ABSTRACT
Liraglutide is an analog of glucagon⁃like peptide⁃1 and plays an important role in the treatment of diabetes. But the specific mechanism of liraglutide in improving the function of pancreatic β cells is still unclear. Therefore, high glucose (33 mmol/ L) was used to induce islet MIN6 cells for 48 hours to establish a high glucose injury model in this study. The CCK⁃8 assay was used to detect cell viability, and the results showed that the viability of MIN6 cells in the high glucose group decreased (P<0. 05) compared with the control group, and the cell viability increased after liraglutide treatment (P<0. 05). Using the mouse insulin detection kit and ATP content detection kit, we found that both the insulin re⁃ lease (P<0. 01) and ATP content decreased (P<0. 001) in the high glucose group compared with the control group, and after liraglutide treatment both insulin release (P < 0. 05) and ATP content (P < 0. 001) increased. We used the mitochondrial membrane channel pore (MPTP) fluorescence detection kit in MIN6 cells and found that the green fluorescence intensity of the high glucose group was significant⁃ ly decreased (P<0. 001) compared with the control group, and after liraglutide treatment the green fluo⁃ rescence intensity was significantly increased (P<0. 001). The DCFH⁃DA probe combined with flow cy⁃ tometry was used to detect the level of reactive oxygen species (ROS). We found that compared with the control group, the ROS level in the high glucose group was significantly increased (P<0. 001), and de⁃ creased by liraglutide treatment (P<0. 01). Intracellular malondialdehyde (MDA) contents, superoxide dismutase (SOD) and catalase (CAT), and lactate dehydrogenase (LDH) activities in the cell superna⁃ tant were measured, and the levels of MDA and LDH were significantly increased (P<0. 05), and the levels of SOD and CAT were significantly decreased (P<0. 01) in the high glucose group compared with the control group. After liraglutide treatment, the levels of MDA and LDH were decreased (P<0. 05), and the levels of SOD and CAT were increased (P<0. 05). The results of Western blotting showed that the expression of UCP2 was upregulated in the high glucose group (P<0. 01) compared with the control group, and after liraglutide treatment the expression of UCP2 decreased (P<0. 05). The results indica⁃ ted that liraglutide has significant effects in high⁃glucose induced mitochondrial damage, oxidative stress and insulin secretion in MIN6 cells, which will provide a theoretical basis for clinical utilization of liraglu⁃ tide.
ABSTRACT
Genipin (Gen) is an important antioxidant that plays a crucial role in the process of intracellular resistance to oxidative stress. In order to study the effect of genipin on MIN6 cells injured by high glucose, the CCK-8 method was used to detect the cell survival rate. The cell viability of the high-glucose injury group decreased (P <0. 05). But after genipin acted on the cells injured by high glucose, the cell viability increased (P <0. 05). The mouse insulin detection kit and ATP content detection kit found that the insulin release in the high glucose injury group decreased (P < 0. 001) and the ATP content decreased (P <0. 001). After genipin was given, the insulin release increased (P <0. 01), ATP content increased (P <0. 01). The fluorescent probe DCFH-DA detected the intracellular reactive oxygen species (ROS) level and found that the ROS content in the high glucose-injury group was significantly increased (P <0. 01). After genipin was administered, ROS content decreased (P < 0. 05). Glutathione(GSH) / oxidized glutathione (GSSG), intracellular malondialdehyde (MDA), superoxide dismutase(SOD) and catalase (CAT) and lactate dehydrogenase (LDH) activity in the cell supernatant revealed that the GSH / GSSH ratio, SOD and CAT levels in the high glucose injury group decreased (P <0. 05), and the intracellular MDA and LDH levels were significant increased (P<0. 001) .After administration of genipin, the GSH / GSSH ratio, SOD and CAT levels increased (P <0. 01), MDA and LDH levels were significantly reduced (P <0. 01). Mitochondrial membrane potential (MMP) levels decreased in the highglucose injury group (P <0. 01). After genipin acted on the cells injured by high glucose, the MMP level increased (P < 0. 05). Western blot detected uncoupling protein 2 (UCP2), antioxidative proteinsglutathione reductase (GR) and glutaredoxin 1 (Grx1) contents. The results showed that UCP2 contents in the high glucose injury group increased (P <0. 01) and the content of oxidized protein decreased (P < 0. 01). After genipin was administered, the expression of UCP2 decreased (P < 0. 05), and the expression of antioxidative protein increased (P < 0. 05). Experiments suggest that genipin has anantioxidant protective effect on MIN6 cells damaged by high glucose and maintains the function of MIN6cells to promote insulin secretion. This experiment provides experimental data for the antioxidation of genipin on MIN6 cells injured by high glucose, and also provides new ideas for the follow-up study of genipin in the treatment and prevention of diabetes.
ABSTRACT
OBJECTIVE@#To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD).@*METHODS@#The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA).@*RESULTS@#The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05).@*CONCLUSIONS@#As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.
ABSTRACT
Objective To investigate the relationship between the levels of plasma adrenaline and norepinephrine and gene polymorphism of β1 adrenergic receptor G1165C in children with enterovirus 71 (EV71) infection in hand foot and mouth disease (HFMD). Methods The polymerase chain reaction (PCR) was used to detect the expression of gene polymorphism of β1 adrenergic receptor G1165C in vitro. The levels of plasma adrenaline and norepinephrine were measured by enzyme-linked immunosorbent assay (ELISA). Results The plasma norepinephrine level of severe group was significantly higher than the mild group in children with EV71 infection in HFMD (P 0.05); There was no significant difference in the distribution of β1 adrenergic receptor G1165C genotype and allele between EV71 infection group and healthy control group (P > 0.05). Further analysis of EV71 infection group by dividing it into mild and severe groups showed that there was no significant difference in the distribution of genotype and allele between these two groups as well (P > 0.05). There was no significant difference in the levels of epinephrine and norepinephrine in different genotypes of EV71 infection group (P > 0.05), and in the levels of plasma epinephrine and norepinephrine in the mild and severe groups (P > 0.05). Conclusions As the disease gets worse, the plasma norepinephrine level has a rising trend in children with EV71 infection in HFMD, which is an important indicator to evaluate the progress of the disease. However, the gene polymorphism of β1 adrenergic receptor G1165C have no significant correlation, not only with the susceptibility and severity of EV71 infection in hand, foot and mouth disease, but also with the levels of catecholamine.
ABSTRACT
<p><b>OBJECTIVE</b>To study the effects of Geniposide on SNP(sodium nitroprusside)-induced apoptosis of chondrocyte in vitro and cell cycle.</p><p><b>METHODS</b>The chondrocyte of three-week-old SD rats were separated and cultivated. The second generation of chondrocyte cells were involved in experiment. Chondrocyte proliferation was measured by assay; flow cytometer were adopted to observe cell cycle and apoptosis rate; NO examination adopted nitrate reductase method.</p><p><b>RESULTS</b>Geniposide could significantly decrease the percentage of SNP-induced chondrocytes in G0/G1 phase and increased percentage in S phase and G2/M phase. The apoptosis of chondrocyte and the concentration of NO in the culture supernatants was reduced significantly (r=0.917, P<0.01).</p><p><b>CONCLUSION</b>Geniposide could impact SNP-induced apoptosis of chondrocyte by reducing the concentration of NO in the culture supernatants, promoting proliferation of chondrocytes, which is a probable and important mechanism of Geniposide preventing osteoarthritis.</p>