| Background Parkinson's disease(PD)is the second most common neurodegenerative disorder.With the coming of the old-aged society,the incidence of PD is rising all over the world.According to investigation reference of the world Parkinson's Disease Association,there has been up to four millions of PD patients in the world and our country have 1.72 millions,which occupys the first place.In addition,the total amounts of PD patients are still rising about 100 thousand every year.Therefore,how to deal with PD has become an important medical and social problem.Previous researches demonstrated that cytoplasmic Inclusions known as Lewy bodies are formed within the dopaminergic neurons of the substantia nigra pars compacta in PD.These inclusions contain not only ubiquitin,but also a-synuclein, subunits of the proteasome,and other proteins.It's suggested that polyubiquitinated proteins could accumulate in inclusions because of a dysfunction at the level of degradation by the proteasome.The opinion-above is supported by more and more research data those show us the decreased proteasomal activity in PD nigral tissue.At present,the L-dopamine(L-DA)is worldwide applied to therapy PD.Although it alleviates the symptoms,it can also result in a series of negative effect,and can't effectively hold back the development of PD.Recently researches demonstrated that the therapeutic effect has been significantly improved with Chinese medicine-treated, which not only delays the process of PD,but also relieves the negative effect of some chemicals,and can also control other symptoms besides movement disorders.At present,researchers in our country commomly aimed at the exploration of the mitochondrial dysfunction,increased oxidative stress and cell apoptosis mechanism, However,there is fewer study on the mechanism of Chinese medicine's therapeutic effect of ubiquitin proteasome system in PD.Chinese medicine "AnChanLing" has a beneficial effect on treating with PD, which has been proved by the many years of clinical application.We had established the PD rat model by injecting lactacystin,and observed the AnChanLing effect on the UPS function and anti-apoptosis by immunohistochemistry and TUNEL.In the present study,We are intent to provide AnChanLing with some in-vitro experimental evidence on treating PD,and it would be shed-light on the therapy of PD patients as well.Objective To Explore the effect of AnChanLing on UPS desfunction in PC12 cells.Methods We used pharmacological inhibitors of the proteasome to create a cellular model of proteasomal dysfunction in PC12 cells by lactacystin.PC12 cells were divided into three groups:lactacystin group,Anchanling-treated group and control group,We used MTT assay to evaluate cellular viability,and tyrosine hydroxylase (TH)immunohistochemical staining to investigate the function of PC12 each group.TUNEL staining was employed to observe the apoptosis of nerve cells.The expressions ofα-synuclein,ubiquitin proteins were measured by immunofluorescence.The research data were expressed as mean±SD.The significance of inter-group difference was evaluated by one-way analysis of variance (ANOVA:LSD for post hoc comparisons).The level of significance was accepted at P＜0.05.Results The AnChanLing middle dose group showed the highest cellular viability of the three dose group(P＜0.01).In the contrary of the control group,the proportion of apoptosis in lactacystin group was increased significantly(P＜0.01),and a-synuclein, ubiquitin positive cells were also more than control group,but TH positive cells were less than that(P＜0.01).The proportion of apoptosis in the group of Chinese medicine trated were significantly lower than the lactacystin group(P＜0.01),and a-synuclein, ubiquitin positive cells were also less than that group,but TH positive cells was more than that(P＜0.01).Conclusions AnChanLing could obviously lower the neurotoxicity induced by lactacystin in PC12 Cell,which maybe relate to inhibiting the apoptosis of PC12 cell and ameliorating effectively the function of UPS. |
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