[Calcitonin gene-related peptide induces phenotypic transformation of corpus cavernosum smooth muscle cells in diabetic rats with erectile dysfunction].

OBJECTIVE: 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). METHODS: 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). RESULTS: 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). CONCLUSION: CGRP can transform the phenotype of CCSMs in diabetic ED rats from contractile to synthetic type.

[Role of miR-145 in erectile dysfunction in diabetic rats].

OBJECTIVE: To investigate the role of miR-145 in the corpus cavernosum smooth muscle tissue in the pathogenesis of erectile dysfunction (ED) in diabetic rats. METHODS: The total RNA was extracted from the corpus cavernosum of a diabetic rat model with ED, diabetic rats with normal erectile function and normal rats, and the expression levels of miR145 were compared between the groups. RESULTS: The expression of miR-145 was decreased in the corpus cavernosum of diabetic rats with ED. CONCLUSION: Diabetes mellitus can cause ED in rats, in which process decreased expression of miR145 in the corpus cavernosum may play a role.

[Microarray data analysis of genes related with erectile dysfunction in diabetic rats].

OBJECTIVE: To investigate the changes of gene expression profiles associated with erectile dysfunction in diabetic rats. METHODS: Affymetrix Gene Chip arrays from the Gene Expression Omnibus (GEO) were used to examine the alterations in the gene expression profiles between streptozotocin-induced diabetic rats and littermate controls, and the data were analyzed with GeneSifter microarray analysis software. RESULTS: A total of 661 differentially expressed genes were identified, including 280 up-regulated and 381 down-regulated ones. Among the differentially expressed genes, kruppel-like factor 5 (klf5) was upregulated by 4.01 folds and ceruloplasmin(cp) by 5.14 folds; collagen, type XI, alpha1 was down-regulated by 5.84 folds and collagen, type I, alpha1 by 5.77 folds. The 661 differentially expressed genes involved such functional processes as glycoprotein biosynthesis, collagen fibril organization, angiogenesis in wound healing, triglyceride metabolism, cell proliferation and other important biological processes, and some pathways also involved such as fatty acid metabolism, neurodegenerative disorders, and ECM-receptor interactions. CONCLUSION: Some genes such as klf5, cp, and collagen play important roles in the pathophysiology of diabetes-induced erectile dysfunction. Bioinformatic approaches offer a new means for identifying candidate genes and pathways relevant to the pathophysiology of diabetes-induced erectile dysfunction, highlighting also the potential complexity of this disorder.

[Retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy for treatment of renal and ureteral calculi].

OBJECTIVE: To evaluate the feasibility of retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy through the pelvis for treatment of renal and ureteral calculi. METHODS: In February 2010, 2 patients with renal and ureteral calculi underwent retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy through the pelvis. RESULTS: The operation time in these two cases was 70 and 80 min, and the volume of intraoperative blood loss was about 20 ml. The exposure was excellent, and the patient recovered rapidly without complications or residual calculi. CONCLUSION: Retroperitoneal laparoscopic surgery combined with ureteroscopic lithotomy through the pelvis is feasible for treatment of renal and ureteral calculi.

[A modified tissue culture method for culturing corpus cavernosum smooth muscle cells from the penis of Sprague-Dawley rats].

OBJECTIVE: To culture rat corpus cavernosum smooth muscle cells in vitro using a modified tissue culture method. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Establishment of a cell culture model of corpus cavernosum smooth muscle cells derived from diabetic rats with erectile dysfunction].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.