Effect Of HGF On Podocyte Apoptosis Induced By Adriamycin In Rats

Objective：Hepatocyte growth factor（HGF）is a pleiotropic growth factor which is composed of multiplepolypeptide，and mainly distributed in liver, kidney, lung and pancreas. HGF play an important role as aninteractional medium between glomerular epithelial cell, GBM and vascular endothelial cell, The HGFreceptor (c-Met) all expressed at mesangial cells, glomerular epithelial cell and vascular endothelial cell,HGF can specifically bound to c-Met to activate many signal transduction pathways and act on theoccurrence and development of kidney disease, then maintain the normal structure and function of thekidney. When the kidney damage, HGF can promote it rebuild normal structure and function. Meanwhile，HGF include multiple activities in promoting cell regeneration，which also can retard the development ofglomerular fibrosis and dysfunction. In present study, we injected adriamycin to rats to induce glomerularinjury，then gave the rats HGF for intervention, and observed expression intensity of nephrin after andbefor intervention, The expression intensity and podocyte apoptosis proteinuria and glomerular podocyteprotein nephrin related factors were observed before and after the intervention, to investigate the effect ofHGF on podocyte apoptosis.Methods：1. Adriamycin-rat model:114male SD rats were randomly divided into three groups: controlgroup(42), model group(42) and intervention group(30). The rats in model group and intervention groupwere single intravenous injected to adriamycin (2mg/ml,7.5mg/kg) to induce Adriamycin-rat model.Meanwhile，rats in control group were single intravenous injected to0.9%Nacl （3.25ml/kg）as control.2.After perfect induction, the rats in intervention group were gave HGF（30ug/kg d）through intraperitonealinjection the third week after induction until the end of the experiment. We killed6rats separately fromcontrol group and model group at days7,14,21,28,42,56,72after injection, and killed6rats fromintervention group at days21,28,42,56,72after intervention.3. Collected24hours’ urine of all rats withmetabolism cages on the day before killed, determine the volume of proteinuria. Determine the podocytepathological variation of renal tissue samples through light microscopy and electronic microscopy，anddetecte the expression intensity of nephrin by immunofluorescence stain，observed the expression ofBcl-2/bax、Caspase-3by immunohistochemistry.Result:1. The rats in model group and intervention group were perfect induction ofAdriamycin-rat model.2. compared with the control group, model group, serum albumin declined from seventh days totwenty-first days, the lowest, were lower than the control group, the difference was statistically significant(p<0.01). Serum albumin HGF intervention were significantly higher than those in the model group, thedifference was statistically significant (p<0.05).3. Proteinuria change: compared with control group, theproteinuria of model group increased significantly (p＜0.01), the proteinuria of intervention groupdecreased significantly (p＜0.01).4. Morphology and pathological changes of renal tissue: In control group,we observed macroscopically that the renal tissue were normal-sized，dark red and clear demarcationbetween the cortex and medulla. Compared with control group, the renal tissue in model group was slightlylarger size at early phase，the colour turned gray and white after6weeks, balloon adhesions after8weeks,and mild-to-moderate glomerulosclerosis at the end of12weeks. In intervention group, the renal tissue was normal at early phase, the colour turned gray and white at later stage, balloon adhesions and diffuseeffacement of podocyte foot processes after8weeks, and glomerulosclerosis and podocyte detachment atthe end of12weeks. From the electronic microscopy, in model group, we observed that the podocyte wereswelling after4weeks, podocyte detachment and diffuse effacement of podocyte foot processes after8weeks, but the intervention group turned lighter compared with intervention group.5. The expression ofnephrin: between1and4weeks, compared with control group, the expression of nephrin was increased inrats of model group (p＜0.01), and the expression of nephrin was decreased in rats of intervention group(p＜0.01). Between5and12weeks, compared with control group, the expression of nephrin was increased inrats of model group (p＜0.05), and the expression nephrin was decreased in rats of intervention group(p＞0.05). Moreover, in model group, the expression of nephrin was increased in rats of early stage than the ratsof late stage.6. The expression of Bcl-2/bax and caspase-3: The results showed that rare caspase-3expressed in rats of all groups. The expression of Bcl-2/bax in rats of control group and model group wasincreased compared with intervention group.Conclusion:1.Asingle intravenous injected to adriamycin can perfect induceAdriamycin-rat model.2.HGF can inhibit the apoptosis of podocyte in Adriamycin-rat model and decreased the proteinuria of rats.3,adriamycin rats apoptosis did not occur through apoptosis of classical pathway, but through other apoptoticpathway.