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Objective To summarize the clinical , pathological and genetic characteristics of three patients with caveolin-3 associated myopathy and review the literatures .Methods The clinical data of three patients with caveolin-3 associated myopathy were investigated .With informed consent , we performed muscle biopsy and genetic analysis of CAV 3 and PTRF genes.Results All the three patients presented with percussion/pressure-induced rapid contraction , percussion-induced muscle mounding and mechanically induced muscle rippling.Besides, case 1 had weakness and atrophy of hand muscles .Case 2, who manifested with muscle hyperexcitability at onset , developed weakness and atrophy of distal part of lower limbs.Case 3 showed normal muscle strength and tone .All of them had myalgia or tenderness .Muscle biopsy revealed mild myogenic changes in two patients and a muscular dystrophic pattern in one . Immunohistochemical staining of caveolin-3 revealed an even deficiency in case 1 and a mosaic deficiency in cases 2 and 3.Gene analysis revealed a missense mutation ( c.80G>A, p.R27Q) in CAV3 gene of case 1. No mutations were identified in cases 2 and 3.Conclusions There is phenotypic variability in patients with caveolin-associated myopathy , including limb-girdle syndrome , rippling muscle disease , distal myopathy , muscle hypertrophy , idiopathic hyperCKemia and cardiomyopathy .Muscle biopsy and caveolin-3 staining should be performed for the above patients with muscle rippling .
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OBJECTIVE:To study the effects of Astragalus granules on the expression of Caveolin-3(Cav-3)and Smad family member 3 (Smad3) in the myocardial cells of rats with viral myocarditis. METHODS:90 rats were randomly divided into a nor-mal group,a model group,a Shenmai injection group [positive drug,0.2 g/(kg·d)] and the groups of low,medium and high-dose Astragalus granules [0.42,0.84,1.68 g/(kg·d)],with 15 rats in each group. The rats in all groups except for the normal group were given CVB3 ip for the establishment of viral myocarditis model. Meanwhile,the rats in the drug administration groups were given corresponding drugs ig,while those in the normal group and the model group were given normal saline ig,for 15 consecu-tive days. 5 rats were selected from each group respectively on the 3rd,9th and 15th days of drug use to take an experiment. For the rats,the pathological change of the cardiac muscle tissue was observed and scored,and the mRNA and protein expression of Cav-3 and Smad3 in the myocardial cells were detected. RESULTS:After 15 days of drug use,compared to the normal group,the rats of the model group had hyperplasia of a large number of cardiac muscle fibers,obvious lesions at cardiac muscle fibers, and significantly higher pathological score and levels of the mRNA and protein expression of Cav-3 and Smad3 in the myocardial cells (P<0.05). Compared to the model group,the rats in the drug administration groups had cardiac muscle tissue lesions improved and had obviously lower pathological score and levels of the mRNA and protein expression of Cav-3 and Smad3 in the myocardial cells(P<0.05). CONCLUSIONS:Astragalus granules can markedly downregulate the gene expression of Cav-3 and Smad3 in the myocardial cells of rats with viral myocarditis,which is inferred as a prevention and treatment mechanism of viral myocarditis.
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OBJECTIVE@#To explore the mechanism of Profilin-1 in regulating eNOS/NO pathway and its role in the development of myocardial hypertrophy.@*METHODS@#Spontaneously hypertensive rats (SHR) aged 5 weeks were injected with different adenovirus vectors to induce Profilin-1 expression knockdown (SHR-I) or over express (SHR-H) or to use as control (SHR-C). All these treatment were compared with Wistar-Kyoto rats (SKY) treated with control adenovirus vectors (WKY-C). The same injection was executed at the sixth week during the experiment of 12 weeks. After experiment, the left ventricular weight-to-heart weight ratio (LVW/HW) and left ventricular long axis (LVLA) were measured. Meanwhile, NO contents in blood and myocardium, Profilin-1, eNOS and Caveolin-3 mRNA and protein levels and phosphorylated eNOS (P-eNOS) protein level in myocardium were determined.@*RESULTS@#Compared with WKY-C group, the SHR-C group was statistically higher in LVW/HW (0.79±0.03), LVLA (11.82±0.58 mm) and Profilin-1 mRNA and protein level (P<0.05), but lower in NO content [(18.63±6.23) μmol/L] in blood and [(2.71±0.17) μmol/L] in myocardium), eNOS activity and Caveolin-3 expression (P<0.05). The over expressing Profilin-1 led SHR-H group to a higher value of LVW/HW [(0.93±0.03) mm and LVLA (14.17±0.69) mm] in comparison with SHR-C group (P<0.05), and to a lower value of NO content (in myocardium), eNOS activity and Caveolin-3 expression (P<0.05); however, this phenomenon was reversed by the knockdown Profilin-1 expression (SHR-I group).@*CONCLUSIONS@#Profilin-1 expression, being negative in regulating Caveolin-3 expression and eNOS/NO pathway activity, promotes the development of myocardial hypertrophy which can be reversed by Profilin-1 silencing.
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Objective:To explore the mechanism of Profilin-1 in regulating eNOS/NO pathway and its role in the development of myocardial hypertrophy.Methods: Spontaneously hypertensive rats (SHR) aged 5 weeks were injected with different adenovirus vectors to induce Profilin-1 expression knockdown (SHR-I) or over express (SHR-H) or to use as control (SHR-C). All these treatment were compared with Wistar-Kyoto rats (SKY) treated with control adenovirus vectors (WKY-C). The same injection was executed at the sixth week during the experiment of 12 weeks. After experiment, the left ventricular weight-to-heart weight ratio (LVW/HW) and left ventricular long axis (LVLA) were measured. Meanwhile, NO contents in blood and myocardium, Profilin-1, eNOS and Caveolin-3 mRNA and protein levels and phosphorylated eNOS (P-eNOS) protein level in myocardium were determined. Results:Compared with WKY-C group, the SHR-C group was statistically higher in LVW/HW (0.79±0.03), LVLA (11.82±0.58 mm) and Profilin-1 mRNA and protein level (P<0.05), but lower in NO content [(18.63±6.23) μmol/L] in blood and [(2.71±0.17) μmol/L] in myocardium), eNOS activity and Caveolin-3 expression (P<0.05). The over expressing Profilin-1 led SHR-H group to a higher value of LVW/HW [(0.93±0.03) mm and LVLA (14.17±0.69) mm] in comparison with SHR-C group (P<0.05), and to a lower value of NO content (in myocardium), eNOS activity and Caveolin-3 expression (P<0.05); however, this phenomenon was reversed by the knockdown Profilin-1 expression (SHR-I group).Conclusions:Profilin-1 expression, being negative in regulating Caveolin-3 expression and eNOS/NO pathway activity, promotes the development of myocardial hypertrophy which can be reversed by Profilin-1 silencing.
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Objective To report the clinical,myopathological and genetic features of a patient with distal myopathy caused by caveolin-3 (CAV3) deficiency.Methods The patient was a 27-year-old female.She had an onset symptom of asymmetric lower extremities weakness.The proximal limb-girdle muscles were involved subsequently.Clinical data of this patient were collected.The leg muscle magnetic resonance imaging (MRI) and an open biopsy of left tibialis anterior muscle were performed.In addition to histological,enzyme histochemical staining and ultrastructural examination,immunohistochemical staining with antibody against CAV3 was done.CAV3 gene was analyzed in the patient and her parents.Results Tl-weighted enhanced skeletal muscle MRI of the lower limbs showed the abnormal signal in distal and proximal muscles.Muscle biopsy showed moderate dystrophic changes and immunostaining for CAV3 showed reduced plasmalemma in the muscle fibers.Gene analysis disclosed a heterozygous c.136G ＞ A (p.Ala46Thr)mutation in the CAV3 gene,and the patient's parents did not have this mutation.Conclusions We report a distal myopathy case caused by c.136G ＞ A (p.Ala46Thr) mutation in the CAV3 gene,who had an onset symptom of asymmetric lower extremities weakness.The proximal limb-girdal muscles were also involved.This would help clinical doctors to know more about this rare myopathy.
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Objective: To explore the mechanism of Profilin-1 in regulating eNOS/NO pathway and its role in the development of myocardial hypertrophy. Methods: Spontaneously hypertensive rats (SHR) aged 5 weeks were injected with different adenovirus vectors to induce Profilin-1 expression knockdown (SHR-I) or over express (SHR-H) or to use as control (SHR-C). All these treatment were compared with Wistar-Kyoto rats (SKY) treated with control adenovirus vectors (WKY-C). The same injection was executed at the sixth week during the experiment of 12 weeks. After experiment, the left ventricular weight-to-heart weight ratio (LVW/HW) and left ventricular long axis (LVLA) were measured. Meanwhile, NO contents in blood and myocardium, Profilin-1, eNOS and Caveolin-3 mRNA and protein levels and phosphorylated eNOS (P-eNOS) protein level in myocardium were determined. Results: Compared with WKY-C group, the SHR-C group was statistically higher in LVW/HW (0.79±0.03), LVLA (11.82±0.58 mm) and Profilin-1 mRNA and protein level (P<0.05), but lower in NO content [(18.63±6.23) μmol/L] in blood and [(2.71±0.17) μmol/L] in myocardium), eNOS activity and Caveolin-3 expression (P<0.05). The over expressing Profilin-1 led SHR-H group to a higher value of LVW/HW [(0.93±0.03) mm and LVLA (14.17±0.69) mm] in comparison with SHR-C group (P<0.05), and to a lower value of NO content (in myocardium), eNOS activity and Caveolin-3 expression (P<0.05); however, this phenomenon was reversed by the knockdown Profilin-1 expression (SHR-I group). Conclusions: Profilin-1 expression, being negative in regulating Caveolin-3 expression and eNOS/NO pathway activity, promotes the development of myocardial hypertrophy which can be reversed by Profilin-1 silencing.
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Objective To observe the damage degree and expression pattern of Caveolin-3 mRNA by ischemia-reperfusion injury in rabbits of skeletal muscle cell at different phases.Methods In this study,from April 2013 to December 2013,30 lower limbs of 15 Chinese White Rabbits were used and divided into two groups:all the left lower limbs were experimental group,which were made as an experimental model of ischemia-reperfusion injury by occluding left common iliac artery using noninvasive vascular.All the right lower limbs without surgical treatment were the control group.Gastrocnemius samples were obtained at 4h and 8h after reperfusion and handled by HE staining and observed by optical microscopy.By Real-time PCR,Caveolin-3/GAPDH mRNA were detected.Results HE stain showed:in control group,there was no edema,degeneration and inflammatory cell infiltration; in experi-meatal group,muscle cell degeneration had occured at ischemic 5 h.The edema was aggravated,a large number vacuole were formed and inflammatory cell were infiltrated at 4 h reperfusion.Reperfusion injury at 8h significantly reduced compared to 4 h.The Caveolin-3/GAPDH mRNA expression levels by SPSS 19.0 showed:Control group:1.026 ± 0.065,1.004 ±0.037,1.022 ±0.051,experimental group:1.159 ±0.073,1.445 ±0.053,1.208 ±0.058 at ischemic 5 h,4 h and 8 h reperfusion,respectively.On-line analysis of variance cases of ischemic 5 h and 4 h reperfusion and 8 h reperfusion,the experimental group than the control group were increased,with statistical significance (P ＜ 0.05).The experimental group of ischemic 5 h and 8 h reperfusion was no significant difference (P ＞ 0.05).It showed Caveolin-3 mRNA expression levels in ischemia-reperfusion 8 h group returned to normal.There was significant statistical difference between the ischemic 5 h and 4 h reperfusion (P ＜ 0.05).There was significant statistical difference between the 4 h reperfusion and 8 h reperfusion (P ＜ 0.05).Conclusion The expression of Caveolin-3 in experimental group showed a trend of first increased and then decreased.The expression levels of Caveolin-3 mRNA in skeletal muscle cells after ischemia-reperfusion injury is consistent with the development and progression of muscle cell damage.The results indicate that Caveolin-3 may play a control role in the injury and recovery of skeletal muscle cell.
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Objective To observe the difference of caveolin-3(CAV3) gene polymorphism between normal people and diabetic patients in Chinese Han population. Methods Exon gene polymorphism in 50 normal people and 50 T2DM patients were detected by PCR-SSCP. Results The cumulative incidence rate of electrophoretic variation in T2DM patients was 48%, while cumulative incidence rate of normal people was 7%(P<0.001). It was proved that in the variant bands, there were base variant. Conclusions The variant base number of CAV3 gene in human T2DM samples are significantly more than the normal which can be preliminary detected by PCR-SSCP. It indicates that CAV3 gene polymorphism may be one of the genetic backgrounds for the occurence of Chinese T2DM.
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Objective To evaluate the effects of sevoflurane preconditioning on caveolin-3 expression during myocardial ischemia-reperfusion (I/R) in rats.Methods Healthy male Wistar rats,aged 6-8 weeks,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital 30 mg/kg and heparin 1 000 IU/kg.Their hearts were excised and perfused with K-H solution in a Langendorff apparatus.Thirty isolated rat hearts were randomly assigned into 3 groups (n =10 each) using a random number table:control group (group C),I/R group and sevoflurane preconditioning group (group SP).After 30 min of equilibration,group C was continuously perfused with K-H solution for 90 min,group I/R underwent 30 min of ischemia followed by 60 min of reperfusion,and group SP was perfused with K-H solution saturated with 2％ sevoflurane for 10 min followed by 5 min washout with K-H solution,then underwent 30 min of ischemia followed by 60 min of reperfusion.HR,left ventricular enddiastolic pressure (LVEDP),+ dp/dtmax and-dp/dtmax were recorded at the end of equilibration,immediately before ischemia and at 30 and 60 min of reperfusion.Myocardial specimens were obtained from the cardiac apex for microscopic examination.Myocardial specimens were obtained from the left ventricle for determination of caveolin3 expression.Results Compared with group C,+ dp/dtmax and-dp/dtmax were significantly decreased immediately before ischemia,and HR,LVDEP,+ dp/dtmax and-dp/dtmax were decreased at 30 and 60 min of reperfusion in groups I/R and SP,and caveolin-3 expression was down-regulated in group I/R and up-regulated in group SP (P ＜ 0.05).Compared with group I/R,HR,LVDEP,+ dp/dtmax and-dp/dtmax were significantly decreased immediately before ischemia and increased at 30 and 60 min of reperfusion,and caveolin-3 expression was up-regulated in group SP (P ＜ 0.05).The pathological changes were significantly attenuated in group SP as compared with group I/R.Conclusion The mechanism by which sevoflurane preconditioning attenuates myocardial I/R injury in rats may be related to up-regulation of myocardial caveolin-3 expression.
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Membrane repair is a conserved cellular process, where intracellular vesicles translocate to sites of plasma membrane injury to actively reseal membrane disruptions. Such membrane disruptions commonly occur in the course of normal physiology, particularly in skeletal muscles due to repeated contraction producing small tears in the sarcolemmal membrane. Here, we investigated whether prolonged exercise could produce adaptive changes in expression levels of proteins associated with the membrane repair process, including mitsugumin 53/tripartite motif-containing protein 72 (MG53/TRIM72), dysferlin and caveolin-3 (cav3). Mice were exercised using a treadmill running protocol and protein levels were measured by immunoblotting. The specificity of the antibodies used was established by immunoblot testing of various tissue lysates from both mice and rats. We found that MG53/TRIM72 immunostaining on isolated mouse skeletal muscle fibers showed protein localization at sites of membrane disruption created by the isolation of these muscle fibers. However, no significant changes in the expression levels of the tested membrane repair proteins were observed following prolonged treadmill running for eight weeks (30 to 80 min/day). These findings suggest that any compensation occurring in the membrane repair process in skeletal muscle following prolonged exercise does not affect the expression levels of these three key membrane repair proteins.
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Animals , Carrier Proteins/metabolism , Caveolin 3/metabolism , Gene Expression Regulation , Male , Membrane Proteins/metabolism , Mice , Muscle, Skeletal/cytology , Muscle, Skeletal/physiology , Myocardium/cytology , Physical Conditioning, Animal , Protein Transport , Rats , Sarcolemma/metabolism , Time FactorsABSTRACT
We investigated the effect of phenylephrine (PE)- and isoproterenol (ISO)-induced cardiac hypertrophy on subcellular localization and expression of caveolin-3 and STAT3 in H9c2 cardiomyoblast cells. Caveolin-3 localization to plasma membrane was attenuated and localization of caveolin-3 to caveolae in the plasma membrane was 24.3% reduced by the catecholamine-induced hypertrophy. STAT3 and phospho-STAT3 were up-regulated but verapamil and cyclosporin A synergistically decreased the STAT3 and phospho-STAT3 levels in PE- and ISO-induced hypertrophic cells. Both expression and activation of STAT3 were increased in the nucleus by the hypertrophy. Immunofluorescence analysis revealed that the catecholamine-induced hypertrophy promoted nuclear localization of pY705-STAT3. Of interest, phosphorylation of pS727-STAT3 in mitochondria was significantly reduced by catecholamine-induced hypertrophy. In addition, mitochondrial complexes II and III were greatly down-regulated in the hypertrophic cells. Our data suggest that the alterations in nuclear and mitochondrial activation of STAT3 and caveolae localization of caveolin-3 are related to the development of the catecholamine-induced cardiac hypertrophy.
Subject(s)
Animals , Rats , Catecholamines/pharmacology , Caveolae/metabolism , Caveolin 3/metabolism , Cell Line , Hypertrophy/metabolism , Mitochondria/metabolism , Myocardium/cytology , Myocytes, Cardiac/cytology , STAT3 Transcription Factor/metabolismABSTRACT
OBJECTIVE: To exhibit the caveolin-3 immunoreactivities (IRs) in the peripheral nerve, which was previously known to be present only within the muscle and to be a causative agent of myopathy METHOD: The sciatic nerves of the rat were removed after the perfusion and frozen after cryoprotection by sucrose. The tissue specimens were cut on cryostat and immunostained with anti-caveolin-3 and growth associated protein-43 (GAP-43) antibodies. The sections were observed with a fluorescence microscope. RESULTS: We detected caveolin-3 IRs in myelin sheath of the peripheral nerves, while GAP IRs were detected in the axon. Caveolin-3 IRs were active in the rat of postnatal 1 week, but they were reduced in the rat of postnatal 3 week and disappeared in that of 5 week. CONCLUSION: We detected caveolin-3 IRs in the myelin sheath of peripheral nerve. Caveolin-3 might play roles in the early myelination of peripheral nerve.
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Animals , Rats , Antibodies , Axons , Caveolin 3 , Fluorescence , Immunohistochemistry , Muscular Diseases , Myelin Sheath , Perfusion , Peripheral Nerves , Regeneration , Sciatic Nerve , SucroseABSTRACT
We investigated glucose uptake and the translocation of Akt and caveolin-3 in response to insulin in H9c2 cardiomyoblasts exposed to an experimental insulin resistance condition of 100 nM insulin in a 25 mM glucose containing media for 24 h. The cells under the insulin resistance condition exhibited a decrease in insulin-stimulated 2-deoxy[3 H]glucose uptake as compared to control cells grown in 5 mM glucose media. In addition to a reduction in insulin-induced Akt translocation to membranes, we observed a significant decrease in insulin-stimulated membrane association of phosphorylated Akt with a consequent increase of the cytosolic pool. Actin remodeling in response to insulin was also greatly retarded in the cells. When translocation of Akt and caveolin-3 to caveolae was examined, the insulin resistance condition attenuated localization of Akt and caveolin-3 to caveolae from cytosol. As a result, insulin-stimulated Akt activation in caveolae was significantly decreased. Taken together, our data indicate that the decrease of glucose uptake into the cells is related to their reduced levels of caveolin-3, Akt and phosphorylated Akt in caveolae. We conclude that the insulin resistance condition induced the retardation of their translocation to caveolae and in turn caused an attenuation in insulin signaling, namely activation of Akt in caveolae for glucose uptake into H9c2 cardiomyoblasts.
Subject(s)
Animals , Rats , Biological Transport , Caveolae/drug effects , Caveolins/metabolism , Cell Membrane/metabolism , Cells, Cultured , Cytosol/metabolism , Enzyme Activation/drug effects , Glucose/metabolism , Heart/embryology , Insulin/pharmacology , Insulin Resistance , Myocytes, Cardiac/drug effects , Phosphorylation , Protein Transport , Protein Serine-Threonine Kinases/metabolism , Proto-Oncogene Proteins/metabolismABSTRACT
OBJECTIVE: Caveolae are the microdomain of the plasma membrane that have been implicated in signal transduction and caveolin is a principal component of the caveolae. Caveolin-3, a family of caveolin related protein, is expressed only in muscle tissue. Here we examined the expression of caveolin-3 in the course of myobalst differentiation and within the muscle tissue. METHOD: L6 cell, rat skeletal myoblast, was cultured in the low mitogen medium and caveolin-3 expression was observed both by immunocytochemistry and western blot analysis. Localization of caveolin-3 within the muscle tissue was investigated and compared to that of dystrophin. RESULTS: While caveolin-3 was not expressed in the proliferating myolast, caveolin-3 was expressed in the differentiated myoblast. Caveolin-3 and dystrophin were co-expressed in the membrane of muscle tissue and integrated density of caveolin-3 was elevated in the area of muscle injury. In the Duchenne muscular dystrophy, caveolin-3 was expressed in the membrane of muscle tissue, but dystrophin was not. CONCLUSION: Caveolin-3 was induced during the myobalst differentiation and its expression was increased during the muscle regeneration. Caveolin-3 was physically associated with dystrophin as a complex, but not absolutely required for the biogenesis of dystrophin complex.