Effects Of Curcumin And Ferulie Acid Inhibiting Of Prion Replication

Prions protein (PrPSc) is a misfolded protein rich inβforms, is tansformed bynormal prion protein. Both of them have the same primary structure and encoded bythe same gene Prnp. Recent studies show that, prion diseases are not sensitive toantibiotics and interferons, no significant immune response, it can not be cured byconventional antibiotics and interferons, and conventional methods can not becontaminated with prions sterilized items. Therefore, for diffusion and prionreplication mechanism, drugs used to reduce the accumulation of prions, inhibit prionprotein conformational changes and destruction of β structure of pathogenic proteinsand other methods to achieve the purpose of the prevention and treatment of nervedegeneration. More research reveal that Curcumin and ferulic acid have potentialeffects on Alzheimer’s disease, Parkinson’s disease and other prion likeneurodegenerative diseases, but so far, few studies illustrate whether these twotraditional Chinese medicine have therapeutic effect on prion diseases.In this study, prion infected cell (ScN2a) model is established to researchmonomer ferulic acid and curcumin inhibit the replication of prions, and to investigatethe molecular mechanisms of ferulic acid and curcumin inhibit the prion replication,and settle the foundation for further exploration of prevention and treatment for priondiseases. Results show that,0.1,0.5,1,5μg/mL of curcumin can significantly inhibitthe PrPCand PrPScexpression in ScN2a cells (P <0.01), after PK digest the totalprotein extracted from ScN2a cells, curcumin can significantly inhibit the expressionlevels of the PrPSc.0.5,1,5μg/ml Ferulic acid are able to significantly inhibit theexpression of PrPCand PrPScin ScN2a cells (P <0.01), However, when the totalprotein extracted from ScN2a cells digested by PK, PrPScprotein expression was notsignificantly changed. Using real-time fluorescence quantitative techniques, we foundthat curcumin did not affect prion protein mRNA transcription. In addition, throughthe observation of cell ultrastructure found that N2a cells vaccinaed prion strains, its Golgi apparatus, endoplasmic reticulum, mitochondria and other organelles havedifferent degrees changes, among which the most obvious changes arise inmitochondria, its mitochondria almost disappeared. After treated ScN2a cells with5μg/ml curcumin, mitochondria have obvious recovery. Above findings show thatcurcumin monomer can inhibit the replication of prions, playing its anti-amyloidprotein aggregation precipitation characters.We use two-dimensional gel electrophoresis analysis changes in proteinexpression levels in ScN2a cells treated with monomer curcumin. Compared with thecontrol group, the expression levels of a large number of proteins have changed aftertreated with curcumin. Though SDS-PAGE, We found20protein points which havelarge differences in proteins expression levels,6upregulated, and14downregulated.By mass spectrometry, we got the information of these20points.We detected these protein spots by Western blotting, and found two proteins thatremarkable different than others, ubiquitin C-terminal hydrolase L1(UCHL1) andisocitrate dehydrogenase3(IDH3A) protein. The results consisted with thetwo-dimensional gel electrophoresis. After curcumin treated, these two proteins wereupregulated compared with the control group, especially UCHL1protein, wassignificantly increased (P<0.05). UCHL1protein is a key protein spot study ofneurodegenerative diseases, and negatively correlated with the incidence of thisdisease. We thought that the protein may play a critical role in prion protein diseaseresearch.