Effect Of Epimedium On Expression Levels Of BMP-2 And BMP-4 Genes In Cartilage Tissues Of Rabbit Model Of Lumbar Intervertebral Disc Degeneration

BACKGROUND: Intervertebral Disc Degeneration(IVDD)is a common orthopaedic disease.It refers to the pathological and physiological processes of natural aging and degeneration of the lumbar spine.It often manifests itself as low back pain and acute lower extremity nerve root lesions,which seriously affect people's lives and work..At present,the mechanism of occurrence of lumbar intervertebral disc degeneration is not yet clear.In recent years,it has been found that inflammatory factors,abnormal expression of growth factors and so on are related to disc degeneration.Among them,bone morphogenetic proteins(BMPs)are a class of acidic peptides that induce osteogenesis and stimulate cell growth,belonging to transforming growth factor beta(TGF-?)superfamily members play an important regulatory role in the formation of bone tissue and actively promote intervertebral disc synthesis and metabolism.At present,many sub-members of BMPs have been found to be associated with osteoporosis,of which BMP-2 and BMP-4 have become hot spots in clinical research.Epimedium is a plant of the genus Eupatorium,Eupatorium japonicum and Epimedium sagittatum.The main chemical constituents are flavonoids,icariin,and polysaccharides,which are traditionally used to strengthen the kidneys and strengthen the yang.Bone drugs can antagonize osteoporosis.Animal experiments have confirmed that total flavonoids of Epimedium can promote the proliferation and differentiation of rat osteoblasts in vitro,which provides the possibility to study the effects of Epimedium on BMPs in cartilage tissue of patients with lumbar intervertebral disc degeneration.OBJECTIVE: To establish a rabbit model of lumbar intervertebral disc degeneration,administer Epimedium and normal saline respectively,and detect the expression of BMP-2 and BMP-4 in cartilage.The aim is to determine the morphology of osteoarthritic cartilage in lumbar intervertebral discs.The effect of protein production provides the basis for the clinical application of Epimedium.METHODS: Thirty New Zealand white rabbits were divided into 10 groups: sham operation group,model group,and epimedium group.All of them except the sham operation group were established with the degenerative disc model.The epimedium group was given 10mg/kg·d fornication.The formula was given intragastrically by gavage,the model group was given the same dose of normal saline,and the lumbar cartilage endplate specimens were collected on the 90 th day to observe the changes of histomorphology.The expression of fas receptor was detected by immunohistochemistry and real-time fluorescence was used.Finally,western blot was used to detect the expression of BMP-2 and BMP-4 in sham-operated group,model group and epimedium group.RESULTS: 1.Under the inverted microscope,the color of the nucleus and cytoskeleton is deeper after staining,and dark purple heterochromatic particles are distributed around the cell nucleus.The cytoskeletal portion is uniformly blue.This cell can be identified as the cartilage endplate cell;see the different bands under the inverted fluorescence microscope.Under the excitation of light,the cytoskeletal portion showed green fluorescence,and no green fluorescent labeling was observed in the nucleus.This cell could be identified as a cartilage endplate cell.2.The sham-operated group had intact endplate structure,smooth surface and regular arrangement.The cartilage cells were plump,round or elliptical,arranged in columns,and the cartilage matrix was evenly colored.The cartilage surface of the model group was rough,there was partial defect,and the tissue structure was incomplete.The chondrocytes were unevenly distributed and disordered.In some areas,the chondrocytes showed clonal proliferation,clustered distribution,the cartilage matrix was light and irregular,and the tide lines were distorted and incomplete.The cartilage cells in the Epimedium group were still intact and the cell arrangement was still regular.,Less apoptosis,more uniform cartilage matrix staining,tide line is still complete.3.The positive rates of fas receptors in the model group and epimedium group were higher than those in the sham operation group(P<0.05).The positive rate of fas receptors in the epimedium group was lower than that in the model group,and the difference was statistically significant(P<0.05)..4.The results of Western Blot showed that the relative expression of BMP-2 and BMP-4 mRNA protein in the sham-operated group was significantly higher than that in the other two groups(P<0.05);The relative expression of BMP-4 and BMP-2 protein in the epimedium group was significantly higher than that of the model group,and the difference was statistically significant(P<0.05).CONCLUSION: Epimedium can increase the expression of BMP-2 and BMP-4 in cartilage tissues of rabbit model of lumbar intervertebral disc degeneration,and promote the proliferation and differentiation of osteoblasts.