The Proteomics Research On The Hypolipidemic Effect Of NRE In The Experimental Hyperlipemia Rat

ObjectiveThis study works on the target of NRE in order to illuminatethe proteinic mechanism , approach the regulation of lipidmetabomlism disorder,seek a proteinic regulation target for lipidmetabomlism,direct the development of descendens blood fatdrugs.Method:1. 80 Wistar rats are provided by the Experimental AnimalCenter of Jilin University. Divide the rats randomly into 5 groups:blank group,model group,low-dose-NRE group,large-dose-NREgroup,positive drug control group. Detect the concentration ofplasma TC,TG,HDL,LDL in 5 groups respectively. Take the liverconstitution of rats in 5 groups to observe the state of fattydeposition in liver.2. Extract the total protein of liver in 10 subjects in 4 groups( blank group , model group , low-dose-NRE group ,large-dose-NRE group ) . Using IEF/2-DE PAGE andMALDI-TOF-MS, we compare the abumdamces of protein in4groups. A total number of 54 protein spots with altered abundancesare detected and 11 of them were successfully identified byMALDI-TOF-MS.Result:1. Changes of blood fat:The plasma TG,TC,LDL-c in the low-dose-NRE group andthe large-dose-NRE group are lower than it in experimentalhyperlipemia model group,plasma HDL-c is higher than it in modelgroup and positive group p<0.05. low and large dose of NRE canreduce the LDL-c/HDL-c , TC/HDL-c , AI in experimentalhyperlipemia model group.2. Pathematology result:⑴Blank group:The structure of hepatic lobules is distinct. Thehepatocytes enclose the central vein of lobules radially. There are afew of leukomonocyte in the convergant canal area. inflammatorycell infiltrate obviously in the convergant canal area and necrosisfocus,inflammatory cell infiltration in hepatic lobules in several rats.Congestion can be seen in some hepatocytes.⑵Model group:Cell degeneration and necrosis lesions areobserved in hepatocyte. Hepatocytes are in significant hydropic andfatty degeneration. The inflammatory cells infiltrate obviously innecrosis lesion,bleeding and effusion of the hemaleucin are also canbe seen.⑶Positive control group:Widespread adipose degeneration ,kytoplasm puffing ,cell degeneration lesionly and inflammatorycell infiltration are observed in part of the hepatocytes.⑷Low-dose-NRE group:Significant fatty degeneration andkytoplasm puffing are observed in the freat mass of the hepatocytes.Necrosis lesion and inflammatory cell infiltration can be seen in partof the hepatocytes.⑸Large-dose-NRE group:Hepatocyte fatty degeneration andkytoplasm puffing are observes in only small part of thehepatocytes,while necrosis and inflammatory cell infiltration lesionin several hypatocytes. Merely mild fatty degeneration andkytoplasm puffing can be seen in most of the hypatocyres,and theinflammatory cell infiltration is not obvious.3.The result of proteomics research:The proteins below are obtained using mass spectrographicanalysis:⑴Catalase,CAT:Catalase is a kind of antioxidant enzymewhich has the ability to discard the free radical. It is observed thatthe use of NRE can increase the expression of CAT. So it isconjectured that CAT can inhibit the oxidative stress,keep thebalance of oxidation and anti-oxidation.⑵glucose regulated protein, GRP:GRP is a kind of stressprotein and a member of HSP family which express inceasingly instringent state. In this study,GRP expression increase significantlyafter using NRE. It could conclude that NRE improve the resistability of stress,protect hepatocytes and the protein molecule in thecells,recover the normal structure and function.⑶Selenium binding protein2:Selenium binding protein2 is amember of selenium-cantaining protein family,participate thestorage,transportation and utilization of Se in vivo. The increasedexpression of this protein indicates the increased utilization of Se.Se is the active center of GSHPx,PHGPx which blocks the radicalreaction and discards LPO. In this study,the use of NRE raise theexpression of selenium. So we infer that NRE could increase theutilization of Se,enhance the recovery of impaired hepatocytes,decompose the poisonous peroxide,promote the antibody providedby lymphocytes,block the radical reaction,discard LPO,prevent thehepatocytes from impairment.⑷Endoplasmic retuclum protein 29, ERP-29:ERP-29 is adissoluble protein located in eucaryon endoplasmic retuclum. Itpromotes the folding of the protein and/or participate the secretionof the endoplasmic retuclum secretory protein. In our study, theERP29 expression decreases in model group compared with blankgroup. The expression increases after using NRE,but still lowerthan the blank group. So it is concluded that NRE relieve the degreeof adipose degeneration in fatty liver cell,recuperate the structureand function of hepatocytes gradually,maintain the activity ofhepatocytes simultaneously , keep and promote the speed ofhepatocyte growth,enhance the anti-stress ability .X⑸CAIII is the major soluble protein in liver and muscle. Itsfunction is probably to protect proteins of these tissues fromoxidation catalyzed by iron-containing degradation products ofhaemoglobin and myoglobin. It is proved that the expression ofCA-III has the positive correlation with leptin which has the abilityto regulate the fat metabolism , inhibit the happen ofhyperlipoidemia. In this study,the use of NRE can improve thefunction of liver cells , lessen the hyperlipoidemia , add theexpression of CA-III. So it is suggested that the increasedexpression of CA-III cause the raised leptin level,increase theactivity of leption,promote the fat metabolism,play an importantrole in anti-hyperlipoidemia.⑹Phosphatidylethanolamine binding protein,PEBP:It isproved that PEEP family is a kind of important regulateion moleculewhich inhibit the Raf21/MEK/ERK regeneration signal transmissionthoroughfare. In our study,ny using NRE the hyperlipoidemia islessened,the recovery of impaired performs gradually. So thedemand of regeneration decreases,while the required amount ofPEEP increases propotionally to inhibit the regeneration. It is just inthis way to maintain the constancy of liver cell.Conclusion1.This study comfirmed that NRE has effect to reduce bloodlipids which can decrease the contents of TC,TG and LDL-C, andincrease the HDL-C content and reduce the content of . TC/HDL-Cand the ratio of LDL-C/HDL-C.2. NRE has prevent the fatty liver formation(see pathologicphotoes).3. NRE may possibly play a role by the interaction of CAT,GRP, selenium binding protein2, endoplasmic retuclum protein29(ERP-29) , carbonic anhydrase 3(CA-III) and Phosphatidy-lethanolamine binding protein(PEBP), et al.