Influences Of Yiqi Huoxue Method And Bushen Shengsui Method On Genetic And Protein Expressions Of Notch Signaling Pathway In Rats With Focal Cerebral Ischemia/Reperfusion

ObjectThis study is based on "blood-brain","essence-brain" theory, use the proliferationand differentiation of neural stem cells regulate Notch signaling pathway as the startingpoint, make MCAO/R rat model and intervened by YiqiHuoxue fang andBushenShengsui fang, observe MCAO/R rats differentially expressing genes and theprotein differences in Notch signaling pathway, by investigate the mechanismYiqiHuoxue method and BushenShengsui method to provide experimental basis forclinical treatment of ischemic stroke.Method60SD rats, male,4months, weight280-320g. Randomly divided into sham group,model group, YiqiHuoxue group and BushenShengsui group. Suture method makeMCAO/R rat model, while use YiqiHuoxue Fang-Naoluoxintong and BushenShengsuiFang taking moderate doses, respectively8.54g/kg and12.87g/kg, YiqiHuoxue groupand BushenShengsui group were administrated of a second30minutes before makingMCAO/R model, then continuous administration for7days, sham group and modelgroup were given the same amount of saline in the same way. The gene expressionchanges in Notch signaling pathway in frontoparietal cortex were detected by Real-timequantitative PCR gene chips in all groups. The protein expression changes of β-cateninand Krt1in frontoparietal cortex were detected by Envision two step method. Theprotein expression changes of Nr4a2in frontoparietal cortex were detected byWestern-Blot. Data were statistically analyzed using SPSS17.0. Statistics diagrammingsoftware using EXCEL2007.Result91genes were detected by Notch signaling pathway PCR gene chips. Comparedwith the model group,14kinds of genes (Axin1, Ctnnb1, Fzd2, Gsk3b, Krt1, Lmo2,Lrp5, Map1b, Ncstn, Notch4, Ptcra, Smo, Supt6h, Tead1) expression were significantly different in sham group, which Notch4gene expression was significantly lower (P<0.05),the other13kinds of genes expression were significantly increased (P<0.05). Comparedwith the model group,17kinds of genes (Cflar, Chuk, Ctnnb1, Fosl1, Fzd4, Hes1, hr,Krt1, Map2k7, Notch3, Notch4, Nr4a2, Pdpk1, Psen1, Rbpjl, Rfng, Zic2) expressionwere significantly different in YiqiHuoxue group, which Ctnnb1, Krt1and Nr4a2genesexpression were significantly reduced (P<0.05), the other14kinds of genes expressionwere significantly increased (P<0.05). Compared with the model group,20kinds ofgenes (Aes, Cflar, Chuk, Ctnnb1, Fos, Fzd1, Fzd2, Fzd4, Hey1, hr, Map2k7, Mycl1,Notch4, Nr4a2, Numb, Pdpk1, Rfng, Sel1l, Stat6, Tead1) expression were significantlydifferent in BushenShengsui group, which Ctnnb1, Fzd1, Fzd2, Hey1, Mycl1, Nr4a2,Sel1l and Tead1genes expression were significantly lower (P<0.05), the other12kindsof genes expression were significantly increased (P<0.05). Compared withBushenShengsui group,7genes (Cflar, Ctnnb1, Fzd2, Fzd5, Hes1, Mycl1, Zic2)expression were significantly different, which Fzd2, Fzd5, Hes1, Mycl1and Zic2genesexpression were significantly increased (P<0.05), Cflar, Ctnnb1gene expression wassignificantly reduced (P<0.05).The protein expression changes of β-catenin and Krt1in frontoparietal cortex weredetected by immunohistochemical method: β-catenin and Krt1protein showed noexpression in negative group, and showed visible expression in sham group, mainlyexpressed in the cytoplasm of nerve cells. Compared with sham group, the proteinexpression of β-catenin and Krt1were significantly increased (P<0.05) in model group;compared with model group, the protein expression of β-catenin and Krt1weresignificantly lower (P<0.05) in YiqiHuoxue group, the protein expression of β-cateninwas significantly lower (P<0.05) in BushenShengsui group; compared withBushenShengsui group, the protein expression of β-catenin was significantly lower(P<0.05) in YiqiHuoxue group.Compared with sham group, the protein expression of Nr4a2was significantlyincreased in model group (P<0.01); compared with model group, the protein expression of Nr4a2was significantly reduced in YiqiHuoxue group and BushenShengsui group(P<0.01); compared with BushenShengsui group, the protein expression of Nr4a2wassignificantly reduced Yiqihuoxue party group (P<0.01).ConclusionEarly studies confirmed that YiqiHuoxue method can promote neural stem cellproliferation and differentiation after focal cerebral ischemia, BushenShengsui methodcan promote synaptic remodeling after cerebral ischemia and neuronal axons regenerate,this study found that YiqiHuoxue method and BushenShengsui method and theirrepresentative of compounds can change related genes expression in Notch signalingpathway after cerebral ischemia and reperfusion, and by reducing cortical β-catenin,Krt1and Nr4a2protein expression, generated on the regulation of Notch signalingpathway, and then affecting the endogenous NSCs’ proliferation and differentiation.