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OBJECTIVE@#To investigate the effects of Naoluo Xintong Decoction (NLXTD) on pyroptosis and angiogenesis of brain microvascular endothelial cells (BMECs) and explore the possible mechanisms in rats with oxygen-glucose deprivation/ reperfusion (OGD/R).@*METHODS@#Rat BMECs with or without caspase-1 siRNA transfection were cultured in the presence of 10% medicated serum from NLXTD-treated rats (or blank serum) and exposed to OGD/R. CCK-8 assay, Transwell chamber assay, and tube formation assay were used to assess proliferation, migration, and tube-forming abilities of the cells. The activity of lactate dehydrogenase (LDH) in the culture supernatant was determined using a commercial assay kit, and the levels of inflammatory factors IL-1β and IL-18 were detected with ELISA. The cellular expressions of pro-caspase-1, caspase-1, NLRP3, Gasdermin D, and angiogenesis-related proteins VEGF and VEGFR2 were detected using Western blotting.@*RESULTS@#The BMECs showed obvious injuries after OGD/R exposure. Compared with the blank serum, the medicated serum significantly improved the cell viability, migration ability, and lumen-forming ability (P < 0.01) and lowered the levels of IL-1β and IL-18 and the LDH release (P < 0.01) of the cells with OGD/R exposure. Western blotting showed that in the BMECs exposed to OGD/R, the medicated serum strongly upregulated the expression of VEGF and VEGFR2 proteins (P < 0.01) and reduced the protein expressions of pro-caspase-1, caspase-1, NLRP3, and Gasdermin D (P < 0.01), and transfection of the cells with caspase-1 siRNA further promoted the expressions of VEGFR2 protein in the cells (P < 0.01).@*CONCLUSION@#NLXTD can improve the proliferation, migration, and tube- forming ability and promote angiogenesis of BMECs with OGD/R injury probably by inhibiting the caspase-1/Gasdermin D pathway in pyroptosis, alleviating cell injury, and upregulating the expressions of VEGF and VEGFR2.
Subject(s)
Animals , Rats , Endothelial Cells , Caspase 1 , Gasdermins , Interleukin-18 , NLR Family, Pyrin Domain-Containing 3 Protein , Vascular Endothelial Growth Factor A , Reperfusion Injury , Brain , Angiogenic Proteins , GlucoseABSTRACT
Aim To investigate the effect of salvianolic acid B (Sal B)on c-Jun N-terminal kinase (JNK)ac-tivation and apoptosis of INS-1 cells induced by inter-mittent high glucose.Methods INS-1 cells were pre-incubated with Sal B for 24 h,followed by exposure to intermittent high glucose (IHG,11.1 mmol·L-1 12 h,33. 3 mmol·L-1 12 h)for 72 h.Cell viability was assessed by MTT assay and cell apoptosis was evalua-ted by flow cytometry.Glucose induced insulin secre-tion capacity and intracellular reactive oxygen species (ROS)contents were measured by enzyme linked im-munosorbent assay (ELISA)and a fluorescent probe DCFH-DA,respectively.Levels of JNK activation and PDX-1 protein expression were determined by Western blot analysis.Results Sal B significantly alleviated IHG-induced cell injury and apoptosis,with glucose induced insulin secretion capacity improved evidently (P<0.05 or P<0.01).Preincubation with Sal B no-tably decreased intracellular ROS and JNK activation in INS-1 cells,while the level of PDX-1 protein was in-creased markedly (P<0.05 or P<0.01 ).Conclu-sion Sal B is capable of ameliorating IHG-induced cell injury and apoptosis in INS-1 cells,which might be derived from suppression of JNK activation and up-regulation of PDX-1 protein expression.
ABSTRACT
<p><b>OBJECTIVE</b>To establish a blood glucose fluctuation model in diabetic rats.</p><p><b>METHODS</b>Male SD rats were randomly divided into 2 groups, namely normal group and diabetes group. Rat model of diabetes was established by single intraperitoneal injection of streptozotocin (STZ) and then divided into sustained high blood glucose group and blood glucose fluctuation group. Rat model of blood glucose fluctuation was established by subcutaneous injection with regular insulin or gavaging of glucose twice daily in diabetic rats. The general condition, body weight, daily blood glucose levels of 5 different times, daily average blood glucose (MBG), standard deviation of daily average blood glucose (SDBG), the maximum amplitude of glycemic excursions (LAGE), fasting serum insulin (FINS) and pancreatic tissue pathology were observed.</p><p><b>RESULTS</b>Rats in blood glucose fluctuation group and sustained high blood glucose group developed symptoms of polyphagia, polyuria and polydipsia. Though significant differences in body weight were observed at different time (P<0.01), no significant differences were found among the three groups (P>0.05). After 6 weeks of blood glucose fluctuation, MBG, SDBG and LAGE in blood glucose fluctuation group and sustained high blood glucose group were increased significantly than those in normal group (P<0.01), the level of FINS decreased markedly (P<0.05). SDBG and LAGE in blood glucose fluctuation group were higher than those in sustained high blood glucose group (P<0.01). Islet of diabetic rat became atrophy, irregular shape, sparse distribution, and decreased in number, and the changes were more obvious in blood glucose fluctuation group.</p><p><b>CONCLUSION</b>Rat model of blood glucose fluctuation can be successfully established by subcutaneous injection of regular insulin of gavage of glucose twice daily in diabetic rats.</p>
Subject(s)
Animals , Male , Rats , Blood Glucose , Body Weight , Diabetes Mellitus, Experimental , Fasting , Glucose , Insulin , Rats, Sprague-Dawley , StreptozocinABSTRACT
To explore the effects of exercise and Danzhi Jiangtang Capsule (DJC), a compound traditional herbal medicine, on the JNK signaling pathway in pancreatic tissues of diabetic rats and to investigate the possible mechanisms of exercise and DJC in treating diabetes.