| Objectives Explore PI3K/AKT/m TOR signal for Parkinson’s disease in rats induced by rotenone,using LY294002 and Rapamycin(channel blockers)to observe the content of channel proteins and oxidative stress related index SOD,GSH,GSH – XP and MDA to explore the PI3K/AKT/m TOR signal associated protein in the substantia nigra of Parkinson’s disease model rats areas.Methods 1 The sham group,Parkinson disease model group,LY294002 group and Rapamycin group are composed of 96 PD Healthy male Sprague wrote-Dawley(SD)rats.The fore group male rats are randomly divided into 4 day and 8 day two subgroups(n=12)after the success of Parkinson’s disease model preparation.2 The model of Parkinson disease was established by subcutaneous injection of rotenone in rats.3 Dosing method Lateral ventricle injection: link sterilized glass micro pipettes with stereotaxis by Pipe rack,downwarding the cutting-edge(drag)vertical,using the scale standard by 0.9 mm and 1.5 mm,Tiping penetration into the rats skull 2-3 mm,LY294002 and Rapamycin group injecting 10 kg/L LY294002 solution(Rapamycin 80 ng/ml,4 mu L)with in 5 min,pull the needle out slowly.4 Useing immunohistochemistry assay and western blotting observeing the content of PI3K、p-AKT and p-m TOR.5 Useing spectrophotometric method to detecte the expression of SOD,GSH,,MDA,GSH-XP.6 We use microscope camera BX53 to observe the tissue slices and analyzeing them by Motic Med 8.0.Then useing Image J detection the stripe of Western Blot.7 All the reselts use statistical procedures analysis,Own-way ANOVA to deal with multiple sets of values.Results 1 Immunohistochemistry assay and western blotting results of PI3K: Compared with sham operation group,the expression of PI3 K protein content are increased significantly at(8 d 、4 d)two time points in the substantia nigra of PD group(P < 0.05),The expression levels of PI3 K are hihger at 8 d than at 4 d;the expression of PI3 K protein content in the group of LY294002 and Rapamycin(4 d 、8 d)has no obvious change compared with those in PD group(P>0.05).2 The immunohistochemistry assay and western blotting results of p-AKT: Compared with sham group,the content of pAKT is increased significantly at(8 d、4d)two time points in the substantia nigra of PD group rat(P < 0.05);the expression of p-AKT decrease significantly at(8 d、4 d)two time points in LY294002 group(P < 0.05);the content of p-AKT has no obvious change at(4 d 、 8 d)two time points in rapamycin group(P > 0.05).3 The immunohistochemistry assay and western blotting of p-m TOR: Compared with sham operation group,the content of p-m TOR is increased significantly at(8 d 4d)two time points in the substantia nigra of PD group(P < 0.05),The levels of p-m TOR is hihger at 8 d than at 4 d;the content of m TOR decrease significantly at(8 d、4 d)two time points of LY294002 group and rapamycin group compared(P < 0.05).4 Oxidative stress indicators SOD detection results :Compared with control group,the vigor of SOD is reduced significantly at each time point in PD model group(P < 0.05),The levels of SOD is lower at 8 d than at 4 d group;The content of SOD is increased activitly at anytime point in LY294002 group and rapamycin group.5 Oxidative stress indicators GSH detection results : Compared with control group,the vigor of GSH is reduced significantly at each time point in PD model group(P < 0.05),The levels of GSH is lower at 8 d than at 4 d group;The content of GSH vigor are increased activitly at anytime point in LY294002 group and rapamycin group.6 Oxidative stress indicators GSH-XP detection results : Compared with control group,the vigor of GSH is reduced significantly at each time point in PD model group(P < 0.05),The vigor of GSH-XP is lower at 8 d than at 4 d;The content of GSH-XP is increased activitly at anytime point in LY294002 group and rapamycin group.7 Oxidative stress indicators MDA detection results : Compared with control group,the vigor of MDA is increasesed significantly at each time point in PD model group(P < 0.05),The vigor of MDA is higher at 8 d than at 4 d group;The content of GSH-XP is reduced at anytime point in LY294002 group and in rapamycin group.Conclusions 1 The high content of PI3K、Akt and m TOR protein in substantia nigra density of Parkinson’s disease rats are related with its occurrence and development.2 PI3K/AKT/m TOR signaling pathway play a promoting effect to oxidative stress at Parkinson’s disease model rats which is induced by rotenone. |
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