| Objectives:Through the observation of CRJD extract water provided liquid of Parkinson’s di sease (PD) model mice brain tissue GSK-3 beta and related protein expression, to exp lore mechanism of Kidney nourishing compound CRJD extract of PD, so as to provid e a theoretical basis for the clinical treatment of PD.Methods:PD model was established by intraperitoneal injection of (30mg/kg/day)MPTP in 8-week-old healthy C57BL/6 male mice for 7 days. Successful model male mice were divided randomly into the following groups:(1) MPTP (30mg/kg/day, i.p.,7days), (2) MPTP+low dose of CRJD(4g/kg/day, i.g.,14days, CRJD-L), (3) MPTP+middle dose of CRJD(8g/kg/day, i.g.,14days, CRJD-M), (4) MPTP+high dose of CRJD(16g/kg/day, i.g., 14days, CRJD-H);And randomly selected healthy C57BL/6 male mice 10 as the normal group.The behavioral changes of mice were observed by swimming experiment and independent activity test in 14days. After the behavioral test, the mice were sacrificed and the mice were sacrificed according to the requirements,the level of DA in Mouse substantia nigra was analyzed through high performance liquid chromatography(HPLC). We also analyzed the protein expression of GSK-3 beta,tau,alpha synaptic(a-synuclein),Bcl-2 and Bax to clarify the mechanism with western blot (WB).Results:We found that the swimming immobility time in mice model group were increased, the number of rearing of PD model mice were reduce,and there was significant difference (P<0.01).The PD model mice were appeared behavioral disorder. Compared with model group mice, high dose of CRJD group mice swimming immobility time is shortened obviously, locomotor activity was significantly enhanced, there was significant difference (P <0.01) and behavioral barriers to improvement.HPLC levels of DA detection showed that compared with the normal group, the model group mouse brain DA content was significantly reduced, there were significant differences (P<0.01). desert CRJD extract low dose group and model group mice compared, brain DA content had no obvious difference, but CRJD extract middle and high groups of mice brainDA content showed a dose dependent increase, CRJD extract in the high dose group of DA content was significantly higher than that of model group,a significant difference (P<0.01).Expression of GSK-3 beta, Tau and alpha-synuclein protein.The results showed that the mice in the model group compared with normal group, model group of GSK-3 beta and tau expression had no significant difference, but the expression of alpha synuclein protein and phosphorylation of GSK-3 beta and tau protein were significantly increased, with significant difference (P<0.01);compared with the model group, CRJD water extract of intervention, Each dosing group cell GSK-3 beta and tau expression had no significant difference, but alpha synuclein and phosphorylation of GSK-3 beta, tau protein expression were significantly lower. There was significant difference (P<0.01).Compared with the normal group, the expression of Bcl2 protein in the model group was significantly decreased, and the expression of Bax protein was significantly increased, and there was significant difference (P<0.01).high dosage group of CRJD compared with the model group mice in the midbrain of the BCL2 protein expression was significantly increased and Bax protein expression were significantly decreased, there was significant difference (P< 0.01).Conclusion:1.CRJD can increase the content of DA in substantia nigra in MPTP induced PD model mice. CRJD can improve the behavioral disorder.2.CRJD extract has anti apoptotic effects, can reduce MPTP induced PD model mice brain GSK-3 beta activity, to dopaminergic neurons play a protective role.3. CRJD extract can by reducing the activity of GSK-3 beta, inhibition of tau protein excessive phosphate, reduce the expression of alpha synuclein, so as to reduce damage to dopaminergic neurons, which may is clinical Cistanche extract on PD therapy effective mechanism. |
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