Effects of needle knife interventions on lumbar disc herniation rats by regulating the TGF-β/BMP signaling pathway
Objective To investigate the effects of needle knife interventions on intervertebral disc degeneration of rats with lumbar disc herniation(LDH)and the involvement of the Transforming growth factor-beta/bone morphogenic protein(TGF-β/BMP)signaling pathway.Methods The LDH rat model was established.Rats were randomly divided into blank group,sham operation group,model group,celecoxib group(0.34 g/kg/d celecoxib solvent),needle knife group,TGF-β/BMP inhibitor group(needle knife+100 μL of SB431542 every 2 days),and TGF-β/BMP agonist group(needle knife+10 mg/kg SRI-011381 hydrochloride every 2 days),with 20 rats in each group.The general behavioral status of rats in each group was observed.Hematoxylin and eosin(H&E)staining was applied to observe the morphological changes in intervertebral disc cartilage tissue.Real-time reverse transcriptase-polymerase chain reaction(RT-qPCR)was applied to detect the mRNA expressions of TGF-β1,BMP-7,Smad2/3,Smad1/5/8,Smad4 and TIMP-3.Enzyme-linked immunosorbent assay(ELISA)was applied to detect relative levels of TNF-α,IL-6 and MMP-3.Immunohistochemistry was applied to detect the positive expression of TIMP-3 in intervertebral disc cartilage tissues.Western blot was applied to detect protein levels of TGF-β1,BMP-7,Smad2/3,Smad1/5/8,BMPR-1A,ALK5,Sox9 and COL2A1.Results Compared with those of blank group and sham operation group,rats in the model group presented abnormal behaviors,intervertebral disc rupture,matrix loss,decreased chondrocytes,disordered boundary between cartilage endplates and nucleus pulposus,significantly lower mRNA levels of Smad4 and TIMP-3,mRNA and protein levels of TGF-β1,BMP-7,Smad2/3 and Smad1/5/8,protein levels of BMPR-1A,ALK5,Sox9 and COL2A1 and mean optical density of TIMP-3(P＜0.05),but significantly higher levels of TNF-α,IL-6 and MMP-3(P＜0.05).Compared with those of model group,rats in needle knife group and celecoxib group presented reduced abnormal behaviors,alleviated intervertebral disc damages,significantly higher mRNA levels of Smad4 and TIMP-3,mRNA and protein levels of TGF-β1,BMP-7,Smad2/3 and Smad1/5/8,protein levels of BMPR-1A,ALK5,Sox9 and COL2A1 and mean optical density of TIMP-3(P＜0.05),but significantly lower levels of TNF-α,IL-6 and MMP-3(P＜0.05).Compared with those of needle knife group,rats in the TGF-β/BMP inhibitor group presented aggravated abnormal behaviors and intervertebral disc rupture,significantly lower mRNA levels of Smad4 and TIMP-3,mRNA and protein levels of TGF-β1,BMP-7,Smad2/3 and Smad1/5/8,protein levels of BMPR-1A,ALK5,Sox9 and COL2A1 and mean optical density of TIMP-3(P＜0.05),but significantly higher levels of TNF-α,IL-6 and MMP-3(P＜0.05).Rats in the TGF-β/BMP agonist group presented relieved abnormal behaviors and intervertebral disc rupture,significantly higher mRNA levels of Smad4 and TIMP-3,mRNA and protein levels of TGF-β1,BMP-7,Smad2/3 and Smad1/5/8,protein levels of BMPR-1A,ALK5,Sox9 and COL2A1 and mean optical density of TIMP-3(P＜0.05),but significantly lower levels of TNF-α,IL-6 and MMP-3(P＜0.05).Conclusion Needle knife interventions delay the development of intervertebral disc degeneration in LDH rats by activating the TGF-β/BMP signaling pathway,inhibiting extracellular matrix degradation,and relieving inflammation in intervertebral disc tissue.
万方数据
维普
