Effects Of Radix Astragali And Hirudo Extracts On NF-κB、IκB、MMP-9、PDGF-BB Expression Of Glomerular Mesangial Cells In Rats

In our country there are many kinds of pathological types of primary glomerular disease, mesangial proliferative glomerulonephritis (MsPGN) is a common pathological type, mesangial cell proliferation and extracellular matrix deposition is the main pathogenesis, these two kinds of pathological factors can result in glomerular sclerosis, development of renal impairment, and end-stage renal disease. At present, immune inhibitors, antiplatelet agents, glucocorticoid and symptomatic treatment are commonly used drugs, and achieved a certain effect in the clinical treatment, but its side effects and easy recurrence significantly. In recent years, studies have shown that invigorating kidney and promoting blood circulation to remove blood stasis t2dm and other traditional Chinese medicine has obvious curative effect in the disease, especially represented by astragalus root, leech, replenishing qi blood drug, has a large number of literature reports Chinese medicine can reduce the cell proliferation, regulation of cell apoptosis, degradation of deposition in the extracellular extra cellular matrix and improve the renal fibrosis, slow disease progression as glomerular sclerosis.Domestic and foreign scholars study showed that mesangial cell proliferation and growth of nuclear transcription factor (nf-kappa B) caused by the activation of the release of inflammatory mediators and extracellular matrix formation is closely related with the imbalance of degradation, and the activation of the nf-kappa B and nuclear growth factor inhibiting factor (I kappa B predominate) there is a close relationship between phosphorylation and degradation; Matrix metalloproteinases (MMPs) is a matrix of cell formation and degradation of balanced system, once the balance is broken, the formation and degradation of imbalance, matrix deposition.This experiment from the nf-kappa B, I kappa B predominate, MMP-9, PDGF-BB active ingredients, to analyze factors of astragalus root, leech inhibit mesangial cell proliferation mechanism.Objectives:Exploring different effective components of astragalus root, leech on after lipopolysaccharide (LPS) in rats induced by MC, the influence of the nf-kappa B, I kappa B predominate, PDGF-BB, MMP-9 factors as the breakthrough point, to explore different effective components of astragalus root, leech mechanisms play a role.Methods:The proliferation of mesangial cells is induced by LPS and intervened by the astragalus polysaccharides, astragalus glycosides, calycosin separately, and hirudin, collecting cells after 24 hours, CCK-8 method are used to observe the monomer and the drug-containing serum of HBEH proliferation inhibition rate, WesternBlotting and Real time PCR method to determine the expression of NF-κB、IκB、MMP -9and PDGF-BBchange.Results:1. Shown by WesternBlotting:afte the Proliferation of mesangial cells induced by LPS, model group NF-κB protein expression increased significantly compared with the normal group (P<0.05), and astragalus polysaccharides, astragalus glycosides, calycosin separately, and hirudin in the NF-κB protein expression levels were significantly lower than model group (P<0.05). As can be seen, each drug group can be different degrees of inhibition of NF-κB protein expression level; there was no obvious difference between the drug group, had no statistical significance(P>0.05).2. Shown by WesternBlotting:afte the Proliferation of mesangial cells induced by LPS, model group IκB protein expression decreased significantly compared with the normal group (P<0.05), and astragalus polysaccharides, astragalus glycosides, calycosin separately in the IκB protein expression levels were significantly higher than model group (P<0.05), but the hirudin does not increased too much. As can be seen, each drug group except hirudin can be different degrees of promotion of IκB protein expression level; there was no obvious difference between the drug group, had no statistical significance(P>0.05).3. Shown by Real time PCR:after the Proliferation of mesangial cells induced by LPS, the expression of MMP-9mRNA in model group compared with normal group significantly increased (P<0.01), and astragalus polysaccharides, astragalus glycosides, calycosin separately, and hirudin in the MMP-9mRNA expression levels were significantly lower than model group (P<0.01). As can be seen, the drug groups can inhibit the MMP-9mRNA expression level in different degree; there was no obvious difference between each drug serum group, had no statistical significance(P>0.05).4. Shown by Real time PCR:after the Proliferation of mesangial cells induced by LPS, the expression of PDGF-BBmRNA in model group compared with normal group significantly increased (P<0.01), and astragalus polysaccharides, astragalus glycosides, calycosin separately, and hirudin in the PDGF-BBmRNA expression levels were significantly lower than model group (P<0.01). As can be seen, the drug groups can inhibit the PDGF-BBmRNA expression level in different degree; there was no obvious difference between each drug serum group, had no statistical significance(P>0.05)Conclusion:Astragalus polysaccharides, astragalus glycosides, calycosin separately, and hirudin can decrease the high expression of NF-κB、MMP-9、PDGF-BB after the proliferation of HEBH induced by LPS,astragalus polysaccharides, astragalus glycosides, calycosin separately can increase the high expression of IκB. The study show that effective constituent of astragali and hirudo can reduce the proliferation of mesangial cells and extracellular matrix deposition and delay the progression of glomerular sclerosis by influencing the expression of NF-κB、IκB、MMP-9、PDGF-BB.