IL-1? On Rat Endplate Chondrocytes Study On The Effct And Mechanism Of Proliferation

Objective: Through the experiments of cultured rat endplate cartilage cells,to detect whether the expression level of ADAMTS-5 and IL-1? was associated with IL-1?,inflammatory factor intervention,through the detection of phosphorylated p65 protein,p65 protein content change,to explore whether IL-1? by promoting the expression of ADAMTS-5 NF-KB signal pathway.In order to promote the endplate chondrocyte degeneration,thus providing new theories and methods for the prevention and treatment of intervertebral disc degeneration.Methods: The 4-6 week old rat vertebral endplate chondrocytes were purified by primary and subculture.The cell morphology was observed,the 3-generation cells with better growth were selected and the cell growth curve was depicted.1.The effects of different concentrations of IL-1? on the proliferation of rat endplate chondrocytes were studied by MTT.The light absorption values of each hole were measured on the ultraviolet spectrophotometer,and the time was the transverse coordinate and the absorption value was the ordinate,and the cell proliferation was observed.2.The effects of different concentrations of IL-1?(5 ng/ml,10 ng/ml,20 ng/ml,40 ng/ml)on the expression of ADAMTS-5 protein in the rat endplate cartilage cells were studied.3.Ten ng/mL IL-1? was added to the Petri dish to observe the changes in the expression of ADAMTS-5 protein in the vertebral lamina chondrocytes at 24,48 and 96 hours.4.Ten ng/mL IL-1? was added into Petri dishes,and blank control was established.By detecting the changes of phosphorylation of p65 protein and p65 protein content,we explored whether IL-1? promoted ADAMTS-5 expression through activating NF-?B signaling pathway.5.Rat endplate cartilage cells were divided into 4 groups,the first group was the blank control group,10 ng/mL with second IL-1? 50 ug/mlNF-,B pathway blocker SN50 group with third,10 ng/mL in fourth group IL-1? and 50 ug/ml SN50,to observe the expression of ADAMTS-5 variation,in order to verify IL-1? promotes ADAMTS-5 expression through activation of NF-?B signaling pathway.Results:1.With the increase of IL-1? concentration,the number of endplate chondrocytes in the rat gradually decreased,and the difference between each group was statistically significant(P < 0.05).2.Different concentrations of IL-1? increased the expression of ADAMTS-5 protein in rat endplate chondrocytes compared with blank control group,and the difference between groups was statistically significant(P < 0.05).The IL-1? effect of 10 ng/ml is the strongest.3.At different time points,the expression of ADAMTS-5 of 10 ng/ml IL-1? on rat endplate chondrocytes increased compared with the blank control group,and the difference between groups was statistically significant(P < 0.05),and reached the peak at 48 hours.4.Compared with the blank,the concentration of p-p65 protein increased significantly after 48 hours of 10 ng/ml IL-1? endplate chondrocytes,while the concentration of p65 protein remained unchanged.The difference between the two groups was statistically significant(P < 0.05).5.Four groups of different intervention proteins acted on rat endplate chondrocytes respectively.The expression level of ADAMTS-5 protein from high to low was IL-1? group,IL-1? + SN50 group,blank control group and SN50 group.The difference between each group was statistically significant(P < 0.05).Conclusions:1.IL-1? may inhibited the proliferation of endplate chondrocytes in rats.2.The inhibitory effect of IL-1? on the fine increment of rat endplate cartilage may be partly achieved by affecting NF-?B signal pathway.