| ObjectiveDesminopathy is a kind of hereditary protein aggregate myopathy caused by mutations in the desmin gene. Mitochondrial abnormality in the myocytes is one of early pathological characteristics of desminopathy, while its underlying pathogenesis is not completely clear and additional studies are required. Voltage-dependent anion channel 1(VDAC1) is one of important subunits constructing mitochondrial permeability transition pore(m PTP). When the permeability of mPTP increased, multiple mitochondrial apoptogenic factors would be released into the cytosol, leading to the activation of mitochondrial induced-apoptosis cascade. There are no studies clearly articulated whether there is a clear relationship between the VDAC1 and desminopathy. In this study, we observed the expression of VDAC1 and its related apoptosis protein in the animal model and desmin myopathy, which aim to explore the possible mechanism of mitochondrial abnormalities in the muscle cells of the desminopathy. Methods1. Producted adenovirus packaged eukaryotic plasmid of mutant and wild-type desmin gene.2. In the experimental group, the concentration of 1.0 x 1011pfu/ml packaging mutant type desmin gene plasmid of adenovirus by Squared Up method divided into nine microinjection into the 28-35 days old male SD rat's left posterior thigh muscle, each rat total injection is 100?l. Injection of the same volume of the packaging wildtype desmin gene plasmid of adenovirus in to the right posterior thigh corresponds to the point as the control group. 2 rats were sacrificed at the second, seventh,fourteenth,twenty-first,twenty-eighth day after transfection respectively, and the muscle biopsy at the injection point was performed, and the routine staining and desmin immunohistochemical staining were performed. Comparative observation of the days after transfected appears typical desminopathy pathology changes, to determine the model conditions.3. In order to observe the stability of the model. The remained packaging mutant type desmin gene plasmid of adenovirus microinjection of four 28-35-day-old male SD rats. The methods are the same as the above. The muscle biopsy at the injection point was performed, and the routine staining and desmin immunohistochemical staining were performed at the days after transfected appears typical desminopathy pathology changes has been observed above.4. Skeletal muscle routine staining and immunofluorescence double staining of the desmin myopathy patient and the successful animal model was performed, and comparative observation of VDAC1 and its related apoptosis protein expression. Results1. After the first time, the results show that the fourteenth day after transfection that there was a typical desminopathy pathological change in the muscle fibers of 2 SD rats. Take 4 SD rats were repeated modeling, we observed 3 rats appear the typical desminopathy pathological changes in the injection point muscle fibers.2. The skeletal muscle fibers of desmin protein accumulation in desmin myopathy patients and desminopathy animal model were found VDAC1, Bax and ATF2 gather high signal, and the anti apoptotic protein(Bcl-2, Bcl-xl, HK?) was slightly lower or no significant change compared with the normal signal. The Bcl-2, Bcl-xl in the skeletal muscle fibers of desmin protein accumulation in desmin myopathy patients were no significant change compared with the normal signal, and the sign of HK? was slightly lower. The HK?,Bcl-xl in the skeletal muscle fibers of desmin protein accumulation in desminopathy animal model were no significant change compared with the normal signal, and the sign of Bcl-2 was slightly lower.Conclusion1. Packaging mutant type desmin gene plasmid of adenovirus microinjection into the SD rat muscle can successfully simulate pathological of desminopathy.2. Desminopathy may be the result of activation of mitochondrial related pro-apoptotic protein, inhibit the expression of anti-apoptotic protein, stimulate the VDAC1 open, high expression, and then induce apoptosis. |
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