抗氧化肽SS-31对高糖诱导的小鼠肾小球系膜细胞凋亡的影响

目的观察抗氧化肽SS-31对高糖诱导小鼠肾小球系膜细胞凋亡的影响。方法将体外培养小鼠肾小球系膜细胞分为正常糖组、正常糖+甘露醇组、高糖组及高糖+SS-31组。采用原位末端转移酶标记技术(TUNEL)及流式细胞术检测细胞凋亡;流式细胞术检测细胞ROS水平;Western blot检测caspase-3、cleaved caspase-3、Bax、Bcl-2、p38 MAPK、p-p38 MAPK及细胞色素C的表达。结果与正常糖组相比,高糖组系膜细胞ROS产生和细胞凋亡明显增加,cleavedcaspase-3和p-p38 MAPK表达增高,Bax/Bcl-2比率明显升高以及细胞色素C易位。SS-31干预能够明显抑制高糖诱导的系膜细胞凋亡和ROS产生,下调cleaved caspase-3和p-p38 MAPK的表达,减少Bax/Bcl-2比率和细胞色素C易位。结论 SS-31能够抑制高糖诱导的系膜细胞凋亡可能是通过减少ROS产生,保护线粒体功能,抑制p38MAPK信号通路激活而实现的。     

异丙酚预处理对大鼠海马神经元凋亡的影响

目的探讨异丙酚预处理对缺氧/复氧后大鼠海马神经元凋亡的影响。方法取离体培养的大鼠海马神经元,随机分成5组:正常组(A)组。CoCl2组(B)组:加入300μmol·L-1(下同)CoCl2处理4h,然后更换正常的培养基培养24h,之后更换无血清的培养基培养。脂肪乳组(C)组:加入10%脂肪乳剂90μl预处理1h后同B组。异丙酚组(D)组:加入100μmol·L-1异丙酚1h后同B组。7-NI组(E)组:如D组,但加入CoCl2的同时加入3·3μl7-NI(25g·L-1)处理4h。MTT法测定细胞增殖,流式细胞技术测定细胞的凋亡。RT-PCR技术检测神经型一氧化氮合酶和凋亡相关基因的表达情况。结果脂肪乳组与CoCl2组比较,各项指标相似(P>0·05);异丙酚可增加神经元的增殖能力,减少凋亡(P<0·05或P<0·01),上调神经型一氧化氮合酶和凋亡抑制基因bcl-2的表达,下调促凋亡基因cyclinD1的表达。而这些调节作用可被神经型一氧化氮合酶抑制剂7-NI抑制(P<0·05或P<0·01)。结论100μmol·L-1异丙酚预处理对缺氧/复氧后大鼠海马神经元有保护作用,神经型一氧化氮合酶和凋亡相关基因在其中起重要作用。     

寿尔智胶囊对老年性痴呆与海马神经元细胞凋亡的影响

寿尔智胶囊以中医“化痰开窍，交通心肾”立法组方，经多年临床应用治疗AD疗效较好。本课题在前期药效学、急性毒性实验、神经细胞培养等研究基础上，进一步从细胞、分子水平上探索AD发病及治疗机理，阐明化痰开窍、交通心肾法在治疗AD上的作用和意义，从而获取中医防治AD的新思路。     

硫氧还蛋白对缺氧/复氧海马神经元生长与凋亡的影响

本研究诱导原代培养10d的新生SD大鼠海马神经元,在神经元缺氧／复氧前,先用硫氧还蛋白处理,检测在不同时点的神经元存活率及Caspase-3蛋白酶活性,探讨硫氧还蛋白在神经元生长及凋亡中的可能作用机理。     

雌激素对NMDA诱导离体大鼠海马神经元凋亡的作用

目的观察雌激素对NMDA诱导离体海马神经元凋亡的作用,探讨雌激素的神经保护作用机制。方法用形态学鉴定和细胞死亡率的测定以及Western blot方法分析雌激素对NMDA诱导神经元凋亡的作用。结果培养的神经细胞经形态学鉴定绝大多数是神经元,占总细胞数目的81·3%,凋亡细胞百分数为15·1%,NMDA(300μmol·L-1+甘氨酸5μmol·L-1)能明显诱导神经元凋亡,凋亡发生数为31·6%(与对照组相比P<0·05),雌激素(300μmol·L-1)能明显拮抗NMDA的凋亡毒性作用,凋亡发生数为18·2%(与对照组相比P>0·05)。MTT实验测定细胞生存率:NMDA组为70·2%(与对照组相比P<0·05),NMDA+雌激素组为89·7%(与对照组相比P>0·05)。蛋白印迹也显示,caspase-3的活性能被雌激素所抑制。结论雌激素具有神经保护作用,作用机制可能通过抑制凋亡相关酶的活性来实现。实验结果为开发雌激素的临床应用和寻找神经保护作用药物提供实验参考依据。     

铅对大鼠海马神经元培养细胞ERK活性的影响

目的 观察不同浓度和不同时间染铅对大鼠海马神经元培养细胞细胞质ERK活性的影响。方法 出生4－8d Wistar大鼠海马神经元原代细胞培养,抑制剂PD098059预处理培养细胞,不同浓度及不同时间染铅,应用改良的Takai法测定ERK活性的变化。结果 0．1、0．5、1．0、5．0μmol染铅,ERK活性依浓度依赖方式被激活,比活性倍数分别为2．36、3．16、7．00、3．19（P＜0．05）;PD098059抑制铅激活ERK活性,1．0μmol染铅30min时ERK活性最高,为2．66（P＜0．05）,120min时降至正常,为1．72（P＞0．05）。结论 低浓度染铅可短暂激活大鼠海马神经元原代培养细胞ERK。     

纳洛酮对海马神经元的预防保护作用

目的为观察缺氧对大鼠海马神经元的影响及纳络酮的保护作用。方法采用细胞培养方法获取10～14d的海马神经元,将含药血清加入到培养液中,4h后建立95%N2＋5%CO2混合气体急性缺氧模型,胎盘兰染色细胞计数并做流式细胞仪检测以及酶法测定抗氧化酶（超氧化物歧化酶）的抗氧化能力及丙二醛、NO的含量。结果离体条件下纳络酮可显著增强海马神经元的活性,降低其凋亡率及死亡率;提高细胞上清夜中超氧化物歧化酶的活性,降低丙二醛及NO的含量,且成量效比例关系。结论纳络酮在体外有抗缺氧损伤的作用。其作用机制可能是通过抑制细胞凋亡,提高抗氧化酶的活力,清除自由基,而发挥作用。     

缺血预适应对大鼠海马神经元凋亡的影响

目的 探讨缺血预适应对缺氧复氧后大鼠海马神经元的影响.方法 用300μmol/L氯化钴预处理大鼠海马神经元2 h,然后更换正常的培养基培养24 h,之后用无血清的培养基培养,建立预适应的细胞模型.四甲基偶氮唑蓝(MTT)法测定细胞增殖,流式细胞术测定细胞凋亡,反转录-聚合酶链反应(RT-PCR)检测凋亡相关基因的表达情...     

休克对缺血性兔脑海马CA1区神经元凋亡的影响

目的探讨休克对缺血性兔脑海马CA1区神经元凋亡的影响。方法选取30只健康新西兰兔,随机分成三组:对照组(C组);实验组(E组),即平均动脉压(MAP)降至基础值的50%,30 min后给予等量林格氏液回输;处理组(S组),即MAP降至基础值的50%,30 min后给予等量血液回输。每组各10只。6 h后取材,光镜观察脑海马CA1区神经元病理学变化。免疫组化SP法检测海马CA1区c-fos蛋白表达。原位末端标记技术(TUNEL法)检测CA1区神经元细胞凋亡情况并计算凋亡指数。结果病理学HE染色C组及S组可见海马CA1区神经元形态排列较规则,细胞核呈椭圆形增大,核仁明显,细胞间隙清楚;E组可见神经元损伤更重,细胞排列紊乱,神经细胞脱失严重,胞核固缩明显,出现大量凋亡细胞。E组的c-fos平均光密度值较其他两组低(P<0.01),S组也低于C组(P<0.01)。C、S组凋亡指数较E组低(P<0.01)。结论休克可导致兔脑海马CA1区神经元细胞发生凋亡,可降低c-fos的蛋白表达。     

二苯乙烯苷对缺糖缺氧致海马神经元凋亡的影响*

目的观察中药何首乌有效成分2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(2,3,5,4'-tetrahydrox-ystilbene-2-O-β-D-glucoside,简称二苯乙烯苷,TSG)对缺糖、缺氧诱导的大鼠海马神经元细胞凋亡的影响.方法采用新生Wistar大鼠进行海马神经元细胞的原代培养,用连二亚硫酸钠合并无糖培养基处理细胞建立体外缺糖、缺氧模型,用酶标仪测定细胞乳酸脱氢酶(LDH)的漏出率,TUNEL法检测细胞凋亡,并观察TSG对缺糖、缺氧诱导的细胞凋亡的拮抗作用.结果模型组与对照组相比,LDH漏出率明显升高、出现了凋亡细胞,胞核致密并有凋亡小体产生.与模型组相比,TSG 10,50和100μmol·L-1能够明显抑制LDH的漏出,降低神经细胞的凋亡率,凋亡细胞数量、面积及灰度均下降.结论TSG可以抑制缺糖、缺氧诱导的神经元细胞凋亡,对缺血性脑病的治疗具有一定的应用前景.     

Neuroprotective and Antioxidant Effects of Novel Benzofuran-2-Carboxamide Derivatives

In the present study, we synthesized a series of novel 7-methoxy-N-(substituted phenyl)benzofuran-2-carboxamide derivatives in moderate to good yields and evaluated their neuroprotective and antioxidant activities using primary cultured rat cortical neuronal cells and in vitro cell-free bioassays. Based on our primary screening data with eighteen synthesized derivatives, nine compounds (1a, 1c, 1f, 1i, 1j, 1l, 1p, 1q and 1r) exhibiting considerable protection against the NMDA-induced excitotoxic neuronal cell damage at the concentration of 100 μM were selected for further evaluation. Among the selected derivatives, compound 1f (with -CH₃ substitution at R2 position) exhibited the most potent and efficacious neuroprotective action against the NMDA-induced excitotoxicity. Its neuroprotective effect was almost comparable to that of memantine, a well-known NMDA antagonist, at 30 μM concentration. In addition to 1f, compound 1j (with -OH substitution at R3 position) also showed marked anti-excitotoxic effects at both 100 and 300 μM concentrations. These findings suggest that -CH₃ substitution at R2 position and, to a lesser degree, -OH substitution at R3 position may be important for exhibiting neuroprotective action against excitotoxic damage. Compound 1j was also found to scavenge 1,1-diphenyl-2-picrylhydrazyl radicals and inhibit in vitro lipid peroxidation in rat brain homogenate in moderate and appreciable degrees. Taken together, our structure-activity relationship studies suggest that the compound with -CH₃ substitution at R2 and -OH substitution at R3 positions of the benzofuran moiety might serve as the lead exhibiting potent anti-excitotoxic, ROS scavenging, and antioxidant activities. Further synthesis and evaluation will be necessary to confirm this possibility.     

乌司他丁对脓毒症患儿炎性反应和肾功能的影响

目的:研究鸟司他丁对脓毒症患儿炎性反应和肾功能的影响.方法:42例脓毒症患儿随机数余数分组法分为实验组21例和对照组21例,实验组给予乌司他丁1.2万U/kg一次性加入生理盐水100 mL中静脉滴注,连用7d,对照组以等量生理盐水代替,7d后检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、白介素-10(IL-10)和尿素氮(BUN)、肌酐(Cr)的变化.结果:实验组血清TNF-α、IL-6浓度明显低于对照组(P＜0.01),IL-10水平高于对照组(P＜0.05);实验组BUN、Cr水平降低(P＜0.05),对照组BUN、Cr水平无明显变化(P＞0.05).结论:鸟司他丁能降低脓毒症患儿TNF-α、IL-6水平,升高IL-10,减轻炎性反应,同时对肾功能有一定保护作用.     

槲皮素对糖尿病视网膜病变小鼠氧化应激损伤和炎症反应的影响

目的：探讨槲皮素对糖尿病视网膜病变（DR）的保护作用及其抗氧化和抗炎机制。方法： 腹腔注射链脲佐菌素（STZ）建立小鼠糖尿病模型,随机分为正常对照组、造模组、槲皮素治疗组（n = 10）。治疗后定期监测小鼠精神状态、血糖、体重;检测小鼠视网膜结构变化、超氧化物歧化酶（SOD）活性、8-羟基脱氧鸟苷（8-OHdG）、丙二醛（MDA）、肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）、白介素-β（IL-β）的mRNA等水平。结果：与造模组相比,槲皮素降低小鼠血糖（P ＜ 0.05）,小鼠体重无明显差异（P ＞ 0.05）,视网膜细胞排列整齐,厚度增加;视网膜组织中8-OHdG、MDA、TNF-α mRNA、IL-6 mRNA、IL-β mRNA、Bax及p-p38 MAPK蛋白表达指数显著降低（P ＜ 0.05）,T-SOD活力和Bcl-2蛋白表达水平明显升高（P ＜ 0.05）,但各组p38 MAPK蛋白表达水平无明显差异（P ＞ 0.05）。结论： 槲皮素改善2型糖尿病小鼠视网膜组织氧化应激损伤和炎症反应,其机制可能与p38 MAPK信号通路有关。     

Protective effect of polypeptides from larva of housefly (Musca domestica) on hydrogen peroxide-induced oxidative damage in HepG2 cells

Housefly (Musca domestica) is an important medical insect and its larva is an ideal high protein food source. We isolated from housefly larvae the polypeptides hydrolyzed by neutral protease (PHNP), and investigated the protective effect of PHNP on hydrogen peroxide (H₂O₂)-induced oxidative damage in HepG2 cells. Cells exposed to H₂O₂ showed a marked decrease in proliferation and intracellular superoxide dismutase (SOD) activity, and a significant increase in reactive oxygen species (ROS) level and malondialdehyde (MDA) content. H₂O₂ also caused apoptosis and mitochondrial dysfunction including mitochondrial fragmentation and the loss of mitochondrial membrane potential. Pretreatment with PHNP at concentrations of 2.5, 5, 10 μg/mL blocked these H₂O₂-induced cellular events in a dose-dependent manner. The effect of PHNP at 10 μg/mL is equal to that of ascorbic acid at 10 μM. In summary, PHNP has a protective effect against H₂O₂-induced oxidative injury in cells due to its ability to decrease intracellular ROS and elevate antioxidant enzyme activities.     

氟西汀对卒中后抑郁大鼠行为学及海马神经元凋亡的影响

目的 探讨以氟西汀为代表的SSRIs对卒中后抑郁(post-stroke depression,PSD)大鼠行为学和神经元凋亡的影响。方法 用大脑中动脉线栓法(MCAO)建立大鼠局灶性脑缺血模型,再结合慢性不可预见的温和性应激(chronic unpredictable mild stress,CUMS)和孤养法建立PSD模型,加以10 mg?kg^-1氟西汀干预。比较各组大鼠体质量与糖水消耗量;旷野实验测定直立活动和水平活动得分;流式细胞仪分析神经元细胞的凋亡率。结果 应激14 d后,与PSD组大鼠比较,氟西汀干预组大鼠体质量和糖水消耗比例低(P<0.05或P<0.01),水平和垂直试验得分下降(P<0.01),海马神经元凋亡率较高(P<0.01)。结论 氟西汀对PSD有一定的治疗作用。     

Antioxidant Effect of CoQ10 on N-nitrosodiethylamine-induced Oxidative Stress in Mice

The antioxidant effect of CoQ₁₀ on N-nitrosodiethylamine (NDEA)-induced oxidative stress was investigated in mice. Food intake and body weight were similar in both CoQ₁₀ and control groups during the 3-week experimental period. NDEA significantly increased the activities of typical marker enzymes of liver function (AST, ALT and ALP) both in control and CoQ₁₀ groups. However, the increase of plasma aminotransferase activity was significantly reduced in the CoQ₁₀ group. Lipid peroxidation in various tissues, such as heart, lung, liver, kidney, spleen and plasma, was significantly increased by NDEA, but this increase was significantly reduced by 100 mg/kg of CoQ₁₀. Superoxide dismutase activity increased significantly upon NDEA-induced oxidative stress in both the control and CoQ₁₀ groups with the effect being less in the CoQ₁₀ group. Catalase activity decreased significantly in both the control and CoQ₁₀ groups treated with NDEA, again with the effect being less in the CoQ₁₀ group. The lesser effect on superoxide dismutase and catalase in the NDEA-treated CoQ₁₀ group is indicative of the protective effect CoQ₁₀. Thus, CoQ₁₀ can offer useful protection against NDEA-induced oxidative stress.     

Cathepsins: key modulators of cell death and inflammatory responses

Apoptosis is a key mechanism in the build up and maintenance of both innate and adaptive immunity as well as in the regulation of cellular homeostasis in almost every organ and tissue. Central to the apoptotic process is a family of intracellular cysteine proteases with aspartate-specificity, called caspases. Nevertheless, there is growing evidence that other non-caspase proteases, in particular lysosomal cathepsins, can play an important role in the regulation of apoptosis. In this review, the players and the molecular mechanisms involved in the lysosomal apoptotic pathways will be discussed as well as the importance of these pathways in the immune system and the pathogenesis of diseases.     

二甲双胍对糖基化终末产物诱导的成骨细胞氧化应激和凋亡的影响

目的:观察二甲双胍对糖基化终末产物(AGEs)诱导的大鼠颅骨成骨细胞内氧化应激产物活性氧簇(ROS)、细胞早期凋亡的影响.方法:分离培养大鼠颅骨成骨细胞,2 '7'-二乙酰二氯荧光素(DCFH-DA)作为荧光探针,流式细胞检测技术检测细胞内ROS水平,膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/ PI)双染色分析细胞早期凋亡率.结果:与未经葡萄糖处理的牛血清白蛋白(BSA)组对比,500 μg/mLAGEs显著促进成骨细胞内ROS形成和细胞凋亡(均P＜ 0.01);给予二甲双胍(100～500μmol/L)呈浓度依赖性抑制BSA组及AGEs组成骨细胞内ROS形成和细胞凋亡,浓度分别为500、400 μmol/L时,对细胞内ROS形成和细胞凋亡的抑制作用达到最强.结论:AGEs显著诱导原代成骨细胞内ROS的形成和细胞凋亡,而二甲双胍能够呈浓度依赖性抑制AGEs诱导的成骨细胞内ROS的形成和细胞凋亡,减轻AGEs对成骨细胞的损害.