Johnson B D, Sieck G C
Department of Anesthesiology, Mayo Clinic and Foundation, Rochester, Minnesota 55905.
J Appl Physiol (1985). 1993 Dec;75(6):2689-95. doi: 10.1152/jappl.1993.75.6.2689.
In this study, we examined whether exhaustive activation reduces succinate dehydrogenase (SDH) activity in diaphragm muscle fibers. In adult male rats (approximately 300 g), the costal diaphragm was excised and positioned in a chamber perfused with mammalian Ringer solution kept at 26 degrees C and oxygenated with 95% O2-5% CO2. The muscle was stimulated directly at 10 or 75 Hz in trains of 500 ms duration (1/s) for 8 min. An adjacent unstimulated segment of muscle served as control. The two muscle segments were frozen, and serial sections were stained for myofibrillar adenosinetriphosphatase activity after alkaline and acid preincubation to classify type I, IIa, and IIb fibers. The extent of glycogen utilization was also examined histochemically to confirm exhaustive activation of muscle fibers. SDH activity was quantified using a microdensitometric procedure implemented on an image-processing system. Exhaustive activation at both 10 and 75 Hz caused a significant decrease in SDH activity of all fiber types, with the decrease after 10-Hz stimulation being greater than that after 75-Hz stimulation. At both stimulation frequencies, type IIb fibers demonstrated the greatest decrease in SDH activity (36% after 10-Hz and 27% after 75-Hz stimulation), whereas type I and IIa fibers both displayed reductions of approximately 27 and approximately 19% after 10- and 75-Hz stimulation, respectively. The greater reduction of SDH activity in type IIb fibers indicates an inverse relationship between activation-induced reductions in SDH activity and fiber oxidative capacity.(ABSTRACT TRUNCATED AT 250 WORDS)
在本研究中,我们检测了彻底激活是否会降低膈肌肌纤维中的琥珀酸脱氢酶(SDH)活性。选用成年雄性大鼠(约300克),切除其肋膈膜并置于一个用哺乳动物林格氏液灌注的腔室中,该溶液保持在26℃,并用95% O₂ - 5% CO₂ 进行氧合。以10或75赫兹的频率、500毫秒的时长(1次/秒)对肌肉进行直接刺激,持续8分钟。相邻的未受刺激的肌肉段作为对照。将这两个肌肉段冷冻,在碱性和酸性预孵育后对连续切片进行肌原纤维三磷酸腺苷酶活性染色,以区分I型、IIa型和IIb型纤维。还通过组织化学方法检测糖原利用程度,以确认肌纤维的彻底激活。使用图像处理系统上实施的微量光密度测定程序对SDH活性进行定量。10赫兹和75赫兹的彻底激活均导致所有纤维类型的SDH活性显著降低,10赫兹刺激后的降低幅度大于75赫兹刺激后的降低幅度。在两种刺激频率下,IIb型纤维的SDH活性降低幅度最大(10赫兹刺激后降低36%,75赫兹刺激后降低27%),而I型和IIa型纤维在10赫兹和75赫兹刺激后分别降低约27%和约19%。IIb型纤维中SDH活性的更大降低表明激活诱导的SDH活性降低与纤维氧化能力之间呈负相关。(摘要截断于250字)