Activation of G-protein-coupled receptor 39 reduces neuropathic pain in a rat model

Activated G-protein-coupled receptor 39(GPR39)has been shown to attenuate inflammation by interacting with sirtuin 1(SIRT1)and peroxisome proliferator-activated receptor-γcoactivator 1α(PGC-1α).However,whether GPR39 attenuates neuropathic pain remains unclear.In this study,we established a Sprague-Dawley rat model of spared nerve injury-induced neuropathic pain and found that GPR39 expression was significantly decreased in neurons and microglia in the spinal dorsal horn compared with sham-operated rats.Intrathecal injection of TC-G 1008,a specific agonist of GPR39,significantly alleviated mechanical allodynia in the rats with spared nerve injury,improved spinal cord mitochondrial biogenesis,and alleviated neuroinflammation.These changes were abolished by GPR39 small interfering RNA(siRNA),Ex-527(SIRT1 inhibitor),and PGC-1αsiRNA.Taken together,these findings show that GPR39 activation ameliorates mechanical allodynia by activating the SIRT1/PGC-1αpathway in rats with spared nerve injury.

基于Kisspeptin/GPR54系统探讨新加二甲地黄汤对PCOS模型大鼠卵泡发育的影响

目的探讨经新加二甲地黄汤干预后的多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型大鼠的卵泡发育情况及其可能存在的效应机制。方法筛选28只拥有规律动情周期的雌性SD大鼠,随机分为正常对照组、模型组、中药组及西药组,每组7只。除正常对照组外,其余三组均连续予来曲唑-羧甲基纤维素混悬液0.1mg/(kg·d)灌胃,以构建PCOS大鼠模型。自第22天起,中药组以新加二甲地黄汤5.268g/(kg·d)灌胃,西药组以炔雌醇环丙孕酮片0.286mg/(kg·d)灌胃,正常对照组及模型组均以10ml/(kg·d)蒸馏水灌胃。3周后比较各组大鼠卵巢系数,苏木精-伊红染色观察各组大鼠卵巢组织形态学改变,对各组大鼠血清激素均采用酶联免疫吸附试验进行检测,蛋白质印迹法检测卵巢亲吻素(kisspeptin,Kp)、G蛋白偶联受体54(G-protein-coupled receptor 54,GPR54)蛋白表达水平。结果与正常对照组比较,模型组大鼠卵巢内呈现为囊状扩张和闭锁的卵泡增多,其颗粒细胞层数变少,卵巢系数增大;血清睾酮(testosterone,T)、黄体生成素(luteinizing hormone,LH)、Kp水平升高;血清卵泡刺激素(follicle stimulating hormone,FSH)、雌二醇(estradiol,E2)水平及大鼠卵巢组织中Kp、GPR54蛋白表达均显著降低(P<0.05)。予中药干预后,与模型组比较,中药组大鼠卵巢囊样扩张卵泡数量变少,颗粒细胞的层数增多,存在近成熟的卵泡,并见少量黄体存在;大鼠血清LH、T、Kp水平下降,血清FSH、E2水平及卵巢Kp、GPR54蛋白表达显著升高(P<0.05)。结论PCOS模型大鼠的卵泡发育情况经新加二甲地黄汤干预后得以改善,该治疗机制可能为通过调控Kisspeptin/GPR54系统影响FSH和LH的释放,以此影响激素含量,调整卵巢功能,使卵泡发育和排卵能力得以改善。

MicroRNA-760 acts as a tumor suppressor in gastric cancer development via inhibiting G-protein-coupled receptor kinase interacting protein-1 transcription

BACKGROUND Gastric cancer(GC)has become a serious threat to people's health.Accumulative evidence reveals that dysregulation of numerous microRNAs(miRNAs)has been found during malignant formation.So far,the role of microRNA-760(miR-760)in the development of GC is largely unknown.AIM To measure the expression level of miR-760 in GC and investigate its role in gastric tumorigenesis.METHODS Real-time quantitative polymerase chain reaction and Western blot analysis were used to measure the expression of miR-760 and G-protein-coupled receptor kinase interacting protein-1(GIT1).Cell growth was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)and cell colony formation assays.Apoptosis was assessed by flow cytometric analysis.The relationship between miR-760 and GIT1 was verified by luciferase reporter assay.RESULTS The results showed that the expression of miR-760 was decreased in GC and associated with poor clinical outcomes in GC patients.Furthermore,miR-760 restrained cell proliferation and cell colony formation and induced apoptosis in GC cells.In addition,miR-760 directly targeted GIT1 and negatively regulated its expression in GC.GIT1 was upregulated in GC and predicted a worse prognosis in GC patients.We also found that upregulation of GIT1 weakened the inhibitory CONCLUSION In conclusion,miR-760 targets GIT1 to inhibit cell growth and promote apoptosis in GC cells.Our data demonstrate that miR-760 may be a potential target for the treatment of GC.

G-protein-coupled estrogen receptor as a new therapeutic target for treating coronary artery disease

Coronary heart disease(CHD) continues to be the greatest mortality risk factor in the developed world. Estrogens are recognized to have great therapeutic potential to treat CHD and other cardiovascular diseases; however,a significant array of potentially debilitating side effects continues to limit their use. Moreover,recent clinical trials have indicated that long-term postmenopausal estrogen therapy may actually be detrimental to cardiovascular health. An exciting new development is the finding that the more recently discovered G-protein-coupled estrogen receptor(GPER) is expressed in coronary arteries-both in coronary endothelium and in smooth muscle within the vascular wall. Accumulating evidence indicates that GPER activation dilates coronary arteries and can also inhibit the prolif-eration and migration of coronary smooth muscle cells. Thus,selective GPER activation has the potential to increase coronary blood flow and possibly limit the debilitating consequences of coronary atherosclerotic disease. This review will highlight what is currently known regarding the impact of GPER activation on coronary arteries and the potential signaling mechanisms stimulated by GPER agonists in these vessels. A thorough understanding of GPER function in coronary arteries may promote the development of new therapies that would help alleviate CHD,while limiting the potentially dangerous side effects of estrogen therapy.

Mechanisms of regulation and function of G-protein-coupled receptor kinases

G-protein-coupled receptor kinases (GRKs) interact with the agonist-activated form of G-protein-coupled receptor (GPCR) to affect receptor phosphorylation and to initiate profound impairment of receptor signaling, or desensitization. GPCR forms the largest family of cell surface receptors, and defects in GRK function have the potential consequence to affect GPCR-stimulated biological responses in many pathological situations.

Desensitization of G-protein-coupled receptors induces vascular hypocontractility in response to norepinephrine in the mesenteric arteries of cirrhotic patients and rats

BACKGROUND: The increased β-arrestin-2 and its combination with G-protein-coupled receptors (GPCRs) lead to GPCRs desensitization. The latter may be responsible for decreased contractile reactivity in the mesenteric arteries of cirrhotic patients and rats. The present study is to investigate the machinery changes of α-adrenergic receptors and G proteins and their roles in the contractility of mesenteric arteries of cirrhotic patients and animal models. METHODS: Patients with cirrhosis due to hepatitis B and cirrhotic rats induced by CCl 4 were studied. Mesenteric artery contractility in response to norepinephrine was determined by a vessel perfusion system. The contractile effect of G protein-coupled receptor kinase-2 (GRK-2) inhibitor on the mesenteric artery was evaluated. The protein expression of the α 1 adrenergic receptor, G proteins, β-arrestin-2, GRK-2 as well as the activity of Rho associated coiled-coil forming protein kinase-1 (ROCK-1) were measured by Western blot. In addition, the interaction of α 1 adrenergic receptor with β-arrestin-2 was assessed by co-immunoprecipitation. RESULTS: The portal vein pressure of cirrhotic patients and rats was significantly higher than that of controls. The doseresponse curve to norepinephrine in mesenteric arteriole was shifted to the right, and EC 50 was significantly increased in cirrhotic patients and rats. There were no significant differences in the expressions of the α 1 adrenergic receptor and G proteins in the cirrhotic group compared with the controls. However, the protein expressions of GRK-2 and β-arrestin-2 were significantly elevated in cirrhotic patients and rats compared with those of the controls. The interaction of the α 1 adrenergic receptor and β-arrestin-2 was significantly aggravated. This interaction was significantly reversed by GRK-2 inhibitor. Both the protein expression and activity of ROCK-1 were significantly decreased in the mesenteric artery in patients with cirrhosis compared with those of the controls, and this phenomenon was not shown in the cirrhotic rats. Norepinephrine significantly increased the activity of ROCK-1 in normal rats but not in cirrhotic ones. Norepinephrine significantly increased ROCK-1 activity in cirrhotic rats when GRK-2 inhibitor was used. CONCLUSIONS: β-arrestin-2 expression and its interaction with GPCRs are significantly upregulated in the mesenteric arteries in patients and rats with cirrhosis. These upregulations result in GPCR desensitization, G-protein dysfunction and ROCK inhibition. These may explain the decreased contractility of the mesenteric artery in response to vasoconstrictors.

Role of doublecortin-like kinase 1 and leucine-rich repeat-containing G-protein-coupled receptor 5 in patients with stage Ⅱ/Ⅲ colorectal cancer:Cancer progression and prognosis

BACKGROUND Cancer stem cells(CSCs)are a subpopulation of cancer cells with the potential of self-renewal and differentiation.CSCs play critical roles in tumorigenesis,recurrence,metastasis,radiation tolerance and chemoresistance.AIM To assess the expression patterns and clinical potential of doublecortin-like kinase 1(DCLK1)and leucine-rich repeat-containing G-protein-coupled receptor 5(Lgr5),as prognostic CSC markers of colorectal cancer(CRC).METHODS The expression of DCLK1 and Lgr5 in CRC tissue sections from 92 patients was determined by immunohistochemistry.Each case was evaluated using a combined scoring method based on signal intensity staining(scored 0-3)and the proportion of positively stained cancer cells(scored 0-3).The final staining score was calculated as the intensity score multiplied by the proportion score.Low expression of DCLK1 and Lgr5 was defined as a score of 0-3;high expression of DCLK1 and Lgr5 was defined as a score of≥4.Specimens were categorized as either high or low expression,and the correlation between the expression of DCLK1 or Lgr5 and clinicopathological factors was investigated.RESULTS DCLK1 and Lgr5 expression levels were significantly positively correlated.CRC patients with high DCLK1,Lgr5 and DCLK1/Lgr5 expressions had poorer progression-free survival and overall survival.Moreover,high expression of DCLK1 was an independent prognostic factor for recurrence and overall survival in patients with CRC by multivariate analysis(P=0.026 and P=0.049,respectively).CONCLUSION DCLK1 may be a potential CSC marker for the recurrence and survival of CRC patients.

Correlation of serum P2X7 receptor, CD64 and CD54 expression with infection process in children with bacterial pneumonia

Objective:To investigate the correlation of serum P2X7 receptor, CD64 and CD54 expression with infection process in children with bacterial pneumonia.Methods: A total of 164 children with bacterial pneumonia hospitalized in this hospital between June 2016 and February 2018 were selected as bacterial pneumonia group, and 100 healthy children who received vaccination in this hospital during the same period were selected as normal control group. The expression levels of P2X7 receptor, CD64 and CD54 as well as the contents of inflammatory factors, acute phase proteins and immunoglobulins in serum of the two groups were detected. Results: Immediately after admission, serum P2X7 receptor, CD64 and CD54 expression of bacterial pneumonia group were higher than those of normal control group, inflammatory cytokines TNF-α, sTREM-1, IL-2 and IL-6 contents were higher than those of normal control group, acute phase proteins 1-AGP, CRP, CP and HP contents were higher than those of normal control group, and immunoglobulins IgA, IgM and IgG contents were higher than those of normal control group;serum P2X7 receptor, CD64 and CD54 expression in children with bacterial pneumonia were positively correlated with TNF-α, sTREM-1, IL-2, IL-6, 1-AGP, CRP, CP, HP, IgA, IgM and IgG contents.Conclusion:The serum P2X7 receptor, CD64 and CD54 expression increase in children with bacterial pneumonia, and they are positively correlated with the degree of infection.

G蛋白偶联受体54 mRNA在雌性性早熟大鼠下丘脑中的表达及其意义

目的检测GPR54在雌性性早熟大鼠下丘脑中的表达,探讨其在真性性早熟发生发展过程中的作用。方法正常5日龄雌性大鼠40只。随机分为4组,每组10只。实验1组和2组大鼠皮下一次注射300μg达那唑以诱导真性性早熟;检查阴道涂片、体质量及子宫、卵巢质量,用化学发光法测其血清黄体生成素水平;半定量RT-PCR法检测下丘脑GPR54 mRNA表达水平。结果GPR54 mRNA在青春前期组、性早熟青春早期组及性早熟青春中期组大鼠中的表达呈逐渐上升趋势,各组差异具有显著性(F=467.55 P<0.01)。性早熟青春早期组与正常青春早期组GPR54表达无显著性差异(P>0.05)。结论GPR54在性早熟启动和发展过程中可能起重要作用。

Kisspeptin-GPR54-GnRH神经元轴在雌性大鼠中枢性性早熟中的作用

目的探讨Kisspeptin-GPR54-GnRH神经元轴在雌性大鼠中枢性性早熟(CPP)中的作用。方法选择雌性SD大鼠50只,随机分为对照1组(正常青春前期)、对照2组(正常青春早期)、对照3组(正常青春中期)、实验1组(性早熟青春早期)、实验2组(性早熟青春中期)各10只。实验组皮下注射N-甲基-DL-天冬氨酸(NMA)建立CPP模型。观察各组阴道开放时间及性周期,测量其子宫指数、卵巢指数、卵巢黄体出现个数、子宫壁厚度和血清黄体生成素;用Real-Time RT-PCR法检测下丘脑中的KISS-1 mRNA、GPR54 mRNA、促性腺激素释放激素(Gn-RH)mRNA表达;在电镜下观察各组下丘脑内分泌神经元的超微结构。结果实验组性发育起始时间早于对照组,实验组各检查指标明显高于对照1组(P均<0.05),实验1组与对照2组、实验2组与对照3组比较均无统计学差异。随着青春期发育,实验组和对照组大鼠下丘脑中KISS-1 mRNA、GPR54 mRNA、GnRH mRNA表达均逐渐升高,下丘脑内分泌神经元代谢逐渐活跃,分泌旺盛。结论应用NMA可建立理想的雌性大鼠CPP模型,随着大鼠青春期发育,其下丘脑中的KISS-1 mRNA、GPR54 mRNA、GnRH mRNA表达逐渐升高;提示Kisspeptin-GPR54-Gn-RH神经元轴在CPP的发生、发展中起重要作用。

前列腺癌患者ARA54和Apaf的表达及临床意义

目的探讨前列腺癌患者雄激素受体辅助因子(ARA)54和凋亡蛋白酶活化因子(Apaf)的表达及临床意义。方法选取2015年1月~2018年5月我院收治的76例前列腺癌患者设为前列腺癌组,选取同期36例良性前列增生设为前列腺增生组,采用免疫组化法检测与应用实时定量聚合酶链反应(RT-PCR)(SYBR染色法)技术检测两组前列腺标本中ARA 54与Apaf的表达。采用Gleason评分法对患者临床分期进行评估,运用Spearman相关性分析各临床分期与ARA54、Apaf表达水平的相关性。结果前列腺癌组ARA 54阳性、Apaf阳性表达水平均明显低于前列腺增生组(P<0.05),转化生长因子β(TGF-β)阳性表达水平明显高于前列腺增生组(P<0.05);ARA 54与Apaf阳性表达在病理学分级、临床分期、有无远处转移存在明显差异(P<0.05),其中,病理学分级越低、临床分期越高、有远处转移的前列腺癌患者中ARA 54与Apaf阳性表达水平明显降低(P<0.05);Spearman相关性分析结果提示,ARA 54表达水平与Gleason评分分级呈正相关性(r值分别为0.724、0.742、0.768、0.750、0.784,P<0.001);Apaf表达水平与Gleason评分分级呈正相关性(r值分别为0.549、0.488、0.425、0.460、0.562,P<0.05);TGF-β表达水平与Gleason评分分级呈负相关性(r值分别为-0.674、-0.789、-0.856、-0.724、-0.764,P<0.05)。结论 ARA 54与Apaf表达异常与前列腺癌的发生发展有关,ARA 54与Apaf的低表达可促进癌细胞的增殖与生长,因此,调节前列腺癌患者前列腺细胞中ARA 54与Apaf表达具有重要意义。

Kisspeptins和GPR54对青春期发育启动机制的研究进展

青春期发育的启动标志着人在身体上的全面成熟,关系到每个人生活、工作、心理、婚姻以及生育能力,研究青春期发育启动机制为揭开人类发育的进程有着重大的意义。本综述通过回顾最近在青春期发育启动机制的重大发现——kisspeptins和GPR54信号通路在激活促性腺激素释放激素(GnRH)神经元的关键作用,为今后青春期发育研究的取向提出建议。

Effect of organophosphorus insecticides on phosphorylation of the M_2 muscarinic acetylcholine receptor

BACKGROUND： Organophosphorus insecticides may promote the accumulation of acetylcholine at synapses and the neuromuscular junction by inhibiting acetylcholinesterase activity to cause disturbance of neural signal conduction and induce a toxic reaction. Organophosphorus insecticides may act on M2 muscarinic acetylcholine receptors, whose combination with G proteins is regulated by phosphorylation of G protein-coupled receptor kinase 2. OBJECTIVE： To investigate the effects of organophosphorus insecticides on the phosphorylation of G protein-coupled receptor kinase 2-mediated M2 muscarinic acetylcholine receptors and to reveal other possible actions of organophosphorus insecticides. DESIGN, TIME AND SETTING： An observational study, which was performed in the Central Laboratory of Shenyang Medical College, and Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University from June 2002 to December 2004. MATERIALS： Paraoxon, parathion, chlorpyrifos, and chlorpyrifos oxon were provided by Chem Service Company, USA, [γ -p^32] ATP and [^35S]GTP γ S by New England Nuclear Life Science Products, and recombinant β 2-adrenergic receptor membrane protein by Sigma Company, USA. METHODS： The M2 muscarinic acetylcholine receptor was extracted and purified from pig brain using affinity chromatography. Subsequently, the purified M2 muscarinic acetylcholine receptor, G protein-coupled receptor kinase 2, and [γ -p^32] ATP were incubated with different concentrations of paraoxon and chlorpyrifos oxon together. The mixture then underwent polyacrylamide gel electrophoresis, and the gel film was dried and radioactively autographed to detect phosphorylation of the M2 muscarinic acetylcholine receptor. Finally, the radio-labeled phosphorylated M2 receptor protein band was excised for counting with an isotope liquid scintillation counter. MAIN OUTCOME MEASURES： Effects of chlorpyrifos oxon, paraoxon, chlorpyrifos, and parathion in different concentrations on the phosphorylation of the M2 muscarinic acetylcholine receptor; effects of chlorpyrifos oxon on the phosphorylation of the β -adrenergic receptor. RESULTS： Chlorpyrifos oxon could completely inhibit the phosphorylation of the M2 muscarinic acetylcholine receptor, and its IC50 was 70 μ mol/L. Chlorpyrifos could also inhibit the phosphorylation of the M2 muscarinic acetylcholine receptor. However, paraoxon and parathion could not inhibit the phosphorylation of the M2 muscarinic acetylcholine receptor. Chlorpyrifos oxon in different concentrations could also not inhibit the phosphorylation of the β 2-adrenergic receptor catalyzed by G protein-coupled receptor kinase 2. CONCLUSION： Different kinds of organophosphorus insecticides have different effects on the phosphorylation of the G protein-coupled receptor kinase 2-mediated M2 muscarinic acetylcholine receptor. Organophosphorus insecticides possibly have different toxic effects.

p54nrb, a PSF Protein Partner, Contributes to Meningitic <i>Escherichia coli</i>K1-Mediated Pathogenicities

IbeA is an important invasion determinant contributing to Escherichia coli K1 entry into brain microvascular endothelial cells (BMEC) that is a key step in the pathogenesis of E. coli meningitis. Our previous studies have shown that IbeA-induced signaling and E. coli K1 invasion is mediated by two IbeA-binding proteins, vimentin, which is constitutively present in the surface of human BMECs (HBMECs), and PSF, which is inducibly expressed in both mesenchymal (endothelium) and non-mesenchymal (epithelium) cells. However, it is unknown whether p54nrb, a PSF partner protein, could contribute to the pathogenesis of E. coli K1 meningitis. Here, we reported that a 54-kDa protein was identified by copurification with PSF through IbeA-affinity chromatography as an IbeA-binding protein, which is identical to p54nrb. Both p54nrb and PSF are RNA-binding proteins and share significant sequence homology. The specific interaction between IbeA and p54nrb was confirmed by Western blot and ligand overlay assays. Recombinant p54nrb blocked E. coli K1 invasion of human BMEC very effectively. Overexpressed p54nrb as a GFP fusion protein in the transfected 293T cells significantly enhanced E. coli K1 invasion. Furthermore, higher levels of surface p54nrb in the transfected 293T cells were detected by flow cytometry. These results suggest that the IbeA invasion protein of E. coli K1 interacts with p54nrb for bacterial invasion of human BMEC.

Molecular regulation of calcium-sensing receptor(CaSR)-mediated signaling

Calcium-sensing receptor(CaSR),a family C G-protein-coupled receptor,plays a crucial role in regulating calcium homeostasis by sensing small concentration changes of extracellular Ca^(2+),Mg^(2+),amino acids(e.g.,L-Trp and L-Phe),small peptides,anions(e.g.,HCO_(3)^(-)and PO_(4)^(3-)),and pH.CaSR-mediated intracellular Ca^(2+)signaling regulates a diverse set of cellular processes including gene transcription,cell proliferation,differentiation,apoptosis,muscle contraction,and neuronal transmission.Dysfunction of CaSR with mutations results in diseases such as autosomal dominant hypocalcemia,familial hypocalciuric hypercalcemia,and neonatal severe hyperparathyroidism.CaSR also influences calciotropic disorders,such as osteoporosis,and noncalciotropic disorders,such as cancer,Alzheimer's disease,and pulmonary arterial hypertension.This study first reviews recent advances in biochemical and structural determination of the framework of CaSR and its interaction sites with natural ligands,as well as exogenous positive allosteric modulators and negative allosteric modulators.The establishment of the first CaSR protein-protein interactome network revealed 94 novel players involved in protein processing in endoplasmic reticulum,trafficking,cell surface expression,endocytosis,degradation,and signaling pathways.The roles of these proteins in Ca^(2+)-dependent cellular physiological processes and in CaSR-dependent cellular signaling provide new insights into the molecular basis of diseases caused by CaSR mutations and dysregulated CaSR activity caused by its protein interactors and facilitate the design of therapeutic agents that target CaSR and other family C G-protein-coupled receptors.

呼和浩特市地区蒙古与汉族寻常型银屑病患者皮损细胞间粘附分子及表皮生长因子受体免疫组化研究

目的 :探讨蒙汉族银屑病皮损细胞间粘附分子 (CD54 )及表皮生长因子受体 (EGFR)的表达情况。方法 :采用免疫组化染色方法研究银屑病患者皮损部位CD54 、EGFR的表达。结果 :蒙汉族银屑病患者皮损部位CD54 、EGFR的表达均阳性。结论 :CD54 、EGFR与银屑病发病机理有关。CD54 、EGFR在蒙。

大补阴丸对真性性早熟模型大鼠的治疗作用

目的探讨大补阴丸对真性性早熟大鼠的治疗作用,评价大补阴丸对真性性早熟的疗效并探讨可能的作用机制。方法 5日龄SD雌性大鼠一次性sc给予达那唑30μg.g-1建立性早熟模型。15日龄时开始ig给予大补阴丸0.81,1.62和3.24 g·kg-1,每天1次,至模型组大鼠阴门开启数超过50%。21日龄起,密切观察各组大鼠阴门开启情况,并记录开启时的日龄。对阴门已开启的大鼠,于每日早晨进行阴道脱落细胞涂片,显微镜下观察性周期的变化。当模型组阴门开启大鼠数超过该组的50%时(33日龄),所有大鼠股动脉放血处死,取子宫和卵巢计算子宫和卵巢系数,并常规制作子宫和卵巢组织切片测定子宫壁厚度和卵巢黄体生成数;半定量逆转录(RT)-PCR法测定下丘脑促性腺激素释放激素(GnRH)、G蛋白偶联受体54(GPR54)和Kiss-1 mRNA的表达水平。结果大补阴丸3.24 g·kg-1组子宫系数为115±12,较模型组154±14显著降低(P<0.05);大补阴丸3.24 g·kg-1组子宫壁厚度为(166±27)μm,较模型组(477±71)μm显著降低(P<0.05),并使卵巢黄体生成个数显著减少;大补阴丸能明显降低下丘脑GnRH,GPR54和Kiss-1mRNA的表达水平,卵巢系数则无明显变化。结论大补阴丸可能通过抑制下丘脑Kiss-1和GPR54基因表达,抑制下丘脑GnRH的合成和释放,从而抑制下丘脑-垂体-性腺轴的启动,发挥治疗真性性早熟的作用。

Altered profiles of fecal bile acids correlate with gut microbiota and inflammatory responses in patients with ulcerative colitis

BACKGROUND Gut microbiota and its metabolites may be involved in the pathogenesis of inflammatory bowel disease.Several clinical studies have recently shown that patients with ulcerative colitis(UC)have altered profiles of fecal bile acids(BAs).It was observed that BA receptors Takeda G-protein-coupled receptor 5(TGR5)and vitamin D receptor(VDR)participate in intestinal inflammatory responses by regulating NF-ĸB signaling.We hypothesized that altered profiles of fecal BAs might be correlated with gut microbiota and inflammatory responses in patients with UC.AIM To investigate the changes in fecal BAs and analyze the relationship of BAs with gut microbiota and inflammation in patients with UC.METHODS The present study used 16S rDNA sequencing technology to detect the differences in the intestinal flora between UC patients and healthy controls(HCs).Fecal BAs were measured by targeted metabolomics approaches.Mucosal TGR5 and VDR expression was analyzed using immunohistochemistry,and serum inflammatory cytokine levels were detected by ELISA.RESULTS Thirty-two UC patients and twenty-three HCs were enrolled in this study.It was found that the diversity of gut microbiota in UC patients was reduced compared with that in HCs.Firmicutes,Clostridium IV,Butyricicoccus,Clostridium XlVa,Faecalibacterium,and Roseburia were significantly decreased in patients with UC(P=3.75E-05,P=8.28E-07,P=0.0002,P=0.003,P=0.0003,and P=0.0004,respectively).Proteobacteria,Escherichia,Enterococcus,Klebsiella,and Streptococcus were significantly enriched in the UC group(P=2.99E-09,P=3.63E-05,P=8.59E-05,P=0.003,and P=0.016,respectively).The concentrations of fecal secondary BAs,such as lithocholic acid,deoxycholic acid,glycodeoxycholic acid,glycolithocholic acid,and taurolithocholate,in UC patients were significantly lower than those in HCs(P=8.1E-08,P=1.2E-07,P=3.5E-04,P=1.9E-03,and P=1.8E-02,respectively)and were positively correlated with Butyricicoccus,Roseburia,Clostridium IV,Faecalibacterium,and Clostridium XlVb(P<0.01).The concentrations of primary BAs,such as taurocholic acid,cholic acid,taurochenodeoxycholate,and glycochenodeoxycholate,in UC patients were significantly higher than those in HCs(P=5.3E-03,P=4E-02,P=0.042,and P=0.045,respectively)and were positively related to Enterococcus,Klebsiella,Streptococcus,Lactobacillus,and pro-inflammatory cytokines(P<0.01).The expression of TGR5 was significantly elevated in UC patients(0.019±0.013 vs 0.006±0.003,P=0.0003).VDR expression in colonic mucosal specimens was significantly decreased in UC patients(0.011±0.007 vs 0.016±0.004,P=0.033).CONCLUSION Fecal BA profiles are closely related to the gut microbiota and serum inflammatory cytokines.Dysregulation of the gut microbiota and altered constitution of fecal BAs may participate in regulating inflammatory responses via the BA receptors TGR5 and VDR.

Sphingosine-1-phosphate signaling in vasculogenesis and angiogenesis

Blood vessels either form de novo through the process of vasculogenesis or through angiogenesis that involves the sprouting and proliferation of endothelial cells in pre-existing blood vessels. A complex interactive network of signaling cascades downstream from at least three of the nine known G-protein-coupled sphingosine-1-phosphate (S1P) receptors act as a prime effector of neovascularization that occurs in embryonic development and in association with various pathologies. This review focuses on the current knowledge of the roles of S1P signaling in vasculogenesis and angiogenesis, with particular emphasis on vascular cell adhesion and motility responses.

基于Kisspeptin/GPR54系统探讨七子益肾理冲汤对多囊卵巢综合征模型大鼠卵泡发育的影响

目的探讨七子益肾理冲汤对多囊卵巢综合征(PCOS)卵泡发育的影响及可能作用机制。方法将17只动情规律雌性SD大鼠随机分为空白组5只、模型组6只、七子益肾理冲汤组6只。模型组及七子益肾理冲汤组大鼠每日予来曲唑-羧甲基纤维素混悬液1 mg/(kg·d)连续灌胃21天构建PCOS大鼠模型。第22天开始,七子益肾理冲汤组以七子益肾理冲汤34.02 g/(kg·d)灌胃,每日1剂,空白组、模型组以10 ml/(kg·d)无菌饮用水灌胃。15天后比较各组大鼠体质量、卵巢系数,体视显微镜观察各组大鼠卵巢外观,HE染色观察各组大鼠卵巢组织形态学改变,放射免疫法检测各组大鼠血清激素卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、睾酮(T)、孕激素(P)水平、抗苗勒管激素(AMH)、抑制素B(INHB)水平,Western blot法检测下丘脑及卵巢Kisspeptin(Kp)、G蛋白偶联受体54(GPR54)、细胞外信号调节激酶1/2(ERK1/2)蛋白表达水平。结果各组大鼠的体质量、卵巢系数比较,差异均无统计学意义(P>0.05)。与空白组比较,模型组大鼠卵巢外观较苍白,囊状扩张卵泡及闭锁卵泡增加,卵泡颗粒细胞层变薄;血清FSH、LH、AMH水平,下丘脑及卵巢Kp、GPR54、ERK1/2蛋白表达显著升高(P<0.05或P<0.01),血清E2、P水平显著降低(P<0.01)。与模型组比较,七子益肾理冲汤组大鼠卵巢外观较红润,囊状扩张卵泡减少,卵泡颗粒细胞层增加,出现近成熟卵泡,可见少量黄体;大鼠血清FSH、LH、AMH水平,下丘脑、卵巢Kp、GPR54蛋白表达显著下降,下丘脑ERK1/2蛋白表达显著下降(P<0.05或P<0.01)。结论七子益肾理冲汤可以改善PCOS模型大鼠卵泡发育,其机制可能是通过激活Kp/GPR54系统,调节下游ERK信号通路,进而调控下丘脑-垂体-卵巢轴,改善激素水平,发挥调节卵巢功能、促进卵泡发育的作用。