| Objective:Intervertebral disc degeneration is the primary cause of causing chronic low back pain,prevention and treatment of lumbar intervertebral disc degeneration is still a difficult problem in the world.Macrophages are an important part of the innate immune system,the CD163molecule is a glycoprotein in the macrophage surface,it can mediate endocytosis and anti-inflammatory cytokine release LPS,play an anti-inflammatory effects.HMGB1is a kind of inflammatory response which plays an important role in proinflammatory mediators.In view of this,we establish the animal model of intervertebral disc degeneration,to explore the expression and correlation of CD163and HMGB1in lumbar intervertebral disc degenerationMethod:Selecting three-month-old42Japanese big ear rabbits,whose weight were2.3-3.Okg,regardless of gender,were randomly divided into experimental group and control group,each group of21only,all of the rabbits were randomly divided into4weeks,8weeks,12weeks,3time points,each time point killed14rabbits.From the retroperitoneal exposure the intervertebral disc,with16#epidural puncture needle L2-3,L3-4,L4-5intervertebral disc,intervertebral disc injury model caused by acupuncture,the control group only exposed without puncture.According to the time point killed the animal,to completely removed the entire spine,for gross observation,HE dyeing,and remove the cartilaginous endplate of intervertebral disc detected by RT-PCR.Result.1.General observation:the intervention disc visible traces not healing in4weeks experimental group,the color,texture thickness of the intervertebral disc has no obvious changes,anterior edge of smooth.The8weeks experimental group visible the pinprick disappeared,intervertebral disc compared to the control group.color gray,texture hard and the disc height has lose mildly,12weeks experimental group visible the interference disc height loss.fiber ring marked hyperplasia of fibrous tissue,osteophyte formation obviously.Each time point of the control group showed no obvious degeneration of intervertebral disc2.The results of the tissue sections HE dyeing:4week experimental group histologically visible fiber ring structure disorder,endplate cartilage cell level is unclear,arranged in disorder;8week experimental group visible nucleus pulposus shrunken,fibrous ring rupture,cartilage endplate calcification ayer thickening,cartilage lacuna chapped.tidal line forward,the number of cells decreased;12week experimental group intervertebral disc degenerative changes obviously,disorder of cartilaginous endplate calcification ayer thickening,ossification,cartilage lacuna disappeared.nuclear pyknosis,fragmentation and dissolution,Tidal line thickening and fuzzy,the nucleus pulposus adjacent the cartilage endplate has degeneration and fibrosis.Each time point of the control group,the structure of cartilaginous endplate is good,tide line is clear.the nucleus pulposus and annulus structure clear and full,has no obvious fissure.3.The result of UV spectrophotometer and electrophores:the OD value of all cartilage plate showed higher purity of RNA.The result of electrop--horesis also prove that there has no RNA degradation in the whole process4.RT-PCR results:the expression of CD163mRNA on experimental group at each time point showed a rising trend,the difference has statistically significant(P<0.05),the experimental group were significantly increased(P<0.05)than the control group.Experimental group at each time point,the expression of HMGB1mRNA has no significant difference(P>0.05),but the difference at each time point in the experimental group and control group has statistically significant(P<0.05).The correlation analysis of CD163and HMGB1showed no significant correlation of the two indicators(Pearson Correlation=0.75)Conclusion: l.The animal mode of intervertebral disc degeneration from retroperitoneal by acupuncture can simulate better human intervertebral disc degeneration process,has the advantages of simple operation,repeatability and controllability,and cost benefit principle2.1n the animal models of intervertebral disc degeneration,each time point of the experimental group and the control group had significant difference in CD163mRNA expression.there was a significant difference between the experimental group,suggesting that CD163in degenerative intervertebral discs maybe derived from the cartilage endplate,and plays an important role in intervertebral disc degeneration3.In the animal models of intervertebral disc degeneration,each time point of the experimental group and the control group had significant difference in HMGB1mRNA expression.but the differences between each experimental groups was not significant,suggesting that HMGB1in degenerative intervertebral discs maybe derived from the cartilage endplate.and may play an important role in the pathogenesis of disc degeneration,but its content is not increased with increasing degeneration4In the disc degeneration process,CD163and HMGB1levels higher than that of health disc.but it is no obvious correlation between the two indexes. |
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