ABSTRACT
Objective To observe the effects of high fat dietinduced elevation of blood glucose on the microvascular function of testis and male reproduction in C57BL/6 mice. Methods A total of 40 male C57BL/6 mice were randomly divided into control group and high fat diet (HFD) group (n =20). The mice in HFD group were fed with high fat diet for 20 weeks. Blood glucose and body weight were measured weekly. The permeability of bloodtestis barrier was evaluated by intraperitoneal injection of Evans blue. The blood flow of testicular microcircu-lation and the frequency and amplitude of microvascular vasomotion were detected by laser Doppler blood flow ima-ging system. The morphology of testicular tissue was observed by HE staining. The expressions of platelet- endothelial cell adhesion molecule-1 (PECAM-1/CD31) in testicular microvascular endothelial cells and prolifera-ting cell nuclear antigen (PCNA) in spermatogenic cells were detected by immunohistochemistry. The apoptosis of spermatogenic cells was observed by TUNEL staining. Results The body weight and blood glucose of HFD group were significantly higher than those in control group (P＜0.01). Evans blue staining showed that the integrity of blood-testis barrier of HFD group was damaged, and increased permeability was observed in seminiferous tubules. In HFD group, the mean blood flow of testis and the frequency and amplitude of microvascular vasomotion were sig-nificantly lower than those in control group (P＜0.01). The number of spermatogenic epithelial cells and the thickness of seminiferous epithelium decreased. The expressions of CD31 in microvascular endothelial cells and PCNA in spermatogenic cells were significantly lower in HFD group than those in control group (P＜0.01). The apoptosis level of spermatogenic cells was higher than that in the control group (P <0.01). Conclusions Increased blood glucose level induced by high fat diet in mice can impair the testicular microvasculature and damage the integrity of blood-testis barrier and injure the structure of seminiferous epithelium in mice.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide(W7) on the differentiation from human adipose-derived mesenchymal stem cells (hADSCs) to endothelial cells.</p><p><b>METHODS</b>hADSCs were cultured with serum-free differential medium containing 40 ng/ml vascular endothelial growth factor (VEGF) and 10ng/ml basic fibroblast growth factor (bFGF). Cells were divided into control group (differential medium without W7), high-dose group (containing 30 μmol/L W7), medium-dose group (containing 20 μmol/L W7), and low-dose group ( containing 10 μmol/L W7). The hADSCs were cultured for 8 days, and then the changes in the phenotypes of von Willebrand factor (vWF) and vessel-selective cadherin (VE-Cadherin) were detected by flow cytometry (FCM). The intracellular Ca(2+) labeled with Fluo-3 was detected by laser confocal microscopy. After hADSCs planting on Matrigel, their angiogenic potentials were observed under the inverted phase contrast microscope, and the expression of extracellular regulated kinase (ERK) and phosphorylated extracellular regulated kinase (p-ERK) were evaluated by Western blot.</p><p><b>RESULTS</b>After the hADSCs were cultured for 8 days, compared with the control group, the expressions of vWF and VE-Cadherin significantly increased along with the decrease of W7 level and the intracellular Ca(2+) also significantly increased (Pü0.01). Lumina-like vascular structure was formed in W7 treatment groups, but not in the blank control group. Compared with the blank control group, the expression of ERK showed no significant in W7 treatment groups (high-, medium-, and low-dose groups)(P>0.05); however, along with the decrease of W7 levels, the expression of p-ERK significantly increased(P<0.05).</p><p><b>CONCLUSION</b>W7 in proper levels can effectively induce the differentiation from hADSCs to endothelium by increasing the intracellular Ca(2+) level and thus activating the ERK/MAPK pathway.</p>
Subject(s)
Humans , Adipose Tissue , Cell Biology , Cell Differentiation , Cells, Cultured , Endothelial Cells , Cell Biology , Metabolism , Mesenchymal Stem Cells , Cell Biology , Metabolism , Sulfonamides , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To study the number of CECs in patients with acute myocardial infarction (AMI) and unstable angina (UA), and to investigate its relationship with inflammatory related cytokines.</p><p><b>METHODS</b>37 patients with AMI, 12 patients with UA, and 42 health controls were studied. CECs were isolated from peripheral blood by using of immunomagnetic beads coated with antibodies against CD146. Their endothelial origin was confirmed by the positive labelling of von Willebrand Factor (vWF), CD31 and electron microscope. Annexin V-FITC/PI kit was used to measure the apoptosis of CECs. Inflammatory related cytokines were analyzed turbidimetrically or ELISA using of commercially available testing kit.</p><p><b>RESULTS</b>CECs number was significantly higher in AMI and UA patients [medians (interquartile range) were 52 (28 approximately 81.5) cells/ml and 29 (18 approximately 61) cells/ml respectively] compared with health control [10.5 (6-16.5)cells/ml, P < 0.001]. After excluding diabetes patients, the number of CECs and CRP in AMI and UA group (n = 26) were still significantly higher than controls. The necrotic rate of CECs in AMI and UA was significantly higher than controls (P < 0.01). Correlation analysis revealed a significant positive correlation between CECs and CRP, or IL-6 (r = 0.677, 0.316, P = 0.000, 0.002). The multivariate linear regression analysis showed that CRP and Diabetes increased the number of CECs significantly (OR = 0.620, 0.164, 95% CI 3.985-6.751, 0.301-21.877, P = 0.000, 0.044).</p><p><b>CONCLUSION</b>The mechanism responsible for the increase of CECs in acute coronary disease may be due to the vessel injury caused by inflammation.</p>