北大核心
2005年第28期211-213,共3页
Chinese Journal of Clinical Rehabilitation
