超低分子量肝素抑制神经细胞内钙释放保护谷氨酸损伤原代培养大鼠神经元

目的探讨超低分子量肝素(ultra low molecular weight heparin,ULMWH)对谷氨酸(Glu)诱导原代培养大鼠大脑皮层神经细胞损伤的保护作用及其作用机制。方法采用体外培养大鼠大脑皮层神经细胞,建立谷氨酸(Glu)诱导损伤模型,采用MTT法检测细胞活力、Hoechest33258染色法观察凋亡细胞形态改变和检测凋亡细胞数。同时,采用Fura-2/AM双波长荧光分光光度法测定神经细胞内钙离子浓度([Ca2+]i)。结果提前应用ULMWH可提高Glu损伤神经细胞的生存能力,降低Glu诱导的凋亡细胞数。同时,ULM-WH不论是在含Ca2+测量介质还是在无Ca2+测量介质中均可降低神经细胞[Ca2+]i。结论ULMWH对Glu损伤神经细胞具有保护作用,该作用可能与其抑制神经细胞内Ca2+释放而降低胞内[Ca2+]i有关。     

L—焦谷氨酸对抗从氨酸钠诱发的大鼠皮层神经元损伤

目的：在大鼠皮层神经元研究Ｌ－吡咯烷酮羧酸（Ｌ－ＰＧＡ）对谷氨酸钠（Ｇｌｕ）诱发神经毒性的拮抗作用。方法：原代培养的皮层神经元取自１６ｄ龄的胎鼠,与Ｇｌｕ作用３０分钟,２４后测定神经元的存活及增益昌质中亚硝酸盐的浓度;以Ｆｕｒａ ２－ＡＭｏ ｘｌｅｑｍｗ 〖Ｃａ＾２＋〗;荧光探针,,ＡＲ－ＣＭ－ＭＩＣ阳离子测定系统测定〖Ｃａ＾２＋〗ｉ。结果：Ｌ－ＰＧＡ１０－８０βｍｏｌ．Ｌ＾－１浓度依赖地抑制Ｇ     

β—内啡肽增强谷氨酸神经毒性

探讨β－内啡肽对谷氨酸单钠诱导的神经毒性的影响。方法：形态学观察、神经元面积图象分析、线粒体膜蛋白结合钙和单细胞内游离钙浓度测定。结果：β－内啡肽在０．５－５．０ｍｇ·ｋｇ＾－１范围内以剂量依赖方式加剧谷氨酸单钠诱导的下丘脑弓状核神经元损伤，谷氨酸单钠诱导的线粒体膜蛋白结合钙增多可以被β－内啡肽２ｇ·Ｌ＾－１加强，谷氨到单钠诱导的单细胞内钙浓度升高也被β－内啡肽２ｇ·Ｌ＾－１从３２０±８４提高至５     

β-内啡肽对谷氨酸神经毒性作用的影响

采用形态学观察、β-内啡肽(β-End)放射免疫测定及单细胞内游离钙浓度——[Ca²⁺]i检测等方法,观察了β-End对谷氨酸单钠(MSG)诱发神经元损伤的影响,分析了可能的作用机制。结果表明,β-End可以明显加重MSG诱发的下丘脑弓状核神经元的损伤;β-End和MSG诱发的[Ca²⁺]i增高可被维拉帕米部分逆转。另外,吗啡可以进一步加剧MSG诱导的各脑区β-End含量的变化。提示β-End可以明显地加剧MSG的神经毒性作用,其机制与MSG能诱发脑内β-End的含量的增多及β-End可进一步破坏MSG引起的胞内钙稳态失衡有关。     

金丝桃苷对分离的新生大鼠脑细胞内游离钙浓度的影响

AIM: To study the effects of hyperin (Hyp) on free intracellular calcium concentration ([Ca2+]i) of brain cells. METHODS: The neonatal rat brain cells were dissociated. [Ca2+]i in presence and absence of extracellular high K+, L-glutamic acid (Glu), 5-hydroxytryptamine (5-HT), and norepinephrine (NE) were assayed with Fura 2-AM. RESULTS: The resting [Ca2+]i in Hanks' solution (CaCl2 1.3 mmol.L-1) was (208 +/- 12) nmol.L-1 (n = 17). Hyp had no significant effects on the resting [Ca2+]i. Hyp 1.0, 4.0, and 16.0 mumol.L-1 markedly inhibited the increase of [Ca2+]i evoked by K+ 50 mmol.L-1 in a concentration-dependent manner. Hyp 16.0 mumol.L-1 inhibited the increases of [Ca2+]i induced by NE 1, 2, 4, and 8 mumol.L-1. Hyp (16.0 mumol.L-1) also markedly attenuated 5-HT and Glu-induced increase of [Ca2+]i. CONCLUSION: Hyp possessed inhibitory effects on influx of Ca2+ in the neonatal rat brain cells.     

TMB—8抑制组胺,5—羟色胺和谷氨酸引起的培养乳牛基底动脉平滑肌细 …

AIM: To study the effects of 8-(N,N-diethylamino)-n-octyl-3,4,5- trimethoxybenzoate (TMB-8) on intracellular free calcium ([Ca2+]i) in cultured calf basilar artery smooth muscle cells. METHODS: [Ca2+]i was examined by a system of measurement of AR-CM-MIC, using Fura 2-AM as a fluorescent indicator. RESULTS: In the presence of extracellular Ca2+ 1.3 mmol.L-1, histamine (His), serotonin (5-HT), and sodium glutamate (Glu) markedly increased the [Ca2+]i which was attenuated by TMB-8. In Ca2+ free Hanks' solution containing egtazic acid 0.1 mmol.L-1, TMB-8 not only reduced the resting [Ca2+]i, but also inhibited the elevation of [Ca2+]i evoked by His and 5-HT. CONCLUSION: TMB-8 reduced the resting [Ca2+]i and attenuated His-, 5-HT-, and Glu-induced increases of [Ca2+]i in basilar artery smooth muscle cells.     

褪黑激素对小鼠脑细胞内游离钙浓度的抑制作用

目的 研究褪黑激素（Ｍｅｌ）对老年小鼠大脑皮层突触体内钙含量以及激动剂动诱发的新生小鼠脑细胞（Ｃａ＾２＋）升高的影响,以探讨Ｍｅｌ抗衰老的作用机理。方法 钙离子荧光染料Ｆｕｒａ－２ＡＭ负载已制备的突触体或细胞,ＲＦ－５０００型双波长荧光分光光度计测定（Ｃａ＾２＋）ｉ,结果：长期使用Ｍｅｌ抑制老年小鼠大脑皮层Ｃａ＾２＋超负荷,Ｍｅｌ也降低钙通道激活剂Ｂａｙ－Ｋ８６４４,高浓度氯化钾（ＫＣｌ）和谷氨酸     

大蒜新素对分离大鼠脑细胞内游离Ca^2＋的影响

观察大蒜新素对不同刺激剂所致分离大鼠脑细胞内游离钙的影响。方法：以Ｆｕｒａ２－ＡＭ为细胞内游离钙的荧光指示剂,用ＡＲ－ＣＭ－ＭＩＣ阳离子测定系统,直接测定了分离新生大鼠脑细胞内游离钙值,观察了大蒜新素的影响。结果;大蒜新素对脑细胞静息「Ｃａ＾２＋」ｉ无明显影响,大蒜新素１－１００μｍｏｌ．Ｌ＾－１能剂量依赖性地抑制高Ｋ＾＋和谷氨酸引起的「Ｃａ＾２＋」ｉ升高,其中ＩＣ５０分别为５９．７和６９．９μｍ     

TMB—8抑制神经递质引起的单个脑细胞内游离钙的升高

AIM: To study the effects of TMB-8 on [Ca2+]i elevation induced by neurotransmitters in dissociated brain cells. METHODS: The brain cell suspension was made using a gentle trituration for 1 min with a polished pipette. The changes of [Ca2+]i were detected by the fluorescent indicator, Fura 2-AM. RESULTS: In the presence of extracellular Ca2+ 1.3 mmol.L-1, sodium glutamate (Glu), histamine (His), and serotonin (5-HT) markedly increased the [Ca2+]i which were reduced by TMB-8 30 mumol.L-1. TMB-8 3 mumol.L-1 produced inhibitory effects on the increase of [Ca2+]i by His and 5-HT in a Ca(2+)-free Hanks' solution. The increase of [Ca2+]i by His and 5-HT was reduced to control level by TMB-8 10 mumol.L-1. CONCLUSION: TMB-8 inhibited the [Ca2+]i elevation induced by Glu, 5-HT, and His in brain cells.     

槲皮素降低大鼠心率和心肌钙振荡频率和抑制小鼠心肌肥厚

目的 研究槲皮素对心肌兴奋收缩耦联及构型重建的影响。方法 经动脉插管记录大鼠血流动力学,缩窄小鼠腹主动脉致心肌肥厚;检测Ｆｕｒａ２－ＡＭ负载的培养大鼠心肌细胞内游离钙（Ｃａ＾２＋）ｉ及钙振荡。结果：槲皮素剂量相关地降低大量窦性心率,而动脉血压,左室压及其微分改变轻微;１０－２５０μｍｏｌ．Ｌ＾－１时浓度依赖性降低培养心肌血发钙振荡频率,１００μｍｏｌ．Ｌ＾－１时预防异丙肾上腺素及哇巴因加速钙振荡频     

三七总皂苷的抗炎作用

研究三七总皂苷抗炎作用。方法;用角叉菜胶复制大鼠气囊滑膜炎模型。蛋白含量,ＰＬＡ２活性,Ｄｉｎ含量及炎细胞内游离钙水平分别用Ｌｏｗｒｙ法,微量酸滴定法,放射法及Ｆｕｒａ２－ＡＭ荧光分析法进行测定。结果：ＰｎＳ（６０－２４０ｍｇ．ｋｇ＾－１）明显抑制角叉菜胶诱导的白细胞数升高和蛋白渗出;     

三七皂苷对抗顺铂导致的肾毒性(英文)

目的:研究三七皂苷(PnS)对顺铂肾毒性的防护作用。方法:采用小鼠和原代兔肾近端小管细胞培养(FTC)建立体内外顺铂肾毒性模型。用双乙酰、苦味酸、溴乙锭、台酚蓝、~(125)碘标记和Fura 2-AM方法分别测血尿素氮、血清肌酐、细胞存活率、DNA链间交联、DNA-蛋白交联和细胞内游离钙离子。结果:预先2d给PnS(100,200mg·kg~(-1)·d~(-1))使顺铂导致的小鼠血尿素氮下降到83％和31％,血清肌酐下降到86％和42％(P<0.01)。提前24h PnS(10,100mg/L)与PTC孵育,细胞存活率从顺铂组的78％提高到81％和89％,DNA链间交联下降到47％和40％,DNA-蛋白交联下降到77％和42％,细胞内游离钙下降到70％和63％(P<0.01)。结论:PnS可预防顺铂的肾毒性,其机制是降低顺铂导致的DNA链间交联、DNA-蛋白交联和钙离子超载。     

Regulation of alpha 1-adrenoceptor on rat hepatocyte apoptosis induced by D-galactosamine and lipopolysaccharide

AIM: To study the regulation of alpha 1-adrenoceptor on the biochemical changes involved in hepatocyte apoptosis induced by D-galactosamine (D-GalN) and lipopolysaccharide (LPS). METHODS: Prazosin (Pra) 4 mg.kg-1 ig was administrated 1 h before i.p. D-GalN 600 mg.kg-1 and LPS 1 microgram.kg-1. The morphological changes of hepatocytes were observed. The concentrations of intracellular free ion calcium ([Ca2+]i), the expression of secretory phospholipase A2 (sPLA2) and cytosolic phospholipase A2 (cPLA2) in hepatocytes, and the levels of alanine aminotransferase (ALT), sPLA2, and tumor necrosis factor-alpha (TNF-alpha) in serum were all assayed. RESULTS: The amounts of apoptotic bodies and apoptotic hepatocytes were reduced by the treatment of Pra (P < 0.01). The levels of [Ca2+]i, ALT, sPLA2, and TNF-alpha were (605 +/- 95) nmol.L-1, (214 +/- 141) u.L-1, (11.2 +/- 1.1)%, and (314 +/- 93) ng.L-1, respectively. Pra could decrease all the above biochemical changes to (364 +/- 61) nmol.L-1, (157 +/- 99) u.L-1, (7.9 +/- 1.6)%, and (156 +/- 62) ng.L-1 (all P < 0.05), respectively. The expression of cPLA2 was reduced by Pra treatment, too, but Pra had no significant effect on the expression of sPLA2. CONCLUSION: The apoptosis in rat hepatocytes is regulated by alpha 1-AR signal transduction pathway including changes in [Ca2+]i, sPLA2 secretion, and cPLA2 expression. TNF-alpha is also involved in rat hepatocyte apoptotic regulation.     

汉防己甲素抑制三种神经递质引起的新生大鼠脑细胞内游离钙的升高

应用Ｐｕｒａ－２技术测定游离新生大鼠脑［Ｃａ２＋］ｉ浓度的技术、研究了Ｔｅｔ对静息脑［Ｃａ２＋］ｉ和３种递质引起的脑［Ｃａ２］ｉ变化的影响。Ｔｅｔ（１，１０和２０μｍｏｌ·Ｌ－１）对静息脑［Ｃａ２＋］ｉ无明显影响。Ｔｅｔ（１０μｍｏｌ·Ｌ－１）可降低Ｌ－Ｇｌｎ（０．１、１．０和１０μｍｏｌ·Ｌ－１）引起的脑（Ｃａ２＋）ｉ的升高。在Ｈａｎｋ＇ｓ液Ｃａ２＋为１．３ｍｍｏｌ·Ｌ－１时，Ｔｅｔ１０μｍｏｌ·Ｌ－１可降低Ｈｉｓ（５０和１００μｍｏｌ·Ｌ－１）和５－ＨＴ（０．１、１．０、１０和１００μｍｏｌ·Ｌ－１）引起的脑［Ｃａ２＋］ｉ的升高。但不能降低Ｈａｎｋ＇ｓ液无Ｃａ２＋时Ｈｉｓ和５－ＨＴ引起的脑［Ｃａ２＋］ｉ的升高。研究表明Ｔｅｔ可阻滞Ｌ－Ｇｉｎ、Ｈｉｓ和５－ＨＴ受体调控的钙通道。但对Ｈｉｓ和５－ＨＴ引起的细胞内贮存钙的释放并无明显影响。Ｔｅｔ的这种降低脑［Ｃａ２＋］ｉ的作用可能是其治疗脑缺血性疾病的机理之一。Ｐ＜０．０１在Ｔｅｔ１０μｍｏｌ·Ｌ－１作用下，相同浓度的细胞外液钙和Ｈｉｓ（０、５０和１００μｍｏｌ·Ｌ－１），脑［Ｃａ２＋］ｉ分别是２２１±５、２４５±５和３０２±６ｎｍｏｌ·Ｌ－１。增加了１１．８?     

前胡丙素对高血压大鼠血管肥厚、细胞内钙、胶原及NO的影响

目的　研究前胡丙素对自发性高血压大鼠SHR及肾型高血压大鼠RHR的血管肥厚、细胞内钙、胶原、NO及血管收缩的反应性影响。方法用显微测微仪测定血管中膜层厚度,细胞大小,用Fura-2/AM为荧光指示剂,测定单细胞内［Ca²⁺］i,以测定羟脯氨酸含量反映胶原含量,用Griess法测定NO含量,以血管环观察收缩反应。结果　前胡丙素抑制血管中膜层增厚,维持细胞内［Ca²⁺］i稳态。减少胶原形成,增加SMCs释放NO。抑制血管环高反应状态。结论　前胡丙素抑制高血压血管肥厚,降低胶原含量及血管异常反应。     

糖基化终产物对培养的大鼠主动脉平滑肌细胞增殖及胞浆游离钙含…

目的：研究糖基化终产物（ＡＧＥＰ）对主动脉平滑肌细胞增殖的影响及其与［Ｃａ＾２＋］ｉ的关系。方法：采用同位素掺入法分别测定ＤＮＡ和蛋白质合成；Ｆｕｒａ２－ＡＭ测定［Ｃａ＾２＋］ｉ。结果：ＡＧＥＰ以浓度、时间相关的方式促进［＾３Ｈ］ＴｄＲ与［＾Ｈ］Ｌｅｕ掺入细胞，随ＡＧＥＰ作用时间、糖化时间延长，掺入率增加明显，ＡＧＥＰ增加［Ｃａ＾２＋］ｉ，与时间、浓度相关，但随ＡＧＥＰ作用时间延长（４０分钟后）而     

内钙动员调控凝血酶诱导的血小板Na^＋／H^＋交换

目的：研究凝血酶诱导的血小板活化中细胞内钙动员和Ｎａ＾＋／Ｈ＾＋交换的关系。方法Ｆｕｒａ－２负载测［Ｃａ＾２］ｉ和ＢＣＥＣＦ负载测ｐＨｉ。结果：凝血酶０．１ＩＵ·Ｌ＾－１引起［Ｃａ＾２＋］ｉ和ｐＨｉ，［Ｃａ＾２］ｉ增加先于ｐＨｉ增加。在无钠溶液中，Ｎａ＾＋／Ｈ＾＋交换被抑制而［Ｃａ＾２］ｉ增加不受影响；用尼日利亚菌素（１ｍｇ·Ｌ＾－１）使胞内酸化可抑制［Ｃａ＾２＋］ｉ增加，用依他酸（ＢＧＴＡ）阻断     

小檗胺对高钾,去甲肾上腺素及咖啡因引起大鼠心肌细胞内钙动拮？…

AIM: To study the effects of berbamine (Ber) on intracellular calcium concentration ([Ca2+]i) mobilized by KCl depolarization, norepinephrine (NE), and caffeine. METHODS: [Ca2+]i was measured with fluorescent intensity (FI) by confocal microscope in single cultured cardiomyocytes of newborn rats loaded with Fluo 3-AM 2 mumol.L-1. RESULTS: FI value of [Ca2+]i in control level was 248 +/- 70 in the presence of extracellular calcium 1.5 mmol.L-1 and was not changed by Ber 3-30 mumol.L-1. KCl (60 mmol.L-1)- and NE (30 mumol.L-1)-induced [Ca2+]i mobilizations were inhibited (P < 0.01) by Ber 30 mumol.L-1, similar to that of verapamil (Ver). The inhibitory effect of Ber on [Ca2+]i induced by KCl was further increased (P < 0.05) in the presence of egtazic acid 3 mmol.L-1, but that on [Ca2+]i induced by NE was not changed. The [Ca2+]i mobilized by caffeine 80 and 160 mumol.L-1 in D-Hanks' solution was not affected (P > 0.05) by Ber and Ver. CONCLUSION: Ber possessed the antagonistic effects on [Ca2+]i increases via voltage-dependent Ca2+ channel and receptor-operated Ca2+ channel in newborn rat cardiomyocytes, but without effect on intracellular Ca2+ release.     

血小板释放的ADP通过稳定细胞内钙浓度而稳定PAF诱导的兔血小板 …

AIM: To examine whether platelet-released adenosine diphosphate (ADP) would contribute to the stabilization of rabbit platelet aggregation induced by platelet activating factor (PAF). METHODS: Rabbit platelet aggregation induced by PAF was measured turbimetrically. ADP release from rabbit platelets stimulated by PAF was determined by HPLC. Intracellular Ca2+ was measured using Ca(2+)-sensitive fluorescent indicator Fura 2-AM. RESULTS: PAF > or = 1 nmol.L-1 induced full platelet aggregation, which did not deaggregate over 5 min after aggregation reached peak. Platelet aggregation was deaggregated in a concentration-dependent manner by subsequent addition of ADP scavenger ATP-diphosphohydrolase (apyrase) at 5-100 mg.L-1. PAF 3 nmol.L-1 stimulated release of ADP (29% vs 6% of control), and elicited a rapid rise in intracellular calcium ([Ca2+]i) which peaked at approximately 15 s. Then the [Ca2+]i gradually decayed from 585 +/- 80 nmol.L-1 within 100 s to a low level (364 +/- 82 nmol.L-1). Apyrase 100 mg.L-1, added 2 min after PAF, reduced [Ca2+]i to a lower level (171 +/- 29 nmol.L-1). CONCLUSION: Platelet-released ADP stabilizes PAF-induced rabbit platelet aggregation by stabilizing [Ca2+]i at elevated level.     

前胡香豆素对肾型高血压大鼠左室肥厚及心肌胞内钙、Na+,K+-ATP酶和Ca2+,Mg2+-ATP酶活性的影响

目的研究前胡香豆素组分对肾型高血压左室肥厚的预防和逆转作用及机制。方法用两肾一夹肾型高血压左室肥厚大鼠(RHR)模型,测定前胡香豆素组分对其血压、左室湿重、心肌细胞面积、胞内静息钙及胞膜和线粒体ATP酶活性的影响。结果前胡香豆素组分(30 mg·kg^-1·d^-1,ig)预防组及逆转组大鼠血压、左室湿重/体重均较肥厚组明显降低;左室心肌细胞面积、胞内静息钙均较肥厚组降低;对KCl致钙浓度升高亦明显低于肥厚组;两组均可增加心肌细胞膜及线粒体Na⁺,K⁺-ATP酶和Ca²⁺,Mg²⁺-ATP酶活性。结论前胡香豆素组分可预防及逆转RHR左室肥厚,减少心肌细胞内钙含量,增加ATP酶活性。