首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
Biochemical changes in the creatine kinase isoenzyme compositions in single muscle fibres of different types in rats were induced by endurance running training. Single muscle fibres were dissected from the soleus and extensor digitorum longus muscles of Wistarstrain male rats trained on a motor-driven treadmill for 16 weeks. Each fibre was typed histochemically (SO, slow-twitch oxidative; FOG, fast-twitch oxidative glycolytic; FG, fast-twitch glycolytic), and the activities of total creatine kinase and its four isoenzymes (CK-MM, -MB,-BB, and mitochondrial creatine kinase) were measured. The endurance training did not affect the total creatine kinase activity, but resulted in significantly increased activities of CK-MB and CK-BB in SO and FOG fibres, and the mitochondrial enzyme activity in FOG and FG fibres. Endurance training induced biochemical changes in the isoenzyme compositions, specifically in FOG fibres. These results suggest that changes in creatine kinase isoenzymes with endurance training reflect changes in the energy metabolism in the different muscle fibres, supporting the hypothesis that the different isoenzymes play different roles in energy transduction.  相似文献   

2.
The effects of training by running (sprint or endurance) on the mitochondrial volume, number, and succinate dehydrogenase (SDH) activity of different types of single muscle fibers, and on the elemental composition of soleus and extensor digitorum longus muscles were studied employing histochemical, electron microscopic, and electron probe-micro analysis (EPMA). Newly weaned male Wistar rats were trained on a motor-driven treadmill endurance exercise for 14 weeks. The relative mitochondrial volume per single muscle fiber of slow-twitch oxidative (SO) fiber was significantly increased following endurance training (p less than 0.01). There was no significant correlation between mitochondrial volume and number of SO fibers following endurance exercise training. Following sprint training, both mitochondrial volume and number of fast-twitch oxidative glycolytic (FOG) fibers increased significantly (p less than 0.01), with significant correlation (r = 0.69) between these parameters. SDH activity was higher in the order of SO, FOG, and fast-twitch glycolytic (FG) fibers following endurance training; however, the activity was higher in the order FOG, SO, and FG fibers following sprint training. The potassium concentration in cytoplasm of the soleus muscle showed a tendency to decrease following both types of training. These results suggest that the oxidative capacity of each type of muscle fiber does not always increase equally following training. Changes in mitochondrial number and volume in response to the two different types of training differed according to the type of fiber.  相似文献   

3.
Summary Single muscle fibres separated from extensor digitorum longus (EDL) as well as soleus (SOL) in the Wistar strain male rat in relaxing solution were typed histochemically, then glycolytic and oxidative enzyme activities were determined on the same fibres. Glycolytic enzyme lactate dehydrogenase (LDH), phosphofructokinase (PFK), pyruvate kinase (PK) and creatine kinase (CK) showed highest activities in fast-twitch glycolytic (FG), lower in fast-twitch oxidative glycolytic (FOG) and lowest in slow-twitch oxidative (SO) fibres. Also oxidative enzyme succinate dehydrogenase (SDH) and malate dehydrogenase (MDH) showed highest activities in SO, lower in FOG and lowest in FG fibres. The activities of LDH, PFK, PK and CK in FOG fibres separated from EDL showed higher activity compared to those separated from SOL, whereas the opposite result was obtained for the activities of SDH and MDH. Enzyme activities in a single muscle fibre type were not distinguishable from those of another type, and the activity profiles overlapped over a considerable range. The correlations among the separate enzyme activities of CK, LDH and MDH obtained from the same single fibre overlapped over a considerable range.  相似文献   

4.
Summary Single muscle fibres from soleus (SOL) as well as extensor digitorum longus (EDL) muscles from Wistar male rats in relaxing solution were divided into three types by their histochemical features — slow-twitch oxidative (SO), fast-twitch oxidative glycolytic (FOG), or fast-twitch glycolytic (FG) fibres. The relationship between ultrastructural profiles (mitochondrial volume, number, and Z-line width) and metabolic profiles (glycolytic and oxidative enzymes' activity) were analysed using the same types of fibres dissected from different SOL and EDL muscles using stereological and biochemical techniques. The Z-line width is specialized in different fibre types. Fast-twitch (FG and FOG) fibres have narrow Z-line width compared to slow-twitch (SO) fibre in SOL and EDL muscles. A significant difference was observed between SOL muscle SO and FOG fibres and EDL muscle SO and FOG fibres. All glycolytic (lactate dehydrogenase (LDH), phosphofructokinase (PFK), pyruvate kinase (PK) and creatine kinase (CK) activities in FOG fibres from EDL muscles were significantly higher (p<0.01) than those dissected from SOL muscles. The oxidative enzyme (succinate dehydrogenase (SDH) and malate dehydrogenase (MDH) activity in SO and FOG fibres dissected from SOL muscle were significantly higher (p<0.01) than those dissected from EDL muscles. Mitochondrial volume and number in SO fibres dissected from SOL muscle were significantly higher (volume,p<0.01, number,p<0.01) than those dissected from EDL muscles. A significant difference was not observed in mitochondrial volume of FOG fibres between SOL and EDL muscles. Significant positive correlation was observed in FOG (p<0.05) and FG (p<0.01) fibres between mitochondrial volume and number dissected from EDL muscle.The results suggest that the same type of single muscle fibres in different muscles have different ultrastructural and metabolic profiles, and these profiles resembled those of the fibre types primarily constituting those muscles.  相似文献   

5.
Summary Creatine kinase (CK) isoenzyme compositions of different types of single muscle fibres dissected from soleus (SOL) and extensor digitorum longus (EDL) muscles from rats were examined. CK isoenzymes were separated into cytoplasmic (CK-MM, CK-MB, CK-BB) (muscle, brain and hybrid types, respectively) and mitochondrial (m-CK) isoenzymes. Total CK and CK-MM activities showed the highest activities in fast-twitch glycolytic fibres (FG), lower in fast-twitch oxidative glycolytic (FOG) and the lowest in slow-twitch oxidative (SO) fibres. Conversely, the activity of m-CK was highest in SO, lowest in FG and intermediate in FOG fibres. The activity of CK-MB was highest in SO and lower in FOG and FG fibres. However, the activities of total CK and CK isoenzymes in a single muscle fibre type were not distinguishable from those of another type, and the profiles of CK isoenzyme compositions from the same type of single muscle fibres overlapped over a considerable range. The relationships between the four CK isoenzymes activities in single muscle fibres of different types were not similar. These results suggest that CK isoenzymes of single muscle fibres of different types play different roles in intracellular energy metabolism. Therefore, it is supposed that the CK isoenzyme compositions of single muscle fibres are suitable for their contractive and metabolic properties.  相似文献   

6.
The relationships between ultrastructural and metabolic profiles in different types of single muscle fiber after hindlimb suspension in rats were examined. Glycolytic (lactate dehydrogenase, LDH; phosphofructokinase, PFK) and oxidative (succinate dehydrogenase, SDH; malate dehydrogenase, MDH) enzyme activities in extensor digitorum longus (EDL) and soleus (SOL) muscles were measured. Relative mitochondrial and lipid droplet volumes were also measured in single muscle fiber of different types. Glycolytic enzyme activity in EDL muscle and oxidative enzyme activity in soleus muscle decreased following suspension for 2 weeks. LDH and PFK activities in fast-twitch (FG, fast-twitch glycolytic; FOG, fast-twitch oxidative glycolytic) fibers and oxidative enzymes in FOG and FG fibers decreased following suspension. Relative mitochondrial volume decreased significantly in all types (SO, slow-twitch oxidative; FOG, and FG) of fibers following suspension. The mitochondrial volume in SO fiber of the control group was significantly (p less than 0.01) higher than that of suspended group; however, SDH and MDH activities were not different between the control and suspended groups. The structural and metabolic changes following hindlimb suspension were influenced by different factors, respectively. Changes in ultrastructural and metabolic profiles in response to the hindlimb suspension differed according to the type of fibers.  相似文献   

7.
Muscle fibre type composition and distribution in the biceps brachii (long head) and triceps brachii (long head) of the rat and rabbit were investigated using the following histochemical techniques: myosin ATPase, with preincubation at pH 10.4 and 4.35; succinate dehydrogenase (SDH) and glycogen phosphorylase. The muscle fibres were classified into slow-twitch (SO), fast-twitch glycolytic (FG), fast-twitch oxidative glycolytic (FOG and FOg) and fast-twitch oxidative fibres (FO). Significant differences in the regional distribution of muscle fibre types have been observed between the rat and the rabbit. In the rat, SO fibres were restricted to the deep regions of both biceps and triceps brachii, whereas FG fibres were located in the intermediate and superficial regions (the superficial regions contained the highest percentages of FG fibres). In the rabbit, SO and FG fibres were spread over the entire muscle, although SO and FG fibres were most abundant in the deep and superficial regions respectively. These findings indicate that the biceps and triceps brachii are more regionalised in the rat than in the rabbit.  相似文献   

8.
Summary Twenty 4-week-old Wistar rats exercised voluntarily in running wheels each day for 45 days. Fibre type composition, fibre cross-sectional area and the number of capillaries around a fibre of the slow-twitch soleus and fast-twitch plantaris muscles were examined and compared with animals which had no access to running wheels. The exercise group had a higher percentage of fast-twitch oxidative glycolytic (FOG) fibres and a lower percentage of fast-twitch glycolytic (FG) fibres in the deep portion of the plantaris muscle. The area of FOG fibres in the surface portion of the plantaris muscle was also greater in the exercise group. In the exercised animals, there was a positive relationship between the running distance and the area of FOG fibres in both the deep and surface portions of the plantaris muscle. In addition, the running distance correlated positively with the percentage of FOG fibres and negatively with that of FG fibres in the deep portion of the plantaris muscle. There were no relationships between the running distance and fibre type composition, or fibre area and capillary supply in the soleus muscle. These results suggested that the increase in the percentage and area of FOG fibres in the fast-twitch muscle was closely related to voluntary running.  相似文献   

9.
Summary The effects were investigated of high intensity short duration exercise and anabolic steroid treatment on the medial gastrocnemius muscle of female rats. Twelve rats were divided equally into four groups, exercise with and without steroid administration and sedentary with and without steroid administration. Animals were made to swim for 5 weeks, 6 days·week–1. Muscle fibres were classified as slow-twitch (ST), fast-twitch oxidative glycolytic (FOG) and fast-twitch glycolytic (FG). Muscle fibre size was measured as the equivalent circle diameter. Exercise (P<0.001) and steroid (P<0.05) treatments alone, significantly elevated FOG and decreased FG fibre proportions. Overall proportions of fast-twitch and ST muscle fibres did not vary with any of the treatments. Significant differences in the proportion of muscle fibres were found to exist between different areas within the gastrocnemius muscle (P<0.05). Exercise and steroid treatments alone did not alter muscle fibre diameters. Combined exercise and steroid treatments did significantly increase ST fibre diameters (P<0.05). Exercise only treatment resulted in significant increases in the number of capillaries surrounding ST fibre (P<0.05) and FOG fibre (P<0.01) types. In conclusion the main finding of this study indicated that anabolic steroids in conjunction with high intensity swimming instigated ST fibre hypertrophy. Exercise and steroid only treatments significantly elevated FOG fibre proportions while FG fibre proportions diminished. Exercise only treatment resulted in significant increases in the number of capillaries surrounding both ST and FOG fibre types.  相似文献   

10.
Three muscles were analyzed, Longissimus dorsi, Semimembranosus and Caput longum Tricipitis brachii taken from nine cow buffaloes, by examining the histochemical and morphometrical characteristics of different muscle fibres types and their distribution inside the examined muscles. Cross sectional area, perimeter, maximum and minimum diameter of about 200 fibres were measured for each muscle, and fast-twitch glycolytic fibres (FG), fast-twitch oxidative-glycolytic fibres (FOG), slow-twitch oxidative fibres (SO) were histochemically differentiated. The data have been elaborated with the SPSS software. The variance analysis indicates that there are not significant differences about dimensions between FG and FOG fibres, while the average values of transversal section area and perimeter are greater than the oxidative fibres in all examined muscles. The Semimembranosus muscle in comparison to the Longissimus dorsi and to the Caput longum Tricipitis brachii muscles has muscle fibres with the smallest value of transversal section area and perimeter. The balanced distribution and intense myofibrillar adenosine triphosphatose and succinic dehydrogenase activities of the three fibres types in Caput longum Tricipitis brachii muscle can be justified by the function performed by this muscle which, together with the other heads of the Triceps brachii acts essentially as extensor of the forearm in fact, differences in the dimensions of the different fibre types inside the three examined muscles have been underlined; this fact can be justified for every muscle performs different motor functions.  相似文献   

11.
Vastus medialis muscles of patients with chronic anterior instability of the knee after anterior cruciate ligament rupture were analysed to investigate changes in defined muscle fibres of the diseased leg in comparison to the healthy leg of the same patient. Metabolic and morphological parameters were obtained by cytophotometrical measurements of the activities of succinate dehydrogenase (a marker of oxidative metabolism) and glycerol-3-phosphate dehydrogenase (a marker of glycolytic metabolism) of slow-oxidative (SO), fast-oxidative glycolytic (FOG) and fast-glycolytic (FG) fibre types in serial sections and by measuring the minimal fibre diameters of type I (slow) and type II (fast) fibres. We found decreased glycolytic activity and a shift to more oxidative metabolism in each fibre type suggesting diminished fast force and shift to endurance force development. The latter was interpreted as a sign of active compensation for the knee instability. Significantly decreased minimal fibre diameters to 85.9% in type I fibres, and to 88.7% in type II fibres of the diseased muscle were measured, indicating the fibre atrophy. Our findings suggest that the atrophied muscle fibres of the affected vastus medialis muscle adapt to the altered conditions by changing their metabolic profile. Muscle fibres of different types were found to be affected similarly.  相似文献   

12.
Differentiation of rat skeletal muscle fibres during development and ageing   总被引:1,自引:0,他引:1  
The purpose of the present study was to determine at which point in the period from embryonic day 21 up to postnatal day (PD) 75, the different fibre types and subtypes are detectable in rat extensor digitorum longus, soleus and gastrocnemius muscles using immunohistochemical, enzyme histochemical and cytophotometrical methods. Moreover, fibre type-specific changes in metabolic profile and changes in fibre type population during postnatal development were analysed. Before birth, no clear differentiation of fibre types was found. At PD 1, slow and fast fibres were typed by antibodies against neonatal, slow and fast myosin heavy chains (MHCs). At PD 8, the different ATPase activities of slow and fast MHCs after alkaline preincubation were detected histochemically. At PD 21, differences in acid stability of ATPase activity of fast MHC isoforms revealed the fast subtypes IIA and IIB (including IIX). At this age, also differences in metabolic properties (oxidative and glycolytic enzyme activities) of fibres were detected for the first time by cytophotometry classifying the fibres into SO, FOG I, FOG II and FG. Before the age of 21 days, the fast fibres were metabolically undifferentiated. During further development and ageing, the population of FG fibres with high glycolytic activity increased at the expense of FOG fibres suggesting FOG to FG transformation. Cytophotometrical measurements revealed that the muscle fibres developed their highest contractile, oxidative and glycolytic activity at PD 21, the time of weaning. In this way, muscle fibres may be prepared for the higher demands for posture and mobility after leaving the nest.  相似文献   

13.
Proximal (vastus lateralis) and distal (gastrocnemius) muscles of 100-day-old normal and myopathic BIO TO-2 hamsters were analysed to study the effects of myopathy on the different muscle fibre types: SO (slow oxidative), FOG (fast oxidative glycolytic) and FG (fast glycolytic). Cytophotometric measurements of enzyme activities (myofibrillic adenosine triphosphatase, succinate dehydrogenase and glycerol-3-phosphate dehydrogenase), Western blot analysis of nitric oxide synthase (NOS) I, II, III isoforms and NOS II immunohistochemistry were performed. The following alterations were found in myopathic muscle fibres: all fibre types of both proximal and distal myopathic muscles showed decreased myofibrillic adenosine triphosphatase activity indicating depressed contractility. This was associated with depressed oxidative activity of the muscle fibres. A shift to more glycolytic metabolism was observed, mainly in FG fibres of proximal muscle. We found an increased NOS II expression in both myopathic muscle types investigated. It means that increased NO production inhibits force generation in myopathic muscle. NOS II immunoreactivity was found mainly in the cytoplasm of FG fibres. NOS I and NOS III expression was not significantly effected by this form of myopathy. Our findings demonstrate that muscle fibres of proximal and distal skeletal muscles of 100-day-old cardiomyopathic BIO TO-2 hamsters are altered with respect to contractility, metabolism and NOS II expression. FG fibres of the proximal muscle were effected most strongly.  相似文献   

14.
Summary In rabbits blood flow was measured in 19 muscles with the radioactive microsphere technique. Fibre type composition (SO, slow-twitch oxidative; FOG, fast-twitch oxidative-glycolytic; FG, fasttwitch glycolytic) was determined histochemically for the same muscles. While no significant changes occur in the transition from quiet wakefulness (QW) to synchronized sleep (SS), in desynchronized sleep (DS) blood flow decreases in SO and increases in FOG and FG fibres. These changes may be related to the changes in motor activity characteristic of DS: muscle atonia and twitches, respectively.  相似文献   

15.
Male Wistar rats were made hyperthyroid by intraperitoneal injections on alternate days for 1–6 weeks, of 200 g/kg triiodothyronine (T3). The effects of this treatment on the contractile properties of the soleus, a slow twitch and the extensor digitorum longus (EDL), a fast twitch skeletal muscle, were studied in vivo in the anaesthetized animal. Post mortem, serial frozen sections of both muscles were stained histochemically for myosin ATPase, succinic dehydrogenase and phosphorylase. Muscle fibres were classified as either slow twitch (SO), fast twitch oxidative glycolytic (FOG) or fast twitch glycolytic (FG).Elevation of plasma T3 levels is associated with progressive alterations in the muscle fibre populations of both muscles. In the soleus there is conversion of SO to FOG fibres while in the EDL the main effect is FG to FOG conversion. There are also marked changes, mainly confined to the soleus muscle, in contractile properties; progressive increases in isometric twitch and tetanic tension and in the rates of contraction and relaxation during both twitch and tetanus.The effect of T3 on slow muscle contractility may be related to its effect on muscle phenotype. However, changes in the former precede detectable fibre population changes. T3 may influence properties such as the Ca2+ binding activity of sarcoplasmic reticulum of existing slow twitch fibres before the later changes associated with the interconversion of fibre types occur.  相似文献   

16.
Mammalian skeletal muscle fibers are classified into three basic types based on their enzyme histochemical profiles: fast-twitch glycolytic (FG), fast-twitch oxidative glycolytic (FOG), and slow-twitch oxidative (SO) types. The type-shift of fibers from FOG to SO in the slow soleus muscle of rats occurs during postnatal growth. Our previous studies have demonstrated that hypoxic exposure inhibits a growth-related type-shift of fibers from FOG to SO in the rat soleus muscle, irrespective of the duration and age at which the animals are exposed to hypoxia. Our previous studies have also revealed that a high percentage of FOG fibers in the soleus muscle of the hypoxia-adapted rats is found only under hypoxic conditions. Furthermore, we have found that these adaptations in fibers of the rat soleus muscle correspond well with those in motoneurons at the ventral horn of the spinal cord that innervate the muscle fibers.  相似文献   

17.
Single fibres from tibialis anterior, extensor digitorum longus, gastrocnemius and soleus muscles in young (4–week–old) and adult (35–week–old) Wistar male rats were classified into three types on the basis of their enzyme–histochemical features: slow–twitch oxidative (SO), fast–twitch oxidative and glycolytic (FOG) and fast–twitch glycolytic (FG) fibres. Ultrastructural (volume density of mitochondria: Vmt and Z line width) and metabolic (phosphofructokinase: PFK and succinate dehydrogenase: SDH activities) profiles were measured. PFK activity in all types of fibres1 was higher in adult rats, and the difference between the two age–groups (adult/young) was largest between FG, FOG and SO fibres respectively. SDH activity and Vmt were lower in adult rats in a similar way in all fibres. A significant positive correlation was observed between the Vmt and SDH activity in both age–groups. This positive correlation was very specific in fast–twitch and slow–twitch fibres. Changes in the Vmt did not relate directly to the changes in fibre cross–sectional area. The overall pattern indicates that glycolytic capacity of fast–twitch fibres in flexor muscles (TA and EDL) is higher than in extensor muscles (GC and SOL), and that oxidative capacity of all types of fibre in extensor muscles is higher than in flexor muscles. These profiles were changed by growth, and may be related to the specific differences in pattern of activity of each skeletal muscle, and may reflect differences in the recruitment order of different muscles.  相似文献   

18.
The alterations in activity patterns of representative enzymes in energy metabolism were investigated in the superficial (white) and deep (red) portions of the fast vastus lateralis muscle of the adult rat in response to prolonged endurance training. It was found that following 15 weeks of extreme training (final running duration: 210 min per day, 27 m/min at 15 degree grade), increases in the activities of marker enzymes of the citric acid cycle (citrate synthase), β-oxidation (3-hydroxyacyl CoA dehydrogenase), and ketone body utilization (3-ketoacid CoA transferase) as well as of glutamate pyruvate transaminase occurred in both regions of the muscle, with the geatest increase being observed in the superficial portion (2.6–4.2-fold). Pronounced increases were also seen for hexokinase which showed highest activities after 7 weeks of training. Conversely, decreases were noted for various glycogenolytic, glycolytic and gluconeogenic enzymes (phosphorylase, glyceraldehydephosphate dehydrogenase, pyruvate kinase, lactate dehydrogenase and fructose-1,6-diphosphatase). Reduction in the activities of these enzymes was most pronounced in the deep portion of the muscle. These results demonstrate a fundamental rearrangement of the energy metabolism of the muscle in response to prolonged, high intensity training. In the case of the deep portion of the vastus lateralis muscle, which has been shown to be composed of a large percentage of fast oxidative-glycolytic fibres (FOG), the enzyme profile becomes similar to the slow oxidative (SO) fibre. In the superficial portion which contains predominantly fast glycolytic fibres (FG), the enzyme profile becomes similar to FOG fibres. The magnitude of the observed changes in enzyme activities was greater than in previous animal studies. This suggests that there might be no limit to the metabolic adaptability of skeletal muscle to increased contractile activity. In this context, the adaptations observed here qualitatively resemble the metabolic transformations reported for chronic low frequency electrical stimulation.  相似文献   

19.
Comparative analyses of the fibre content (FG, FOG, and SO fibres) and the capillary density (the number of capillaries surrounding individual fibres and the capillary/fibre ratio) were performed in hind limb muscles of the cat. Cross-sections from the tenuissimus, the biceps femoris, the lateral head (LG) and the medial head (MG) of the gastrocnemius and the soleus were cut in a cryostat. The sections were stained histochemically for the NADH2-diaphorase and alkaline (pH 9.4) actomyosin ATPase activity, which enables differentiation of different types of fibres. The endothelium of the capillaries was identified via staining for unspecific alkaline ATPase activity. The number of capillaries surrounding each individual muscle fibre had a positive correlation, first to the oxidative capacity and secondly to the average diameter of the fibres. The thin tenuissimus muscle did not differ in this respect from the thicker muscles. The highest proportion of SO fibres was found in the soleus and the MG muscles. FG fibres of two different types were dominating the fibre mass in the biceps femoris and the LG muscles, while the tenuissimus contained more FOG fibres than these muscles. In general the FG fibres had a larger diameter than the FOG and the SO fibres. The soleus and the MG muscles contained larger fibres than the other examined muscles. FG fibres were surrounded by fewer capillaries than FOG and SO fibres. The soleus and the MG muscles, with a higher percentage of SO fibres and also larger fibres, had the largest number of capillaries around the fibres and the highest capillary/fibre ratio.  相似文献   

20.
The enzyme activities of intra- and extrafusal fibers in the tibialis anterior muscle of rats during postnatal development have been investigated. Muscle fibers 1 day after birth showed a uniform reaction for adenosine triphosphatase (ATPase), succinate dehydrogenase (SDH), and alpha-glycerophosphate dehydrogenase (alpha-GPD) activities. Fast-twitch (F) and slow-twitch (S) fibers with ATPase activity were found at 9 and 11 days. Thereafter, the type shift of muscle fibers from S to F was observed in the deep and middle portions. Fast-twitch oxidative glycolytic (FOG), fast-twitch glycolytic (FG), and slow-twitch oxidative (SO) fibers with ATPase, SDH, and alpha-GPD activities were found at 15 (the superficial portion) and 17 days (the deep and middle portions). The histochemical differentiation of intrafusal muscle fibers (7 and 9 days) was found earlier than that of extrafusal muscle fibers.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号