胞内游离钙离子浓度增加抑制大鼠离体比目鱼肌肌梭传入放电（英文）
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摘要
目的探讨大鼠离体比目鱼肌胞内游离钙离子浓度增加对肌梭传入放电的影响。方法 12只大鼠随机分为3组,2.0ion组、5.0ion组、二甲基亚砜(dimethyl sulfoxide,DMSO)组。分离大鼠比目鱼肌单一肌梭,2.0ion组、5.0ion组分别给予2和5μmol/L钙离子导入剂伊吾诺霉素(ionomycin)孵育,采用激光扫描共聚焦显微镜观察肌梭内游离钙离子浓度的改变。采用空气隔绝法记录大鼠比目鱼肌单一肌梭传入放电活动,观察3组肌梭传入放电活动的变化。结果伊吾诺霉素(2和5μmol/L)能显著增加大鼠比目鱼肌梭內肌静息钙的含量,且呈浓度依赖性(P<0.01)。DMSO对大鼠比目鱼肌单一肌梭传入放电活动无明显影响。2和5μmol/L伊吾诺霉素可明显抑制比目鱼肌单一肌梭传入放电(P<0.05)。5μmol/L伊吾诺霉素可延长单一肌梭传入放电周期间隔(P<0.05),在此基础上给予50mg/L氯化琥珀酰胆碱(Sch)诱发肌梭传入发电,可见肌梭传入放电无明显改变。结论大鼠比目鱼肌梭內肌游离钙离子浓度的增加可抑制肌梭传入放电。
Objective To investigate the effects of the increase of intracellular free calcium on afferent discharges of isolated soleus muscle spindle in rats.Methods Twelve rats were randomly divided into three groups:the 2.0 ion group,the 5.0 ion group,and the dimethyl sulfoxide(DMSO)group.The isolated soleus muscle spindle of rats in 3 groups were separated.After incubation with 2μmol/L ionomycin in 2.0 ion group and 5μmol/L ionomycin in 5.0 ion group respectively,the changes of free calcium concentration in soleus muscle spindle were observed by laser scanning confocal microscopy.The air-gap technique was used to observe the afferent discharge activities of rats' isolated soleus muscle spindle,and the changes of the afferent discharge activities in 3 groups were observed after treated with DMSO,2μmol/L,or 5μmol/L Ionomycin respectively.Results Ionomycin(2 and 5μmol/L)could significantly increase the level of resting calcium in intrafusal muscle of rats' soleus muscle,and the effect was concentration dependent(P<0.01).Ionomycin(2 and 5μmol/L)could significantly reduce the afferent discharges of single muscle spindle of soleus muscle(P<0.05).5μmol/L ionomycin could prolong the periodic interval of spontaneous discharge of single muscle spindle(P <0.05),and there was no significant changes on evoked discharges of muscle spindle after the adding of Succinylcholine chloride(Sch)with concentration of 50 mg/L.Conclusion Increased free calcium concentration in rat soleus muscle spindle can inhibit afferent discharges of muscle spindle.
Objective To investigate the effects of the increase of intracellular free calcium on afferent discharges of isolated soleus muscle spindle in rats.Methods Twelve rats were randomly divided into three groups:the 2.0 ion group,the 5.0 ion group,and the dimethyl sulfoxide(DMSO)group.The isolated soleus muscle spindle of rats in 3 groups were separated.After incubation with 2μmol/L ionomycin in 2.0 ion group and 5μmol/L ionomycin in 5.0 ion group respectively,the changes of free calcium concentration in soleus muscle spindle were observed by laser scanning confocal microscopy.The air-gap technique was used to observe the afferent discharge activities of rats' isolated soleus muscle spindle,and the changes of the afferent discharge activities in 3 groups were observed after treated with DMSO,2μmol/L,or 5μmol/L Ionomycin respectively.Results Ionomycin(2 and 5μmol/L)could significantly increase the level of resting calcium in intrafusal muscle of rats' soleus muscle,and the effect was concentration dependent(P<0.01).Ionomycin(2 and 5μmol/L)could significantly reduce the afferent discharges of single muscle spindle of soleus muscle(P<0.05).5μmol/L ionomycin could prolong the periodic interval of spontaneous discharge of single muscle spindle(P <0.05),and there was no significant changes on evoked discharges of muscle spindle after the adding of Succinylcholine chloride(Sch)with concentration of 50 mg/L.Conclusion Increased free calcium concentration in rat soleus muscle spindle can inhibit afferent discharges of muscle spindle.
引文
[1]Smith SM,Heer MA,Shackelford LC,et al.Benefits for bone from resistance exercise and nutrition in long-duration spaceflight:Evidence from biochemistry and densitometry[J].Journal of Bone and Mineral Research,2012,27(9):1896-1906．
[2]Chen S,Wang C,Chen X,et al.Study on Changes of Human Performance Capabilities in Long-duration Spaceflight[J].Space Medicine&Medical Engineering,2015,28(1):1-10.(in Chinese）
[3]Zhao X,Zhang W,Zhou X,et al.Changes in muscle spindle afferent discharge activities in rat soleus following hindlimb immobilization[J].J South Med Univ,2015,35(2):252-255.(in Chinese）
[4]Zhao X,Gao Y,Fan X.Effects of muscle vibration on the electrophysiological activity of isolated muscle spindles in rat following hind limb immobilization[J].J Biol,2015,32(2):17-19．
[5]Zhu Y,Fan X,Li X,et al.Effect of hindlimb unloading on resting intracellular calcium in intrafusal fibers and ramp-and-hold stretches evoked responsiveness of soleus muscle spindles in conscious rats[J].Neurosci Lett,2008,442(3):69-73．
[6]Ito F,Komatsu Y.Transduction and encoding mechanisms in muscle spindle[J].Nagoya J Med Sci,980,42(3):37-48．
[7]Liu C,Herman T.Characterization of ionomycin as a calcium ionophore[J].J Biol Chem,1978,253(17):5892-5894．
[8]Wheeler J,Veiro J,Cullis PR.Ionophore-mediated loading of Ca2+into large unilamellar vesicles in response to transmembrane pH gradients[J].Mol Membr Boil,1994,11(3):151-157．
[9]Sun GB,Sun H,Meng XB,et al.Aconitine induced Ca2+overload causes arrhythmia and triggers apoptosis through p38 MAPK signaling pathway in rats[J].Toxicol Appl Pharmacol,2014,279(1):8-22．
[10]Sun Z,Han J Zhao W,Zhang Y,et al.TRPV1activation exacerbates hypoxia/reoxygenation induced apoptosis in H9C2cells via calcium overload and mitochondrial dysfunction[J].Int J Mol Sci,2014,15(10):18362-18380．
[11]Chang G,Zhang D,Liu J,et al.Exenatide protects against hypoxia/reoxygenation induced apoptosis by improving mitochondrial function in H9C2cells[J].Exp Biol Med,2014,239(4):414-422．
[12]Wang ZM,Qian F,Lu G,et al.Regulation of nAChRγ-Promoter activity by electrical stimulation-induced calcium oscillation in mice skeletal muscle cell lines[J].Chin JBiochem Mol Biol,2003,9(3):283-289.(in Chinese）
[13]Brini M,Carafoli E.The plasma membrane Ca2+ATPase and the plasma membrane sodium calcium exchanger cooperate in the regulation of cell calcium[J].Cold Spring Harbor Perspectives in Biology,2011,3(2):a004168．
[14]Jiang T,Grant RL,Acosta D.A digitized fluorescence imaging study of intracellular free calcium,mitochondrial integrity and cytotoxicity in rat renal cells exposed to ionomycin,a calcium ionophore[J].Toxicology,1993,85(1):41-65．
[15]Zhao XH,Fan XL,Song XA.Influence of 14-day hind limb unloading on isolated muscle spindle activity in rats[J].J Muscle Res Cell Motil,2010,31(3):155-161．
[16]Zhu YJ,Wu SD,Fan XL.Effects of hindlimb unloading on diameter of intrafusal fibers and capsule at equatorial region of rat soleus muscle spindle[J].Space Medicine&Medical Engineering,2006,19(4):273-276．
[17]Li J,Gu C.Study on the relationship of Ca2+transport system in skeletal muscle and muscle function[J].J Shenyang Inst Phys Ed,2004,23(3):301-303.(in Chinese）
[18]Tate CA.Enhanced calcium uptake of cardiac Sar coplasmic reticulum in exercise-trained old rats[J].Am J Physiol,1990,258(2):431-435．
[19]Murphy RM,Dutka TL,Horvath D,et al.Ca2+-dependent proteolysis of junctophilin-1and junctophilin-2in skeletal and cardiac muscle[J].J Physiol,2013,591(3):719-729．
[20]Gerard K,Hipkiss A,Schneider D.Degradation of intracellular protein in muscle.Lysosomal response to modified proteins and chloroquine[J].J Biol Chem,1988,263(35):18886-18890．
[21]Lukyanenko V,Muriel JM,Bloch RJ.Coupling of excitation to Ca2+release is modulated by dysferlin[J].J Physiol,2017,595(15):5191-5207．
[22]Naparus A,Hofer S,Cahoon N,et al.Assessment of mitochondrial calcium(MCa2+)overload and function in ex vivo human skeletal muscle undergoing ischemia and reperfusion(I/R)[J].Plastic Reconstruct Surg,2011,127:59．
[23]Cully TR,Launikonis BS.Leaky ryanodine receptors delay the activation of store overload-induced Ca2+release,a mechanism underlying malignant hyperthermia like events in dystrophic muscle[J].Am J Physiol Cell Physiol,2016,310(8):C73-80．
[24]Logan CV,Szabadkai G,Sharpe JA,et al.Loss-of-function mutations in MICUl cause a brain and muscle disorder linked to primary alterations in mitochondrial calcium signaling[J].Nat Genet,2014,46(2):88-93．
[25]Fu ML,Shi WH,Li ZL,et al.Activation of mPTP-dependent mitochondrial apoptosis pathway by a novel Pan HDAC inhibitor resminostat in hepatocellular carcinoma cells[J].Biochem Biophys Res Commun,2016,477(4):27-33．
[26]Macintosh BR,Holash IU,Renaud JM.Skeletal muscle fatigue regulation of excitation-contraction coupling to avoid metabolic catastrophe[J].J Cell Sci,2012,125(9):5-14.
[2]Chen S,Wang C,Chen X,et al.Study on Changes of Human Performance Capabilities in Long-duration Spaceflight[J].Space Medicine&Medical Engineering,2015,28(1):1-10.(in Chinese）
[3]Zhao X,Zhang W,Zhou X,et al.Changes in muscle spindle afferent discharge activities in rat soleus following hindlimb immobilization[J].J South Med Univ,2015,35(2):252-255.(in Chinese）
[4]Zhao X,Gao Y,Fan X.Effects of muscle vibration on the electrophysiological activity of isolated muscle spindles in rat following hind limb immobilization[J].J Biol,2015,32(2):17-19．
[5]Zhu Y,Fan X,Li X,et al.Effect of hindlimb unloading on resting intracellular calcium in intrafusal fibers and ramp-and-hold stretches evoked responsiveness of soleus muscle spindles in conscious rats[J].Neurosci Lett,2008,442(3):69-73．
[6]Ito F,Komatsu Y.Transduction and encoding mechanisms in muscle spindle[J].Nagoya J Med Sci,980,42(3):37-48．
[7]Liu C,Herman T.Characterization of ionomycin as a calcium ionophore[J].J Biol Chem,1978,253(17):5892-5894．
[8]Wheeler J,Veiro J,Cullis PR.Ionophore-mediated loading of Ca2+into large unilamellar vesicles in response to transmembrane pH gradients[J].Mol Membr Boil,1994,11(3):151-157．
[9]Sun GB,Sun H,Meng XB,et al.Aconitine induced Ca2+overload causes arrhythmia and triggers apoptosis through p38 MAPK signaling pathway in rats[J].Toxicol Appl Pharmacol,2014,279(1):8-22．
[10]Sun Z,Han J Zhao W,Zhang Y,et al.TRPV1activation exacerbates hypoxia/reoxygenation induced apoptosis in H9C2cells via calcium overload and mitochondrial dysfunction[J].Int J Mol Sci,2014,15(10):18362-18380．
[11]Chang G,Zhang D,Liu J,et al.Exenatide protects against hypoxia/reoxygenation induced apoptosis by improving mitochondrial function in H9C2cells[J].Exp Biol Med,2014,239(4):414-422．
[12]Wang ZM,Qian F,Lu G,et al.Regulation of nAChRγ-Promoter activity by electrical stimulation-induced calcium oscillation in mice skeletal muscle cell lines[J].Chin JBiochem Mol Biol,2003,9(3):283-289.(in Chinese）
[13]Brini M,Carafoli E.The plasma membrane Ca2+ATPase and the plasma membrane sodium calcium exchanger cooperate in the regulation of cell calcium[J].Cold Spring Harbor Perspectives in Biology,2011,3(2):a004168．
[14]Jiang T,Grant RL,Acosta D.A digitized fluorescence imaging study of intracellular free calcium,mitochondrial integrity and cytotoxicity in rat renal cells exposed to ionomycin,a calcium ionophore[J].Toxicology,1993,85(1):41-65．
[15]Zhao XH,Fan XL,Song XA.Influence of 14-day hind limb unloading on isolated muscle spindle activity in rats[J].J Muscle Res Cell Motil,2010,31(3):155-161．
[16]Zhu YJ,Wu SD,Fan XL.Effects of hindlimb unloading on diameter of intrafusal fibers and capsule at equatorial region of rat soleus muscle spindle[J].Space Medicine&Medical Engineering,2006,19(4):273-276．
[17]Li J,Gu C.Study on the relationship of Ca2+transport system in skeletal muscle and muscle function[J].J Shenyang Inst Phys Ed,2004,23(3):301-303.(in Chinese）
[18]Tate CA.Enhanced calcium uptake of cardiac Sar coplasmic reticulum in exercise-trained old rats[J].Am J Physiol,1990,258(2):431-435．
[19]Murphy RM,Dutka TL,Horvath D,et al.Ca2+-dependent proteolysis of junctophilin-1and junctophilin-2in skeletal and cardiac muscle[J].J Physiol,2013,591(3):719-729．
[20]Gerard K,Hipkiss A,Schneider D.Degradation of intracellular protein in muscle.Lysosomal response to modified proteins and chloroquine[J].J Biol Chem,1988,263(35):18886-18890．
[21]Lukyanenko V,Muriel JM,Bloch RJ.Coupling of excitation to Ca2+release is modulated by dysferlin[J].J Physiol,2017,595(15):5191-5207．
[22]Naparus A,Hofer S,Cahoon N,et al.Assessment of mitochondrial calcium(MCa2+)overload and function in ex vivo human skeletal muscle undergoing ischemia and reperfusion(I/R)[J].Plastic Reconstruct Surg,2011,127:59．
[23]Cully TR,Launikonis BS.Leaky ryanodine receptors delay the activation of store overload-induced Ca2+release,a mechanism underlying malignant hyperthermia like events in dystrophic muscle[J].Am J Physiol Cell Physiol,2016,310(8):C73-80．
[24]Logan CV,Szabadkai G,Sharpe JA,et al.Loss-of-function mutations in MICUl cause a brain and muscle disorder linked to primary alterations in mitochondrial calcium signaling[J].Nat Genet,2014,46(2):88-93．
[25]Fu ML,Shi WH,Li ZL,et al.Activation of mPTP-dependent mitochondrial apoptosis pathway by a novel Pan HDAC inhibitor resminostat in hepatocellular carcinoma cells[J].Biochem Biophys Res Commun,2016,477(4):27-33．
[26]Macintosh BR,Holash IU,Renaud JM.Skeletal muscle fatigue regulation of excitation-contraction coupling to avoid metabolic catastrophe[J].J Cell Sci,2012,125(9):5-14.