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缺氧对乳鼠耳蜗螺旋神经节神经元外向电流的影响及其环磷酸腺苷、环磷酸鸟苷含量变化的研究 预览
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作者 王艳萍 朱贺 +3 位作者 刘欢 马克涛 司军强 李丽 《中国现代医学杂志》 CAS 北大核心 2016年第13期12-17,共6页
目的观察缺氧时间对体外培养的耳蜗螺旋神经节神经元(SGNs)外向电流的影响及其细胞内第二信使环磷酸腺苷(c AMP)、环磷酸鸟苷(c GMP)含量变化。方法体外原代培养新生1~3 d SD大鼠SGNs并进行免疫荧光鉴定。用膜片钳全细胞记录模式... 目的观察缺氧时间对体外培养的耳蜗螺旋神经节神经元(SGNs)外向电流的影响及其细胞内第二信使环磷酸腺苷(c AMP)、环磷酸鸟苷(c GMP)含量变化。方法体外原代培养新生1~3 d SD大鼠SGNs并进行免疫荧光鉴定。用膜片钳全细胞记录模式观察缺氧外液灌流SGNs时,外向电流的变化趋势和特点。用酶联免疫吸附实验检测缺氧5、15和20 min时SGNs内c AMP和c GMP的浓度变化。结果 1酶解分离可获得生存状态良好的神经元,经免疫荧光鉴定为SGNs。2当电压钳制在-60 m V时,急性缺氧增强SGNs外向电流,主要增强0~+60 m V电压区间的激活电流幅度。缺氧5 min时,+60 m V激活电流幅度从(971.2±50.3)p A增强到(2361.0±207.4)p A,差异有统计学意义(P〈0.01)。随后出现下降趋势,11~15 min时,增强的电流幅度与对照组基础电流比较,差异无统计学意义(P〉0.05)。3分别检测缺氧5、15和20 min后SGNs中c AMP和c GMP的含量变化。缺氧5 min组与正常对照组比较,c AMP浓度升高,差异有统计学意义(P〈0.01)。缺氧15 min组与正常对照组比较,c AMP浓度变化差异无统计学意义;缺氧20 min组与正常组对照组比较,c AMP浓度水平下降(P〈0.01)。c GMP浓度随着缺氧时间的延长,总体呈下降趋势,但在20 min后有部分回升(缺氧15 min vs缺氧20 min,P〈0.01)。结论在SGNs急性缺氧损伤过程中,外向电流增强,增强幅度随缺氧时间变化先升后降,推测在短期缺氧时细胞通过降低兴奋性,影响听觉信息的传导,且该过程与胞内第二信使c AMP和c GMP浓度的改变相关。 展开更多
关键词 缺氧 SGNs 环磷酸腺苷 环磷酸鸟苷
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Differential Effect of Calcium-Activated Potassium and Chloride Channels on Rat Basilar Artery Vasomotion 预览
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作者 李丽 王蕊 +5 位作者 马克涛 李新芝 张传林 刘卫东 赵磊 司军强 《华中科技大学学报:医学英德文版》 SCIE CAS 2014年第4期482-490,共9页
Spontaneous, rhythmical contractions, or vasomotion, can be recorded from cerebral vessels under both normal physiological and pathophysiological conditions. We investigated the cellular mechanisms underlying vasomoti... Spontaneous, rhythmical contractions, or vasomotion, can be recorded from cerebral vessels under both normal physiological and pathophysiological conditions. We investigated the cellular mechanisms underlying vasomotion in the cerebral basilar artery(BA) of Wistar rats. Pressure myograph video microscopy was used to study the changes in cerebral artery vessel diameter. The main results of this study were as follows:(1) The diameters of BA and middle cerebral artery(MCA) were 314.5±15.7 μm(n=15) and 233.3±10.1 μm(n=12) at 10 mmHg working pressure(P<0.05), respectively. Pressure-induced vasomotion occurred in BA(22/28, 78.6%), but not in MCA(4/31, 12.9%) from 0 to 70 mmHg working pressure. As is typical for vasomotion, the contractile phase of the response was more rapid than the relaxation phase;(2) The frequency of vasomotion response and the diameter were gradually increased in BA from 0 to 70 mmHg working pressure. The amplitude of the rhythmic contractions was relatively constant once stable conditions were achieved. The frequency of contractions was variable and the highest value was 16.7±4.7(n=13) per 10 min at 60 mmHg working pressure;(3) The pressure-induced vasomotion of the isolated BA was attenuated by nifedipine, NFA, 18β-GA, TEA or in Ca2+-free medium. Nifedipine, NFA, 18β-GA or Ca2+-free medium not only dampened vasomotion, but also kept BA in relaxation state. In contrasts, TEA kept BA in contraction state. These results suggest that the pressure-induced vasomotion of the isolated BA results from an interaction between Ca2+-activated Cl- channels(CaCCs) currents and KCa currents. We hypothesize that vasomotion of BA depends on the depolarizing of the vascular smooth muscle cells(VSMCs) to activate CaCCs. Depolarization in turn activates voltage-dependent Ca2+ channels, synchronizing contractions of adjacent cells through influx of extracellular calcium and the flow of calcium through gap junctions. Subsequent calcium-induced calcium release from ryanodine-sensitive stores activates KCa channe 展开更多
关键词 动脉血管 基底动脉 钙通道 氯通道 激活 大鼠 血管平滑肌细胞 工作压力
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神经病理性疼痛大鼠不同大小DRG神经元兴奋性分型的研究 预览
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作者 谭朝阳 周颖 +8 位作者 屈祖卫 张萌 陈沁怡 田俊杰 许珍珍 邓诗瑜 李丽 马克涛 司军强 《中国现代医学杂志》 CAS 2019年第11期1-7,共7页
目的研究神经病理性疼痛状态下,不同大小背根神经节(DRG)神经元兴奋性分型的特点,探讨不同大小DRG神经元兴奋性分型与神经病理性疼痛的关系。方法将SD大鼠随机分成正常组(Control组)、假手术组(Sham组)和坐骨神经分支选择性损伤模型组(... 目的研究神经病理性疼痛状态下,不同大小背根神经节(DRG)神经元兴奋性分型的特点,探讨不同大小DRG神经元兴奋性分型与神经病理性疼痛的关系。方法将SD大鼠随机分成正常组(Control组)、假手术组(Sham组)和坐骨神经分支选择性损伤模型组(SNI组)。运用热和冷刺激实验观察大鼠疼痛行为学变化;运用膜片钳技术记录不同大小DRG神经元兴奋性分型变化。结果与Control组比较,SNI组大鼠热缩足潜伏期无改变,但冷刺激抬足时间增加(P<0.05);与Control组比较,SNI组大鼠DRG神经元的1型和2型细胞构成比增加,3型细胞构成比减少(P <0.05);在Control组内,与大细胞比较,中、小细胞3型细胞的构成比较低,而1型和2型细胞的构成比较高(P <0.05);在SNI组内,与大细胞比较,中细胞3型细胞构成比较低,1型和2型细胞构成比较高(P <0.05),而小细胞与大细胞的构成比差异无统计学意义(P>0.05)。结论不同大小的DRG神经元兴奋性分型的改变是产生神经病理性疼痛的机制之一。 展开更多
关键词 神经痛 神经节 膜片钳术 细胞大小 兴奋性分型
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硫化氢对自发性高血压大鼠心肌重构的作用及机制研究 预览
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作者 张晓景 王童 +5 位作者 孔繁秀 刘宇航 陈宇鑫 张亮 李玲 司军强 《中国现代医学杂志》 CAS 2019年第13期1-6,共6页
目的探讨硫化氢H2S对自发性高血压大鼠(SHR)间隙连接蛋白40(Cx40)、43(Cx43)表达的调控作用,以及与心肌重构的关系。方法取8周龄雄性SHR16只,随机分为SHR对照组(n=8)、SHR+硫氢化钠NaHS组(n=8);取同周龄的正常Wistar京都(WKY)雄性大鼠8... 目的探讨硫化氢H2S对自发性高血压大鼠(SHR)间隙连接蛋白40(Cx40)、43(Cx43)表达的调控作用,以及与心肌重构的关系。方法取8周龄雄性SHR16只,随机分为SHR对照组(n=8)、SHR+硫氢化钠NaHS组(n=8);取同周龄的正常Wistar京都(WKY)雄性大鼠8只,设为WKY组。SHR+NaHS组腹腔注射NaHS56μmol/(kg·d),SHR对照组和WKY组每日腹腔注射等量生理盐水,持续8周。用分光光度计检测各组大鼠外周血和心肌组织中H2S含量;通过HE染色及Masson三色染色观察H2S对心肌的病理学改变;通过免疫组织化学检测3组大鼠心脏组织中Cx40、Cx43表达位置的变化;运用Westernblotting检测3组大鼠心脏组织中Cx40、Cx43及α-平滑肌肌动蛋白(α-SMA)、骨桥蛋白(OPN)表达量的变化。结果与WKY组大鼠比较,SHR组大鼠外周血及心肌组织中H2S含量减少(P<0.05),NaHS干预后SHR外周血及心肌组织中H2S含量增加(P<0.05);与WKY组大鼠比较,SHR组大鼠心肌纤维结构排列较为紊乱,NaHS干预后SHR心肌纤维排列较为整齐;SHR组大鼠心肌中的Cx40、Cx43表达增加且分布紊乱,SHR+NaHS组大鼠心肌中的Cx40、Cx43分布较规整;且SHR组大鼠心肌中Cx40、Cx43、α-SMA、OPN表达升高(P<0.05),SHR+NaHS组大鼠心肌中Cx40、Cx43、α-SMA、OPN表达降低(P<0.05)。结论H2S可能通过调控Cx40、Cx43的表达来改善SHR心肌重构。 展开更多
关键词 心肌重构 大鼠 近交SHR 硫化氢 连接蛋白类
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细胞外信号蛋白调节激酶5在异氟醚后处理减轻大鼠海马脑片缺氧无糖损伤中的作用 预览
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作者 崔迪 王胜 +4 位作者 葛明月 王芹 殷姜文 代志刚 司军强 《中国现代医学杂志》 北大核心 2017年第21期1-6,共6页
目的 探讨异氟醚后处理大鼠海马脑片缺氧无糖(OGD)损伤神经保护机制中细胞外信号蛋白调节激酶5(ERK5)的作用。方法 使用雄性SD大鼠,将其麻醉后取出海马组织,制成离体脑片,然后随机进行正常对照组(Con组)、损伤组(OGD组)和药物... 目的 探讨异氟醚后处理大鼠海马脑片缺氧无糖(OGD)损伤神经保护机制中细胞外信号蛋白调节激酶5(ERK5)的作用。方法 使用雄性SD大鼠,将其麻醉后取出海马组织,制成离体脑片,然后随机进行正常对照组(Con组)、损伤组(OGD组)和药物处理组的处理。采用花粉活力染色(TTC)法评价各组脑片损伤程度,采用碘化丙啶(PI)染色法检测海马CA1区神经细胞凋亡程度,采用实时荧光定量聚合酶链反应(q RT-PCR)测定细胞外调节蛋白激酶信使核糖核酸(ERK5 m RNA)的表达,采用Western blot法检测ERK5蛋白的表达与磷酸化水平。结果 与Con组比较,OGD组脑片损伤程度升高、海马CA1区凋亡细胞增多,海马组织内ERK5 m RNA和p-ERK5表达升高(P〈0.05);XMD组阻断ERK5 m RNA和p-ERK5表达,脑片损伤程度升高、海马CA1区凋亡细胞增多(P〈0.05)。与OGD、1.5%ISPOC和4.5%ISPOC组比较,3.0%ISPOC组脑片损伤程度减轻、海马CA1区凋亡细胞减少,海马组织内ER K5 m R N A和p-ER K5表达升高(P〈0.05)。与3.0%ISPO C组比较,XMD阻断ISPOC海马组织内ERK5 m RNA和p-ERK5表达,脑片损伤程度升高、海马CA1区凋亡细胞增多(P〈0.05)。结论 一定浓度的异氟醚后处理大鼠海马脑片对抗O GD损伤的保护性机制可能与上调ER K5 m R N A和ERK5磷酸化水平有关。 展开更多
关键词 麻醉药 异氟醚后处理 细胞外信号调节MAP激酶类 脑缺血 海马
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PKCε Mediates Substance P Inhibition of GABA_A Receptors-Mediated Current in Rat Dorsal Root Ganglion 预览
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作者 李丽 赵磊 +4 位作者 王洋 马克涛 石文艳 王英姿 司军强 《华中科技大学学报:医学英德文版》 SCIE CAS 2015年第1期1-9,共9页
The mechanism underlying the modulatory effect of substance P(SP) on GABA-activated response in rat dorsal root ganglion(DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clamp te... The mechanism underlying the modulatory effect of substance P(SP) on GABA-activated response in rat dorsal root ganglion(DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clamp technique was used to record GABA-activated current and sharp electrode intracellular recording technique was used to record GABA-induced membrane depolarization. Application of GABA(1–1000 μmol/L) induced an inward current in a concentration-dependent manner in 114 out of 127 DRG neurons(89.8 %) examined with whole-cell patch-clamp recordings. Bath application of GABA(1–1000 μmol/L) evoked a depolarizing response in 236 out of 257(91.8%) DRG neurons examined with intracellular recordings. Application of SP(0.001–1 μmol/L) suppressed the GABA-activated inward current and membrane depolarization. The inhibitory effects were concentration-dependent and could be blocked by the selective neurokinin 1(NK1) receptors antagonist spantide but not by L659187 and SR142801(1 μmol/L, n=7), selective antagonists of NK2 and NK3. The inhibitory effect of SP was significantly reduced by the calcium chelator BAPTA-AM, phospholipase C(PLC) inhibitor U73122, and PKC inhibitor chelerythrine, respectively. The PKA inhibitor H-89 did not affect the SP effect. Remarkably, the inhibitory effect of SP on GABA-activated current was nearly completely removed by a selective PKCε inhibitor epilon-V1-2 but not by safingol and LY333531, selective inhibitors of PKCα and PKCβ. Our results suggest that NK1 receptor mediates SP-induced inhibition of GABA-activated current and membrane depolarization by activating intracellular PLC-Ca2+-PKCε cascade. SP might regulate the excitability of peripheral nociceptors through inhibition of the "pre-synaptic inhibition" evoked by GABA, which may explain its role in pain and neurogenic inflammation. 展开更多
关键词 背根神经节 激活电流 受体介导 PKC P物质 全细胞膜片钳技术 大鼠 DRG神经元
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Differential Expression of Alpha-Adrenoceptor Subtypes in Rat Dorsal Root Ganglion after Chronic Constriction Injury 预览
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作者 成洪聚 马克涛 +3 位作者 李丽 赵磊 王洋 司军强 《华中科技大学学报:医学英德文版》 SCIE CAS 2014年第3期322-329,共8页
mRNAs of alpha-adrenoceptor(α-AR) subtypes are found in neurons in dorsal root ganglion(DRG) and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was studied in L5 DRG o... mRNAs of alpha-adrenoceptor(α-AR) subtypes are found in neurons in dorsal root ganglion(DRG) and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was studied in L5 DRG of normal rats and rats with chronic constriction injury of sciatic nerve(CCI). Using immunofluorescence technique, it was found that α1A-, α1B-, and α2A-AR proteins were expressed in large, medium, and small size neurons in normal DRG, and significantly increased in all size neurons 14 days after CCI. α1D- and α2C-AR was also expressed in all size neurons in normal DRG. However, α1D-AR was significantly increased and α2C-AR was decreased in small size neurons 14 days post CCI. α2B-AR neurons were not detectable in normal and CCI DRG. Co-expression of α1A- and α2A-AR in the same neuron was observed in normal DRG and increased post CCI. Collectively, these results indicated that there is distinct distribution of α-AR subtypes in DRG neurons, and the distribution and levels of expression of α-AR subtypes change differently after CCI. The up-regulation of α-AR subtypes in DRG neurons may play an important role in the process of generating and transmitting neuropathic pain. 展开更多
关键词 肾上腺素能受体 周围神经损伤 背根神经节 受体亚型 大鼠 DRG神经元 慢性 差分
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Modulatory effect of substance P on GABA-activated currents from rat dorsal root ganglion 被引量:3
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作者 Jun-qiangSI Zhi-qinZHANG +3 位作者 Chun-xiaLI Li-fengWANG Yun-leiYANG Zhi-wangLI 《中国药理学报:英文版》 SCIE CAS CSCD 北大核心 2004年第5期 623-629,共7页
To explore the modulatory effect of substance P (SP) on GABA-activated current of dorsal root ganglion (DRG) neurons in rat. METHODS: The whole-cell patch-clamp technique was used to record SP- and GABA activated curr... To explore the modulatory effect of substance P (SP) on GABA-activated current of dorsal root ganglion (DRG) neurons in rat. METHODS: The whole-cell patch-clamp technique was used to record SP- and GABA activated currents in neurons freshly dissociated from rat DRG neurons. Drugs were applied by rapid solution exchange. RESULTS: Application of SP (28/41, 68.5 %) and GABA (36/41, 88.2 %) could induce concentrationdependent inward current in some cells. SP-(10 ~unol/L) and GABA (100 ~unol/L)-activated inward currents were (244+83) pA (n=9) and (1.8+0.5) nA (n=13), respectively. The majority of GABA-activated current had obvious three processes, the peak value (lp), the steady state (lss) and the desensitization (ld). The desensitization of GABA-activated current was a biphasic process, including fast and slow desensitization. However, pre-application of SP (0.001-1 ~unol/L) could inhibit the GABA-activated inward current which was identified to be GABAA receptormediated current. The inhibitory effects were concentration-dependent. The inhibitory effect of SP on the peakvalue of GABA-activated current was more than the steady state of GABA-activated current. The inhibition ofGABA-activated current by SP (0.1 prnol/L) was related to the time after application of SP, the inhibition of GABAactivated currents by SP reached the peak at about 4 min (49.8 %+7.2 %, n=7, P<0.01) and took about 12 min to get a full recovery. The inhibition of GABA-activated currents by SP was almost completely removed after blockade of PKC by H-7 with the re-patch clamp. CONCLUSION: Pre-application of SP exerts a more strong inhibitory effect on the peak value of GABA-activated current than the steady state of GABA-activated current. 展开更多
关键词 脊神经根 神经兴奋 动物实验 P物质 膜片钳技术
80株鲍曼不动杆菌的耐药性分析 预览 被引量:2
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作者 何文军 罗强 +1 位作者 易四维 徐宁 《中国实用医药》 2007年第13期 43-45,共3页
目的 了解鲍曼不动杆菌的耐药性,为临床合理选用抗菌药物提供参考.方法 从临床标本分离出的80株鲍曼不动杆菌的药敏试验结果进行统计分析.结果 鲍曼不动杆菌对美洛培南无耐药菌株产生;对亚胺培南和哌拉西林/他唑巴坦的耐药率分别为8.8%... 目的 了解鲍曼不动杆菌的耐药性,为临床合理选用抗菌药物提供参考.方法 从临床标本分离出的80株鲍曼不动杆菌的药敏试验结果进行统计分析.结果 鲍曼不动杆菌对美洛培南无耐药菌株产生;对亚胺培南和哌拉西林/他唑巴坦的耐药率分别为8.8%、11.3%;对其他抗菌药物均有一定程度的耐药,多重耐药率高.结论 加强对鲍曼不动杆菌的耐药性监测,规范抗菌药物的应用,对保持敏感抗菌药物的抗菌活性具有重要意义. 展开更多
关键词 鲍曼不动杆菌 抗菌药物 耐药性
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