Association between gene polymorphism of calcium/calmodulin-dependent kinase 4 and efficacy of amlodipine in the treatment of hypertension in Chinese Han nationality / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the association between single nucleotide polymorphisms of calcium/calmodulin-dependent kinase 4 (CAMK4) and the therapeutic effect of amlodipine in essential hypertensive patients in Chinese Han nationality.
@*METHODS@#A total of 108 mild-to-moderate essential hypertension patients in Chinese Han nationality were treated with amlodipine for 8 weeks at a dosage of 5 mg/d. Polymerase chain reaction-restriction fragment length polymorphism was performed to detect the genotypes (rs10491334). Blood pressure was measured and analyzed.
@*RESULTS@#The result of rs10491334 polymorphism of CAMK4 was consistent with Hardy-Weinberg equilibrium distribution and the frequencies of C allele and T allele were 88.89% and 11.11%, respectively. The systolic blood pressure and diastolic blood pressure before amlodipine treatment were not statistically different among different genotype carriers (P>0.05). The blood pressure was significantly reduced in all patients after amlodipine treatment (P<0.05). Systolic blood pressure was significantly decreased in patients with rs10491334 CC genotype and CT genotype compared with those patients with rs10491334 TT genotype. Total effective rates of CT and TT carriers were higher than those of the CC genotype carriers (P<0.01).
@*CONCLUSION@#The CAMK4 gene polymorphism might be associated with the efficacy of calcium channel blocker in treating mild-to-moderate essential hypertension patients.

Effect of Pinggan Qianyang recipe on the expression of Tpx II HSP27 and ANXA1 in the hypothalamus of spontaneously hypertensive rats with hyperactivity of liver-YANG syndrome / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect Pinggan Qianyang recipe on expression of Tpx, HSP27 and ANXA1 in the hypothalamus of spontaneously hypertensive rats (SHRs) with the hyperactivity of liver-YANG syndrome.@*METHODS@#A total of 30 SHRs were subjected to administration of Aconiti Praeparatae Decoction to establish the model of SHR with liver-YANG hyperactivity first, then they were randomly divided into three groups: the control group, the model group and the treatment group (n=10 per group). A total of 10 SD rats were served as the normal group. The rats in control group and treatment group were given Enalapril plus Pinggan Qianyang recipe for four weeks. The change of behavior and blood pressure of rats were monitored. RT-PCR and Western-blot were performed to detect the expression of Tpx II, HSP27 and ANXA1 mRNA and protein in the hypothalamus, respectively.@*RESULTS@#Compared with the normal SD rats, the heart rate, blood pressure and grade of irritability were significantly increased while rotation endurance time was dramatically reduced in the SHR model with liver-YANG hyperactivity (P<0.01), these changes were reversed by the application of Enalapril plus Pinggan Qianyang recipe. Compared with the normal SD rats, the protein and mRNA expression of Tpx II and ANXA1 in the model group were significantly upregulated (P<0.01) while the HSP27 was significantly downregulated (P<0.01). Compared with the model group, the protein and mRNA expression of Tpx II and ANXA1 in the control group or treatment group were significantly decreased (P<0.05 or P<0.01) while HSP27 was significantly increased (P<0.05 or P<0.01). Compared with the control group, the expression of Tpx II and ANXA1 protein in treatment group were significantly reduced (P<0.05 or P<0.01).@*CONCLUSION@#Pinggan Qianyang recipe can improve the blood pressure and behavior in SHRs with hyperactivity of Liver-YANG syndrome, which might be related to the regulation of Tpx, HSP27 and ANXA1 expression in hypothalamuses.

Herbs for calming liver and suppressing yang in treatment of hyperthyroidism with hyperactive liver yang: herbal effects on lymphocyte protein expression / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the herbal effects on hyperthyroidism patients with syndrome of hyperactivity of liver-Yang by method for calming the liver and suppressing Yang and investigate its effects on the lymphocyte protein expression. This approach may lay a foundation for the further investigation of the curative mechanisms of calming the liver and suppressing Yang treatment.</p><p><b>METHOD</b>A total of 48 hyperthyroidism patients with syndrome of hyperactivity of liver-Yang were randomly divided into treatment group and control group. The treatment group was treated by method for calming the liver and suppressing Yang in accordance with traditional Chinese medicine (TCM) and the control group with thiamazole tablets for three periods of treatment The therapeutic effects, the score of TCM symptom, electrocardiogram (P wave), thyroid hormones and ultrasound were observed in both groups before and after the treatment. The side effects in the treatment course were observed in both groups. The level of differential protein expression was analyzed by two-dimensional electrphoresis and matrix assisted laser desorption/ionizaton time-of-flight mass spectrometry.</p><p><b>RESULT</b>The treatment group has the effect on stepping down the heart rate, cutting down the P wave amplitude changes, regulating the level of thyroid hormones and decreasing the volume of thyromegaly. There are not statistically significant between the treatment group and control group. However, the treatment group has obviously better effect on regulating TCM symptom and decreasing the side reaction than the control group (P<0.05). There are not statistically significant on the total effective between the treatment group and control group. The average spots in lymphocyte for normal people, before and after treating hyperthyroidism patients with syndrome of hyperactivity of liver-Yang were (429 +/- 31), (452 +/- 28) and (437 +/- 36) spots respectively. Eight down-regulated protein expressions and 11 up-regulated protein expressions were obtained in the hyperthyroidism patients with syndrome of hyperactivity of liver-Yang and normal people. Five strengthened expressions of protein were also obtained in 8 down-regulated expressions of protein and 8 lower expressions of protein in 11 up-regulated expressions of protein before and after treating the migraine patients with syndrome of hyperactivity of liver-Yang. Ten of the total 8 differential protein spots were successfully identified by MALDI-TOF-MS. The functions of these proteins were involved in metabolism associated, transportation, antioxidation, sigal transduction and immume associated protein, etc. according to information provided by NCBI and MSDB database.</p><p><b>CONCLUSION</b>In this study, the TCM complex prescription with herbs for calming the liver and suppressing Yang can regulate the thyroid hormones, improves TCM symptoms, and decrease the adverse reaction. It can possibly regulate lymphocyte protein expression.</p>

Changes of protein expression profile in vascular tissues of spontaneously hypertensive rats treated by a compound Chinese herbal medicine / 中西医结合学报

Objective: To investigate the effects of a Chinese herbal formula for calming liver and suppressing yang on the protein expressions of vascular tissues in spontaneously hypertensive rats (SHRs), and to explore the mechanism of efficacy. Methods: Twenty SHRs were randomly divided into model group and treatment group. Another 10 Wistar-Kyoto rats were selected as a normal control. SHRs in the treatment group were administered with the formula for calming liver and suppressing Yang for 4 weeks. During the course of treatment, blood pressure and heart rates were monitored every week and the ethology of rats, including irritability and rotation endurance was also evaluated. After treatment, thoracic aorta was obtained and its proteins were separated by 2-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and database query. Results: The formula for calming liver and suppressing yang not only decreased the systolic blood pressure and heart rate, but also improved irritability degree and rotation endurance time of SHRs. This experiment had established the 2-DE graph of protein expressions of vascular tissues in SHRs. Compared with the normal group, the expressions of 15 proteins were down-regulated, and 12 proteins were up-regulated in vascular tissues of the model group. The formula for calming liver and suppressing yang treatment up-regulated expressions of 10 proteins in the 15 down-regulated proteins, and down-regulated 8 proteins in the 12 up-regulated proteins in vascular tissues of SHRs. After analysis, 16 obviously differentially expressed proteins were found, and 13 of them were identified. Conclusion: The formula for calming liver and suppressing yang can improve the ethology of SHRs. The mechanism is probably concerned with regulating the protein expressions of vascular tissues.

Clinical efficacy study on calming liver and restraining Yang formula in treating patients with mild or moderate degree of essential hypertension / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of calming the liver and restraining the Yang formula in treating patients with mild or moderate degree of essential hypertension (syndrome of hyperactivity of liver-Yang), and to explore its mechanism in lowering blood pressure.</p><p><b>METHOD</b>The 348 patients with EH of stage I , II were randomly divided into two groups, the 174 patients in the treated group were treated with the calming the liver and restraining the Yang formula, and the 174 patients in the control group were treated with amlodipine. The treatment course for them all was 12 weeks. The related clincial symptoms score and quality of life score estimated before and after treatment at 4th week, 8th week and 12th week were observed. Before and after treatment, the ambulatory blood pressure (AMBP), heart rate, blood lipid, serum livels of high-sensitivity C-reactive protein (Hs-CRP), Angiotensin-II (Ang II) and calcitonin gene-related peptide (CGRP) were measured respectively in 40 patients of the treared group and 40 patients of the control group.</p><p><b>RESULT</b>After treatment, the treatment in the treated group showed an effect better than that in the control group in terms of nigh-time blood pressure reducing rate (P < 0.05). The reducing blood pressure variability and total effective rate in the treated group were no significant than that in the control group. In respect of reducing symptomatic scores on dizzy, soreness and weakness of the waist and knees, disturbed and dry and bitter of mouth, ameliorating quality of life score, decreasing the levels of heart rate, total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol(HDL-C) in the treated group were showing marked improvement as compared with that in the control group (P < 0.05 or P < 0.1). The improvement in the level of Ang II , Hs-CRP and CGRP before treatment in two groups were more significant than that after treatment (P < 0.05). However There were no difference in after treatment between the treated group and the control group.</p><p><b>CONCLUSION</b>The calming the liver and restraining the Yang formula shows favorable efficacy in lowering blood pressure on the patients with mild or moderate degree of essential hypertension. It can reduce the clincial symptoms, improve the quality of life, regulate blood lipid metabolism. Its mechanism may be related to the functional relieving inflammatory reaction and inhibition the activity of renin-angiotensin-aldosterone system (RAAS).</p>

Effect of the formulae for calming the liver and suppressing YANG on lymphocyte proteome in migraine rats with syndrome of hyperactivity of liver-YANG / 中南大学学报(医学版)

Objective To explore the molecular mechanism of the formulae for calming the liver and suppressing YANG in migraine rat model with syndrome of hyperactivity of the liver-YANG.Methods A rat model of migraine with hyperactivity of liver-YANG was established through electrical trigeminal ganglion stimulation and syndrome of oral administration of Fuzi decoction. The total proteins of the lymphocyte in the rats were separated by immobilized pH gradient-based 2-dimensional gel electrophoresis(2-DE), and the 2-DE image was analyzed by PDQuest 7.0 software. Matrix-assisted laser desorption/ionzation-time of flight mass spectrometry (MALDI-TOF-MS) and the SWISS-PORT and MSDB database were used to identify differential proteins.Results The formulae for calming the liver and suppressing YANG could also improve headache. Well-resolution and reproducible 2-DE patterns of rat lymphocyte from normal, model, and therapy tissues were obtained. Eleven of the total 13 differential protein spots were successfully identified by MALDI-TOF-MS. These proteins were alcohol dehydrogenase 3(ADH3), glycogen phosphorylase, ATP synthase D chain, annexin-3, ubiquitin, neutrophil defensin 4 precursor, melanoma-associated antigen E2, heat shock protein-27, annexin-A1, peroxirdoxin-Ⅱ, MU class glutathione S-transferase (Fragment)(GSH). Conclusion Differences occur in the expression of lymphocyte proteins in migraine rats with syndrome of hyperactivity of liver-YANG after treatment with the formulae for calming the liver and suppressing YANG, and the 11 identified protein spots may be associated with its mechanism.

mRNA expression of neuroglobin in thalamus of DAI rats / 中国医师杂志

Objective To study mRNA expression of neuroglobin (NGB) in thalamus of DAI rats and explore the relation between NGB and heed injury. Method 46 male SD rats were randomly divided into four groups. The DAI model was established according to modi-fied Mamamu method. The mRNA expression of NGB in thalamus at different time point after injury was detected by RT-PCR method. All data were analyzed with statistical software. Result The mRNA expression of NGB in thalamus increased obviously from 30 minutes after in-jury while decreased at the 24th hour after injury. But the expression increased again and reached peak at the 48th hour after injury. Conclu-sion The mRNA expression of NGB in thalamus increased after DM. Those changes suggested NGB might participate in the response mech-anism of stress, secondary ischemia and hypoxic brain injury after neuron injury.

Differential proteomic analysis of thalamus in rats with diffuse axonal injury / 中南大学学报(医学版)

OBJECTIVE@#To examine the differential expression of protein of thalamus in rats with diffuse axonal injury.@*METHODS@#Twenty-five rats were randomly divided into a normal group (n=10) and a trauma group (n=15). Total proteins of brain trauma tissue and normal brain tissue were extracted separately, and then proteins were separated by two dimensional gel electrophoresis and stained with Coomassie brilliant blue. The differentially expressed protein spots were identified with biospectrometry. Images were analyzed by PDQuest 7.0.@*RESULTS@#The distribution of protein spots in the trauma group was similar to that of the normal group, the matching rate was 95%, and the repeatability was good. Proteins were mainly displayed at pI 3-8, with relative molecular mass 14.4-75.0 kD. Compared with the normal group, 16 spots of proteins increased and 18 spots of proteins decreased in the trauma group.@*CONCLUSION@#There is some difference in protein expression between the normal group and the trauma group. Brain trauma may lead to changes of proteins in the thalamus.

Herbs for calming liver and suppressing liver-yang in treatment of migraine with hyperactive liver-yang syndrome and its effects on lymphocyte protein expression: a randomized controlled trial / 中西医结合学报

To observe the efficacy of herbs for calming liver and suppressing liver-yang in treatment of migraine patients with hyperactivity of liver-yang syndrome and to investigate its effects on the lymphocyte protein expression. This approach may lay a foundation for the further investigation of pathogenic mechanisms in migraine with hyperactive liver-yang syndrome and the curative mechanisms of calming liver and suppressing liver-yang treatment.

Adrenal protein expressions after Pinggan Qianyang Formula treatment in hypertensive rats with liver-yang hyperactivity: a comparative proteomic analysis / 中西医结合学报

OBJECTIVE: To explore the pathogenic mechanism of liver-yang hyperactivity type of hypertension and to observe the effects of Pinggan Qianyang Formula (PGQYF), a compound of traditional Chinese herbals for calming the liver and suppressing yang, so as to provide experimental evidence for new marker proteins of drug therapy. METHODS: A rat model of liver-yang hyperactivity was prepared with spontaneous hypertensive rats (SHRs) by administration of Aconiti Praeparatae Decoction. Adrenal proteins were separated by 2D gel electrophoresis (2-DE). The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database analysis. RESULTS: The rat model of liver-yang hyperactivity was successfully reproduced, and the PGQYF could decrease the grades of irritability, conjunctival congestion and systolic blood pressure of the rats (P<0.05, P<0.01). After analysis, twelve obviously differentially expressed proteins were found, eight of which were identified. The expression levels of isocitrate dehydrogenase and steroidogenic acute regulatory protein in the untreated group were up-regulated as compared with those in the normal control group, and down-regulated in the treatment group. The expression levels of ferritin light chain, elongation factor Tu, Rho GDP disassociation inhibitor 1, flavin reductase and basic transcription factor 3 in the untreated group were down-regulated as compared with those in the normal control group, and up-regulated in the treatment group. CONCLUSION: Differentially expressed adrenal proteins in SHRs with live-yang hyperactivity are successfully identified. This approach may lay a foundation for the further investigation of pathogenic mechanisms in hypertension with liver-yang hyperactivity and the mechanisms of PGQYF treatment.

Effects of ping-gan qianyang therapy on adrenal gland proteome in migraine rat model with hyperactivity of Hver-yang / 中国医师杂志

Objective To explore the molecule mechanisms of ping-gan qianyang therapy on migraine rat model with the hyperactivity of liver-yang.Methods An animal model of migraine with hyperactivity of liver-yang was established through electrical trigeminal ganglion stimulation and administering orally with Fuzi decoction.The total proteins of adrenal#and in the rats were separated by immobilized PH gradient-based two-dimensional gel electrophoresis, and the 2-DE images Was analyzed by PDQuest 7.0 software.Matrix-Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectrometry(MALDI-TOF-MS)and SWISS-PORT and MSDB database were used to identify differential proteins.Result The well-resolution and reproducible 2-DE patterns of rat adrenal glands in normal group,model group and therapy group were obtained.A total of 8 differentially expressed proteins were identified.Conclusion Ping-gan qianyang therapy may effect on migraine by regulating the expression of proteins related to energy metabolism and ubiquitin.

Comparative Proteomics Analysis on Splenic Lymphocytes in Migraine Rat Model with Hyperactivity of Liver-yang / 中国中医药信息杂志

Objective To explore the molecule mechanisms of migraine with hyperactivity of liver-yang related proteins. Methods Animal model of migraine with hyperactivity of liver-yang was established through electrical trigeminal ganglion stimulation,and by administering orally with Fuzi decoction. The total proteins of splenic lymphocytes in the rats were separated by immobilized pH gradient-based two-dimensional gel electrophoresis. With coomassie blue staining,the 2-DE images were analyzed by PDQuest 7.0 software. Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS) and database were used to identify the differential proteins. Result Ten differential spots protein were identified,and they were chloride intracellular channel 1,thioredoxin peroxidase Ⅰ,thioredoxin peroxidase Ⅱ,sequence 7 from patent WO9709348,Rho GDP dissociation inhibitor (GDI) alpha,actin related protein 2/3 complex neurofilament light polypeptide,proteasome subunit beta type and heat shock protein-27,annexin A1. Conclusion These protein may be involved in the development of migraine with hyperactivity of liver-yang. These data provide useful clues for elucidating the mechanisms of migraine with hyperactivity of liver-yang.

Analysis of hypothalamuses in the spontaneous hypertension rats with hyperactivity of liver-yang using two-dimensional electrophoresis / 中国医师杂志

Objective To establish two-dimensional electrophoresis(2-DE) profiles with high resolution and reproducibility in analysis of hypothalamuses in the spontaneous hypertension rats(SHR) with hyperactivity of liver-yang to analyze the difference on the total proteins of hypothalamuses between the SHR and normal rats.Methods The total proteins of hypothalamuses in the rats were separated by immobilized pH gradient-based two-dimensional gel electrophoresis.With coomassie blue staining,the 2-DE image was analyzed by PDQuest 7.0 software.Results The average matching rate of the 2-DE images of hypothalamuses in the two kinds rats was 92% with good reproducibility.Proteins were mainly displayed at range of pI3-8,Mr(14.4～75) kD.The average spots in the SHR with hyperactivity of liver-yang and in the normal rats were 777?27 and 780?31 respectively.There were 22 spots of proteins up-regulated more than two times and 25 spots down-regulated more than two times in the SHR with hyperactivity of liver-yang than those of the normal rats.Conclusion There are 22 spots of proteins up-regulated and 25 spots of proteins down-regulated in the SHR with hyperactivity of liver-yang,which may be involved in the development of hyperactivity of liver-yang.

Effect of Therapy of Soothing Liver and Subsiding Yang on Hypothalamic Protein Expression in Hypertensive Rats / 广州中医药大学学报

Objective To observe the effect of therapy of soothing liver and subsiding yang on hypothalamic protein expression in hypertensive rats with hyperactivity of liver yang,and to explore its therapeutic molecular mechanism for hypertension.Methods Spontaneous hypertensive rats (SHR)were given gastric gavage of Aconiti Praeparatae Decoction (APD)to induce hypertensive rat model with hyperactivity of liver yang,and SD rats served as the normal control.The treatment group received modified Tianma Gouteng Decoction (TGD,0.1g/mL).Two-dimension gel electrophoresis (2-DE)was used to separate the hypothalamic proteins,and the differential expressed proteins were identified by matrix-associated laser desorption-ionization time-of-flight massspectrometry (MALDI-TOF-MS)and database analysis.Results Hypertensive rat model with hyperactivity of liver yang was established successfully.The irritation degree,in-take water volume and systolic pressure were increased in the model group(P