High calcium enhances the expression of double‐stranded RNA sensors and antiviral activity in epidermal keratinocytes

Double‐stranded RNA (dsRNA) sensors including TLR3, MDA5 and RIG‐I are expressed in epidermal keratinocytes and play an important immunological role by enhancing various innate and adaptive immune responses. Although the role of elevated extracellular calcium concentration in keratinocyte differentiation is well understood, the effect of high calcium on dsRNA sensors is not well studied. We investigated alterations in dsRNA sensor expression and antiviral activity induced by a high extracellular concentration of calcium in epidermal keratinocytes. Normal human epidermal keratinocytes (NHEKs) were stimulated with high calcium and/or synthetic dsRNA, poly (I:C). TLR3, IFIH1 (MDA5) and DDX58 (RIG‐I) expression were measured via qPCR, and IFN‐β and human beta‐defensin 2 (HBD2) levels were measured using ELISA. TLR3 localization was evaluated with immunocytofluorescence. Antiviral activity was quantified with virus plaque assays using herpes simplex virus type 1 (HSV‐1). High calcium significantly upregulated mRNA expression of TLR3, IFIH1 and DDX58 in NHEKs. In addition, high calcium significantly enhanced poly (I:C)‐induced anti‐HSV‐1 activity in NHEKs. The antiviral molecule HBD2 but not IFN‐β induction by poly (I:C) was enhanced by high calcium. Our findings indicate that high levels of extracellular calcium enhance the expression of dsRNA sensors and augment antiviral activity in epidermal keratinocytes.     

Chemoprevention of liver cancer by plant polyphenols

Primary liver cancer or hepatocellular carcinoma (HCC) is one of the most frequent tumors representing the fifth commonest malignancy worldwide and the third cause of mortality from cancer. Currently, the treatments for HCC are not so effective and new strategies are needed for its fight. Chemoprevention, the use of natural or synthetic chemical agents to reverse, suppress or prevent carcinogenesis is considered an important way for confronting HCC. Many of the chemopreventive agents are phytochemicals, namely non-nutritive plant chemicals with protective or disease preventive properties. In this review, we focus on plant polyphenols, one of the most important classes of phytochemicals, their chemopreventive properties against HCC and discuss the molecular mechanisms accounting for this activity.     

银杏叶提取物EGb761对铜与高胆固醇共处理家兔主动脉病变的影响

目的 观察银杏叶提取物EGb761对铜和高胆固醇诱导的家兔主动脉病变的保护作用.方法 采用Sparks的方法复制铜和高胆固醇家兔模型,EGb761以50 mg·kg-1·d-1的剂量灌胃3 w后,苏丹Ⅲ和HE染色观察主动脉的大体和镜下形态改变,图像分析测量动脉粥样硬化面积占主动脉面积的百分比,酶法检测血清总胆同醇(TC)、甘油三酯( TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)水平,硫代巴比妥酸( TBA)法测定血清丙二醛(MDA)水平,实时定量RT-PCR法检测肝组织LDLR和LR PI mRNA表达.结果 模型组主动脉可见明显的动脉粥样硬化改变,EGb761组动脉粥样硬化病变较模型组减轻,图像分析结果显示,EGb761组斑块面积百分比显著降低,与模型组相比差别具有统计学意义(P＜0.05).模型组血清TC、TG、HDL-C、LDL-C和MDA水平均显著升高,与正常组相比差别具有统计学意义(P＜0.05);EGb761组血清LDL-C和MDA水平均显著降低,与模型组相比差别具有统计学意义(P＜0.05).EGb761组肝组织LRPI mRNA表达显著升高,与模型组相比差别具有统计学意义(P＜0.05).结论 EGb761具有改善动脉粥样硬化病变的作用,其作用机制与调节血脂水平和抗过氧化损伤有关.     

海马补肾丸对小鼠学习记忆障碍的改善作用

目的考察海马补肾丸对小鼠学习记忆障碍的改善作用,并探讨其作用机制。方法小鼠随机分为对照组、模型组、吡拉西坦组、龟龄集组和海马补肾丸组。采用腹腔注射东莨菪碱制备小鼠记忆获得性障碍模型,计算学习正确率,测定蛋白含量和乙酰胆碱酯酶活力。采用腹腔注射亚硝酸钠制备小鼠记忆巩固性障碍模型,计算训练、测定学习正确率,测定蛋白含量、去甲肾上腺素(NE)、多巴胺(DA)、5-羟色胺(5-HT)以及丙二醛(MDA)、超氧化物歧化酶(SOD)含量。结果海马补肾丸2.20 g/kg组能显著增加小鼠学习记忆的正确率(P0.05),显著降低乙酰胆碱酯酶活力(P0.01)。测定时,与模型组比较,海马补肾丸1.10、2.20 g/kg组能显著增加小鼠学习记忆正确率(P0.05),能不同程度增加小鼠大脑SOD含量,降低MDA含量(P0.05、0.01)。结论海马补肾丸能明显改善小鼠的记忆获得、记忆巩固障碍,推测其作用机制主要是通过降低乙酰胆碱酯酶活性,增加脑内乙酰胆碱的含量,改善记忆获得能力;通过抗脑组织的氧化损伤,增加大脑SOD含量,降低MDA含量,改善记忆巩固能力。     

还原型谷胱甘肽对慢性肝病患者血清MDA、GSH-Px、SOD含量的影响

目的：评价脂质过氧化在慢性乙型肝炎及酒精性肝炎发病中的作用及还原型谷胱甘肽的作用机制及临床疗效。方法：慢性乙型肝炎（CH）27例、慢性酒精性肝炎（ALH）14例分别作为治疗组，给予常规加还原型谷胱甘肽治疗；CH13例、ALH7例组成对照组给予常规治疗（极化液＋脉安定静滴），疗程均为1个月。结果：两组病人治疗前血清丙二（MDA）、超氧化物歧化酶（SOD）水平明显高于正常对照组，谷胱甘肽过氧化物酶（GHS－Px)水平明显低于正常对照组，治疗后两组病人血清MDA、ALT、TBil水平均明显下降，SOD水平无明显变化，GSH－Px水平明显升高。与治疗组比较除ALT水平明显下降外，其余无明显变化。结论：脂质过氧化是CH和ALH共同的发病机制，还原型谷胱甘肽通过有效抑制脂质过氧化，可明显促进病人肝功能的恢复。     

碱性电解水对大鼠抗氧化及抗纤维蛋白原作用的实验研究

目的　探讨碱性电解水抗氧化及抗纤维蛋白原的作用。方法　W istar雄性大鼠 36只 ,分三组 ,每组 1 2只。A组 :空白对照 ,B组 :高脂模型组。 C组 :高脂 +碱性电解水组。实验结束后 ( 30 d) ,乙醚麻醉 ,腹主动脉取血 ,采用硫代巴比妥酸比色法在日本岛津 UV— 36 0分光光度计上测定丙二醛 ( MDA) ;上海斯隆纤维蛋白原测定仪测定纤维蛋白原。结果　 A、B、C三组 M DA分别为 3.39± 0 .50 ( nmol/ m1 )、4 .1 0± 0 .37及 3.6 8±0 .4 6 ;纤维蛋白原分别为 1 94 .6 3± 2 3.6 3( mg/ d1 )、2 0 1 .2 2± 2 4 .32及 1 6 9.50± 1 1 .1 7。 C组比 B组显著下降 ( P<0 .0 5)。结论　碱性电解水具有一定抗氧化及降低纤维蛋白原的功能。     

铅和噪声联合作用对作业工人血SOD、MDA含量的影响

目的 探讨铅和噪声联合作用对工人的血清超氧化物歧化酶（SOD）、丙二醛（MDA）含量的影响.方法 选择年龄、工龄及接触噪声强度相近的A（铅和噪声联合作用组）、B（单纯噪声组）两组工人,测定血SOD、MDA含量,同时测定血铅含量,并比较两组数据差异.结果 A、B两组工人血SOD值分别为（65.2±16.3） U/ml、（63.8±13.9） U/ml,两组工人血SOD值无明显差异（P＞0.05）;A、B两组工人血MDA含量分别为（6.81±3.12）nmol/ml、（5.02±2.36）nmol/ml,两组工人血MDA含量差异有统计学意义（P＜0.05）.结论 铅和噪声联合作用可加重机体自由基损害和脂质过氧化.     

迷迭香酸对5/6肾切除大鼠尿毒素、尿蛋白及血清丙二醛和超氧化物歧化酶的影响

目的 观察迷迭香酸对5/6肾切除大鼠肾功能以及对大鼠尿蛋白和血清丙二醛(MDA)和超氧化物歧化酶(SOD)的影响.方法 制作大鼠慢性肾功能不全模型,实验分组为正常对照组,模型对照组,迷迭香酸组.根据手术后2周血肌酐值随机分组,每组6只.以迷迭香酸作为干预因素,应用丽春红S法等方法测定迷迭香酸对5/6肾切除大鼠尿蛋白、尿肌酐和血肌酐、血清MDA和S0D影响.结果 (1)与对照组及模型组比较,迷迭香酸组血肌酐及内生肌酐清除率均升高(F=38.912、19.968,P＜0.01,P＜0.05);(2)迷迭香酸对慢性肾功能不全大鼠的24h尿蛋白有明显改善作用(F=16.288,P＜0.01);(3)迷迭香酸对大鼠MDA及SOD无明显影响(P均＞0.05).结论 迷迭香酸可以降低尿蛋白量及血肌酐值,提高内生肌酐清除率,延缓肾脏衰竭.但对大鼠MDA及SOD无明显影响.     

盐酸戊乙奎醚对脂多糖诱导的内皮细胞损伤的保护作用

目的 探讨盐酸戊乙奎醚(PHC)对脂多糖(LPS)诱导的内皮细胞损伤的影响及其作用机制.方法 用RPMI 1640培养基将人脐静脉内皮细胞在5%CO2、37℃培养箱中培养,并分为对照组,LPS组及PHC组.检测细胞上清液乳酸脱氢酶(LDH)、细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)和一氧化氮(NO)水平.提取细胞核蛋白,Western blot分析细胞外信号调节激酶1/2(ERK1/2)和c-Jun氨基末端激酶(JNK)磷酸化蛋白的表达.采用SPSS 13.0统计软件进行分析,组间比较采用单因素方差分析,以P<0.05为差异具有统计学意义.结果 与对照组比较.LPS组LDH含量[(1 642±367)U/L vs.(169±33)U/L]、MDA含量[(13.2±1.2)nmol/L vs.(7.2±0.8)nmoL/mL]及NO水平[(143.2±10.3)μmol/L vs.(85.5 4.4.1)μmol/L]明显增多,SOD活性明显下降[(41.2 ±2.7)U/mL vs.(61.1±2.8)U/mL],ERK1/2和JNK磷酸化表达明显增高.PHC可显著降低LDH含量[(392 4.90)U/L],降低MDA[(8.6±1.3)nmol/mL]和NO水平[(92.1 ±6.6)μmol/L],提高SOD活性[(58.0±3.0)U/mL],减弱ERK1/2磷酸化蛋白表达.结论 盐酸戊乙奎醚对脂多糖诱导的内皮细胞损伤具有保护作用,其作用机制可能与抑制ERK1/2磷酸化表达减少过氧化物的产生有关.

Abstract：

Objective To investigate the effects of penehyclidine hydrochloride ( PHC) on lipopolysaccharide (LPS) -induced endothelial cells injury and its mechanism. Methods ECV-304 was cultured in RPMI1640 in a 5% humidified CO2 atmosphere at 37 ℃. Then cultured cells were used to assess the following treatments: control group, LPS group (1 μg/mL) and PHC group(2 μg/mL). At the end of the experiments, supernatant was collected for determination of lactate dehydrogenase ( LDH), and cells were collected for determination of malondialdehyde (MDA), superoxide dismutase (SOD), and nitric oxide (NO) levels. And extracellular regulated kinasel/2( ERKl/2)and JNK MAPK (mitogen-activated protein kinases, MAPK) protein expressions were determined using Western blot technique. Analysis of variance (ANOVA) was used for statistical analysis to compare values among all groups. A significant difference was presumed for a probability value < 0.05. Results Compared with control group, LDH leakage [(1642 ± 367) U/L vs (169±33)U/L], the contents of MDA[(13. 2 ± 1. 2) nmol/L vs (7. 2 ±0. 8)nmol/mL] and NO levels [(143.2 ± 10.3) μmol/L vs(85.5 ±4.1) μmol/L], expressions of ERK1/2 and JNK were remarkably increased and SOD activities[(41.2 ±2.7) U/mL vs (61. 1 ±2.8) U/mL] were obviously decreased in LPS group. PHC markedly decreased LDH leakage [(392 ±90) U/L], MDA contents [(8. 6 ± 1. 3) nmol/ mL] and NO levels [(92.1 ±6.6) μmol/L], ERK1/2 activation and enhanced SOD activities [(58.0± 3.0) U/mL]. Conclusions PHC could protect endothelial cells against LPS-induced cell injury. The effect of PHC is likely mediated through inhibition of ERK1/2 MAPK activation.     

红豆杉多糖对Beagle犬心肌缺血再灌注损伤模型心肌NADPH氧化酶mRNA及SOD、MDA的影响

目的 观察红豆杉多糖对心肌缺血-再灌注损伤（MIRI）模型心肌损伤及心肌还原型辅酶II（NADPH）氧化酶mRNA、超氧化物歧化酶（SOD）、丙二醛（MDA）的影响。方法 将30只Beagle犬随机分为假手术组、模型组、红豆杉多糖低剂量组与高剂量组、卡维地洛对照组,每组6只,给药7 d后,建立MIRI模型。HE染色,光镜观察心肌组织病理改变;原位杂交检测NADPH氧化酶 p47^phox（NCF-47K）mRNA表达;比色法检测心肌SOD活性和MDA水平。结果 光镜下可见,模型组心肌严重紊乱、断裂,核固缩及胞浆嗜酸性变明显,轻至中度充血、水肿,炎细胞浸润;红豆杉多糖高剂量组心肌细胞紊乱、断裂程度较轻,核固缩及胞浆嗜酸性变明显减少,部分组织仍有轻度充血、水肿。模型组、红豆杉多糖低剂量组心肌NCF-47K mRNA表达较假手术组显著升高（P＜0.01）,红豆杉多糖高剂量组NCF-47K mRNA表达显著低于模型组（P＜0.05）。除红豆杉多糖高剂量组心肌SOD活性显著高于模型组（P＜0.01）外,模型组及各治疗组心肌组织SOD活性较假手术组显著降低（P＜0.01）。模型组及红豆杉多糖低剂量组心肌组织MDA水平较假手术组显著增加（P＜0.05）,红豆杉多糖高剂量组心肌MDA水平显著低于模型组（P＜0.05）。结论 红豆杉多糖可能通过降低心肌组织NADPH氧化酶NCF-47K mRNA表达,升高SOD活性,减少O₂^÷等氧自由基对心肌细胞的损伤,减轻缺血再灌注导致的心肌损伤。