The Study Of The Role Of Aquaporins In The Mechanism Of Intervertebral Discs Degeneration And The Modulation Of Aquaporins

Background and Objectives: Intervertebral disc degeneration(IVDD) is the important reason of lumbar disc herniation and low back pain. But up to now, there is no exact anwer to the mechanism of intervertebral disc degeneration. When IVDD is occurred, the water content in the disc is decreased. At present, the impairment of nutrition supply to the disc is thougt to as an initiating agent of IVDD. The water is one of the essential nutrient of the discs, before the aquaporin was discovered, the water entering and venting disc was thought to be mainly mediated by the osmosis, but now the transmembrane transport of water is thought to be mainly mediated by AQPs. The study is to investigate the role of AQPs'in IVDD, to analyze the relation of the expression of AQPs and the water content in the discs, and to understantd the role of IVDD related fators in the modulation of AQPs. Furthermore, a new pathophysiologic mechanism of IVDD is hoped to be advocated.Methods. 1. The animal model of IVDD was established by destroying the rats'spinal instability, then the X-ray changes of the lumar spine was observed and the histological changes of the discs was observed by Hemattoxylin and eosin staining,Toluidine blue staining and Safranin O staining, the degeneration degree of the discs was evaluated. 2. The expression types and space distribution of AQPs in interverbral discs was detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). 3. By the animal model of IVDD, the expression changes of AQP1\3 were observed by western blot and RT-PCR. 4. The expression of AQP1\3 in endplate chondrocyte was detected by immunocytochemistry, it's used to be analyzed the relationship between MRI T2 weight intensity of degenerated discs and the expression. 5. The lumbar endplate chondrocyte was cultivated, it's characteristic of cell biology was observed and it's cell phenotype was identified. 6.The inhibitor of AQPs, mercuric chloride, was used to observed it's effect on chondrocyte proliferation; by the Hemattoxylin and eosin staining and X-ray, the histological and imageological changes caused by mercuric chloride were recorded. 7. The expression vector of AQP3 siRNA was constructed, by western blot and RT-PCR, it's...