Effect of Curcumin on Cholesterol Metabolism of Nonalcoholic Fatty Liver Disease Cells / 浙江中医药大学学报

[Objective]To study the effect of curcumin on the cholesterol metabolism of nonalcoholic fatty liver disease(NAFLD) cells model induced in vitro and its potential mechanism. [ Methods]The cellmodel of nonalcoholic fatty liver disease(NAFLD) was established by oleic acid and treated with curcumin. The method of oil red O staining was used to observe accumulation of intracellular lipid while the intracellular content of TG, FC and TC was detected by enzymatic method. Meanwhile, the mRNA levels of SR-BΙand HMGCR were detected with qPCR.[ Results] The NAFLD cellmodel was successful y established by culturing with 30 μg·mL-1 oleic acid. After curcumin intervention, TG, FC and intracellular lipid accumulation levels were significantly reduced in NAFLD cellmodel. Meanwhile, curcumin can reduce HMGCR mRNA expression and raise SR-BΙ mRNA expression. [Conclusion] Curcumin can decrease FC level in NAFLD cellmodel and the mechanism might be related with its capacity of restraining endogenous cholesterol biosynthesis and promoting foreign cholesterol transfer into the liver cells for metabolism.

Proteomic research on anti-atherosclerosis mechanism of curcumin in RAW264.7 / 中国中药杂志

<p><b>OBJECTIVE</b>Two-dimensional difference gel electrophoresis and mass spectrum were used to study the anti-atherosclerosis mechanism of curcumin.</p><p><b>METHOD</b>The 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.</p><p><b>RESULT</b>The 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.</p><p><b>CONCLUSION</b>The 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.</p>

Effects of Liuwei Dihuang Decoction on hepatic mitochondria proteome of kidney-yin deficiency rat induced by hyperthyrea / 国际中医中药杂志

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.

Effects of warm and tonify kidney-yang herbs on liver mitochondria proteome of kidney-yang deficiency rats / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of warm and tonify kidney-yang herbs on the liver mitochondria proteome of the thyroidectomized kidney-yang deficiency rats.</p><p><b>METHOD</b>Twelve 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.</p><p><b>RESULT</b>The 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.</p><p><b>CONCLUSION</b>The 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.</p>

The Status and Considerations of Study on Kidney-Yang Deficiency / 浙江中医药大学学报

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.

Effects of Warming and Tonifyin Kidney-Yang Drugs on Liver Mitochondria Proteome of Rats with Kidney-Yang Deficiency / 中华中医药杂志

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.

Effect of Siwu Decoction on Liver Mitochondrial Proteome of Blood Deficiency Model Rats / 中医杂志

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