Expression Of VDR And EMP-1 In Nucleus Pulposus Of Human Degenerated Lumbar Intervertebral Disc

Objective: To investigate the expression of vitamin D receptor(VDR)and epithelial membrane protein 1(EMP-1)in human degenerated lumbar disc nucleus pulposus,and to detect the mRNA expression of VDR and EMP-1 in human degenerated disc nucleus pulposus.And to explore the mechanism of human intervertebral disc degeneration in order to provide a new idea for the prevention and treatment of degenerative disc disease caused by disc degeneration.Methods:(1).All the 23 cases of disc nucleus pulposus collected from the Affiliated Hospital of North Sichuan Medical College Orthopaedics January 2016 to August 2016,were divided into control group and experimental group.The control group was diagnosed suffered from lumbar burst fracture,a total of 8 cases,of which 5 males and 3 females,previous health,excluding other chronic diseases,confirmed by imaging no significant disc degeneration.Experimental group diagnosis of lumbar disc herniation,a total of 15 cases,including 10 males and 5 females,previously healthy,the exclusion of other chronic diseases,there are obvious disc degeneration confirmed by imaging,and there is significant low back pain patients.All of these were approved by the Ethics Committee,patients and their families agree and sign.(2).The nucleus pulposus of the same patient was divided into two pieces,one with 4% formaldehyde fixed,conventional dehydrated paraffin embedded,to be HE staining and immunohistochemistry.The other nucleus pulposus tissue was stored in a cryopreserved tube at-80 ° C in the refrigerator,and then subjected to quantitative real-time PCR.The SPSS19.0 software was used to analyze the experimental data.The measured data were expressed as mean ± standard deviation(SD).The t-test was used to compare the two samples,P<0.05 was considered statistically significant.Count data were described by positive rate,Statistical inference using the ?2-test,P<0.05 was considered statistically significant.Results:(1).HE staining and counting of nucleus pulposus cells: In the control group,the nucleus pulposus showed a more uniform distribution of nucleus pulposus cells,irregular lacunae around the nucleus pulposus cells,the cells were oval,more isolated in the cell lacuna.In the experimental group,the cytoplasm of nucleus pulposus cells showed vacuolization,and there were giant clones of nucleus pulposus cells,showing the “nest-like” structure of nucleus pulposus cells.Nucleus pulposus cells in the matrix distribution is uneven,as “nest-like” structure of the nucleus pulposus cells giant clones appear in the disc crack around.The number of nucleus pulposus cells was 88.64 ± 3.16 in experimental group,While it was 174.58 ± 10.23 in control group,it in the experimental group was significantly lower than that in the control group(P<0.05).(2).Immunohistochemistry: The positive cell rates of VDR in the control group and experimental group were 25% and 86.67% respectively.The positive rate of EMP-1 in control group and experimental group was 12.5% and 93.33% respectively.The results showed that the expression of VDR and EMP-1 in the experimental group was significantly higher than that in the control group respectively(P<0.05).(3).Quantitative real-time PCR: The gene amplification and melting curve are favourable.The expression of VDR and EMP-1 mRNA in the experimental group was significantly higher than that in the control group respectively(P<0.05).Conclusions:(1).The number of nucleus pulposus cells in the degenerated lumbar intervertebral disc is significantly reduced.(2).The expression of VDR and EMP-1 in nucleus pulposus of lumbar intervertebral disc was significantly increased.(3).The increase of VDR and EMP-1 expression in the nucleus pulposus of lumbar intervertebral disc may be one of the causes of the imbalance of proliferation and apoptosis of nucleus pulposus in degenerated lumbar intervertebral disc,and it is also one of the important mechanisms of degeneration of lumbar intervertebral disc.