The Role And Mechanism Of Ihh In Degeneration And Abnormal Calcification Of Lumbar Cartilage Endplate

Objective:To investigate the role and mechanism of Ihh in degenerative and abnormal calcification of lumbar cartilage endplate in vitro by chondrocytes and gene knockout mice.Methods:Part ? Effect and mechanism of ihh on abnormal calcification and degeneration of chondrocytes in vitro1.Costal cartilage cells,extraction of C57 mice by HE staining,Saffron O solid green stain and immunohistochemical detection of the expression of collagen type ? to identify chondrocytes.2.The mRNA expression levels of COLII?COLX?Runx2?OCN?ALP?AGG?ANKH>ENPP1 and other factors in ihh high expression group,ihh signaling pathway inhibition group and blank control group were detected by RT-PCR.3.Effects of inhibition of ihh and high expression of ihh on chondrocyte apoptosis By flow cytometry.Part ? Analysis of Abnormal Calcification Degeneration of Lumbar Cartilage Endplate in ihh Conditional Knockout Mice1.Using ihhfl/fl;rosa and prxl-cre;Ihfl/;rosa gene mice bred ihh-/-gene knockout mice as the experimental group,ihhfl/fl gene mice as the control group,respectively to observe the gross morphology of each group of gene nice;X-ray was used to observe the changes of the vertebral bodies.2.micro-CT scanning of the lumbar 1-2 intervertebral discs of mice was performed to observe the changes in the number of bone trabeculae,index of structural model,thickness of bone trabeculae,degree of separation of bone trabeculae,and other indicators.3.Detection of each gene in mice respectively using RT-PCR in lumbar endplate Cartilage COL ??COLX?Runx2?OCN?ALP?AGG?ANKH?ENPP1 and other factors mRNA expression difference.Results:Part ? Effect and mechanism of Ihh on abnormal calcification and degeneration of chondrocytes in vitrol.ihh high expression group compared with blank control group ALP,OCN,Runx2,COLX expression increased,ANKH,AGG,COL ?,ENPP1 expression is reduced,the difference was statistically significant(P<0.05),Ihh signaling pathway inhibition group compared with blank control gro up ANKH,ENPP1,COL ?,AGG expression increased,COLX?Runx2?OCN?ALP expression is reduced,the difference was statistically significant(P<0.05).2,The apoptotic rate of the blank control group was 9.85±0.39,the apoptosis rate of the ihh hMgh expression group was 8.34±0.22,and the apoptosis rate of the ihh signaling pathway inhibition group was 14.48±1.23.The apoptosis rate of the ihh signaling pathway inhibition group was higher than that of the control group,and the difference was Statistically significant,the apoptotic rate of the ihh high expression group was lower than that of the control group,and the difference was statistically significant.Part ? Analysis of Abnormal Calcification Degeneration of Lumbar Cartilage Endplate in ihh Conditional Knockout Mice1.The overall morphology and spinal length of ihh knockout mice were shorter than that of ihhfl/fl mice of the same age.2.The number of trabecular bone in ihh knockout mice was higher than that in the control group,the structural model index and the degree of trabecular separation were lower than that in the control group,and there was no significant difference in the thickness of trabecular bone.3.The experimental group compared with control group ANKH,ENPP1,COL ?,AGG expression increased,COLX,Runx2,OCN,ALP expression is reduced,the difference was statistically significant(P<0.05).Conclusion:Ihh can raise COLX,Runx2,OCN and ALP gene expression,inhibition of ANKH,ENPP1,AGG and COL II gene expression to promote cartilage endplate cells of abnormal calcification and degeneration.