The Protective Effects Of BMSCs On Kidney Podocyte Injury In Rat

Objective: To investigate the protective effects of rat BMSCs on rat kidneypodocyte injury.Methods:1.MSCs were extracted from SD rat bone marrow and werefollowed by in vitro amplification, purification and identification.2.Extractedpodocytes from kidney of SD rat,cultured in vitro,and identification.3.Invitro experiments, divided into three.Normal group:Podocytes;Doxorubicingroup:Podocytes+doxorubicin(2ug/ml);The stem cell group:Podocytes+BMSCs(cultured24h)+doxorubicin(2ug/ml).After all experimental groupsintervention24h,48h,72h then detecte groups podocyte apoptosis rate byflow cytometry.After12h、24h and48h measurement of nephrin andsynaptopodin gene expression by RT-PCR and Real time PCR.Detectedexpression of podocyte cytoskeleton-associated protein of nephrin andsynaptopodin by western bolt after24h、48h and72h.Results:1. Flow cytometry results showed that:24h apoptosis rate of normalgroup is19.1%,24h apoptosis rate of doxorubicin group was45.4%,24hapoptosis rate of Stem cell group was22.7%.24h apoptosis rate of stem cellgroup has a significant reduction compared with doxorubicin group inpodocytes,and it reduce significant (P<0.01).48h apoptosis rate of normal group is22.4%,48h apoptosis rate of doxorubicin group is32.5%,48hapoptosis rate of Stem cell group is26.2%.48h apoptosis rate of stem cellgroup has a reduction compared with doxorubicin group in podocytes,but thereduce is non-statistically significant (P>0.05).72h apoptosis rate of normalgroup is17.7%,72h apoptosis rate of doxorubicin group is29.6%,72hapoptosis rate of Stem cell group is28.7%.72h apoptosis rate of stem cellgroup has a reduction compared with doxorubicin group in podocytes,but thereduce is non-statistically significant (P>0.05).2. RT-PCR electrophoresis showed:Nephrin mRNA expression of normalgroup,idoxorubicin group and stem cells group are no significant differencein24h,48h and72h.Synaptopodin mRNA expression of doxorubicin groupisless than normal group in24h and48h.Synaptopodin mRNA expression ofdoxorubicin group is less than stem cells group in24h and48h.3. Real time PCR: Nephrin mRNA expression of normal group,idoxorubicingroup and stem cells group are no significant difference in24h,48h and72h.Synaptopodin mRNA expression of doxorubicin group is less than normalgroup in48h.Synaptopodin mRNA expression of stem cells group is morethan doxorubicin group in48h and it reduce significant (P<0.05).4. Western bolt: Nephrin protein expression of normal group,idoxorubicingroup and stem cells group are no significant difference in24h,48h and72h.Synaptopodin protein expression of stem cells group is more than doxorubicin group at48h.Synaptopodin protein expression of idoxorubicingroup and stem cells group are no significant difference at24h and72h.Conclusion:1. The rat BMSCs can protect the apoptosis of podocytes induced bydoxorubicin in vitro.2. The rat BMSCs can improve the synaptopodin mRNA of damagepodocyte caused by doxorubicin.3. The rat BMSCs can improve the synaptopodin protern of damagepodocyte caused by doxorubicin.