ABSTRACT
To determine the effect of andrographolide (Andro) on angiogenesis of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were treated with different concentrations of Andro and the cell viability was detected with Cell Counting Kit-8 (CCK-8). HUVECs were treated with half lethal dose (IC50) of Andro. Matrigel was used to make capillary formation of HUVECs and the effect of Andro on capillary formation was evaluated by calculating the percentage of capillary formation. Moreover, the effects of Andro and the supernatant from cultured A549 tumor cells on capillary formation were evaluated by calculating the percentage of capillary formation. The effect of Andro on the expression of matrix metalloproteinase-9 (MMP-9) was determined with Western blot. Results: The cell viability of HUVECs decreased with the increase of Andro concentrations. IC50 was 20 μmol/L. The capillary formation of HUVECs was inhibited when treated with 20 μmol/L Andro for 24 hours. Moreover, Andro was able to antagonize the promotion of the capillary formation induced by the supernatant from cultured tumor cells. Andro could suppress the expression of MMP-9 and antagonize the capillary formation. Conclusion: Andro inhibits the capillary formation of HUVECs and can antagonize the promotion of angiogenesis induced by the supernatant from cultured tumor cells.
Subject(s)
Humans , Capillaries , Cell Survival , Collagen , Culture Media , Diterpenes , Pharmacology , Drug Combinations , Human Umbilical Vein Endothelial Cells , Laminin , Matrix Metalloproteinase 9 , Metabolism , Neovascularization, Pathologic , Proteoglycans , Tumor Cells, CulturedABSTRACT
Objective:To investigate the effect of human growth hormone releasing hormone receptor splice variant type 1 (GHRHR SV1) on the proliferation of human liver cancer HepG2 cells,and to clarify the proliferation effect of GHRHR SV1 on the human cancer cells.Methods:The GHRHR SV1 plasmids were transfected into the human HepG2 cells to construct the HepG2-SV1 cell line.HepG2 group(HepG2 cells),HepG2-empty group(HepG2-pcDNA3.0 cell line) and HepG2-SV1 group(HepG2-SV1 cells) were set up.PCR and Western blotting methods were used to identify the HepG2-SV1 cell line;CCK-8 method was used to detect prolifernation rate of cells;colony formation assay was used to detect the colony formation rate of cells;cell wound healing assay was used to evaluate the migration rate of cells.Results:The PCR and Western blotting results showed the HepG2-SV1 cell line expressed GHRHR SV1 steadily.The CCK-8 results showed that the proliferation rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The colony formation assay results showed that the colony formation rate of HepG2-SV1 cells in HepG2-SV1 group was 3.5 times higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).The cell wound scratch assay results showed that the migration rate of the HepG2-SV1 cells in HepG2-SV1 group was higher than that of the HepG2-pcDNA3.0 cells in HepG2-empty group(P<0.05).Conclusion:GHRHR SV1 could increase the proliferation of HepG2 cells.
ABSTRACT
This research is to construct a basic medical sciences’experimental teaching system in order to cultivate innovative talents. It is guided by cultivating innovative practical ability and post compe-tence and implements a teaching mode with “five combinations”by integrating teaching resources, strength-ening interdisciplinary combination , integrating curriculum and organ systems , and optimizing teaching modules and experiment content. A preliminary personnel training mode and experimental teaching system have been constructed for innovative talent cultivation, and correspondently a diversified experiment exami-nation system and teaching quality monitoring system have been constructed through teaching practice, which aims at continuously improving experiment teaching quality and talent training quality.
ABSTRACT
Growth hormone (GHRH) and pitutary adenylate cyclase activating peptide (PACAP) are the members of the PACAP/Glucagon superfamily,who are related in both sequence and function.Their stimulation of GH secretion and animal growth is concerned.A series of expression plasmid,pIRES1-GHRH-PACAP (P-G-P),plRESI-GHRH (P-G) and plRESI-PACAP(P-P),were constructed,extracted and purified,then transfected into CHO cell line with Lipofectamine.The expression was examined by RT-PCR,dot-ELISA and Western blotting.The biological activity of expression products was detected in rats.At 8 h after injection of transfection supematant,serum IGF-I concentrations in P-G-P group were significantly higher than that in other groups(P ＜ 0.05).PLGA encapsulating plasmid microspheres were prepared and injected intramuscularly into rabbit legs.Growth behavior and IGF-1 level were measured at day 0,15,30 and 45 after injection.Greater body weights gain and higher serum 1GF- [ levels were observed in three plasmid microsphere injection groups,compared with control group.At day 30,the body weight gain in P-G-P group was greater than saline group (81%,P＜ 0.01),P-G mierosphere group (15%,P＜ 0.05) and P-P microsphere group (7%,P＞ 0.05),serum IGF-I concentration in P-G-P microsphere group showed a 16.68% increase to P-G microsphere (P ＞ 0.05),a 17.14% increase to P-P microsphere(P ＞ 0.05) and a 50.46% increase to control (P ＜ 0.05).These results suggest that co-expression of GHRH and PACAP in one expression plasmid might exert an additive stimulation of GH secretion and growth when delivered into rabbit skeletal muscle with PLGA mierosphere.The results may provide a new approach to regulate animal growth.