氯吡格雷联合注射用尤瑞克林对急性脑梗死患者血浆超敏C反应蛋白和溶血磷脂酸的影响

目的观察氯吡格雷联合注射用尤瑞克林对急性脑梗死患者血浆超敏C反应蛋白(hs-CRP)和溶血磷脂酸(LPA)表达的影响。方法将90例急性脑梗死患者随机分为治疗组50例和对照组40例。对照组采用常规治疗,治疗组在常规治疗基础上加用氯吡格雷联合注射用尤瑞克林治疗,连续治疗14d。观察2组临床疗效及治疗前后NIHSS评分、日常生活活动能力(ADL)评分、hs-cRP及LPA水平。结果治疗组总有效率为96.0%高于对照组的82.5%,差异有统计学意义(P<0.05)。2组治疗后NIHSS和ADL评分均较治疗前明显改善,且治疗组改善程度优于对照组(P<0.05)。2组治疗后hs-CRP、LPA水平均较治疗前降低,且治疗组较对照组降低更明显,差异均有统计学意义(P<0.05)。结论氯吡格雷联合注射用尤瑞克林治疗急性脑梗死,可通过减少患者hs-CRP和LPA的表达而改善病情。     

Signalling properties of lysophosphatidic acid in primary human skin fibroblasts: role of pertussis toxin-sensitive GTP-binding proteins

We have investigated the signalling properties of the naturally occuring intercellular signalling molecule lysophosphatidic acid (LPA) in primary human skin fibroblasts. LPA stimulated phospholipase C activity resulting in the formation of inositol 1,4,5-trisphosphate (IP₃) which was accompanied by a concentration-dependent increase in intracelluar calcium concentration ([Ca²⁺]_i). The increase in [Ca²⁺]_i was subject to homologous desensitisation but not to heterologous desensitisation by sphingosine-1-phosphate. The half-maximal effect of LPA on the rise in [Ca²⁺]_i was attained at 7–20 nM. IP₃ formation and Ca²⁺ mobilisation were highly pertussis toxin (PTX)-sensitive (100% and 75%, respectively). LPA also inhibited forskolin-stimulated formation of cAMP, which was partially reversed (51%) when fibroblasts were pretreated with PTX. To directly test the involvement of guanine nucleotide-binding regulatory proteins (G proteins), LPA-induced binding of the stable GTP analogue GTPγS was measured. LPA induced an increase in GTPγS binding, which was completely inhibited by PTX, implicating the involvement of G_i-type G proteins in LPA signalling. Furthermore, LPA increased DNA synthesis and cell proliferation. Finally, LPA induced the migration of human skin fibroblasts, which in conjunction with the stimulation of cell growth strengthens the presumed involvement of LPA in wound healing and tissue regeneration. Both effects (cell growth and migration) were almost completely PTX-sensitive. Overall, these investigations in primary cultures of human skin fibroblasts confirm and extend our knowledge about LPA signalling, suggesting a pivotal role of receptor coupled activation of G_i-type proteins at least in this cell type. Received: 21 June 1996 / Accepted: 19 September 1996     

LPA、CA125与经阴道彩色多普勒超声联合诊断卵巢上皮癌的临床价值

目的:探讨血浆溶血磷脂酸(LPA)在卵巢上皮癌患者血浆中的表达水平,及其与血清CA125和经阴道彩色多普勒超声(TV-CDUS)联合应用诊断卵巢上皮癌的临床价值。方法:术前检测卵巢上皮癌48例,卵巢良性肿瘤30例的LPA、CA125,以20例健康者作为对照,卵巢肿瘤患者同时经阴道超声评分和TV-CDUS检查。结果:卵巢癌患者LPA水平明显高于卵巢良性肿瘤组和健康对照组,差异有统计学意义(P0.05),LPA水平在良性肿瘤组与健康对照组之间无显著差异(P0.05)。单独应用LPA、CA125、TV-CDUS检测诊断卵巢癌的敏感性和特异性分别为87.5%、79.16%、81.25%和80%、70%、86%,各组间敏感性和特异性比较,无显著差异(P0.05)。LPA、CA125、TV-CDUS 3项联合检测诊断卵巢癌的敏感性和特异性为95.80%和94%,与单独应用CA125检测特异性比较,差异有统计学意义(P0.05)。LPA诊断卵巢癌的敏感性和特异性与卵巢癌分期和病理类型无关(P0.05),CA125诊断卵巢癌的敏感性和特异性与卵巢癌的分期和病理类型有关(P0.05)。结论:卵巢上皮癌患者血浆LPA水平明显升高,有望成为卵巢上皮癌诊断的敏感指标,联合检测血浆LPA、血清CA125与TV-CDUS有助于术前卵巢癌的诊断。     

Lysophosphatidic acid stimulation of platelets rapidly induces Ca2+-dependent dephosphorylation of cofilin that is independent of dense granule secretion and aggregation

Cofilin is an actin dynamizing protein and inactivated after Ser3 phosphorylation by LIM-kinases (LIMKs). We studied whether in platelets stimulated by lysophosphatidic acid (LPA), Rho-kinase or p21-activated kinase (PAK) mediates LIMK-1 activation leading to subsequent phosphorylation and inactivation of cofilin and the increase of F-actin. During LPA (0.1 microM)-induced shape change, a rapid Rho-kinase activation and a slower activation of PAK were observed. Rho-kinase activation led to rapid LIMK-1 (Thr508) phosphorylation. Despite of LIMK-1 activation, cofilin net phosphorylation was not increased. Cofilin rapidly associated with F-actin and preceded the F-actin increase. Pretreatment with the Rho-kinase inhibitor Y-27632 inhibited LIMK-1 phosphorylation, unmasked cofilin dephosphorylation and inhibited the reversible F-actin increase during shape change. In the presence of fibrinogen, LPA (10 microM) induced ATP-secretion from dense granules and aggregation, and cofilin was rapidly dephosphorylated and then rephosphorylated in a Rho-kinase/LIMK-1-dependent manner. In the absence of fibrinogen, cofilin de- and rephosphorylation after LPA (10 microM) was unchanged, but secretion and aggregation were absent. Cofilin dephosphorylation was completely blocked by BAPTA-AM indicating that it was mediated by an increase of cytosolic Ca(2+). We conclude that in LPA-stimulated platelets, Rho-kinase-dependent LIMK-1 activation mediates the F-actin increase during shape change without enhancing cofilin net phosphorylation. However, a rapid dephosphorylation of cofilin occurs during secretion and aggregation, which is Ca(2+)-dependent, upstream of secretion and aggregation and might regulate these platelet responses.     

胰腺癌内皮分化基因受体表达

目的 观察胰腺癌组织溶血磷脂酸(LPA)内皮分化基因受体(Edg/LPA)的表达,探讨其临床意义.方法 收集50例胰腺癌及对应癌旁正常胰腺组织标本,采用实时定量PCR、免疫组化和蛋白质印迹法检测3种Edg/LPA受体亚型Edg-2/LPA1、Edg-4/LPA2和Edg-7/LPA3受体mRNA及蛋白的表达,分析其与胰腺癌临床病理参数的关系.结果 胰腺癌组织Edg-2/LPA1、Edg-4/LPA2、Edg-7/LPA3受体mRNA表达量分别为(0.142 ±0.042)％、(0.471 ±0.064)％、(0.231±0.043)％,对应癌旁正常胰腺组织分别为(0.132 ±0.029)％、(0.027±0.015)％、(0.163±0.046)％,其中胰腺癌组织的Edg-4/LPA2受体mRNA表达较对应癌旁组织显著增加(P＜0.05).同样,胰腺癌组织Edg-4/LPA2蛋白表达也显著高于癌旁正常胰腺组织.胰腺癌组织的3种Edg/LPA受体mRNA表达水平与血清CA19-9浓度呈平行关系,差异无统计学意义.胰腺癌组织EDG-4/LPA2受体mRNA表达与肿瘤大小、导管腺癌的分化程度及侵袭转移有关,而Edg-2/LPA1、Edg-7/LPA3受体mRNA表达仅与肿瘤的侵袭转移有关.结论 胰腺癌组织高表达Edg-4/LPA2受体,其高表达反映出肿瘤的恶性生物学行为.     

Propofol and Aminophylline Antagonize Each Other During the Mobilization of Intracellular Calcium in Human Umbilical Vein Endothelial Cells

This study examined whether propofol and aminophylline affect the mobilization of intracellular calcium in human umbilical vein endothelial cells. Intracellular calcium was measured using laser scanning confocal microscopy. Cultured and serum-starved cells on round coverslips were incubated with propofol or aminophylline for 30 min, and then stimulated with lysophosphatidic acid, propofol and aminophylline. The results were expressed as relative fluorescence intensity and fold stimulation. Propofol decreased the concentration of intracellular calcium, whereas aminophylline caused increased mobilization of intracellular calcium in a concentration-dependent manner. Propofol suppressed the lysophosphatidic acid-induced mobilization of intracellular calcium in a concentration-dependent manner. Propofol further prevented the aminophylline-induced increase of intracellular calcium at clinically relevant concentrations. However, aminophylline reversed the inhibitory effect of propofol on the elevation of intracellular calcium by lysophosphatidic acid. Our results suggest that propofol and aminophylline antagonize each other on the mobilization of intracellular calcium in human umbilical vein endothelial cells at clinically relevant concentrations. Serious consideration should be given to how this interaction affects mobilization of intracellular calcium when these two drugs are used together.     

Jaundice associated pruritis:A review of pathophysiology and treatment

To review the underlying pathophysiology and currently available treatments for pruritis associated with jaundice.English language literature was reviewed using MEDLINE,Pub Med,EMBASE and clinicaltrials.gov for papers and trails addressing the pathophysiology and potential treatments for pruritis associated with jaundice.Recent advances in the understanding of the peripheral anatomy of itch transmission have defined a histamine stimulated pathway and a cowhage stimulated pathway with sensation conveyed centrally via the contralateral spinothalamic tract.Centrally,cowhage andhistamine stimulated neurons terminate widely within the thalamus and sensorimotor cortex.The causative factors for itch in jaundice have not been clarified although endogenous opioids,serotonin,steroid and lysophosphatidic acid all play a role.Current guidelines for the treatment of itching in jaundice recommend initial management with biliary drainage where possible and medical management with ursodeoxycholic acid,followed by cholestyramine,rifampicin,naltrexone and sertraline.Other than biliary drainage no single treatment has proved universally effective.Pruritis associated with jaundice is a common but poorly understood condition for which biliary drainage is the most effective therapy.Pharmacological therapy has advanced but remains variably effective.     

LPA经PI3K/Akt信号转导通路抑制顺铂诱导卵巢癌细胞凋亡

 目的 研究PI3K/AKT信号转导通路对LPA保护顺铂诱导的卵巢癌SKOV3细胞凋亡作用的影响。 方法 MTT法检测LPA和LY294002对DDP作用后的SKOV3细胞增殖活性的影响;Hoechst33258荧光染色 观察凋亡细胞;FCM分析细胞凋 亡率;凝胶电泳观察凋亡细胞的DNA“梯状”条带;Western blot检测LPA、LY294002对磷酸 化Akt蛋白表达的影响。 结果 LPA+LY294002+DDP组对SKOV3细胞增殖的抑制作用、凋亡小体的产生及细胞凋亡率高于 LPA+DDP组(P<0.05),而与 LY294002+DDP组差异无统计学意义, DNA片段凝胶电泳示LPA作用后不产生明显的凋亡片段, LPA和LY294002同时作用可出现DNA断裂梯形条带。Western blot结果示LPA作用后磷酸化Akt 蛋白表达升高,而LY294002作用后,磷酸化Akt蛋白表达明显下降。 结论 LPA通过激活PI3K/Akt信号转导通路抑制顺铂诱导的卵巢癌细胞的凋亡。