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1.
National Journal of Andrology ; (12): 913-917, 2011.
Article in Chinese | WPRIM | ID: wpr-305764

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of the calcitonin gene related peptide (CGRP) on the phenotypic transformation of corpus cavernosum smooth muscle cells (CCSM) in diabetic rats with erectile dysfunction (ED).</p><p><b>METHODS</b>Models of diabetes and diabetic ED were established in male Sprague-Dawley rats by administration of streptozotocin, and CCSMs were primarily cultured and subjected to immunocytochemical assay. The cells were divided into a diabetic ED and a normal control group, and exposed to 0, 10, 60 and 100 nmol/L of CGRP for 24 hours. Then the relative expressions of calponin 1 (Cnn1) and osteopontin (OPN) mRNA were determined by real-time fluorescence quantitative RT-PCR (qRT-PCR).</p><p><b>RESULTS</b>The rate of SMalpha-actin positive cells in the CCSMs was (95.94 +/- 0.03) %. The expression of Cnn1 mRNA was significantly lower while that of OPN mRNA remarkably higher in the diabetic ED rats (4.41 +/- 0.29 and 5.28 +/- 0.32) than in the normal controls (10.35 +/- 0.62 and 1.32 +/- 0.24) (P < 0.01). Exposure to 100 nmol/L of CGRP significantly upregulated the expression of Cnn1 mRNA and downregulated that of OPN mRNA as compared with the unexposed rats (6.9 +/- 0.22 vs 4.41 +/- 0.29 and 3.26 +/- 0.31 vs 5.28 +/- 0.32, P < 0.01).</p><p><b>CONCLUSION</b>CGRP can transform the phenotype of CCSMs in diabetic ED rats from contractile to synthetic type.</p>


Subject(s)
Animals , Male , Rats , Calcitonin Gene-Related Peptide , Pharmacology , Calcium-Binding Proteins , Metabolism , Cells, Cultured , Diabetes Mellitus, Experimental , Genetics , Metabolism , Erectile Dysfunction , Genetics , Metabolism , Microfilament Proteins , Metabolism , Muscle, Smooth , Cell Biology , Metabolism , Osteopontin , Metabolism , Penis , Cell Biology , Metabolism , Phenotype , Rats, Sprague-Dawley
2.
Journal of Southern Medical University ; (12): 494-497, 2010.
Article in Chinese | WPRIM | ID: wpr-355092

ABSTRACT

<p><b>OBJECTIVE</b>To culture rat corpus cavernosum smooth muscle cells in vitro using a modified tissue culture method.</p><p><b>METHODS</b>Fifteen male rats were randomized into 3 equal groups, namely enzyme digestion group, tissue culture group, and modified tissue culture group. The penis of the rats was separated carefully and cut into small pieces, and seeded onto culture flasks and cultured in complete medium consisting of DMEM containing 20% fetal calf serum at 37 degrees C; in a humidified atmosphere with 5% carbon dioxide. The cells growth was observed under phase contrast microscope and the smooth muscle cell specific proteins alpha-SM-actin and desmin were identified immunohistochemically.</p><p><b>RESULTS</b>The alpha-SM-actin-positive cell rate was 96.3% in rat corpus cavernosum smooth muscle and 23.8% in the fibroblasts, and the corpus cavernosum smooth muscle contained 74.4% desmin-positive cells while the fibroblasts showed no desmin positivity. Significant difference was found in the positive cell rate for desmin among the 3 groups, with the highest positive cell rate occurred in modified tissue culture group.</p><p><b>CONCLUSION</b>Desmin may serve as a marker for identifying corpus cavernosum smooth muscle cells. The modified tissue culture method can result in highly purified corpus cavernosum smooth muscle cells with intact structure and functions.</p>


Subject(s)
Animals , Male , Rats , Actins , Biomarkers , Cell Proliferation , Desmin , Muscle, Smooth , Cell Biology , Penis , Cell Biology , Rats, Sprague-Dawley , Tissue Culture Techniques
3.
Journal of Southern Medical University ; (12): 351-354, 2010.
Article in Chinese | WPRIM | ID: wpr-269553

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the method for culturing corpus cavernosum smooth muscle cells (CCSMs) derived from diabetic rats with erectile dysfunction (ED) for the study of ED caused by diabetes.</p><p><b>METHODS</b>CCSMs were isolated from the corpus cavernosum of diabetic rats with ED and cultured using a modified method of adherent tissue culture. The cultured cells were identified by immunohistochemistry and the cell morphology and proliferation were observed.</p><p><b>RESULTS</b>The primary culture of CCSM was performed successfully, and the cells were seen to migrate from the small tissue pieces 3 days later, reaching nearly confluence in 16-18 days. A typical "hill-valley" growth pattern was noted in the cell passaging. Immunohistochemical staining for alpha-smooth muscle actin (alpha-SM-actin) and desmin yielded positive results in the cells.</p><p><b>CONCLUSION</b>The modified method for adherent tissue culture is convenient and reliable in establishing the in vitro cell culture model of CCSMs from diabetic rats with ED, and the cultured CCSMs display a faster proliferation than normal CCSMs. No obvious differences in the cell morphology can be found between diabetic and normal CCSMs under light microscope.</p>


Subject(s)
Animals , Male , Rats , Cell Culture Techniques , Cells, Cultured , Diabetes Mellitus, Experimental , Pathology , Erectile Dysfunction , Pathology , Models, Biological , Myocytes, Smooth Muscle , Cell Biology , Physiology , Penile Erection , Penis , Cell Biology , Rats, Sprague-Dawley
4.
Journal of Southern Medical University ; (12): 925-928, 2009.
Article in Chinese | WPRIM | ID: wpr-268809

ABSTRACT

<p><b>OBJECTIVE</b>To validate the hypothesis that the phenotypic transformation occurs in the smooth muscle cells in the corpus cavernosum of hypertensive rat and explore its impact on the erectile function of rats.</p><p><b>METHODS</b>Eighteen 16-week-old male spontaneously hypertensive rats (SHR) and 10 syngeneic normotensive rats (WKY) were used in this experiment. After measurement of systolic blood pressure of the caudal artery and examination of the erectile function with subcutaneous injection of apomorphine (APO), the rats were divided into 3 groups, namely hypertensive with erectile dysfunction (HBP-ED) group (n=6), hypertensive (HBP) group (n=12) and control group (n=10). Immunohistochemical staining and color image analysis system were used to observe expression of calponin 1 and osteopontin (OPN) in rat corpus cavernosum. Real-time quantitative RT-PCR was used to determine the expression of calponin 1 and OPN mRNAs in different groups.</p><p><b>RESULTS</b>The expressions of calponin 1 protein and mRNA were the highest in the control group and the lowest in HBP-ED group, while the expressions of OPN protein and mRNA were the highest in HBP-ED group and the lowest in the control group.</p><p><b>CONCLUSION</b>The smooth muscle cells may transform from the contractile phenotype into synthetic phenotype in the corpus cavernosum of the hypertensive rats, resulting ultimately in erectile dysfunction.</p>


Subject(s)
Animals , Male , Rats , Calcium-Binding Proteins , Genetics , Metabolism , Erectile Dysfunction , Pathology , Hypertension , Pathology , Microfilament Proteins , Genetics , Metabolism , Myocytes, Smooth Muscle , Pathology , Osteopontin , Genetics , Metabolism , Penis , Pathology , Phenotype , RNA, Messenger , Genetics , Metabolism , Rats, Inbred SHR
5.
Journal of Applied Clinical Pediatrics ; (24)1992.
Article in Chinese | WPRIM | ID: wpr-638318

ABSTRACT

Objective To study the difference in therapeutic effects and side effects of ibuprofen versus indomethacin for symptomatic patent ductus arteriosus (PDA) in the premature infants.Methods Meta analysis was used to qualitatively and quantitatively evaluate the data extracted from 6 public papers about comparative study of ibuprofen and indomethacin.Results The rate of ductal closure was similar with the two treatment regimes (intravenous ibuprofen and indomethacin).In side effects on PDA,the incidence of oliguria induced by ibuprofen was significantly lower than that of indomethacin though there were no difference in other side effects.Conclusions The efficacy of ibuprofen for the early treatment of PDA in preterm infants is similar with indomethacin,and has low incidence of oliguria.

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