Naoluo Xintong Decoction activates caspase-1/Gasdermin D pathway to promote angiogenesis of rat brain microvascular endothelial cells after oxygen glucose deprivation/reperfusion injury / 南方医科大学学报

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.

Effect of on proliferation and differentiation of neural stem cells and β-tubulin Ⅲ/GFAP / 南方医科大学学报

OBJECTIVE@#To observe the effects of on the expression of β-tubulin Ⅲ and glial fibrillary acidic protein (GFAP) and the proliferation and differentiation of murine neural stem cells (NSCs) .@*METHODS@#An immortalized murine NSC line was divided into model control (MC) group, 10% drug-containing serum group (NLXT group), and 10% Naoluoxintong drug-containing serum with inhibitor Y27632 group (Y-27632 group) with corresponding treatments. The activity of the NSCs was detected after the treatments using MTT assay, and the migration of the cells was observed with Transwell assay. The expressions of β-tubulin Ⅲ, GFAP and MAP-2 proteins in the cells were detected with immunoblotting, and the expressions of DCX, NEUN, and β-tubulin Ⅲ were also detected with immunofluorescence assay.@*RESULTS@#Compared with that in MC group, the number of migrated cells in NLXT group and Y-27632 group increased significantly at 1 day and 3 days after induction ( < 0.05). The survival rate and the number of migrated cells in NLXT group and Y-27632 group increased significantly on day 7 ( < 0.01). Compared with those in MC group, the expressions of β-tubulin Ⅲ, MAP2 and GFAP protein in NLXT group and Y-27632 group were significantly increased on days 3 ( < 0.01) and 7 ( < 0.05). The numbers of β-tubulinⅢ/ GFAP, BrdU/DCX, and BrdU/NEUN labeled cells in the NLXT group and Y-27632 group were significantly greater than those in the MC group.@*CONCLUSIONS@# promotes the proliferation and differentiation of murine NSCs by regulating the expressions of β-tubulinⅢ/GFAP.

Effects of Naoluo Xintong Decoction on vascular dementia rats' memory and learning, and hippocampal neuronal intracellular calcium concentration / 中成药

AIM To study the effects of Naoluo Xintong Decoction (NLXTD) on vascular dementia (VD)rats' memory and learning,and hippocampal neuronal intracellular calcium concentration.METHODS Rats were divided randomly into model group,NLXTD group (8.54 g/kg),Huperzia-A group (0.06 mg/kg) and sham group.They were made into vascular dementia rats by the improved bilateral carotid artery ligation method (2-VO)thereafter if necessary.After one-month corresponding intragastric administration,the rats were ethologically evaluated by the Morris water maze experiment;their fluorescence intensity of hippocampal neuronal intracellular calcium concentration was determined by flow cytometry,and the expression levels of calcitonin gene related peptide (CGRP) and its hippocampus receptor were detected by enzyme-linked immunosorbent assay (ELISA).RESULTS Compared with the model group,rats in NLXTD group displayed overall superiority to those of the model group in terms of significantly shortened time of escape latency (P ＜0.01),significantly increased number through the platform and the times in the fourth quadrant (P ＜ 0.01),a lower fluorescence intensity indicating a lower hippocampal neuronal intracellular calcium concentration (P ＜ 0.05).CONCLUSION The significant improvement of memory and learning observed among VD rats' due to NLXTD intervention may be attributable to its efficacy in reducing the hippocampal neuronal intracellular calcium concentration by enhancing the expression levels of CGRP and its receptor.