三氧化二砷诱导人脑胶质瘤细胞抑制与活性氧水平的关系

目的研究三氧化二砷（As2O3）对人脑胶质瘤SHG44细胞活性氧浓度及^60Coγ射线照射联合As2O3对细胞DNA损伤中的影响。方法用2，7-二氢二氯荧光素（DCFH）标记细胞内活性氧（ROS），激光共聚焦显微镜扫描检测细胞内荧光强度；用单细胞凝胶电泳法检测DNA彗星尾长及尾部DNA百分含量，比较As2O3、γ线照射单独及联合作用对细胞DNA的损伤程度。结果（1）As2O3可明显提高SHG44细胞内ROS水平，其ROS水平的改变随As2O3作用浓度的增加而升高。（2）^60Coγ射线与As2O3联合作用诱导SHG44细胞DNA的损伤，其损伤程度随照射剂量的增加彗星尾长、尾部DNA百分比递增。两者联合作用后彗星尾长、尾部DNA百分比分别大于相应剂量的单独照射和As2O3组（P〈O．01）。结论As2O3可明显抑制SHG44细胞的增殖，增强细胞的辐射敏感性，其抑制机制与诱导细胞内ROS水平提高具有相关性。     

黄芩素-7-甲醚对缺氧致PC12细胞损伤的保护作用

目的 研究黄芩素-7-甲醚对缺氧PC12细胞的保护作用。方法 将PC12细胞分为正常对照组、缺氧模型组、芦丁组和黄芩素-7-甲醚组,培养24 h后,MTT法检测黄芩素-7-甲醚对细胞活力的影响;酶标仪法检测细胞培养上清液中乳酸脱氢酶（lactate dehydrogenase,LDH）活性以及细胞内丙二醛（malondialdehyde,MDA）、超氧化物歧化酶（superoxide dismutase,SOD）和过氧化氢酶（catalase,CAT）水平;2'',7''-二氯荧光素探针法检测细胞内活性氧簇（reactive oxygen species,ROS）水平;实时荧光定量PCR和Western blot检测核因子E2相关因子2（Nrf2）和血红素氧合酶-1（heme oxygenase-1,HO-1）的mRNA和蛋白的表达。结果 与正常对照组相比,缺氧导致PC12细胞活力显著降低,LDH、MDA和ROS水平显著升高,抗氧化酶SOD和CAT的活力显著降低。黄芩素-7-甲醚预处理能够逆转这些变化。此外,缺氧能够诱导细胞内Nrf2、HO-1的mRNA和蛋白表达增加,而黄芩素-7-甲醚能够进一步提高Nrf2、HO-1的mRNA和蛋白表达。结论 黄芩素-7-甲醚对缺氧致PC12细胞损伤有一定的保护作用,其作用机制可能与激活Nrf2/HO-1通路,抑制氧化应激损伤有关。     

果糖二磷酸钠对2型糖尿病血液流变学、SOD和LPO的影响

目的 :观察果糖二磷酸钠 (FDP)对 2型糖尿病血液流变学 ,SOD ,LPO的影响。方法 :2型糖尿病病人 60例 ,FDP组和常规组各 3 0例 ,FDP组在常规降糖药物 (磺脲类和双胍类 )治疗的同时 ,给予FDP 5 g ,iv ,gtt,bid× 3wk ,常规组只用降糖药物。病人在用药前 1wk测定血液流变学指标、SOD和LPO水平 ,用药 3wk后复查。结果 :FDP组治疗后血液流变学各项指标水平都有显著改善 ,SOD和LPO水平治疗前后分别为 (79± 6)× 10 3NU·L- 1和(5 .2± 0 .7) μmol·L- 1,(98± 10 )× 10 3NU·L- 1和(4.6± 0 .9) μmol·L- 1(P <0 .0 5或P <0 .0 1)。同常规组相比 ,P <0 .0 5或P <0 .0 1。结论 :FDP有改善 2型糖尿病血液粘度、提高SOD和降低LPO水平的作用。     

雷公藤多苷对高糖诱导人肾小管上皮细胞凋亡及CXCL10/CXCR3轴的影响

目的 探讨雷公藤多苷（TG）对高糖诱导人肾小管上皮细胞HK-2凋亡及CXC趋化因子配体10(CXCL10)/CXC趋化因子受体3(CXCR3)轴的影响。方法 体外培养人肾小管上皮细胞HK-2,并分为对照组（含5.5 mmol/L葡萄糖培养基）、高糖组（含25 mmol/L葡萄糖培养基）和12.5、25、50 mg/L TG组（分别用12.5、25、50 mg/L的TG和25 mmol/L葡萄糖培养基）。四甲基偶氮唑盐比色法检测HK-2细胞活力;流式细胞术检测HK-2细胞凋亡情况;2’,7’-二氯二氢荧光素二乙酸酯法检测HK-2细胞活性氧（ROS）水平;黄嘌呤氧化酶法检测HK-2细胞超氧化物歧化酶（SOD）水平;酶联免疫吸附试验检测HK-2细胞炎性因子肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)水平;蛋白免疫印迹法检测HK-2细胞凋亡蛋白[胱天蛋白酶（Caspase）-3、Caspase-9]以及CXCL10、CXCR3蛋白表达。结果 与对照组比较,高糖组HK-2细胞活力、SOD水平显著降低,凋亡率、ROS、TNF-α、TGF-β1水平及Caspase-3、Casp...     

昆布多糖硫酸酯对H2O2诱导乳鼠心肌细胞损伤的保护作用Δ

目的研究昆布多糖硫酸酯(LAPS)对过氧化氢(H2O2)诱导乳鼠心肌细胞氧化损伤的保护作用。方法分次消化法分离原代培养的乳鼠心肌细胞,建立H2O2诱导心肌细胞损伤模型;四甲基偶氮唑盐(MTT)法检测不同浓度LAPS对心肌细胞的保护作用;试剂盒检测LAPS对细胞乳酸脱氢酶(LDH)、肌酸磷酸激酶(CK)、丙二醛(MDA)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)含量的影响;利用荧光探针二氯二氢荧光素-乙酰乙酸酯(DCFH-DA)检测细胞内活性氧(ROS)含量。结果100μmol/L的H2O2能明显造成心肌细胞损伤,LAPS能改善H2O2所造成的心肌细胞损伤;LAPS能够明显降低的过氧化损伤心肌细胞外液中LDH和CK含量、明显降低过氧化损伤心肌细胞内MDA和活性氧(ROS)的含量、明显提高过氧化损伤心肌细胞内SOD、GSH-Px和CAT的活力。结论 LAPS对心肌细胞的过氧化损伤有保护作用,与其降低或阻断脂质过氧化反应,降低细胞内ROS含量,进而阻断因ROS对细胞造成的损害,及时修复受损细胞有着密切关系。     

黄芩苷对香烟烟雾提取物诱导的细胞损伤保护作用研究

目的研究黄芩苷对香烟烟雾提取物（cigarette smoke extract,CSE）诱导的人肺泡上皮细胞损伤的影响,并初步探讨其机制。方法制备CSE,用不同浓度黄芩苷对人肺泡上皮细胞A549进行预处理,再用CSE刺激细胞。用MTT法检测细胞存活率,流式细胞术测定细胞凋亡率,彗星实验观察DNA损伤情况,荧光法测定细胞内活性氧（reactive oxygen species,ROS）含量。结果随着CSE浓度的增加和作用时间的延长,细胞存活率下降,各组之间有统计学差异（P〈0.05）;黄芩苷能减少CSE诱导的细胞存活率下降,抑制CSE诱导的细胞内ROS的产生,并有剂量依赖关系（P〈0.05或P〈0.01）;黄芩苷＋CSE组细胞的彗星尾长、尾部DNA含量、尾距、Olive尾距均小于CSE组（P〈0.05）,并且黄芩苷可以减少CSE导致的细胞凋亡。结论黄芩苷可以拮抗CSE对细胞的损伤,提高细胞的存活率,降低凋亡率,其原因可能与黄芩苷能降低细胞ROS含量,减少DNA损伤有关。     

香烟烟雾对巨噬细胞的氧化损伤及盐酸氨溴索抗氧化作用的研究

目的研究香烟烟雾对体外培养的巨噬细胞的氧化损伤及盐酸氨溴索的抗氧化作用。方法采用黄嘌呤氧化酶法及硫代巴比妥酸法测定香烟烟雾提取物(CSE)和CSE+盐酸氨溴索作用于体外培养的小鼠巨噬细胞后30min、1、3和6h超氧化物歧化酶(SOD)和脂质过氧化物(LPO)的含量。结果加入CSE后3h,CSE组巨噬细胞SOD活性明显低于空白对照组,差异有统计学意义(P<0.05);CSE+氨溴索组SOD活性明显高于CSE组,差异有统计学意义(P<0.05);其余时间点三组SOD活性差异均无统计学意义。加入CSE后1h,CSE组巨噬细胞LPO含量明显高于空白对照组,差异有统计学意义(P<0.05);CSE+氨溴索组LPO含量与CSE组及空白对照组比较差异均无统计学意义;其余时间点三组LPO含量差异均无统计学意义。结论香烟烟雾作用于巨噬细胞产生脂质过氧化反应,造成氧化性损伤。盐酸氨溴索具有抗氧化作用,在一定程度上抑制过氧化反应,减轻氧化反应所致的损伤。     

神经生长因子对小鼠脾淋巴细胞DNA合成、cAMP和cGMP含量的影响

神经生长因子(NGF)是一种营养神经肽,在感觉和交感神经元的生长、维持和存活中起促进作用。NGF 还是一种神经免疫调节剂,可通过多种途径调节免疫反应。鉴于脾细胞表面存在特异性 NGF 受体,作者分别应用~3H-胸苷(~3H-TdR)掺入法及放射免疫法观察2.5s NGF 对小鼠脾淋巴细胞增殖及其细胞内 cAMP 和 cGMP 含量的影响。结果发现,NGF(4×10~(-7)～4×10~(-8)moL/L)在体外可剂量依赖性地抑制~3H-TdR 掺入到脾淋巴细胞 DNA 中,并能提高细胞内 cAMP 水平,但对 cGMP 无影     

香烟烟雾水溶性提取物对肝细胞线粒体的损伤

目的 探讨香烟烟雾水溶性提取物(CSW)对L-02肝细胞线粒体结构与功能的影响.方法 采用噻唑蓝(MTT)检测CSW处理L-02肝细胞24 h后的生存率,以确定后续试验CSW染毒浓度为0、8×10-3、16×10-3、32×10-3、64×10-3支/ml培养基.CSW染毒处理L-02肝细胞24 h后用MTF法检测线粒体总酶活力抑制率,荧光分光光度法检测线粒体膜通透性转运孔(PTP)开放度与细胞线粒体膜电位(△ψm)的变化,透射电子显微镜观察细胞线粒体形态学变化.结果 8×10-3、16×10-3、32×10-3、64×10-3支/ml 4个不同浓度处理组CSW对L-02肝细胞具有一定的细胞毒性,其细胞生存率随CSW处理浓度的增高而降低,且二者呈明显负相关(相关系数r=-0.957);线粒体PTP开放程度则随CSW浓度的增高而增加,32 ×10-3支/ml及其以上CSW浓度组开放度与对照组比较,差异有统计学意义(P＜0.05);线粒体总酶活力与细胞线粒体膜电位(△ψm)随CSW浓度的增加而下降,二者分别在8×10-3、16×10-3支/ml及其以上CSW浓度组与对照组比较,差异均有统计学意义(P＜0.05).透射电子显微镜观察线粒体形态发生显著改变即表现为肿胀、空泡变性、膜和嵴破损.结论 香烟烟雾水溶性提取物可对L-02肝细胞线粒体的结构与功能造成一定程度损伤.     

支气管哮喘氧自由基损伤的实验研究

目的　探讨氧自由基在支气管哮喘发病中的作用。方法　通过支气管哮喘动物模型复制 ,分别测定哮喘组、β2 受体激动剂治疗组 (美喘清组 )、类固醇激素治疗组 (甲强组 )及正常对照组(对照组 )的小鼠肺组织超氧化物歧化酶 (SOD)和脂质过氧化物 (LPO)含量变化。结果　哮喘组、美喘清组、甲强组SOD分别为 ( 92 86± 9 2 9)U/ml、( 94 73± 10 2 5 )U/ml、( 10 1 78± 10 18)U/ml,前二组明显低于正常对照组的 ( 10 8 3 0± 2 1 67)U/ml,甲强组与对照组相比则无差异。三组LPO分别为 ( 3 81± 0 69)nmol/ gwwt、( 3 78± 0 71)nmol/ gwwt、( 3 0 3± 0 64)nmol/gwwt,前二组明显高于正常对照组的 ( 2 69± 0 5 7)nmol/ gwwt ,甲强组与对照组相比亦无差异。美喘清组与哮喘组SOD、LPO之间无显著差异。甲强组SOD明显高于哮喘组 ,LPO明显低于哮喘组。结论　氧自由基损伤参与了支气管哮喘病理过程 ,类固醇激素可以有效地降低氧自由基的产生 ,β2 受体激动剂无此作用。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.