Improvement of Pulsatilla saponin composition on Parkinson's disease and Alzheimer's disease in mice
Objective To investigate the protective effects of a combination of Pulsatilla saponin B4 and hederagenin C(HSC,anemoside B5)with a quality ratio of 4∶1 on Parkinson's disease(PD)and Alzheimer's disease(AD)models in mice and the underlying mechanisms.Methods Sixty healthy male C57BL/6 mice were randomly selected,with 10 mice serving as the control group.The remaining 50 mice were intraperitoneally injected with l-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)to establish a PD model.After the model was successfully established,the mice were randomly divided into the model group,the positive control group(levodopa hydrochloride tablets,30 mgkg-1),and the Pulsatilla saponin B4+HSC high,medium,and low dose groups(20,10,and 5 mg·kg1),with 10 mice in each group.The mice were ig given the drugs for 14 d,once a day.The control and model groups were ig given an equal volume of 0.9％sodium chloride solution.The onset and disappearance times of tremor behavior were observed every two days.Behavioral experiments were conducted and scores were recorded on days 7 and 14 of the treatment.Hematoxylin and eosin(HE)staining was used to observe pathological changes in the substantia nigra of the brain tissue.Immunohistochemistry was used to detect the expression of α-synuclein in the brain tissue.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of dopamine transporter(DAT)and tyrosine hydroxylase(TH)mRNA in the brain tissue.50 APP/PS1 transgenic mice were randomly divided into the model group,the positive drug group(donepezil hydrochloride,1.5 mg·kg-1),and the high,medium,and low dose groups of Pulsatilla saponin B4+HSC(20,10,and 5 mg·kg-1),respectively.Ten APP/PS1 transgenic wild-type mice were taken as the control group.The mice were administered ig once a day for 28 d.The model group and the control group were given the same amount of 0.9％sodium chloride solution.After the administration,behavioral tests were conducted and the scores were recorded.The levels of inflammatory factors interleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),cyclooxygenase-2(COX-2),and reactive oxygen species(ROS)in the serum of the mice were detected by ELISA.The levels of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were detected by kits.Results In the PD model mice,compared with the model group,Pulsatilla saponin B4+HSC significantly reduced the tremor paralysis score(PD-RATS)score(P＜0.01,0.001),significantly reduced the open field test score(P＜0.01,0.001),significantly reduced the climbing rod test score(P＜0.01,0.001),significantly reduced the hanging test score(P＜0.01,0.001),significantly reduced the swimming test score(P＜0.01,0.001),and significantly reduced the comprehensive score of behavioral test(P＜0.001),and significantly improved the structure damage of substantia nigra,significantly reduced the expression of α-synuclein,and significantly increased the levels of DAT and TH mRNA in the brain tissue of mice(P＜0.05,0.01,0.001).In the AD mouse model,compared with the model group,Pulsatilla saponin B4+HSC significantly shortened the escape latency in the water maze spatial navigation experiment(P＜0.001),significantly prolonged the cumulative stay time in the target quadrant(P＜0.05,0.01,0.001),significantly increased the number of accurate crossings of the platform location(P＜0.05,0.01),significantly reduced the serum inflammatory factor level(P＜0.05,0.01,0.001),significantly reduced the level of ROS,and increased the level of SOD and GSH-Px(P＜0.05,0.01,0.001).Conclusion Pulsatilla saponin B4+HSC effectively played protective roles on PD and AD in mice,which may be achieved through anti-inflammatory and antioxidant effects and improvement of brain tissue and neuron damage.
NETL
NSTL
万方数据
