Effect of Decomposed Zuoguiwan Recipes in Improving BMD of Ovariectomized Rats by Regulating OPG/RANKL Pathway Mediated by β2AR / 中国实验方剂学杂志

Objective:To investigate the mechanism of decomposed Zuoguiwan（ZGW） recipes in treating ovariectomized osteoporosis rats. Method:Forty Sprague-Dawley female rats were equally and randomly divided into Sham-operated group, ovariectomized model group, positive group, and low and high-dose ZGW groups. After 12 weeks of administration by gavage, the bone mineral density (BMD) of rats' distal femur was measured by micro-CT, the morphology of bone tissue were observed by hematoxylin-eosin staining (HE), β-cross-linked c-telopeptide of type Ι collagen (β-CTX) and bone-specific alkaline phosphatase (BALP) in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the mRNA and protein expressions of β2AR, OPG and RANKL were evaluated by Western blot analysis and real-time quantitative polymerase chain reaction （PCR）. Result:Compared with Sham-operated group, BMD of rats in ovariectomized model group was decreased (P<0.01), morphology of bone tissue was destroyed, serum BALP was reduced, while β-CTX was boosted (P<0.01)，mRNA and protein expressions of OPG in tibia were reduced, while RANKL were increased, and mRNA and protein expressions of β2AR in the hypothalamus were decreased (P<0.05, P<0.01). Compared with ovariectomized model group, BMDs of rats in low and high-dose ZGW groups were increased (P<0.01), morphology of bone tissue was repaired, serum BALP and mRNA and protein expressions of OPG in tibia were up-regulated (P<0.05, P<0.01), whereas serum β-CTX and mRNA and protein expressions of β2AR in the hypothalamus and RANKL in tibia were down-regulated (P<0.05, P<0.01). Conclusion:Yang-nourishing components in decomposed Zuoguiwan recipes can improve BMD of ovariectomized rats by regulating OPG/RANKL pathway mediated by β2AR. "Seeking Yin in Yang" is a crucial mechanism of Zuoguiwan in treating ovariectomized osteoporosis in rats.

Effect of Zuoguiwan in Improving BMD and Trabecular Bone Microarchitecture in Ovariectomy-induced Osteoporosis Rats by Regulating RANKL/OPG Pathway Mediated by β2AR / 中国实验方剂学杂志

Objective: To investigate the mechanism of Zuoguiwan in treating ovariectomy-induced osteoporosis rats by receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) signaling pathway mediated by β2-adrenergic receptor (β2AR). Method: Forty Sprague-Dawley female rats were randomly divided into Sham-operated group (Sham) and four ovariectomized (OVX) subgroups. Rats in Sham and OVX groups were treated with 17β-estradiol (50 μg·kg-1·d-1), and low and high-dose ZGW (2.3,4.6 g·kg-1 lyophilized powder) for 3 months, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to measure the serum markers of bone turnover. Micro-CT was used to evaluate and measure trabecular bone microarchitecture and bone mineral density (BMD) of the right distal femur. Western blot analysis and Real-time PCR were used to measure mRNA and protein expressions of β2AR, OPG and RANKL. Result: After 12 weeks of treatment with Zuoguiwan, the level of serum β-cross-linked c-telopeptide of type Ι collagen (β-CTX) (PPPβ2AR in the hypothalamus (PPConclusion: The mechanism of Zuoguiwan in alleviating BMD and trabecular bone microarchitecture in ovariectomy-induced osteoporosis rats might be related to the regulation of RANKL/OPG Pathway mediated by β2AR.

Correlation between expression of β2 adrenergic receptors and HER2, K-ras genes in breast cancer / 中华内分泌外科杂志

Objective To investigate the expression of β2 adrenergic receptor (β2 AR) in invasive ductal carcinoma of the breast,and their correlation with HER2 and K-ras genes in breast cancer.Methods The expression of β2 AR,HER2 and K-ras gene in 10 cases of normal breast tissues and 33 cases of invasive ductal carcinoma of the breast were detected by immunohistochemical staining of streptomycin antibiotin protein and peroxidase assay (SP).Such statistical methods as SPSS17.0 statistical software,Mann-Whitney test and Spearman rank correlation test were applied to process and analyze the experimental data.Results β2 AR was expressed in all breast cancer cells,with an overexpression rate of 21/33.The positive expression rate of HER2 in breast cancer cells was 23/33,with an overexpression rate of 20/33.K-ras was positively expressed in breast cancer tissue(28/33),with an overexpression rate of 11/33.All of these three were significantly higher than those in the normal breast tissues.Z value of β-2 AR expression was 4.876,P value was 0.000.Z value of HER2 expression was 2.752,and P value was 0.006.Z value was 3126,and P value was 0.020 for K-ras expression.In the study of the correlation of β2 AR expression with HER2 and K-ras,the expression of β2 AR and HER2 and the expression of β2 AR and K-ras were correlated.The difference in expressions of β2 AR was statistically significant only in groups with or without lymph node metastasis,but not in groups varied in age,tumor size,clinical stage,pathological grading or hormone level.Conclusion The high expression of β2 AR,HER2,and K-ras in breast cancer tissues is associated with the occurrence and development of breast cancer.There is also a greater risk of potential lymph node metastasis in breast cancer with high expression of β-2 AR.β-2 AR may play a role in pathogenesis of breast cancer through HER2 and K-ras.

β2 adrenergic receptor gene polymorphism and coronary atherosclerotic heart disease / 中国生化药物杂志

Objective To investigate the association between β2-adrenergic receptor gene polymorphism and coronary atherosclerotic heart disease, and to provide reference for clinical disease prevention and treatment.MethodsThe clinical data of 200 patients with coronary atherosclerotic heart disease confirmed by coronary angiography were selected and included in the study group, and 200 healthy subjects without coronary atherosclerotic heart disease group.The polymorphism of β2 adrenergic receptor gene was detected and the frequency of each gene was analyzed.ResultsThe genotype frequencies of β2-adrenergic receptor gene were in accordance with Hardy-Weinberg equilibrium, the difference was not statistically significant.According to dominant genetic model, the frequency of AA+AG was 46.0% vs 58.0% lower than that of the control group, and had statistical significance, The genotype frequency of GG genotype in study group was significantly lower than that in control group 14.0% vs 26.0%, χ2=26.20, P=0.00.The frequency of GG genotype in study group was significantly higher than that in control group 54.0% vs 42.0%, χ2=5.76, P=0.01.The frequency of A gene was 38.0% compared with 44.0% in control group, χ2=1.48, P=0.22;the frequency of AA gene in study group was 30.0%, and the frequency of gene A was 38.0%, compared with 56.0% Compared with 32.0% in the control group, χ2=0.18, P=0.66.ConclusionThe A/G polymorphism of β2-adrenergic receptor gene is closely related to the clinical pathogenesis of coronary atherosclerotic heart disease, and the A allele may be a protective factor in patients with coronary atherosclerotic heart disease.

Expression of beta 2 adrenergic receptor in infantile hemangiomas tissue / 中华医学美学美容杂志

Objective To detect the expression of beta 2 adrenergic receptor in the infantile hemangiomas (IH) tissue and to explore its role in the pathological evolution of infantile hemangiomas as well.Methods 48 cases of infantile hemangioma were divided into two groups.29 cases were in the proliferating period,while the other 19 cases were in non-proliferating period.By using immunofluorescence technology,the endothelial nuclei and beta 2 adrenergic receptor of the IH tissue were marked by fluorescent tags,respectively,in two groups.The location of fluorescent labeling was shown in photos.By using the Image-Pro Plus 6.0 software,we analyzed and compared the average fluorescence intensity of endothelial cell nucleus and beta 2 adrenergic receptors in these two groups.Results Beta 2 adrenergic receptors were widely expressed in IH tissue,especially in endothelial cell nucleus shown in fluorescent images.The average fluorescence intensity of endothelial cell core in proliferating IH group was 0.031 ±0.002,which was much higher than that of fading period IH group (0.022±0.002).There was significant statistical different (P＜0.05).The average fluorescence intensity of beta 2 adrenergic receptor in proliferating IH group was 0.035± 0.003,which was much higher than that of fading period IH group (0.028± 0.002).There was significantly statistical different between the two groups (P＜0.05).Conclusions Beta 2 receptors are widely expressed in the endothelial cells of infantile hemangioma.

Borneol promotes catalpol and puerarin crossing through blood-brain barrier in focal cerebral ischemic rats / 中国中药杂志

Previous studies showed that borneol could promote some drugs crossing through the blood-brain-barrier (BBB) at certain conditions. However, the mechanism has not been clarified yet. This study aimed to investigate the effect of bornrol on promoting catalpol and puerarin through BBB and explore the relevant mechanism. The focal cerebral ischemic rats were divided into 7 groups randomly and then were administered corresponding drugs: model group (M, solvent), catalpol-puerarin group (ZG, catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹), catalpol-puerarin-bornrol group(ZGB, catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹), catalpol-bornrol group(ZB, catalpol 45 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹), puerarin-bornrol group(GB, puerarin 200 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹), butoxamine-ZG group(BTX+ZG, butoxamine 1.5 mg•kg⁻¹+ catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹), and butoxamine-ZGB group(BTX+ZGB, butoxamine 1.5 mg•kg⁻¹+ catalpol 45 mg•kg⁻¹+puerarin 200 mg•kg⁻¹ +bornrol 200 mg•kg⁻¹). Another 10 sham-operated rats were set as control (S). Ten minutes after the administration, the cerebrospinal fluid was taken to test the content of catalpol and puerarin, and the brain tissue was taken to test the expression of β2-adrenergic receptor, eNOS, and NO. Compared with the M group, the ZG group showed content of catalpol is 26.673 μg•L⁻¹ and the content of puerarin is below the detection limit;the expression levels of β2-adrenergic receptor, eNOS and the contents of NO in brain tissue are no significant difference. Compared with the ZG group, the ZGB, ZB and GB groups showed significantly increased content of catalpol andpuerarin, as well as the expression of β2-adrenergic receptor, eNOS and NO in the brain tissue (P<0.05). The content of catalpol in BTX+ZG group changed non-significantly. Compared with the ZGB group, the BTX+ZGB group presented significantly decreased content of catalpol and puerarin and reduced expression of eNOS and NO in the brain tissue (P<0.05）.The results demonstrated that borneol could dramatically promote catalpol and puerarin crossing through BBB in the focal cerebral ischemic rats. Moreover, the effect may be related to the up-regulation of β2-adrenergic receptor and the increasing expression of eNOS and NO.

Correlation between the polymorphism ofβ2-AR and the labor progress after labor analgesia / 临床麻醉学杂志

Objective To investigate the distribution ofβ2-adrenergic receptor (β2-AR)genetic polymorphisms among the included parturient women and observe the relationship between the genetic polymorphisms and labor progress after labor analgesia.Methods A prospective observational study was conducted from March to June in 2014 at Nanjing Maternity and Child Health Care Hospi-tal.A total of 1 65 nulliparous women were enrolled in the study,and finally 1 52 patients were includ-ed after exclusion for several reasons.Theβ2-AR gene polymorphic variations at nucleotides encoding amino acids 1 6 and 27 were detected by polymerase chain reaction-restriction fragment length poly-morphism technique.And the demographic data,rate of oxytocin usage,length of labor progress and the incidence of cesarean section after labor analgesia were observed and recorded.Results The allele frequencies ofβ2-AR genetic polymorphism in the parturient women were dominantly presented in the form of Arg1 6Arg (AA)and Arg1 6Gly (AG)at 1 6 position and Gln27Gln(CC)at 27 position respec-tively.The demographic and background characteristics of the subjects didn’t present any significant differences among the groups with differentβ2-AR genetic polymorphism.And the differences of labor progress among β2-AR 27 genetic polymorphisms parturient women were not significant.However, the length of the first stage and the total labor progress in the β2-AR Arg1 6Arg homozygous were both significantly longer than that in the other two groups (P <0.05),and the incidence of cesarean section was also significantly higher (P <0.05 ).Furthermore,we also found that β2-AR Arg1 6Arg homozygous was related to cesarean delivery after labor analgesia through multivariate logistic regres-sion analysis (OR 2.87,95%CI 2.69-3.09).Conclusion The labor progress of the parturient women with β2-AR Arg1 6Arg homozygous is relatively slower,which maybe an important risk for the nullip-arous women transforming to cesarean delivery after labor analgesia.

A Combination of Molecular Simulation and Network Methods for Analysis of Conformation Variation in Activation Process of β2 Adrenergic Receptor / 分析化学

To fill the urgent need of researches on the structure and function of proteins to obtain conformation information, we combined targeted molecular dynamics ( TMD) simulation with protein network methods to analyze conformation variations in the activation process of β2 adrenergic receptor. First, targeted MD was used to obtain the conformation resembles in the activation process, and then the protein network method was applied to identify the key residues and pathways in the activation process. The results indicate that the activa-tion process of β2 adrenergic receptor involves in the cooperation of all regions and the connector region of transmembrane helices is signaling hubs. In addition, the helix ends, including intracellular and extracellular loops, are the core areas. The pathway analysis reveals that there is more than one signaling pathway. All the pathways start from Ser204 of the ligand pocket and finally transmit to NPxxY or ICL2 region, which are depended on the different pathways. While the helix TMIII, TMV, TMVI are the important areas in all the pathways. The observations from the work provide valuable information at molecular level for unraveling the signal transduction mechanism associated with the activation process.

The relationship between β2-adrenergic receptor polymorphisms and early onset Myasthenia Gravis / 中国神经精神疾病杂志

Objective To explore the correlation of β2-adrenergic receptor (β2-AR) polymorphisms (Arg16Gly) with early onset Myasthenia Gravis (MG). Methods Forty-eight with age less than 40 years at disease onset were divided into three groups:normal thymus (13 cases), thymic hyperplasia (22 cases) and thymoma (7 cases) groups according to the thymus histology. These patients were further divided into different subgroups including female (31 cases) and male groups (17 cases) based on the gender, OMG (29 cases) and GMG (19 cases) groups according to the symptom of disease onset and groups associated with (10 cases) or without (33cases) other autoimmune diseases Or with unknown causes (5 cases). The genotypes ofβ2-AR in 48 early onset MG were determined by gene sequencing. Results Arg/Arg was more common in early MG patient with normal thymus ( 53.8%)and thymic hyperplasia(54.6%)whereas Arg/Gly was more common in thymus group(71.4%). The difference in distribution of the genotypes between the three groups was not statis?tically significant (χ2=5.657,P=0.226). Arg/Arg was more common in early female MG patient (58.1%) and Arg/Gly was more common in male MG patients (58.8%). The difference in distribution of the genotypes between the two groups was statistically significant (χ2=6.064,P=0.048). Arg/Arg was more common in early OMG patient (48.3%). Arg/Arg(42.1%) and Arg/Gly(47.4%) were equal common in GMG patients. The difference in distribution of the genotypes between the two groups was statistically significant ( χ2=1.623,P=0.444). Arg/Arg was more common in early MG patient associated with other autoimmune diseases (80.0%). Arg/Gly was more common in MG patients without other autoimmune diseases (39.4%). The difference in distribution of the genotypes between the three groups was statistically significant (χ2=6.394, P=0.041). Conclusionβ2-AR gene polymorphism in position 16 is associated with gender and other autoimmune diseas?es in patients with early onset of MG.

Effect ofβ2-adrenergic receptor expression on migration of early-EPCs in COPD patients / 中国病理生理杂志

AIM:To investigate whether early endothelial progenitor cells (early-EPCs) expressβ2-adrenergic receptor (β2 AR) in the chronic obstructive pulmonary disease ( COPD) patients and the effect of β2 AR expression on the migration of early-EPCs.METHODS:Venous blood samples (20 mL) were obtained from antecubital vein of COPD pa-tients or healthy controls .Peripheral blood mononuclear cells were isolated by standard Ficoll gradient centrifugation , and purified by CD34 positive selection cocktail .The mRNA expression of β2 AR in the early-EPCs was detected by RT-PCR. The protein levels of β2 AR were assessed by Western blotting and flow cytometry .Chemotaxis was studied by Transwell as-say.Cultured early-EPCs were treated with ICI118551, norpinephrine (NE) or monoclonal antibody of β2AR (mAb-β2 AR) for 24 h.The number of migratory cells was counted under a light microscope .RESULTS:The level of β2 AR ex-pression in the COPD patients was higher than that in the controls .The number of migratory early-EPCs to stromal cell-de-rived factor 1αwas significantly improved by ICI 118551 compared with other COPD groups .When early-EPCs from the COPD patients or the controls were treated with different concentrations of mAb-β2 AR for 24 h, the number of migratory early-EPCs from the COPD patients and the controls treated with NE at concentration of 100 nmol/L was significantly re-duced.However, a marked decrease in the number of migratory early-EPCs from the COPD patients treated with NE was observed compared with control group .Before treated with ICI118551 or NE for 24 h, the early-EPCs were co-incubated with mAb-β2 AR for 40 min, and the number of migratory early-EPCs was not significantly different between COPD group and control group .Genetic down-regulation of β2 AR promoted the migration of early-EPCs in COPD group .CONCLU-SION:The level of β2 AR expression in the COPD patients is increased compared with the controls .The down-regulation ofβ2 AR improves the migration of early-EPCs.

Determination of Terbutaline Sulfate and Its Combination with β_2 Adrenergic Receptor in Erythrocyte Membrane by High Performance Liquid Chromatography / 分析化学

A high performance liquid chromatographic method with hypersil C_(18) column, 0.020 mol/L potassium dihydrogen phosphate solution(which contains 0.25% triethylamine, pH 5.1)-methanol(20∶ 80, V/V) as mobile phase and fluorescence detection with 280 nm as excitation wavelength and 320 nm as emission wavelength has been developed for the determination of terbutaline sulfate and propranolol hydrochloride in Erythrocyte suspensions. Erythrocyte surface receptor oscillating reacted 1 h with terbutaline in 37 ℃ water bath. Free terbutaline and propranolol were separated from the membrane receptor-binding complex through 1500 r/min centrifugal in 5 min. The linear range was 0.010-3.000 mg/L in Alsever's solutions and the correlation coefficients were 0.9999 and 0.9998. The relative standard deviations(RSDs) of terbutaline and propranolol were 0.8% and 1.2%, respectively. The limits of detection(LODs) of both were 1.00 and 3.00 mg/L. This simple, safety and sensitive method was suitable for the determination of terbutaline and propranolol in erythrocyte suspensions and for the study of receptor-ligand binding.

Effect of Hot-Spring Hydrotherapy on Lymphocyte Subsets Expressing β₂-Adrenergic Receptors / 日本温泉気候物理医学会雑誌

 We have previously reported that hot-spring hydrotherapy for a short duration can change hormone levels in the blood and quantitatively as well as qualitatively regulate leukocytes and their subpopulations. In this study, we examined the effect of hot-spring hydrotherapy on the lymphocytes expressing β₂-adrenergic receptors (β₂-ARs). We obtained peripheral blood from 9 healthy volunteers (age, 49.7±11.2 years) at the same sampling time before and after (the next day) hot-spring hydrotherapy, and measured the total and differential leukocyte counts and the levels of 3 catecholamines (adrenaline, noradrenaline, and dopamine). We also analyzed the β₂-AR of lymphocyte subsets by combination with fluorescent monoclonal antibodies (CD3, CD4, CD 8, CD 19, and CD56) by using flow cytometry.<br> The number of granulocytes significantly decreased after hot-spring hydrotherapy (p < 0.05). The ratio of β₂-AR⁺ CD4⁺ cells and β₂-AR⁺ CD3⁺ cells was low, but that of β₂-AR⁺ CD56⁺ cells and β₂-AR⁺ CD8⁺ cells was high. The CD8⁺ cell and CD56⁺ cell counts tended to increase after hot-spring hydrotherapy, but no significant variation was found in the β₂-AR⁺ cells of each subset. Adrenaline levels also significantly decreased after hot-spring hydrotherapy (p < 0.05). The rate of change in adrenaline levels before and after hot-spring hydrotherapy was highly correlated with the rate of change in the levels of CD56⁺, CD8⁺, and β₂-AR⁺ CD56⁺ cells. Thus, we found a difference in thepercentage of β₂-AR-expressing cells in the lymphocyte subsets. Therefore, it is possible that a short duration of hot-spring hydrotherapy decreased adrenaline levels and quantitatively influenced the natural killer cells and CD8⁺ cells, which express large amounts of β₂-AR.

Expression of β_2-Adrenergic Receptor and the Receptor Post Signaling Material in Myocardial Infarction Rats / 中国循环杂志

Objective:To investigate the mRNA expression of β_2-adrenergic receptor(β_2-AR) and the receptor post signaling materials cyclic AMP(cAMP) and cAMP-dependent protein kinase (PKA) activity in myocardial infarction(MI)rats.Methods:MI model was made by ligation of rats'left anterior descending coronary artery.A total of 48 adult Wistar rats were randomly divided into four groups:Control group,the rats were sham operated,and 2 weeks post MI group,4 weeks post MI group and 8 weeks post MI group.The myocytes suspensions were untreated or treated in each group by salbutamol,isoproterenol,ICI118,551,atenolol,propranolol and pertussis toxin(PTX)respectively.The mRNA expression of β_2-AR and β_1-AR from left ventricular myocardium were detected by RT-PCR,the concentration of cAMP was determined by competitive enzyme immunoassay and the activity of PKA was determined by non-radioactive protein kinase assay.Results:Compared with Control group,the mRNA expression of β_1-AR was lower in MI groups(P<0.05),and the ratio of β_2-AR to β-AR increased from 19% to 38%(P<0.01)after MI.Compared with the untreated cells,cAMP concentration was increased by salbutamol treatment in Control group and 2 weeks,4 weeks,8 weeks post MI group by 98.1%,133.6%,147.7% and 150.7% respectively(P<0.05).Compared with isoproterenol treatment,ICI118,551 decreased cAMP concentration in 8 weeks post MI group(P<0.05).Atenolol decreased cAMP in Control group,2 weeks and 4 weeks post MI groups(P<0.05);Propranolol decreased cAMP concentration in Control and three MI groups(P<0.05).β-receptor inhibitor inhibited PKA activity in 8 weeks post MI group(P<0.05),Atenolol inhibited PKA activity in Control group and 2 weeks post MI group(P<0.05),Propranolol inhibited PKA activity in Control group and three MI groups(P<0.05,respectively).Conclusion:The ratio of β_2-AR to β-AR increased after MI.β_2-AR agonist increased cAMP concentration and PKA activity in myocytes in post MI rats.