Mechanism Of Yiqi Huoxue Recipe In Promoting Spontaneous Resorption Of Ruptured Nucleus Pulposus

Objective 1.Exploring the gene expression profile of ruptured lumbar disc herniation nucleus,and Screening for differentially expressed genes.2.Exploring the mechanism of Yiqi Huoxue Decoction mediated differential expression gene CCL5 to promote resorption of ruptured lumbar disc herniation nucleus.Methods 1.Collecting ruptured and unruptured lumbar disc herniation nucleus,using gene chip technology,and then preliminary screening of differentially expressed genes in nucleus of ruptured lumbar disc herniation to do bioinformatics analysis.2.Culturing human ruptured lumbar disc herniation nucleus cells by vitro culture,and observing the difference in cell morphology.Detecting the expression of differentially expressed genes CCL5,TNF-? and IL-1? in each group of nucleus cells by real-time quantitative PCR and Western Blotting technology.3.After the intervention of Yiqi Huoxue Decoction,detecting the expression of CCL5,TNF-?and IL-1? in nucleus cells one more time.4.Then collecting data and doing statistical analysis.Resuits 1.According the chip result,there are 75 genes with differential expression more than 2 times and statistical analysis P<0.05.Among them,56 genes were up-regulated and 19 genes were down-regulated.The differential expression of THSD7 A up-regulated by 18-fold is the most obvious among the up-regulated genes.Differentially expressed gene CCL5 is significantly upregulated.Go analysis result showed that the biological functions involved in the differential expression of significant genes mainly include cell migration,cell matrix adhesion,multicellular biological homeostasis,etc.The KEGG pathway analysis showed that the signaling pathways involved in the differentially expressed genes mainly include ECM-receptor interaction,vasopressin-regulated water reabsorption,chemokine signaling pathway,amino acid synthesis and metabolism.2.By observation of cytological morphology,we can see that the unruptured lumbar disc herniation nucleus cells are mostly polygonal and scattered,and the ruptured lumbar disc herniation nucleus cells showa tendency of polymerization,mostly long fusiform.Compared with unruptured lumbar disc herniation nucleus cells,the ruptured lumbar disc herniation nucleus shows obvious morphological degeneration under the microscope.3.Gene expression showed that CCL5,TNF-? and IL-1? expression in ruptured lumbar disc herniation nucleus was significantly higher than unruptured lumbar disc herniation nucleus(P<0.05);expression in nucleus cells of ruptured drug-containing group was significantly higher than that in drug-free group(P<0.05).Conclusion 1.The gene expression of ruptured lumbar disc herniation nucleus changed significantly.THSD7 A can be used as a differentially expressed gene in ruptured lumbar disc herniation nucleus.The expression of CCL5 gene is differentially expressed in ruptured lumbar disc herniation nucleus.2.Yiqi Huoxue Decoction may promote the high expression of proinflammatory cytokines such as TNF-?and IL-1?by mediating the expression of CCL5,and participate in the inflammatory reaction,destroy the homeostasis of extracellular matrix,accelerate degradation and promote reabsorption.