| Objectives：Application Schizandrm Chinensis Polysaccharide intervention rabbit nucleus pulposuscells in vitro, Observation the nucleus pulposus changes after the intervention in thebiological behavior,through the detection of proteoglycan and type II collagen expression,provides an experimental basis for the early treatment of disc degeneration.Methods：Separation and rabbit nucleus pulposus cells in vitro culture, identified byimmunohistochemical methods; when transmitted to the second generation, interventionwhen the highest activity of cell proliferation, The experiment were divided into controlgroup and very low, low, medium, high dose group, Then semi-quantitative detection usingRT-PCR of proteoglycan and type II collagen in the cells after the intervention,andSemi-quantitative analysis of the results with TANON GIS gel image processing system,detected cell proliferation by MTT, growth curve was drawn, observed the impact of the ofSchizandrm Chinensis Polysaccharide intervention on the biological behavior of the nucleuspulposus cells, It can be expected to extend to the maintenance of cell phenotype, long-termexpression of proteoglycan and type II collagen.Results：1. rabbit nucleus pulposus cell growth in good condition and high activity in vitro.Which is more obvious in the first and second strongest, followed by primary activity, thecell activity decreased more significantly by the fourth generation, Primary rabbit nucleuspulposus cells4to6d cell attachment, the exponential growth phase about the first16d afterinoculation, fusion more than90%will take about22d; the first generation of rabbit nucleuspulposus cells of attachment about3days, the exponential growth phase at about14th dayafter inoculation,90%fusion will take about20d.2. RT-PCR semi-quantitative test results show: aggrecan and type II collagen have no significant difference between the blank control group and the extremely low dose group(P>0.05),significant difference between low, middle dose group and control group withstatistical significance(P <0.05, P <0.01), and in dose-dependent manner,The action intensityof the high dose group similar to the middle dose of the experimental group, no significantenhancement.3. MTT The cell growth curve of the nucleus pulposus cell after the interventionshow:in the experimental group, the nucleus pulposus cell proliferation rate was significantlyhigher.Conclusion：1.Successfully cultured rabbit nucleus pulposus cells by monolayer culture method invitro,was identified as the nucleus pulposus cells.2. Schizandrm Chinensis Polysaccharide role in the rabbit nucleus pulposus cells, theincreased expression levels of type II collagen and aggrecan. And presented adose-dependent manner,.3. Schizandrm Chinensis Polysaccharide intervene in the rabbit nucleus pulposus cells,the proliferative capacity of nucleus pulposus cell significantly improve. |
PDF Full Text Request