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>

Advances in Modern Researches on Ligustrazine Prevention of Atherosclerosis / 国际中医中药杂志

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.

Review of the Bioactivity and the Structure-function Relationship of Curcumin / 国际中医中药杂志

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.

Research on Inhibition of Cell Apoptosis and Multidrug Resistance (MDR) / 浙江中医药大学学报

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.

Establishing the SREBP Pathway in the Oocytes Xenopus Laevis and Stuyding the Effects of the Cucurmin / 浙江中医药大学学报

[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.

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.

Effect of curcumin on sterol regulatory report system in HEK-293 / 中国药理学通报

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.

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

Effect of Shenmai Injection on angiogenesis in tumor tissue / 中成药

Objective: To investigate the effect of Shenmai Injection (Radix Ginseng Rubra, Radix Ophiopogonis) on angiogenesis and the expression of proliferating cell nuclear antigen (PCNA) in tumor tissue, and the mechanism of it in the treatment of tumor. Methods: The mouse tumor model was used to investigate the effect of Shenmai Injection on tumor growth on the whole. The expression of von Willebrand factor (vWF) and PCNA in tumor tissue was studied by means of immunohistochemistry. Results: Shenmai inhibited the tumor growth and reduced microvessel density and the expression of PCNA in tumor tissue. Conclusion:Antiangiogenesis is one of the mechanisms of Shenmai Injection in treatment of tumor.

Relation among TCM Syndromes,Sex Hormones and Molybdenum Target Radiography Image of Mammary Hyperplasia / 中医杂志

Objective To explore the correlation between the TCM syndromes of mammary hyperplasia patients and their sex hormones,molybdenum target radiography image to establish the objective indexes for its syndrome differentiation.Methods According to the TCM syndrome differentiation,the 346 patients with mammary hyperplasia were diagnosed liver qi stagnation syndrome,phlegm and blood-stasis syndrome,and chong and ren dysfunction syndrome.Their pituitary gonad stimulating hormones,including estradiol(E2),progesterone(PP),testosterone(PT),prolactin releasing hormone(PRL),follicle-stimulating hormone(FSH),and luteotropic hormone(LH) were measured during ovulatory period.At the same time,the molybdenum target radiography examination was given.Results The level of FSH,LH and E2 in mammary hyperplasia patients during ovulatory period was obviously lower,the level of PRL and PP was obviously higher,and the level of PT was not obviously changed.The comparison among groups showed that the level of FSH in chong and ren dysfunction patients was higher than that in liver qi stagnation patients,but the level of LH of the two was just opposite;the level of E2 was lower in an order of liver qi stagnation,phlegm and blood-stasis,and chong and ren dysfunction patients;the molybdenum target radiography image showed that there was mainly the lobular hyperplasia in liver qi stagnation and phlegm and blood-stasis patients and then the nodular type;but there was mainly nodular hyperplasia in chong and ren dysfunction patients and then the lobular type.Conclusion The pituitary gonad stimulating hormones of mammary hyperplasia patients were disordered in the duration of ovulatory period.The level of sex hormones and change of molybdenum target radiography image were different in different TCM syndromes and they can be used as one of the objective indices in TCM syndrome differentiation for diagnosis.

Effects of poly- N -acetylglucosamine on metabolism of low density lipoprotein and lipoprotein(a) / 中国药理学通报

AIM To study the effects of poly- N -acetylglucosamine on the metabolism of low density lipoprotein and lipoprotein (a). METHODS Poly- N -acetylglucosamine was injected into the body of hedgehogs viathe armpit vein 2 minutes before the 125 I low density lipoprotein ( 125 I-LDL), 125 I lipoprotein(a) 〔 125 I-Lp(a)〕, 125 I desialylated low density lipoprotein ( 125 I-ds-LDL) or 125 I desialylated lipoprotein(a) 〔 125 I-ds-Lp(a)〕 were injected separatively . The animals were put to death in one hour. The radioactivity in the blood, liver, kidney, spleen, gall and adrenal were measured. RESULTS Absorption of ds-LDL and ds-Lp(a) increased in the liver. Later ds-LDL was reutilized and released into blood, but the majority of ds-Lp(a) was excreted out of the body from liver. It was not eliminated that ds-LDL was metabolized via the LDL receptor for the uptake of ds-LDL also increased in the kidney and the adrenal. It is possible that ds-Lp(a) and the part of ds-LDL may be catabolized via the desialylated lipoprotein-related receptor. Poly- N -acetylglucosamine could increase the absorption of LDL in the liver and the adrenal, meanwhile the concentration of LDL decreased in the blood and the absorption of LDL decreased in the spleen. CONCLUSION Poly- N -acetylglucosamine has the effects of lowering lipids through activating LDL receptor.

ANALYSIS OF ESTROGEN AND PROGESTERONE LEVELS DURING MENSTRUAL CY-CLE IN PATIENTS WITH FIBROCYSTIC DISEASE OF BREAST / 中华内分泌代谢杂志

The blood and urine estrogen (E2) and progesterone (P) were measured in 133 patients with fibrocystic disease (FCD) of breast and 17 healthy subsjects during menstrual cycle phases. According to the characteristics of the secretion, the patients were divided into five types. Type A (54 patients): The secretion peak of E2 and P appeared beforehand and secretion of P was prolonged and E2 decreased significantly in ovulatory phase. Type B (16 patients): The secretion peak of E2 disappeared in ovulatory phase although the secretion of P was normal. Type C (16 patients): The secretion peak of P deferred its disappearance and maintained at higher level till pre-menstrual period and sc did E2: the secretion of E2 decreased in ovulatory phase. Type D (41 patients): The secretion peak of E2 in ovulatory phase was normal but could not return to normal level in pre-menstrual period, in the mean time the secretion of P also deferred its disappearance. Type E (6 patients): There was no secretion of P and the blood and urine concentrations of E2 deccreased in all phases. The average value of E2 in the patients with FCD was compared with that of healthy subjects. The concentration of E2 in all the patients increased in follicular and pre-menstrual phases. However, the concentration of E2 in ovulatory phase was much lower in the patients than in healthy subjects.

Effect of ligustrazine on the human scavenger receptor-AI expressed in Xenopus oocytes / 中国药理学通报

Aim To investigate and evaluate the effect of ligustrazine on the expression of scavenger receptor type AI, the human ?-interferon activation site (GAS) elements regulatory systems of SR-AI expression wasfirst established in the Xenopus oocytes. Methods A plasmid, in which the human SR-AI gene was constructed downstream from the GAS elements, was microinjected into the nucleus of Xenopus oocytes. The oocytes then were cultured in the smooth muscle cell conditioned medium (SCM), In which the cell were cultured with the treatment of oxLDL for 2 days. In the other groups, the oocytes were treated with SCM containing the nature medicines ligustrazine, and the positive control drug ?-interferon respectively. Results The results demonstrated that SCM could upregulate the expression of SR-AI in this system, this action was suppressed by ligustrazine and ?-interferon. Conclusion Similar to the action of ?-interferon, ligustrazine can prevent the over-expression of SR-A induced by oxLDL, via their effect on the GAS elements regulatory pathway.

Astragalus polysaccharides(APS) induces the cord blood monocytes into mature dendritic cells(DCs) directly and its impact on T cells proliferation / 中国免疫学杂志

Objective:To observe the effect of Astragalus polysaccharides(APS) on inducing the cord blood monocytes into mature dendritic cells(DCs) in vitro and to investigate their morphous,cellar immunological characteristics,and contribution to T cell proliferation.Methods:①The cord blood monocytes were isolated by lymphocyte isolating solution under axenic condition,and three groups were divided.②Cells cultured with APS in concentration of 100 mg/L as the experiment group, that with the cytokines of GM-CSF/IL-4/TNF-? as the positive control, and another without either GM-CSF/IL-4/TNF-? or APS as the negative control, respectively. The morphology of DCs was identified by inverted optical microscope or scanning electron microscope. The phenotype of 12 days cultures of DCs(CD1a, CD80, CD86 and CD83) were identified by flowcytometry. The DCs preparations from the experiment group were treated with mitomycin for 45 min to remove their proliferative activity as incentive cells in the mixed cultures with allogenic peripheral blood mononuclear cells from healthy volunteers as responders. T cell proliferation induced with the DCs preparations was detected by MTT chromatometry.Results:After cultured for 72 hours, the cell of both the experiment group and the positive control grew dusteringly and began to change from round to irregularin shapes. The longer the cells were cultured, the more obvious the dendritic structure is. The cells of experiment group and the positive control group when cultured for 12 days had the most typical dendritic structure. The negative control group cells had no dendritic structure and became macrophages when cultured for 12 days. The experiment group cells cultured for 10 days showed typical dendritic morphology by SEM. The experiment group cells and the positive group cells cultured for 12 days significantly expressed high level of the phenotypes of DCs(CD1a, CD80, CD86 and CD83) by flowcytometry.And the difference exhibitied statistical significance when compared with the negative control group(P0.05).The mixed lymphocyte reaction showed that the DCs induced by APS trigerred proliferation of allogenic T cells obviously.Conclusion:Both APS and cytokine could induce the cord blood monocytes to differentiate into functional DCs committedly. DCs reduced by APS stimualate proliferation of the allogenic T cells obviously.

Effect of Ginseng Rhizome Saponin by cAMP and cGMP in the Formation of the E~*-Rosette of the Human Lymphocytes / 中国免疫学杂志

The concentration of the cyclic nucleotides and its relation with the E-RFC were determined by using ginseng rhizome saponin in human peripharal leukocytes.Result were compared with the ckange of the E-RFC by using cAMP.cGMP and theophylline. Experiments proved that girseng rhizome saponin mainly affected the lymphocytes and significantly increased the concentration of cAMP and cGMP,especially,the cAMP. Lymphocytes were incubated with ginseng rhizome saponin to form Et-RFC. It is different where lymphocytes were incubated in diffierent times. cAMP and theophylline reduced the Ea-RFC and Et-RFC, cGMP increased the Ea— RFC,but no effect to the Et-RFC. Ginseng rhizome or root saponin could promote the Ea-RFC,but inhibited the Et-RFC. It is consistent with the effect of both cAMP and cGMP. Experiments showed that ginseng rhizome saponin increased the concentration of cAMP and cGMP in human lyphocytes followed by that of E-RFC.It was suggested that the change of cyclic nucleotides in lymphocytes may be a mechanism of adjusting the immunity on cell level.