The Study Of The Resveratrol Inhibited Intervertebral Disc Degeneration Induced By Compressive Stress And It's Correlation To The ERK Expression In The Intervertebral Disc Organ Culturing Model In Vitro

The first part Evaluation of ERK expression in nucleus pulposus cells of intervertebral disc degenerationObjective: Study the expression of ERK in intervertebral disc degeneration(IVDD)tissue.Methods: 10 degenerative(experimental group)and 7 normal(control group)disc nucleus pulposus tissues were taken from inpatients undergoing intervertebral disc surgery.(1)Histological observation of nucleus pulposus in degenerative and normal intervertebral discs tissue.(2)The expression of ERK in degenerative intervertebral disc tissue and normal intervertebral disc tissue was detected by real-time fluorescence quantitative PCR(RT-qPCR).(3)The expression of ERK protein in the two groups was detected by Western blot.Results:(1)The morphology and number of nucleus pulposus cells in degenerative tissue of intervertebral disc changed,and the changes of nucleus pulposus cells advanced synchronized with the progress of IVDD.(2)The expression of ERK gene was significantly increased in IVDD tissue.(3)Western blot showed that ERK protein increased significantly with the aggravation of IVDD.Conclusions:(1)In the degenerative tissue of intervertebral disc,the morphology and number of nucleus pulposus cells change,that is,the change of nucleus pulposus cells synchronizes with the IVDD.(2)The expression of ERK was increased in IVDD,and the expression of ERK was related to the degree of disc degeneration.The second part Expression of ERK in IVDD nucleus pulposus cells under high pressure load in intervertebral disc organ culture modelObjective: Established culture model of intervertebral disc organs,and the expression of ERK in nucleus pulposus cells of IVDD was detected by applying pressure load to promote degeneration.Methods:(1)Established rat intervertebral disc organ culture model and applied high pressure load to promote degeneration;(2)Detected the survival rate of nucleus pulposus cells in cultured model of IVDD organs;(3)Detected the expression of ERK in IVDD tissue cells after organ culture by RT-qPCR;(4)The expression of ERK protein in IVDD tissue cells in the control group and the pressurized group was detected by Western blot on the 7th and 14 th days of culture.Results:(1)With the prolongation of culture time,the tissue degeneration in intervertebral disc culture model was gradually aggravated,and high pressure could accelerate this process;(2)The activity of nucleus pulposus cells in the high pressure load group was lower than that in the low pressure group(P<0.05);(3)The expression of ERK gene in nucleus pulposus cells of intervertebral disc was significantly increased in high pressure load group(P<0.05);(4)The expression of ERK protein in nucleus pulposus cells of intervertebral disc was increased in high pressure loading group(P<0.05).Conclusions:(1)As the pressure is increased and the duration of action increases,the apoptosis of the nucleus pulposus cells of the intervertebral disc is increased;(2)The cell vitality of the group exposed to higher pressure was significantly lower than that of the group with lower pressure.(3)The gene and protein expression levels of ERK in the pressure group were significantly higher than those in the lower pressure group.The third part The relationship between resveratrol inhibition of IVDD induced by high compression stress and expression of ERKObjective: Study the effect of resveratrol(RES)on intervertebral disc culture organs under high pressure load.Method:Established intervertebral disc organ culture model of male SD rats,which were randomly divided into control group,intervertebral disc degeneration group and resveratrol drug intervention group with 10 rats in each group.(1)Compared the effects of RES on tissue changes in intervertebral disc culture organs under high pressure load;(2)Compared the survival rate of cells in the three groups of rat intervertebral disc organs culture at each time point;(3)The apoptosis of the cells in the three groups of rat intervertebral disc organs culture was compared at each time point.(4)Compared the gene expression of ERK in three groups of nucleus pulposus cells;(5)Compared the protein expression of ERK in three groups of nucleus pulposus cells.Results:(1)Under high pressure load,the degeneration of intervertebral disc culture tissues was obvious,and RES drug intervention could reduce the degree of degeneration.(2)The cell survival rate of RES intervention group was between the control group and the IVDD group;(3)The apoptosis of IVDD group was more serious than that of control group,while RES drug intervention group was between the two groups.(4)The mRNA expression of ERK in IVDD model group,RES intervention group and control group decreased in turn,and there was significant difference among the three groups(P<0.05);(5)The protein expression level of ERK decreased gradually in IVDD model group,RES drug intervention group and control group,and there was significant difference among the three groups(P<0.05).Conclusion:RES could reduce the apoptosis induced by pressure load and protect the intervertebral disc,which may be related to the inhibition of ERK overexpression.