Objective The gene expression of skeletal muscle under ischemic condition by direct gene injectionwas observed in order to find a new gene delivery method to treat chronic arterial occlusion disease. Methods Weestabli...Objective The gene expression of skeletal muscle under ischemic condition by direct gene injectionwas observed in order to find a new gene delivery method to treat chronic arterial occlusion disease. Methods Weestablished the rabbit hindlimb ischemic model and used plasmid PSV-β-gal as a reporter gene. We transferedgene intramuscularly and detected the activity of β-galactosidase by histochemistry method. Results Weobserved that the gene expression of skeletal muscle under ischemic condition was higher than normalmuscle. Conclusion The result demonstrated that the direct gene injection was suitable for the chronic peripheralarterial occlusion disease, and might be a novel gene delivery method for this disease.展开更多
Acute interruption of arterial blood flow to the extremities is often associated with significant morbidity and mortality. Broad spectrum mitogenic and non mitogenic activities of FGFs inspired us to study its protect...Acute interruption of arterial blood flow to the extremities is often associated with significant morbidity and mortality. Broad spectrum mitogenic and non mitogenic activities of FGFs inspired us to study its protecting effects on tissue injuries in ischemia reperfusion condition. We found that systemic administration of aFGF after reperfusion onset prevented severe skeletal muscle injuries. In rats treated with aKGF, the tissue edema was reduced significantly, the tissue viability was increased, and the muscle fibers contained more succinate dehydrogenase (SDH) and adenosine triphosphatasc (ATPase). The pathological results supported the concept of improved prevention with aFGF treatment. The possible tissue protection by aFGF may come from its ability to regulate the concentration of evtra- and intracellular calcium ion. Besides, it may moderate other Ca2+ dependent enzyme conversion processes. Also, it may take part in the vascular tone regulation under ischemia and reperfusion conditions. These results suggest further study of tissue ischemia prevention with FGF and its possible mechanisms in the future.展开更多
背景:线粒体活性氧爆发已被证明在骨骼肌缺血再灌注中起着关键作用。3-硝基-N-甲基水杨酰胺(3-nitro-N-methyl salicylamide,3-NNMS)可以有效降低电子传递速度,对肢体缺血再灌注损伤具有潜在的保护作用,但目前尚无明确的研究和临床应用...背景:线粒体活性氧爆发已被证明在骨骼肌缺血再灌注中起着关键作用。3-硝基-N-甲基水杨酰胺(3-nitro-N-methyl salicylamide,3-NNMS)可以有效降低电子传递速度,对肢体缺血再灌注损伤具有潜在的保护作用,但目前尚无明确的研究和临床应用。目的:探讨3-NNMS对肢体缺血再灌注损伤大鼠骨骼肌的保护作用及机制。方法:40只健康8周龄SD大鼠随机分为对照组及3-NNMS的0μg/mL组、25μg/mL组、125μg/mL组,每组10只。除对照组外,其余各组制备肢体缺血再灌注损伤大鼠模型,于再灌注前30 min,向损伤部位注射相应浓度的3-NNMS。再灌注2 h后,心尖取血,取大鼠右下肢股直肌组织进行检测。苏木精-伊红染色观察大鼠股直肌组织病理形态;ELISA检测血清骨骼肌损伤因子肌酸激酶(Creatine Kinase found in the skeletal muscle,CK-MM)、乳酸脱氢酶、髓过氧化物酶水平,并检测股直肌核因子κB、肿瘤坏死因子α、白细胞介素1β、环氧合酶2、丙二醛、活性氧、超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶水平,以及股直肌ATP水平、ATPase活性、线粒体呼吸控制率(RCR)水平。结果与结论:①与对照组相比,缺血再灌注模型大鼠血清CK-MM、乳酸脱氢酶、髓过氧化物酶水平升高,股直肌核因子κB、肿瘤坏死因子α、白细胞介素1β、环氧合酶2、丙二醛及活性氧水平升高,过氧化氢酶、谷胱甘肽过氧化物酶水平下降,ATPase活性、线粒体呼吸控制率水平降低;细胞形态不规则,炎性细胞浸润明显,细胞出现肿胀。②与0μg/mL组相比,25μg/mL组大鼠血清CK-MM、乳酸脱氢酶水平降低,股直肌核因子κB、环氧合酶2水平降低,活性氧减少,超氧化物歧化酶活性升高;细胞形态较规则,炎性细胞浸润较轻,细胞肿胀现象缓解。③与0μg/mL组相比,125μg/mL组大鼠血清CK-MM、乳酸脱氢酶、髓过氧化物酶水平降低,股直肌核因子κB、肿瘤坏死因子α、环氧合酶2量减少,丙二醛、活性氧水平降低,超氧化物歧化酶、谷胱甘肽过氧化物酶活性升高,线粒体呼吸控制率水平升高;细胞排列较整齐,轮廓较清晰完整,炎性细胞浸润较轻。④结果说明:3-NNMS可以减轻肢体缺血再灌注引起的骨骼肌功能损伤,其作用机制可能是通过改善线粒体功能、减少活性氧产生、降低氧化应激和炎症反应,进而减轻组织损伤,修复骨骼肌功能。展开更多
Background Skeletal muscle has recently been recognized as an endocrine organ that can express, synthesize and secrete a variety of bioactive molecules which exert significant regulatory effects. Hydrogen sulfide （H2...Background Skeletal muscle has recently been recognized as an endocrine organ that can express, synthesize and secrete a variety of bioactive molecules which exert significant regulatory effects. Hydrogen sulfide （H2S） iS endogenously produced in mammalian tissues and participates in a number of physiological and pathophysiological processes. We aimed to verify whether H2S could be endogenously generated and released by rat skeletal muscle, and determine the biological effects of H2S in rat skeletal muscle. Methods The study was divided into two parts： detection of endogenous H2S generation and release in rat skeletal muscle and determination of antioxidative activity of skeletal muscle-derived H2S. H2S content and production in tissues were ,detected by sensitive sulfur electrode method. The expressions of H2S producing enzymes cystathionine i^-synthase, cystathionine y-lyase and mercaptopyruvate sulfurtransferase were detected by real-time PCR and western blotting and their tissue distributions were observed by immunohistochemical and immunofluorescent analysis. Rat skeletal muscular ischemia-reperfusion （I-R） injury model was created and evaluated by histological analysis under microscope. The malondialdehyde （MDA） contents, hydrogen peroxide levels, superoxide anion and superoxide dismutase （SOD） activities were detected using spectrophotometer. Results H2S could be endogenously generated and released by skeletal muscle of Sprague-Dawley rats （H2S content： （2.06＋0.43） nmol/mg; H2S production： （0.17＋0.06） nmol.minl.mgl）. Gene and protein expressions of the three H2S producing enzymes ~vere detected in skeletal muscle, as well as the liver and kidney. Endogenous H2S content and production were decreased in skeletal muscles of rats with I-R skeletal muscle injury （P 〈0.05）. Furthermore, H2S significantly protected rat skeletal muscle against I-R injury and resulted in decreased MDA content, reduced hydrogen peroxide and superoxide anion levels, but increased SOD activity and protein expression in skeletal muscles （all P 〈0.01）. Conclusion H2S generation pathway exists in rat skeletal muscle and it acts as an antioxidant in skeletal muscle.展开更多
文摘Objective The gene expression of skeletal muscle under ischemic condition by direct gene injectionwas observed in order to find a new gene delivery method to treat chronic arterial occlusion disease. Methods Weestablished the rabbit hindlimb ischemic model and used plasmid PSV-β-gal as a reporter gene. We transferedgene intramuscularly and detected the activity of β-galactosidase by histochemistry method. Results Weobserved that the gene expression of skeletal muscle under ischemic condition was higher than normalmuscle. Conclusion The result demonstrated that the direct gene injection was suitable for the chronic peripheralarterial occlusion disease, and might be a novel gene delivery method for this disease.
文摘Acute interruption of arterial blood flow to the extremities is often associated with significant morbidity and mortality. Broad spectrum mitogenic and non mitogenic activities of FGFs inspired us to study its protecting effects on tissue injuries in ischemia reperfusion condition. We found that systemic administration of aFGF after reperfusion onset prevented severe skeletal muscle injuries. In rats treated with aKGF, the tissue edema was reduced significantly, the tissue viability was increased, and the muscle fibers contained more succinate dehydrogenase (SDH) and adenosine triphosphatasc (ATPase). The pathological results supported the concept of improved prevention with aFGF treatment. The possible tissue protection by aFGF may come from its ability to regulate the concentration of evtra- and intracellular calcium ion. Besides, it may moderate other Ca2+ dependent enzyme conversion processes. Also, it may take part in the vascular tone regulation under ischemia and reperfusion conditions. These results suggest further study of tissue ischemia prevention with FGF and its possible mechanisms in the future.
文摘背景:线粒体活性氧爆发已被证明在骨骼肌缺血再灌注中起着关键作用。3-硝基-N-甲基水杨酰胺(3-nitro-N-methyl salicylamide,3-NNMS)可以有效降低电子传递速度,对肢体缺血再灌注损伤具有潜在的保护作用,但目前尚无明确的研究和临床应用。目的:探讨3-NNMS对肢体缺血再灌注损伤大鼠骨骼肌的保护作用及机制。方法:40只健康8周龄SD大鼠随机分为对照组及3-NNMS的0μg/mL组、25μg/mL组、125μg/mL组,每组10只。除对照组外,其余各组制备肢体缺血再灌注损伤大鼠模型,于再灌注前30 min,向损伤部位注射相应浓度的3-NNMS。再灌注2 h后,心尖取血,取大鼠右下肢股直肌组织进行检测。苏木精-伊红染色观察大鼠股直肌组织病理形态;ELISA检测血清骨骼肌损伤因子肌酸激酶(Creatine Kinase found in the skeletal muscle,CK-MM)、乳酸脱氢酶、髓过氧化物酶水平,并检测股直肌核因子κB、肿瘤坏死因子α、白细胞介素1β、环氧合酶2、丙二醛、活性氧、超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶水平,以及股直肌ATP水平、ATPase活性、线粒体呼吸控制率(RCR)水平。结果与结论:①与对照组相比,缺血再灌注模型大鼠血清CK-MM、乳酸脱氢酶、髓过氧化物酶水平升高,股直肌核因子κB、肿瘤坏死因子α、白细胞介素1β、环氧合酶2、丙二醛及活性氧水平升高,过氧化氢酶、谷胱甘肽过氧化物酶水平下降,ATPase活性、线粒体呼吸控制率水平降低;细胞形态不规则,炎性细胞浸润明显,细胞出现肿胀。②与0μg/mL组相比,25μg/mL组大鼠血清CK-MM、乳酸脱氢酶水平降低,股直肌核因子κB、环氧合酶2水平降低,活性氧减少,超氧化物歧化酶活性升高;细胞形态较规则,炎性细胞浸润较轻,细胞肿胀现象缓解。③与0μg/mL组相比,125μg/mL组大鼠血清CK-MM、乳酸脱氢酶、髓过氧化物酶水平降低,股直肌核因子κB、肿瘤坏死因子α、环氧合酶2量减少,丙二醛、活性氧水平降低,超氧化物歧化酶、谷胱甘肽过氧化物酶活性升高,线粒体呼吸控制率水平升高;细胞排列较整齐,轮廓较清晰完整,炎性细胞浸润较轻。④结果说明:3-NNMS可以减轻肢体缺血再灌注引起的骨骼肌功能损伤,其作用机制可能是通过改善线粒体功能、减少活性氧产生、降低氧化应激和炎症反应,进而减轻组织损伤,修复骨骼肌功能。
基金The study was supported by the grants from Major Basic Research Development Program of People＇s Republic of China （No. 2011CB503904 and No. 2013CB933801） and National Natural Science Foundation of China （No. 81070212）.
文摘Background Skeletal muscle has recently been recognized as an endocrine organ that can express, synthesize and secrete a variety of bioactive molecules which exert significant regulatory effects. Hydrogen sulfide （H2S） iS endogenously produced in mammalian tissues and participates in a number of physiological and pathophysiological processes. We aimed to verify whether H2S could be endogenously generated and released by rat skeletal muscle, and determine the biological effects of H2S in rat skeletal muscle. Methods The study was divided into two parts： detection of endogenous H2S generation and release in rat skeletal muscle and determination of antioxidative activity of skeletal muscle-derived H2S. H2S content and production in tissues were ,detected by sensitive sulfur electrode method. The expressions of H2S producing enzymes cystathionine i^-synthase, cystathionine y-lyase and mercaptopyruvate sulfurtransferase were detected by real-time PCR and western blotting and their tissue distributions were observed by immunohistochemical and immunofluorescent analysis. Rat skeletal muscular ischemia-reperfusion （I-R） injury model was created and evaluated by histological analysis under microscope. The malondialdehyde （MDA） contents, hydrogen peroxide levels, superoxide anion and superoxide dismutase （SOD） activities were detected using spectrophotometer. Results H2S could be endogenously generated and released by skeletal muscle of Sprague-Dawley rats （H2S content： （2.06＋0.43） nmol/mg; H2S production： （0.17＋0.06） nmol.minl.mgl）. Gene and protein expressions of the three H2S producing enzymes ~vere detected in skeletal muscle, as well as the liver and kidney. Endogenous H2S content and production were decreased in skeletal muscles of rats with I-R skeletal muscle injury （P 〈0.05）. Furthermore, H2S significantly protected rat skeletal muscle against I-R injury and resulted in decreased MDA content, reduced hydrogen peroxide and superoxide anion levels, but increased SOD activity and protein expression in skeletal muscles （all P 〈0.01）. Conclusion H2S generation pathway exists in rat skeletal muscle and it acts as an antioxidant in skeletal muscle.
