Expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in brain tissue of diabetic rats with ischemia reperfusion

OBJECTIVE@#To discuss the expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in the brain tissue of diabetic rats with ischemia reperfusion.@*METHODS@#A total of 60 male Wistar rats were randomly divided into the normal group, sham group, diabetic cerebral infarction group and single cerebral infarction group according to the random number table, with 15 rats in each group. The high sucrose diet and intraperitoneal injection of streptozotocin were performed for the modeling of diabetic rats, while the thread-occlusion method was employed to build the model of cerebral ischemia reperfusion. The immunohistochemical staining was performed to detect the expression of angiostatin, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the brain tissue.@*RESULTS@#The expression of angiostatin after the reperfusion in the brain tissue of rats in the single cerebral infarction group and diabetic cerebral infarction group was increased 6 h after the reperfusion, reached to the peak on 1 d and then decreased gradually. The expression of angiostatin in the diabetic cerebral infarction group 6 h, 1 d, 3 d and 7 d after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). VEGF began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak at 6 h and then decreased gradually. The expression of VEGF in the diabetic cerebral infarction group at each time point after the reperfusion was significantly lower than that in the single cerebral infarction group (P < 0.05). MMP-9 began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak on 1 d and then decreased gradually. The expression of MMP-9 in the diabetic cerebral infarction group at each time point after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05).@*CONCLUSIONS@#The high glucose environment in which the diabetic cerebral infarction is occurred is to induce the formation of MMP-9 at first and then activate and increase the expression of angiostatin. Afterwards, the expression of VEGF is inhibited, resulting in the poor angiogenesis after cerebral infarction, which thus makes the injury of brain tissue after cerebral infarction even worse than the non-diabetes mellitus.

Expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in brain tissue of diabetic rats with ischemia reperfusion

Objective To discuss the expression and significance of angiostatin, vascular endothelial growth factor and matrix metalloproteinase-9 in the brain tissue of diabetic rats with ischemia reperfusion. Methods A total of 60 male Wistar rats were randomly divided into the normal group, sham group, diabetic cerebral infarction group and single cerebral infarction group according to the random number table, with 15 rats in each group. The high sucrose diet and intraperitoneal injection of streptozotocin were performed for the modeling of diabetic rats, while the thread-occlusion method was employed to build the model of cerebral ischemia reperfusion. The immunohistochemical staining was performed to detect the expression of angiostatin, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the brain tissue. Results The expression of angiostatin after the reperfusion in the brain tissue of rats in the single cerebral infarction group and diabetic cerebral infarction group was increased 6 h after the reperfusion, reached to the peak on 1 d and then decreased gradually. The expression of angiostatin in the diabetic cerebral infarction group 6 h, 1 d, 3 d and 7 d after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). VEGF began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak at 6 h and then decreased gradually. The expression of VEGF in the diabetic cerebral infarction group at each time point after the reperfusion was significantly lower than that in the single cerebral infarction group (P < 0.05). MMP-9 began to be increased 1 h after the reperfusion in the single cerebral infarction group and diabetic cerebral infarction group, reached to the peak on 1 d and then decreased gradually. The expression of MMP-9 in the diabetic cerebral infarction group at each time point after the reperfusion was significantly higher than that in the single cerebral infarction group (P < 0.05). Conclusions The high glucose environment in which the diabetic cerebral infarction is occurred is to induce the formation of MMP-9 at first and then activate and increase the expression of angiostatin. Afterwards, the expression of VEGF is inhibited, resulting in the poor angiogenesis after cerebral infarction, which thus makes the injury of brain tissue after cerebral infarction even worse than the non-diabetes mellitus.

Effect of non-contact coculture on bone marrow stromal cells and neural stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of non-contact coculture on bone marrow stromal cells (MSCs) and neural stem cells (NSCs) in neural stem cell culture medium.</p><p><b>METHODS</b>MSCs and NSCs were cultured in non-contact coculture in Transwell plate, and cell morphology and immunocytochemical profile were investigated.</p><p><b>RESULTS</b>In the coculture, the NSCs showed adhering growth and extended long processes, and the migrating cells formed a network of cells within 7 days. The cells differentiated into neurons, astrocytes and oligodendrocytes as shown by immunocytochemistry. Most of the MSCs grew in a non-adherent manner, giving rise to large spherical cell masses which expressed neuronal, astrocyte, and oligodendrocyte phenotypes. In the control group, the NSCs grew in suspension, some of MSCs formed small non-adherent spherical cell masses, while some cells showed adherent growth.</p><p><b>CONCLUSION</b>MSCs and NSCs in the non-contact coculture can mutually promote the cell differentiation into neural cells in neural stem cell culture medium, indicating that both MSCs and NSCs can secrete some neurotrophic factors to provide a microenvironment suitable for survival and differentiation for each other.</p>

Effects of Ciwujia in inducing marrow stromal cell differentiation into neuron-like cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of the traditional Chinese herbal drug Ciwujia in inducing the differentiation of marrow stromal cells (MSCs) into neuron-like cells.</p><p><b>METHODS</b>Rat MSCs isolated from the whole bone marrow were amplified by adherent culture in vitro and induced to differentiate into neuron-like cells using serum-free DMEM/F12 containing Ciwujia. The protein and mRNA expressions of nestin, beta-Tubulin III and glial fibrillary acidic protein (GFAP) in the differentiated cells were detected by indirect immunofluorescence method, Western blotting and RT-PCR.</p><p><b>RESULTS</b>The third-passage MSCs showed positive expression rates for CD44 and CD54 beyond 90% with decreased CD14 expression rate to 2.37%. Induction by Ciwujia of the MSCs resulted in cell body shrinkage and protrusion of the cell processes resembling those of neurons. The differentiated cells were positive for nestin and beta-Tubulin III expression and negative for GFAP as shown by immunofluorescence assay, Western blotting and RT-PCR.</p><p><b>CONCLUSION</b>Ciwujia can induce the differentiation of rat MSCs into neuron-like cells in vitro.</p>

Epidemiological investigation on natural infection to Borna disease virus (BDV) among horses in Yili,Xinjiang / 中华流行病学杂志

Objective To investigate the epidemiological pattern of Borna disease virus (BDV)infection in horses and to analyze the phylogenetic tree of derived BDV in Yili, Xinjiang. Methods We established a modified nested RT-PCR (nRT-PCR) to detect BDV p24 segment in peripheral blood mononuclear cells (PBMCs) and brain tissues of 120 horses in Yili, Xinjiang. Positive products were analyzed by sequencing and homology analysis. Results The positive rate of BDV infection was 2.5% in both PMBCs and brain tissues at the same time. The gene sequence revealed in positive PCR samples was more than 93 % ,identical to that of BDV derived from horses in other countries. We also noticed a high degree of identity ( >98 % ) to standard strain He/80 in gene sequence of positive PCR samples. Conclusion Our study found the presence of BDV natural infection in horses in Yili. The endemic BDV had a high degree of identity to standard strain He/80.

Study of mechanism on differentiation of marrow stroma cells into neuron-like cells induced by Ciwujia injection in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study whether the nuclear factor-kappa B (NF-kappaB) play a role in differentiation of marrow stroma cells (MSCs) into neuron-like cells induced by Ciwujia injection in vitro.</p><p><b>METHOD</b>Rats were randomly divided into control group and Ciwujia induced group. Cell morphology was observed with invert microscope; MSCs phenotype, expression of neural marker protein and nucleus translocation of NF-kappaB subunit RelA (p65) after induction were observed by immunofluorescence; Expression of IkappaBalpha in Ciwujia induced group and control group was detected by Western blot.</p><p><b>RESULT</b>The decreased volume of some MSCs, strengthened three-dimensional structure and shrinkage of cell body with formation of spire or round was observed after induction by Ciwujia for 1 hour. After 5-hour exposure to Ciwujia, changes in the morphology of MSCs were as follows: appearance of more triangular or round shapes cells, retraction of microfilament in MSCs, formation of slender process from pseudopod with reticular structure, exfoliation and necrosis. After 7-day induction, most MSCs became triangular and round shapes, exhibiting a typical neuronal structure. Immunofluorescence method showed that nestin expression displayed intensively positive in Ciwujia group at 1 hour, and still remain positive at 7 day. However, beta-III Tubulin expression was weakly positive at 1 hour and intensive positive at 3 and 5 hour. It was still positive at 7 day. NSE expression was weakly positive at 3 hour and intensive positive at 5 hour and 7 day. GFAP expression was negative. However, in the control group, every expression of marker protein was negative or weakly positive. Expression of p65 was in cytoplasm. Western blot method showed IkappaBalpha protein relative IA was 1.000 before induction. It was 0.4556 +/- 0.1008 after induction in Ciwujia group and decreased significantly than before induction (P < 0.01). It was 0.0296 +/- 0.0099 after induction in control group and decreased significantly than before induction and in Ciwujia group (P < 0.01).</p><p><b>CONCLUSION</b>The data showed that MSCs can induce differentiation into neuron-like cells by Ciwujia, and Ciwujia could inhibit the translocation of NF-kappaB from the cytoplasm to the nucleus, which may be relate to the differentiation of MSCs into neuron-like cells.</p>