Bi Shu Lumbar Disc Degeneration In Rabbit Nucleus Pulposus TNF-α, MMP-1 And TIMP-1 Expression In Effect

Research Objective:Observe the effect of Yaobishu soup on TNF-α, MMP-1and TIMP-1expression by thenucleus pulposus of the degenerated lumbar intervertebral disc, expose whether Yaobishusoup treats lumbar intervertebral disc degeneration by regulating the contents of TNF-α,MMP-1and TIMP-1, and provide the basis for defining the action principle of the Chinesepatent drug, Yaobishu.Materials and Methods:Divide45New Zealand white rabbits into three groups in a random manner, with eachgroup consisting of15rabbits. Perform modeling process to the Group A (Yaobishuintervention group) and B (control group). Inject the rabbits’ lumbar4~6intervertebral disc with0.2ml physiological saline. Perform sham operation to the Group C (blank group). From the8thday after the operation, feed the Group A with Yaobishu in a dose of10ml/kg each day,until they are executed in batches. Feed the Group B and C with mixed feed and let themdrink water freely, until they are executed in batches. After modeling, execute theexperimental animal in batches in the2nd,4thand8thweeks (take X-rays of animal’s lumbarvertebra before the execution) to acquire animal specimens. Each time, execute five rabbits ofeach group in a random manner, and then remove the model of the lumbar intervertebral discto perform morphologic observation, histological observation, MMP-1and TIMP-1immunohistochemistry detection and TNF-α enzyme linked immunosorbent assay. Results:1. Result of medical imaging observation:The X-rays of the modeling lumbar vertebra of the Group A and B show that, theintervertebral space is narrowed down, the shadow of the vertebral edge density is heightenedand osteophyma is generated. The narrowing of intervertebral space and generation ofosteophyma of the Group A is slighter than that of the Group B. The lumbar vertebra andintervertebral space of the Group C have no obvious isabnormalities found.2. Result of morphologic observation:For the Group A, the intervertebral space is narrowed down, and osteophyma is slightlygenerated. The symptoms did not get worse as the extension of the observation period. For the Group B, the intervertebral space is narrowed down, and osteophyma is slightly generated. Asthe extension of the observation period, the modeling lumbar intervertebral disc is hardenedgradually and more osteophyma is generated. The lumbar vertebra and intervertebral space ofthe Group C have no obvious isabnormalities found.3. Result of histological observation:Group C: Under the light microscope, the shape of the lumbar intervertebral disc isregular, the fibrous rings are regularly arrayed regularly and the nucleus pulposus is obvious.Group A: In the2ndweeks after the operation, the fibrous stratum is generous, the fibrousrings are arrayed regularly, the nucleus pulposus existed, and In the4thweek after theoperation, the osteoid hyaline cartilage generated on the rear edge of the lumbar intervertebraldisc. In the8thweek after the operation, the fibrous rings are arrayed disorderly, the fibrousstratum is decreased, part of the fibrous stratum is replaced with the osteoid, and the osteoidhyaline cartilage generated on the rear edge of the lumbar intervertebral disc.Group B: In the2ndweeks after the operation, the fibrous rings are arrayed less regular,the fibrous stratum is decreased, and the osteoid hyaline cartilage generated on the rear edgeof the lumbar intervertebral disc. In the4thweek after the operation, some fibrous tissues arenecrotized, the fibrous stratum is obviously decreased and replaced with the sclerotin. In the8thweek after the operation, the nucleus pulposus is disappeared and completely replaced withthe osteoid and sclerotin.4. Result of immunohistochemistry detection:Group C: The gradation values of MMP-1image in all periods are high, showing that theexpression of MMP-1is not obvious. The gradation values of TIMP-1image in all periods arehigh, showing that the expression of TIMP-1is not obvious.Group A: The gradation values of MMP-1image in the2ndare low, showing that theexpression of MMP-1is not obvious. The gradation values of MMP-1image in the4thand8thare increased, showing that the expression of MMP-1is becoming obvious. There is statisticdifference (p<0.05) between Group A and C. The gradation values of TIMP-1image in allperiods are high, showing that the expression of TIMP-1is not obvious.Group B: The gradation values of MMP-1image in all periods are low, showing that theexpression of MMP-1is obvious. There is statistic difference (p<0.05) between the Group B and C. There is also statistic difference (p<0.05) between the Group B and A. The gradationvalues of TIMP-1image in all periods are low, showing that the expression of TIMP-1exists.There is also statistic difference (p<0.05) between the Group B and Group A as well as C.5. Result of enzyme linked immunosorbent assay:In the8ndweeks after the operation,the TNF-α expression of the Group B is more obviousthan the those of the Group A and C. There are statistic differences (p<0.05). The TNF-αexpression of the Group A is more obvious than the that of the Group C. There is statisticdifference (p<0.05).Conclusion:1. Yaobishu can delay the degeneration of rabbit lumbar intervertebral disc.2. Yaobishu can relief the degeneration of rabbit lumbar intervertebral disc through theinhibited effect on the Inflammatory media TNF-α in the degenerated rabbit lumbarintervertebral disc.3.Yaobishu can relief the degeneration of rabbit lumbar intervertebral disc by depressing theMMP-1expression of the degenerated rabbit lumbar intervertebral disc.4. TIMP-1is the idiosyncracy inhibitor of MMP-1. Its expression is positively correlated withthat of MMP-1, and has no direct relation with Yaobishu.