OBJECTIVETwo-dimensional difference gel electrophoresis and mass spectrum were used to study the anti-atherosclerosis mechanism of curcumin.

METHODThe proteins from RAW264.7 cell and RAW264.7 cell treated with 25 micromol x L(-1) curcumin were labeled with Cy3 or Cy5 randomly. Each Cy3-labeled sample and Cy5-labeled sample was mixed on the same 2-D gel along with a Cy2-labeled mixture of all samples as an internal standard and run on the same gel. The gels were scanned under different wave-length light after electrophoresis. All images were analyzed by DeCyder 6.5 software, and the different proteins were identified by mass spectrum.

RESULTThe expression of ATP synthesis H+ transporting, MHC class II, non-muscle myosin alkali light chain and cytochrome b5 increased in the RAW264.7 cell treated with 25 micromol x L(-1) curcumin, while the expression of phosphodiesterase 4D, elF-3, Hnrpf protein, vimentin, nucleophosminl and Ranbp 1 decreased.

CONCLUSIONThe anti-atherosclerosis mechanism of curcumin is the result of enhancement of the cell inflammation, antioxidant activity and inhibition of cholesterol transport, reduce of the accumulation of intracellular cholesterol and other factors. In addition, curcumin also had the effect of anti-tumor through regulated tumor cell differentiation and apoptosis.

Animals ; Atherosclerosis ; metabolism ; Cell Line ; Curcumin ; pharmacology ; Electrophoresis, Gel, Two-Dimensional ; methods ; Gene Expression ; drug effects ; Major Histocompatibility Complex ; physiology ; Mass Spectrometry ; Mice ; Proteomics ; methods ; Reverse Transcriptase Polymerase Chain Reaction

Objective Two-dimensional electrophoresis and mass spectrum were used to study the effects of Liuwei Dihuang Decoction on hepatic mitochondria proteome of kidney-yin deficiency rat induced by hyperthyrea. Methods The protein of liver mitochondria from kidney-yin deficiency rats and the kidney-yin deficiency rats treated with Liuwei Dihuang Decoction were isolated by two-dimensional electrophoresis, then stained with silver staining method. The gels were respectively imaged by UMAX ImageScanner and spot-features of proteome difference were analyzed by ImageMaster2D6.0 software. MALDI-TOF MS was used to identify the different protein with abundant changes more than 2 folder between normal rats and kidney-yin deficiency rats. Results In the treated group it was found that major urinary protein precursor, peroxiredoxin-6, endoplasmic reticulum protein 29 precursor, chain D rat transthyretin were increased, while 78 kDa glucos eregurated protein precursor, glutathione S-transferase Mu 1, 60 kDa heat shock protein, peptidyl-prolyl cis-trans isomerase, H~+ -transporting ATP synthase, nuclear factor kappa B 2, BNIP1 protein were decreased. Conclusion The mechanism of nourishing kidney-yin action of Liuwei Dihuang Decoction lies in its relieving energy metabolism, regulating inflammatory factorderangement and apoptosis inhibition, antagonizing free radical damage, enhancing gonadal hormone level by the down regulation of NF-κB, H~+-transporting ATP synthase, and the up-regulation of PDX6.

OBJECTIVETo study the effects of warm and tonify kidney-yang herbs on the liver mitochondria proteome of the thyroidectomized kidney-yang deficiency rats.

METHODTwelve rats were divided into normal group, model group and treated group; each group had four rats. The rats of model group and treated group were excised the two side thyroid gland, and the rats of normal group were done the homologous operation, but didn't excise the thyroid gland. After seven days, the rats of model group and treated group appeared the symptoms of kidney-yang deficiency. From the eighth day, the rats of treated group were fed warm and tonify kidney-yang herbs 6.7 g x kg(-1) once daily, and the rats of other two groups were fed the equal normal. All rats of three groups were killed by decollation after six days of treatment, and liver mitochondria proteins were separated. Each liver mitochondria protein sample was labeled with Cy3 or Cy5 randomly, and one Cy3-labeled sample and one Cy5-labeled sample were mixed on the same 2-D gel along with a Cy2-labeled mixture of all samples as an internal standard and run on the same gel. The gels were scanned under different wavelength light after electrophoresis. All images were analyzed by DeCyder 6. 5 software, and the different proteins were identified by mass spectrum.

RESULTThe expression of HSP60, catalase, glutathione peroxidase, carbamoylphosphate synthetase I, ATP synthase, lactotransferrin, H(+)-transporting two-sector ATPase, alpha-ETF and calpain 12 were increased in the thyroidectomized kidney-yang deficiency rats, while the expression of oxoisovalerate dehydrogenase, Acyl-CoA dehydrogenase, ornithine aminotransferase, and GTP-binding regulatory protein were decreased. After the kidney-yang deficiency rats were treated with warm and tonify kidney-yang herbs, the expression of HSP60, catalase, glutathione peroxidase, oxoisovalerate dehydrogenase, Acyl-CoA dehydrogenase, ATP synthase, lacto-transferrin, H(+)-transporting two-sector ATPase, alpha-ETF and GTP-binding regulatory protein were increased, and the expression of carbamoyl-phosphate synthetase I and calpain 12 were decreased.

CONCLUSIONThe warm and tonify kidney-yang herbs perform its therapeutical effect by regulating the metabolism, protecting the stability of mitochondrial membrance and maintaining the signal conduction in the cells.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Kidney ; drug effects ; metabolism ; Male ; Medicine, Chinese Traditional ; Mitochondria, Liver ; drug effects ; metabolism ; Proteome ; drug effects ; metabolism ; Proteomics ; Rats ; Rats, Sprague-Dawley ; Yang Deficiency ; drug therapy ; metabolism

Atherosclerosis is the high risk factor leading to cardiovascular and cerebrovascular diseases at present. In recent years, many clinical and experimental researches have been done by the use of traditional Chinese and modem medical means multi-levelly and multi-anglely. The researches showed notable therapeutic effects of traditional Chinese medicine and the researches on cellular and molecular level has become a. hot spot. Ligustrazine is an effective component of Szechwan Lovage Rhizome. The function of, Lignstrazine in preventing atherosclerosis has provoked an extensive attention. Its in-depth study can provide a more objective and more comprehensive guide for Clinical Practice. The paper tried to make an overview on the researching progress of Ligustrazine in preventing atheroselerosis in terms of parietal cells of blood vessel,hemodynamics, and lipid metabolism.

Curcumin,the active substance of turmeric.has multiple therapeutic activities.However,its water solubility and bio-availability in vivo is very low,which limits the utility of curcumin.This article reviewed recent approaches of the bioactivity and the structure-function relationship of curcumin.The authors suggest that new methods should be tried to modify the structure of cureumin and further studies of therapeutic activities of the homologues are needed.

The inhibition of cell apoptosis could result in NDR,because almost all antitumor durgs are inductor of cell apoptosis. According to the study,the overexpression of mutant p53,Bcl-2 family,c-myc gene,NF-kB,HSPs,or ceramide are important reasons in MDR.

With the development of Chinese medical research,it becomes an important problem how to scientifically explain syndrome in Chinese medicine.In this paper,the progress on deficiency of kidney yang is reviewed from endocrine,immune function and energy metabolism,and it is preferred that the technology of genome and proteome be used to study the deficiency of kidney yang.

[Aim] The expressed system of sterol regulation by SCAP/SREBP Pathway was established in the oocyte of the Xenopus laevis and was applied to investigate the effect of cucurmin on the SREBP pathway.[Methods] The plasmid of PLXRN-4SRE-fpa was restructed and injected into the nucleus of Xenopus Oocytes in phase V or VI. The oocyte was incubated with different concentrations of the curcumin. After three days the fluorescence intensity on the membrane of the Oocytes was measured by Fluoroanalyzer.[Results] The fluoresecence intensity on the membrane of the oocyte increased with increasing of the concentration of the curcumin. The trail group which have all regulation elements of SCAP/SREBP pathway can express more fluorscence protein than the control group. [Conclusion] The experiment provides that the curcumin can upgrade the expression of low density lipoprotein receptor by SCAP/SREBP regulation pathway. The model can be applied for quick screening of the scap ligand drugs and inverstigating the mechanism of the drugs.

Aim To investigate the lipid-lowering mechanism of Chinese Medicine curcumin.A sterol regulatory report system of LDLR gene expression in HEK-293 cell line was established.Method A green fluorescent protein(GFP) gene was constructed downstream of the sterol regulatory element-1(SRE-1) and the constructed plasmid was introduced into HEK-293 cell line.This SRE-GFP transfected 293 cells were treated with different concentration of curcumin for 2 days.The expression of GFP in the cells was measured by a fluorescence microscope and Flow Cytometry.Results The expression of GFP in SRE-GFP transfected 293 cells was obviously induced by curcumin.Conclusion Curcumin may positively affect the expression of LDLR gene by its influence on SRE-1.

Objective To observe the mechanism of Siwu Decoction (Decoction of four ingredients) on blood deficiency syndrome.Methods Wistar rats were randomized into 3 groups: model group, treatment group, and normal group, 6 ones in each. Rats of the model group and the treatment group were injected 2% acetylphenylhydrazine (APH) saline subcutaneously on the first, forth and seventh day. From the forth day, the model group was treated with saline by gavage, the treatment group was treated with Siwu Decoction by gavage, and the normal group was treated with saline for 7 days in succession. Then the amount of peripheral blood cells, wet weight of liver and spleen were detected. Liver mitochondrial proteome of all rats were extracted. Proteomic technology was used to measure the regulated proteins by Siwu Decoction.Results In the model group, the rats had decreased general status significantly such as pale, enlarged liver and spleen, lowered red blood cells (RBC) and hemoglobin (HGB), increased white blood cells (WBC) and reticulocyte (RCT) (P