Experimental Study On The Effect Of Proteomics 70kDa Heat Shock Protein 8 On Human Intervertebral Disc Degeneration

Objective: To investigate the expression of 70 kDa heat shock protein 8(Heat shock 70 kDa protein 8,HSPA8/Heat shock protein 70,HSC70)in normal and different degrees of degeneration lumbar intervertebral disc nucleus pulposus.The role of HSPA8 in the degeneration of human lumbar intervertebral disc was evaluated,and the relationship between the degree of intervertebral disc degeneration and the expre-ssion of HSPA8 was analyzed.Methods: From June 2017 to April 2018,a total of 72 specimens were collected from patients with lumbar disc surgery in our hospital.Among them,18 cases were used as control group(nucleus pulposus tissue from adolescent patients with scoliosis),54 cases were experimental group(taken from the nucleus pulposus of patients with lumbar disc degeneration).According to Spengler and preoperative MRI,lumbar intervertebral disc degeneration was divided into four types: the normal group?protrude group?prolapse group and the sequester group.Inclusion criteria:(1)After hospitalization,the diagnosis was confirmed by MRI in our hospital.(2)In line with the diagnostic criteria of lumbar disc herniation.(3)Patients or their families signed the informed consent voluntarily.Exclusion criteria:(1)Women in lactation and pregnancy.(2)Complicated infection patients.(3)Patients with tumors.(4)Patients with ossification of posterior longitu-dinal ligament and ligamentum flavum.(5)Patients with immune diseases or other systemic diseases.All the specimens involved in this experiment were approved by the patients or his family members and signed the informed consent.The ethics committee of the affiliated hospital of Qingdao University approved it.According to the preoperative MRI,the experimental group were divided into three groups: 18 cases in the protrude group,18 cases in the prolapse group and 18 cases in the sequester group.The nucleus pulposus tissue removed during the operation was washed with normal saline immediately.By using hematoxylin-eosin staining to observe the number of nucleus pulposus cells,morphological differences,and cell density of degenerated intervertebral discs and normal intervertebral discs.The expression of HSPA8 in different degrees of discs degeneration was detected by immunohistochemistry(IHC)technique.Western-blot method for detecting the expression level of HSPA8 in the different degree of intervertebral disc degeneration.In order to make the experimental results more convincing and more reliable,more realistically reflect the changes and the expression levels of HSPA8 in the nucleus pulposus tissue,all specimens were collected in accordance with the required standards,and the influence of other tissues in the spinal canal on the intraoperative nucleus pulposus was excluded and avoided to the extent possible,so that all specimens were obtained from the nucleus pulposus in the central part of the lumbar intervertebral disc,and statistical analysis of the results.P < 0.01 shows that there was a significant difference with statistical significance.Results: Hematoxylin-eosin staining results: There were significant differences in cell morphology and number between the control group and the experimental group.There were significant differences in cell density.(F=936.80,P<0.01).Immunohistochemical results: HSPA8 was expressed in lumbar intervertebral disc nucleus pulposus tissue,and the expression of HSPA8 is gradually decreased with the severity of the disease,and there were significant differences.(F=2110.43,P<0.01).Western-blot results: The expression of HSPA8 in human degenerative nucleus pulposus tissue gradually decreased with the progression of intervertebral disc degeneration,there were significant differences(F= 1841.72,P<0.01).Conclusion: HSPA8 was expressed in normal lumbar intervertebral disc nucleus pulposus and degenerated lumbar intervertebral disc nucleus pulposus.And the expression of HSPA8 in normal intervertebral disc nucleus was significantly higher than degenerative intervertebral disc tissue.The expression of HSPA8 gradually decreased with the increase of lumbar disc degeneration.HSPA8 may be involved in the process of human lumbar disc degeneration,which can be used as one of the biological indicators of the severity of lumbar disc degeneration.However,in the process of lumbar disc degeneration,which signal pathway adjusts the expression of HSPA8 needs to be further studied.