姜黄素调节B淋巴瘤细胞p300和HDAC1的研究

p300是一种具有组蛋白乙酰化酶活性的转录辅助因子，HDAC1是一种组蛋白去乙酰化酶。本研究查明姜黄素对B-NHL细胞株Raji细胞增殖的影响，并探讨这种影响与p300以及HDAC1转录调节表达的关系。以不同浓度（6．25-50μmol/L）的姜黄素作用于体外培养的Raji细胞，用MTT法检测细胞生长抑制率。Annexin．VFITC／PI双标流式细胞术检测细胞的凋亡率和应用RT-PCR和Western blot法检测Raji细胞中HDAC1和p300的mRNA表达和蛋白含量的变化。结果表明：姜黄素对Raji细胞抑制作用呈明显的时效和量效关系。24小时的IC50为25μmol／L；姜黄素能够诱导Raji细胞凋亡，凋亡率为14．38％-61．18％，并呈浓度依赖性。姜黄素明显抑制HDAC1和p300的活性和表达。在IC50浓度时，随着时间的延长，其p300和HDAC1 mRNA表达和蛋白含量逐渐降低，呈时间依赖性，与对照组相比有显著性（P〈0．05）。结论：姜黄素对B细胞淋巴瘤细胞系Raji细胞具有抗肿瘤细胞的增殖作用，并促进其凋亡。姜黄素能够抑制转录共激活因子p300及组蛋白去乙酰化酶HDAC1活性和表达，可能是其作用机制之一。     

重症甲型H1N1流感患者病情危险因素分析及早期预警评分建立

目的:探讨重症甲型H1N1流感的独立危险因素,建立重症甲型H1N1流感患者早期预警评分(H1N1EWS)标准,以筛选潜在的重症患者,为尽早采取有效救治措施提供依据。方法:回顾性分析156例甲型H1N1流感患者的临床资料,并将患者分为轻症组和重症组,先以单因素分析筛选出有统计学意义的危险因素,再通过向前法多元逐步Logistic回归分析确定重症甲型H1N1流感的独立危险因素,并赋予各指标反映病情严重程度的等级分值,据此建立H1N1EWS。应用ROC曲线下面积(AUC)评价其诊断准确性,Kappa检验评估其诊断可靠性。结果:本组患者轻症115例,重症41例,其中死亡4例,单因素分析提示年龄、住院天数、发热病程、咳嗽、既往病史、体温、心率、呼吸频率、白细胞(WBC)、血细胞比容(HCT)、淋巴细胞百分比(LY%)、血红蛋白(Hb)、中性粒细胞百分比(NE%)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、白蛋白(ALB)、肌酸激酶(CK)等17个指标是危险因素,Logistic回归分析提示年龄、NE%、AST、呼吸频率是独立危险因素。每个指标根据其权重分别赋予0～3分,建立H1N1EWS,总分为各独立危险因素评分之和,最高12分。评分标准的AUC为0.905,cutoff值为2.5分,kappa值为0.645;H1N1EWS与重症百分比的关系显示,随着总分的增加,重症百分比增加,对病情的预测效能高于改良早期预警评分系统(MEWS)。结论:H1N1EWS可用于预测甲型H1N1流感患者的病情严重程度。     

姜黄素在体外对氨基酸递质转运体EAAC1及GAT1功能的影响

目的：探讨体外条件下姜黄素对兴奋性氨基酸递质转运体EAAC1及抑制性递质转运体GAT1功能活性的影响。方法：利用爪蟾卵母细胞内源性Na/K-ATPase及外源表达的大鼠Na/K-ATPase、EAAC1及GAT1蛋白,借助双电极电压钳技术,应用姜黄素干预,实时记录并观察姜黄素干预对不同离子转运体介导稳态电流的影响。结果：（1）不同浓度梯度姜黄素（7.5μmol.L^-1、15μmol.L^-1、30μmol.L^-1、75μmol.L^-1）灌流对内源性及外源性Na/K-ATPase介导稳态电流,无显著影响;不同浓度姜黄素（15μmol.L^-1和30μmol.L^-1,）预处理卵母细胞24h,亦未观察到明显效应;借助微量注射方法,将姜黄素（30μmol.L^-1）直接注射到卵母细胞内,对Na/K-ATPase介导稳态电流无显著影响。（2）30μmol.L^-1姜黄素灌流对外源性EAAC1介导稳态电流无明显影响。（3）30μmol.L^-1姜黄素灌流可以轻度抑制外源性GAT1介导稳态电流。结论：姜黄素对细胞膜Na/K-ATPase和EAAC1介导稳态电流无显著影响,对GAT1介导稳态电流有轻度抑制作用。     

MEWS和SMART-COP评分预测重症甲型H1N1流感机械通气临床研究

目的评价改良早期预警评分（MEWS）和SMART-COP评分预测重症甲型H1N1流感患者是否需要机械通气的价值。方法对资料齐全的50例重症甲型H1N1流感患者进行回顾性分析,计算MEWS和SMART-COP评分和评估其是否需要进行机械通气的灵敏度和特异度。结果 MEWS预测患者机械通气灵敏度为92.9%,特异度为83.3%,准确度为86.0%,约登指数为0.76;SMART-COP评分预测患者机械通气灵敏度为71.4%,特异度为97.2%,准确度为90.0%,约登指数为0.68。结论 MEWS预测患者机械通气有很高的灵敏度和特异度,有很好的预测价值,且操作简单。SMART-COP评分灵敏度较低,有很高的特异度,对预测机械通气有较好的价值。     

Heme oxygenase-1 inhibits the proliferation of pancreatic stellate cells by repression of the extracellular signal-regulated kinase1/2 pathway

Activation of pancreatic stellate cells (PSCs) is the key process in the development of pancreatic fibrosis, a common feature of chronic pancreatitis and pancreatic cancer. In recent studies, curcumin has been shown to inhibit PSC proliferation via an extracellular signal-regulated kinase (ERK)1/2-dependent mechanism. In addition, curcumin is a potent inducer of the cytoprotective enzyme heme oxygenase-1 (HO-1) in other cell types. Therefore, the aims of this study were to 1) characterize the effect of curcumin on HO-1 gene expression in PSCs, 2) explore whether HO-1 induction contributes to the inhibitory effect of curcumin on PSC proliferation, and 3) clarify the involvement of the mitogen-activated protein kinase (MAPK) family in this context. Cultured rat PSCs were incubated with curcumin and assessed for HO-1 up-regulation by Northern blot analysis, immunoblotting, and activity assays. The effect of HO-1 on platelet-derived growth factor (PDGF)-induced PSC proliferation and MAPK activation was determined by immunoblotting, cell proliferation assays, and cell count analyses. Curcumin induced HO-1 gene expression in PSCs in a time- and dose-dependent manner and inhibited PDGF-mediated ERK1/2 phosphorylation and PSC proliferation. These effects were blocked by treatment of PSCs with tin protoporphyrin IX, an HO inhibitor, or transfection of HO-1 small interfering RNA. Our data provide evidence that HO-1 induction contributes to the inhibitory effect of curcumin on PSC proliferation. Therefore, therapeutic up-regulation of HO-1 could represent a mode for inhibition of PSC proliferation and thus may provide a novel strategy in the prevention of pancreatic fibrosis.     

姜黄素对干热环境热射病大鼠肾损伤及细胞凋亡的影响

目的探讨姜黄素对干热环境热射病大鼠肾脏损伤的病理变化及细胞凋亡的影响。 方法SPF级成年健康雄性SD大鼠50只，在SPF级环境适应性饲养1周，按随机数字表法分为5组：常温对照组、干热对照组、姜黄素低浓度处理组(50 mg/kg)、姜黄素中浓度处理组(100 mg/kg)、姜黄素高浓度处理组(200 mg/kg)，每组10只。常温及干热对照组给予0.9%生理盐水灌胃，姜黄素组大鼠给予不同浓度姜黄素溶液灌胃，连续7 d。第8天除常温对照组外，其余4组大鼠均转移到模拟干热环境实验舱中。入舱实验的第150 min达到热射病状态，麻醉处死大鼠后留取尿液、肾组织进行分析，肾损伤分子-1(KIM-1)试剂盒检测尿液KIM-1水平变化，苏木精-伊红(HE)染色观察各组大鼠肾组织病理形态变化，进行肾损伤病理学评分，细胞凋亡缺口末端标记技术(TUNEL)检测细胞凋亡并计算凋亡率。 结果常温对照组尿液KIM-1水平、肾损伤病理学评分、肾组织细胞凋亡率分别为[(100.36±5.54)ng/L]、[(35.50±9.52)分]、[(0.55±0.04)%]；干热对照组大鼠各指标分别为[(1 060.57±75.50)ng/L]、[(710.67±74.60)分]、[(5.53±0.48)%]；姜黄素低浓度处理组各指标分别为[(945.73±48.07)ng/L]、[(701.67±64.84)分]、[(5.10±0.37)%]；姜黄素中浓度处理组各指标分别为[(639.17±44.71)ng/L]、[(365.00±34.06)分]、[(2.05±0.35)%]；姜黄素高浓度处理组各指标分别为[(592.67±34.29)ng/L]、[(289.00±35.08)分]、[(1.33±0.20)%]。各项指标水平在各组间比较，均差异有统计学意义(F＝373.70，203.16，289.81；均P<0.01)，干热对照组KIM-1水平和肾损伤病理学评分均明显高于常温对照组(t＝31.04，21.99；均P<0.01)。肾损伤病理学评分和肾组织细胞凋亡率在姜黄素低浓度组和干热对照组之间比较，均差异无统计学意义(t＝0.22，1.74；均P>0.05)，姜黄素中浓度处理组(t＝10.33，14.43)、高浓度处理组(t＝12.53，19.82)与干热对照组之间比较，均差异有统计学意义(均P<0.01)。 结论姜黄素预处理对干热环境热射病大鼠肾损伤具有一定的保护作用，可能通过阻断肾细胞凋亡通路发挥对肾脏损伤的保护作用。     

姜黄素对人子宫内膜癌细胞增殖抑制的初步研究

目的观察姜黄素对体外培养的人子宫内膜癌(HEC-1-B)细胞增殖和细胞周期分布的影响。方法将体外培养的HEC-1-B细胞随机分为5组:对照组、姜黄素Ⅰ组、姜黄素Ⅱ组、姜黄素Ⅲ组和姜黄素Ⅳ组。分别给予不同浓度的姜黄素(0,10,20,40,80)μmol/L培养48 h。采用MTT法和Western blot法检测HEC-1-B细胞的增殖程度;流式细胞仪进行细胞周期时相分析。结果①MTT法显示培养48 h后,姜黄素组的吸光度值A490 nm低于对照组(P<0.01),且在一定浓度范围内呈剂量依赖性。②Western blot法结果显示,与对照组比较,姜黄素作用48 h后,姜黄素各组的PCNA表达减少(P<0.01),且在10～80μmol/L范围内呈剂量依赖性。提示姜黄素可抑制HEC-1-B细胞内PCNA的表达。③流式细胞仪检测显示培养48 h后,姜黄素组的G2/M比例高于对照组(P<0.01),且在一定浓度范围内呈剂量依赖性。结论在一定浓度(10～80μmol/L)范围内,姜黄素可抑制体外培养的HEC-1-B细胞增殖,阻止细胞周期进展。这可能是姜黄素抗HEC-1-B细胞作用的机制之一。     

姜黄素对不稳定型心绞痛患者血清单核细胞趋化蛋白1、超敏C反应蛋白水平的影响

目的观察姜黄素对不稳定型心绞痛（UAP）患者血清单核细胞趋化蛋白1（MCP-1）和超敏C反应蛋白（hsCRP）水平的影响。方法 UAP患者共48例,随机分为对照组（n=23）和治疗组（n=25）。治疗组在常规治疗的基础上加用姜黄素口服,对照组在常规治疗基础上给予辛伐他汀口服。另设健康组（n=20）为周期故城县体检者,年龄、性别等一般资料与上述两组差异无统计学意义。分别用酶联免疫吸附测定法和免疫比浊法测定两组治疗前、第4周血清MCP-1和hsCRP水平的变化。结果与健康组比较,UAP各组血清MCP-1和hsCRP水平显著增高,MCP-1（138±14）ng/L、（144±15）ng/L vs（55±12）ng/L;hsCRP（8.13±1.06）mg/L、（8.83±1.23）mg/L vs（2.92±0.93）mg/L（均P〈0.01）。与治疗前比较,治疗后UAP各组血清MCP-1和hsCRP水平有显著下降,MCP-1（138±14）ng/L vs（99±7）ng/L（、144±15）ng/L vs（76±7）ng/L;hsCRP（8.13±1.06）mg/L vs（5.90±0.68）mg/L、（8.83±1.23）mg/L vs（4.10±0.74）mg/L（均P〈0.01）。与对照组比较,治疗组治疗后血清MCP-1水平和hsCRP水平显著下降,MCP-1（99±7）ng/L vs（76±7）ng/L,hsCRP（5.90±0.68）mg/L vs（4.10±0.74）mg/L（均P〈0.01）。结论 UAP患者应用姜黄素可减轻炎症反应。     

Curcumin ameliorates epithelial-to-mesenchymal transition of podocytes in vivo and in vitro via regulating caveolin-1

AimsEpithelial-mesenchymal transition (EMT) is recognized to play a key role in diabetic nephropathy (DN). Curcumin, the main active component of turmeric extracted from the roots of the Curcuma longa plant, has been reported for its anti-fibrotic effects in kidney fibrosis. The purpose of our study was to investigate the effects of curcumin in reversing epithelial-to-mesenchymal transition (EMT) of podocytes in vivo and in vitro.Materials/methodsIn vivo streptozotocin (STZ)-induced diabetic rats received vehicle or curcumin, and podocytes were treated with high glucose (HG) in the presence or absence of curcumin in vitro. And we investigated the effect of curcumin on HG-induced phosphorylation of cav-1 on the stability cav-1 and β-catenin using immunoprecipitation and fluorescence microscopy analysis.ResultsCurcumin treatment dramatically ameliorated metabolic parameters, renal function, morphological parameters in diabetic rats. We found that HG treatment led to significant down-regulation of p-cadherin and synaptopodin, as well as remarkable up-regulation of α-SMA and FSP-1 in vivo and in vitro. Furthermore, curcumin inhibited HG-induced caveolin-1 (cav-1) Tyr¹⁴ phosphorylation associating with the suppression of stabilization of cav-1 and β-catenin.ConclusionsIn summary, these findings suggest that curcumin prevents EMT of podocytes, proteinuria, and kidney injury in DN by suppressing the phosphorylation of cav-1, and increasing stabilization of cav-1 and β-catenin.