Objective: To study the effects of warming and tonifying kidney-Yang drugs on the liver mitochondria proteome of rats with kidney-yang deficiency using two-dimensional difference gel electrophoresis(2-D DIGE).Methods: Every liver mitochondria protein sample from the normal rats,the kidney-yang deficiency rats and the kidney-yang deficiency rats treated with warm and tonify kidney-yang drugs were labeled with Cy3 or Cy5 randomly,Each Cy3-labeled sample and Cy5labeled 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.All images were analyzed by DeCyder software DIGE technology with internal standard.16 unique proteins were selected,and in which 10 protein were successfully identified.Results: In the kidney-yang deficiency rats,the expressions of carbamoyl-phosphate synthetase 1,dihydrolipoamide branched chain transacylase,ubiquinol-cytochrome c reductase,isovaleryl coenzyme A dehydrogenase,heat shock protein 60,succinyl-CoA synthetase,hydroxyamino-coenzyme A dehydrogenase/ ketoacyl coenzyme acetyl coenzyme A dehydrogenase ?subunit,ornithine transcarbamylase and glutathione peroxidase all increased.While the expression of ornithine aminotransferase decreased.After the treatment of warming and tonifying kidney-yang drugs,the expressions of above-mentioned proteins all had rectification in different degrees. Conclusion: The metabolism of carbohydrate,lipids and protein had been rectified when the kidney-yang deficiency rats were fed with warming and tonifying kidney-yang drugs,which was likely attributed to the improvement of stability of mitochondria membrane.

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.

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.

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

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

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 To observe the effects of Jianpi Qutan Huayu Prescription on oxidative stress and inflammatory response in mini-pigs with atherosclerosis(AS);To explore its mechanism based on the NOX5-ERK1/2 signaling pathway.Methods Twelve Bama mini-pigs were randomly divided into control group,model group,and Jianpi Qutan Huayu Prescription low-and high-dosage groups,with 3 pigs in each group.A high-fat diet was used to feed for 24 weeks to construct an AS model,and the treatment group was also supplemented with Jianpi Qutan Huayu Prescription in the feed.The general condition of mini-pigs(body length,abdominal circumference,body mass,food intake,and fecal water content)was measured at week 0,16,and 24 of administration,HE staining was used to observe the morphology of aortic tissue,while oil red O staining was used to observe lipid deposition in aortic and myocardial tissue,transmission electron microscopy was used to observe the ultrastructure of aortic tissue,and a fully automated biochemical analyzer was used to detect serum contents of TC,HDL-C,and LDL-C.ELISA was used to detect the contents of serum reactive oxygen species(ROS),interleukin(IL)-6,IL-10,tumor necrosis factor(TNF)-α,hypersensitivity-C-reactive protein(hs-CRP),vascular cell adhesion molecule-1(VCAM-1),and intercellular adhesion molecule-1(ICAM-1).Western blot was used to detect the expressions of NADPH oxidase 5(NOX5),extracellular signal regulated kinase 1/2(ERK1/2),p-ERK1/2,VCAM-1,and proliferating cell nuclear antigen(PCNA)proteins.Results Compared with the control group,the abdominal circumference,body mass,and food intake of mini-pigs in the model group increased at 16 and 24 weeks(P<0.01),there was significant thickening of the inner membrane of aorta,destruction of endothelial cells,lipid deposition,edema of smooth muscle cells,and significant swelling of mitochondria,serum TC,LDL-C contents and the contents of ROS,IL-6,IL-10,TNF-α,hs-CRP,VCAM-1,and ICAM-1 increased,while the content of HDL-C decreased(P<0.01);the expressions of NOX5,p-ERK1/2,VCAM-1,and PCNA proteins in aortic tissue increased(P<0.01).Compared with the model group,Jianpi Qutan Huayu Prescription low-and high-dosage groups showed a decrease in abdominal circumference,body mass,and food intake at 16 and 24 weeks(P<0.05,P<0.01),the plaque area and lipid deposition were reduced,and the damage to endothelial cells was alleviated,serum TC,LDL-C contents and the contents of ROS,IL-6,IL-10,TNF-α,hs-CRP,ICAM-1,and VCAM-1 decreased,and the content of HDL-C increased(P<0.01,P<0.05);the expressiond of NOX5,p-ERK1/2,VCAM-1,and PCNA proteins in aortic tissue decreased(P<0.01,P<0.05).Conclusion Jianpi Qutan Huayu Prescription can effectively alleviate AS in mini-pigs,and its mechanism may be related to inhibiting the activation of the NOX5-ERK1/2 signaling pathway and alleviating oxidative stress-induced inflammatory response.

[摘 要] 目的：通过分析非小细胞肺癌顺铂敏感株及耐药株的基因芯片表达数据，筛选差异基因及关键通路，构建蛋白相互作用网络，探讨关键集群功能。方法:从GEO数据库获得基因芯片表达数据，利用GEO2R工具筛选差异基因，通过STRING数据库和Cytoscape软件构建蛋白相互作用网络，经DAVID富集得到相关特征基因与信号通路信息。结果：通过芯片分析共获得481个差异表达基因，相比于敏感细胞株，顺铂获得性耐药细胞株中有418个上调基因和63个下调基因。差异基因功能主要富集在piRNA代谢、DNA甲基化修饰、细胞有丝分裂及细胞周期进程等信号通路。蛋白复合物预测得到主要功能集群6个，分别与细胞趋化性、细胞角化性、piRNA代谢过程、细胞因子受体相互作用、细胞因子分泌调节及染色质沉默相关生物进程相关。结论:本研究利用生物信息学方法，发现顺铂耐药细胞株特征基因及信号通路，其中SAA1、KRT5、TDRD9、BCL2A1、CSF1R和HIST1H1A等显著上调基因及其功能集团可能是非小细胞肺癌顺铂耐药的潜在分子机制，为临床精准治疗提供新的理论依据。