Effects of WIN 55,212-2 on IK Current in Cultured Trigeminal Ganglion Neurons of Rat

Summary To investigate the effects of WIN 55, 212-2 onI _K in cultured rat trigeminal ganglion (TG) neurons, whole-cell patch clamp techniques were used to record theI _K before and after WIN 55,212-2 perfusion at different concentrations. 30 μmol/L WIN 55,212-2 markedly (35.7%±7.3%P<0.01,n=8) inhibitedI _K currents, and the currents were partially recovered after washing. 30 μmol/L WIN 55, 212-2 also induced a significant depolarizing shift in conductance-voltage parameters (control: V_0.5=10.43±4.25 mV, k=16.27±3.86; WIN 55,212-2: V_0.5=24.71±3.91 mV, k=16.69±2.75;n=8,P<0.01 for V_0.5). 0.01 μmol/L WIN 55,212-2 slightly (27.0%±7.9%,P<0.05,n=7) increasedI _K currents, but had no significant change in conductance-voltage parameters (control: V_0.5=10.74±5.27 mV, k=17. 33±2.96; WIN 55, 212-2: V_0.5=11.06±2.05 mV, k=19.69±6.60;n=7,P>0.05 for V_0.5 and k). These results suggested that WIN 55, 212-2 has dual action, which might be through different receptors. MING Zhangyin, male, born in 1968, Doctorial Candidate This project was supported by a grant from National Natural Sciences Foundation of China (No. 30271500).     

多非力特对豚鼠心室肌细胞的电生理作用

目的探讨多非力特对心肌细胞电生理作用的特点。方法（1）应用膜片钳全细胞技术记录单个豚鼠心室肌细胞膜延迟整流性钾流（IK）,观察不同浓度多非力特对IK的影响,并探讨IK的快速激活成分（IKr）的电生理特性。（2）标准微电极技术记录豚鼠右室乳头肌细胞动作电位,观察不同浓度多非力特灌流前后动作电位各参数的变化。结果（1）多非力特0.01～0.5μmol/L呈浓度依赖性抑制IK的时间依赖性外向钾流（IK.time）和尾电流（IK.tail）,但浓度再增大至1μmol/L后,此抑制作用趋于最大。对-20mV钳制电压下IK.time、IK.tail的抑制程度比＋50mV时更明显（P〈0.05或0.01）。（2）通道动力学分析发现,多非力特1μmol/L使IK.tail快失活过程消失,慢失活时间常数无明显变化（P〉0.05）。（3）多非力特敏感的电流成分在-40mV开始呈电压依赖性快速激活,峰值电压为0,开始出现内向整流,其IK.tail也呈电压依赖,无内向整流,但膜电位达＋20mV后达饱和,以上均符合IKr的特点。（4）多非力特对动作电位的影响：0.5和1.0μmol/L浓度下可使动作电位时程（APD30、APD90）均明显延长（P〈0.05或〈0.01）,且呈反转频率依赖性。结论多非力特对IK的作用符合特异性IKr阻滞剂的特点,并呈反转频率依赖性延长APD,是其抗心律失常作用的电生理基础。     

Effect of Interleukin-1βon IA and IK Currents in Cultured Murine Trigeminal Ganglion Neurons

To investigate the effect of interleukin-1β (IL-1β) on IA and IK currents in cultured murine trigeminal ganglion (TG) neurons, whole-cell patch clamp technique was used to record the IA and IKcurrents before and after 20 ng/mL IL-1β perfusion. Our results showed that 20 ng/mL IL-1β inhibited IA currents (18.3±10.7)% (n=6, P＜0.05). IL-1β at 20 ng/mL had no effect on G-V curve of IA but moved the H-infinity curve V0.5 from -36.6 ± 6.1 mV to-42.4 ±5.2 m V (n=5, P＜0.01). However, 20 ng/mL IL-1β had effect on neither the amplitude nor the G-V curve of IK. IL-1β was found to selectively inhibit IA current in TG neurons and the effect may contribute to hyperalgesia under various inflammatory conditions.     

Inhibition of 5-HT3 receptors-activated currents by cannabinoids in rat trigeminal ganglion neurons

This study investigated the modulatory effect of synthetic cannabinoids WIN55,212-2 on 5-HT3 receptor-activated currents (I5-HT3) in cultured rat trigeminal ganglion (TG) neurons using whole-cell patch clamp technique. The results showed that: (1) The majority of examined neurons (78.70%) were sensitive to 5-HT (3-300 μmol/L). 5-HT induced inward currents in a concentration-dependent manner and the currents were blocked by ICS 205-930 (1 μmol/L), a selective antagonist of the 5-HT3 receptor; (2) Pre-application of WIN55,212-2 (0.01-1 μmol/L) significantly inhibited I5-HT3 reversibly in concentration-dependent and voltage-independent manners. The concentra-tion-response curve of 5-HT3 receptor was shifted downward by WIN55,212-2 without any change of the threshold value. The EC50 values of two curves were very close (17.5±4.5) mmol/L vs. (15.2±4.5) mmol/L and WIN55,212-2 decreased the maximal amplitude of I5-HT3 by (48.65±4.15)%; (3) Neither AM281, a selective CB1 receptor antagonist, nor AM630, a selective CB2 receptor antagonist re-versed the inhibition of I5-HT3 by WIN55,212-2; (4) When WIN55,212-2 was given from 15 to 120 s before 5-HT application, inhibitory effect was gradually increased and the maximal inhibition took place at 90 s, and the inhibition remained at the same level after 90 s. We are led to concluded that-WIN55,212-2 inhibited I5-HT3 significantly and neither CB1 receptor antagonist nor CB2 receptor an-tagonist could reverse the inhibition of I5-HT3 by WIN55,212-2. Moreover, WIN55,212-2 is not an open channel blocker (OCB) of 5-HT3 receptor. WIN55,212-2 significantly inhibited 5-HT-activated currents in a non-competitive manner. The inhibition of I5-HT3 by WIN55,212-2 is probably new one of peripheral analgesic mechanisms of WIN55,212-2, but the mechanism by which WIN55,212-2 in-hibits I5-HT3 warrants further investigation.     

氯胺酮对大鼠海马神经元瞬间外向钾电流的抑制作用

目的：观察氯胺酮对大鼠海马锥体神经元瞬间外向钾电流（I_A）的影响。方法：酶消化法急性分离Wistar大鼠海马锥体神经元,采用全细胞膜片钳技术测定I_A。加用不同浓度（10、30、100、300和1 000 μmol/L）氯胺酮后,计算I_A抑制率,建立氯胺酮的浓度-效应曲线,选择30 μmol/L氯胺酮作I_A稳态激活（及失活）曲线。结果：10 μmol/L的氯胺酮对I_A的电流幅度无影响;30、100、300和1 000 μmol/L的氯胺酮对I_A的电流幅度抑制率分别为（11±2）%、（22±3）%、（45±5）%和(53±5)%。IC₅₀为（130±24）μmol/L, Hill系数为1.19±0.56。30 μmol/L氯胺酮使激活曲线的半数最大激活膜电位（V _{1/ 2}）从（-8.70±0.13） mV 移动到 （-11.2±2.10） mV (n=8, P<0.05), K从（15.0±4.6） mV移动到（17.6±5.7）mV (n=8, P>0.05);失活曲线的V _1/2从（-75.53±7.98） mV移动到（-91.94±11.85） mV（n=8, P < 0.05）,K从（10.5±2.2） mV移动到（8.0±1.2） mV（n=8, P>0.05）。30 μmol/L氯胺酮使I_A的激活和失活曲线均向超极化方向明显移动。 结论： 临床浓度的氯胺酮能够同时加速I_A的激活和失活过程,但加速I_A失活的能力大于其加速I_A激活的能力。氯胺酮对中枢神经系统的作用可能与I_A抑制有关。     

大麻素WIN55，212-2对体外培养牛眼小梁细胞cAMP表达的影响

目的通过研究大麻素WIN55，212-2对体外培养牛眼小梁细胞（BTMC）环腺苷酸（cAMP）表达的影响，探讨其增加房水排出而降低眼压的机制。方法对BTMC进行体外原代培养及传代培养，应用免疫组化的方法鉴定细胞。然后用不同浓度的WIN55，212-2作用于BTMC，放射免疫法检测细胞上清液中cAMP的水平。结果WIN55，212-2浓度在0．1-20μmol／L范围内，培养液中cAMP水平随浓度增加而显著性升高，若WIN55，212-2浓度继续增高（40-60btmol／L），cAMP水平不再升高。结论WIN55，212-2可能通过提高BTMC内cAMP水平而起到降低眼压的作用。     

Verapamil modulating arsenic trioxide-induced QT interval rolongation in guinea pig

Objective To investigate therapeutic action of verapamil on QT prolongation induced by arsenic trioxide (As2O3) in guinea pig and to further explore its possible mechanism. Methods Different doses of As2 O3 was infused intravenously to observe the changes of QT interval on the electrocardiogram (ECG) at different times in guinea pig. Patchclamp technique and laser scanning confocal microscopy were utilized to study the action of As2 O3 on action potential duration (APD),L-type calcium current (ICa-L ), rapid delayed rectifier potassium current (IKr) and intracellular calcium concentration ([Ca2+]i) of guinea pig myocytes. At the same time, verapamil was applied preliminarily to evaluate effects of verapamil on changes of the above index induced by As2O3. Results Intravenous administration of As2 O3 at the dose of 1.6mg/kg and 0.8mg/kg prolonged QT interval on ECG obviously in guinea pig hearts dose dependently and time dependently. QTc (corrected QT interval) was progressively prolonged in the 2-hour period of intravenous infusion of 1.6mg/kg As2O3 from (328.5 ± 30.9)ms of control to (388.4 ± 31.3)ms at 2h following As2O3 ( P ＜ 0.01) .When verapamil was pretreated for 5min, then 1.6mg/kg As2 O3 was added, the results showed that QTe was shorter in verapamil-treatment group (357.3±21.4)ms than that in As2O3 group (388.4±31.3)ms (P＜0.05) at 2h. Confocal experiments showed that in normal Tyrode solution, As2 O3 (1μmol/L and 10μmol/L) had no obvious effects on resting [Ca2+]i (P＞0.05) in guinea pig cardiomyocytes, however, 10μmol/L As2 O3 could markedly enhance [Ca2+]i increase induced by KCl 60mmol/L and the peak value increased from 903.4±369.4 to 1674.6±563.2 ( P＜0.05). The action of elevating [Ca2+]icould be blocked by 10μmol/L verapamil incompletely. The patch-clamp studies indicated that As2 O3 at concentration of 10μmol/L prolonged APD50 from (263.6 -±75.2)ms to (523.9±47.8)ms (P＜0.01) and APD90 from (277.5 ± 77.5)ms to (536.3 ±49.6)ms (P ＜0.01) ,and increased ICa-L from (- 6.0±1.5)pA/pF to (-8.7±2.0)pA/pF (P＜0.01) at 0mV and also reduced IKr from (6.7±1.8)pA/pF to (4.5±1.8)pA/pF (P＜0.05). However, 10μmol/L verapamil could modulate prolonging APD50 from (523.9 ± 47.8) ms to (340.4±83.8) ms ( P＜0.01) and APD90 from (536.3 ±49.6)ms to (348.9 ± 85.5)ms (P＜0.01) and correct increasing Ica-L induced by 10μmol/L As2O3 from (-8.7±2.0)pA/pF to ( - 6.6±1.4)pA/pF (P＜0.05) at 0mV. Conclusion As2O3 could induce prolongation of the QT interval on the ECG in guinea pig hearts and the ionic mechanism is associated with increasing Ica-L and inhibiting IKr/HERG. Verapamil may be useful in normalizing QT prolongation during As2 O3 therapy by decreasing Ica-L and [Ca2+]iof ventricular myocytes in guinea pig.     

大麻素受体激动剂预处理对大鼠局灶性脑缺血再灌注损伤的保护作用

目的 探讨大麻素(CB)受体激动剂WIN 55,212-2预处理对大鼠局灶性脑缺血的保护作用.方法采用大鼠大脑中动脉栓塞(MCAO)致局灶性脑缺血模型,将50只雄性SD大鼠随机分为5组:对照组(Con组),行线栓法敛大脑中动脉栓塞(MCAO)前24 h腹腔注射生理盐水(NS)0.3ml;WIN 55,212-2预处理组(WIN1～3),分别于MCAO前24 h腹腔注射WIN 55,212-2 0.3,1和3rag/kg;DMSO组,于MCAO前24 h腹腔注射二甲基亚砜(DMSO,WIN 55,212-2的溶剂)0.3 ml.观察MCAO120 min再灌注24,48和72 h后神经功能评分(NFS)和再灌注72 h脑梗死容积百分比.结果 WIN 55,212-2预处理组大鼠NFS明显高于DMSO组和Con组(P<0.05),脑梗死容积百分比明显小于DMSO组和Con组(P<0.05).WIN2组和WIN3组的脑梗死容积百分比明显小于WIN1组,NFS明显高于WIN1组(P<0.05),WIN2组和WIN3组的NFS和脑梗死容积百分比差异无统计学意义(P=0.928),但3 mg/kg WIN 55,212-2可引起大鼠明显嗜睡和倦怠.结论 CB受体激动剂WIN 55,212-2预处理能诱导脑缺血耐受,对大鼠局灶性脑缺血再灌注损伤具有保护作用,并具有一定的剂量相火性.     

Obestatin抑制胰岛β细胞株INS-1细胞L型钙通道电流

目的 观察obestatin对胰岛β细胞株INS-1细胞L型钙通道电流的影响.方法 应用穿孔全细胞膜片钳技术研究obestatin对培养的胰岛β细胞株INS-1细胞膜L型电压敏感钙通道电流（L-type calcium ion channel current,ICa.L）的影响.结果胰岛β细胞株INS-1细胞钳制膜电位为-20 mV时,分别给予胰岛β细胞株INS-1细胞0nM（对照组）和1、10、100nM obestatin灌流5min后,ICa,L峰值呈浓度依赖性降低,各组为给药前的（97±8）％、（91±7）％、（60±9）％和（45±6）％.其中,10nM和100nM obestatin灌流5min后ICa,L峰值与对照组相比显著减小（P＜0.0I,n=5）.胰岛β细胞株INS-1细胞钳制膜电位分别为-30、-20、-10和0mY时,给予10nM obestatin之前,其ICa,L峰值分别为（-5.48±0.69）、（-5.70±0.68）、（-5.58±0.61）和（-3.90±0.44） pA/pF,灌流10nM obestatin 5 min后,ICa,L峰值分别下降至（-3.11 ±0.68）、（-3.60±0.99）、（-3.26±1.03）和（-2.28±0.71）pA/pF,均明显低于obestatin给药前（P＜0.01,n=5）.结论 obestatin可抑制胰岛β细胞株INS-1细胞L型钙通道电流.     

叶酸治疗短暂性脑缺血发作伴高半胱氨酸血症疗效观察

目的 观察叶酸治疗对合并高半胱氨酸血症(Hcy)的短暂性脑缺血发作(TIA)患者血浆高半胱氨酸(HCA)水平及预后的影响.方法 将129例合并Hcy的TIA患者应用随机数字法随机分为2组,对照组(n=64)只给予常规治疗,观察组(n=65)在常规治疗的基础上给予叶酸治疗,对比分析2组患者治疗前及治疗后3个月HCA水平的变化及其1年内完全缓解率及完全性脑卒中发生率.结果 初发TIA患者Hcy的发生率高达41.4%.治疗3个月后,观察组血浆HCA显著降低[(14.27±6.13)μmol/L,(24.99±6.87)μmol/L,t=2.799,P＜0.01],而对照组血浆HCA无明显变化[(24.68±6.89)μmol/L,(25.68±7.11)μmol/L,t=2.735,P＜0.01].1年后,观察组完全性脑卒中发生率显著低于对照组(9.8%,25.0%,x2=4.849,P＜0.05),TIA症状完全缓解率高于对照组(73.8%,50.0%,x2=7.253,P＜0.01),且无明显不良反应.结论 叶酸治疗可有效降低TIA患者的血浆HCA水平,改善患者的预后.

Abstract：

Objective To observe the therapeutic effect of folic acid on transient ischemic attack(TIA)patients with homocysteinaemia (Hcy ). Methods 129 patients of primary TIA with Hcy were divided into two groups randomly. The observation group ( n = 65 )was administered with conventional therapy and folic acid, and the control group ( n=64 ) was only given conventional therapy. The variances of the plasma HCA level three months later were compared, and remission rate of TIA and complete stroke incidence one year later were analyzed between two groups. Results The Hcy incidence rate of TIA patients was up to 41.4%. Three months later, the plasma HCA level of observation group was lower than control group( ( 14.27 ± 6. 13 ) μmol/L vs (24.99 ± 6.87 )μmol/L, t=2.799, P＜0. 01 ) ,and much lower than that of the control group post-treatment ( ( 14. 27 ±6. 13)μmol/L vs (24.68 ± 6.89) μmol/L, t = 2.735, P ＜ 0.01 ). One year later, the complete stroke incidence of TIA in observation group was lower than that of the control group(9.8% vs 25.0%, P＜0.05 ) ,and complete remission rate was higher than the latter(73.8% vs 50.0%, P ＜ 0. 01 ). Conclusion Folic acid can decrease the plasma HCA level of TIA patients with Hcy efficiently,and improve the prognosis of such patients.     

体质指数与血管重建的冠心病心力衰竭患者预后的关系

目的 了解体质指数(BMI)对接受血管重建治疗的冠心病心力衰竭(心衰)患者预后的影响.方法 药物洗脱支架对血运重建策略影响研究(单中心回顾性注册研究)入选2004年7月1日至2005年9月30日在北京安贞医院接受血管重建治疗的3632例患者,2006年9月1日对患者进行随访.本研究入选其中有体质指数(BMI)资料的慢性心衰患者1010例.将患者按BMI分为3组:BMI<24(正常组),BMI 24～27.9(超重组)和BMI≥28(肥胖组),比较不同组别之间的临床和预后情况.不良心脑血管事件(MACCE)包括全因死亡、非致死性心肌梗死、非致死性卒中和再次血管重建.结果 正常组295例,超重组495例,肥胖组220例.随访中位时间为542 d,超重和肥胖患者较年轻[(59.3±10.1)岁、(58.6±10.3)岁比(62.6.4±9.9)岁,P<0.01],高血压病史(61.2,66.8%比52.5%,P=0.017)和稳定型心绞痛(21.2%,23.7%比17.0%,P=0.005)比例高,甘油三酯[(1.90±1.05)mmol/L,(2.10±1.12)mmol/L比(1.48±0.92)mmol/L,P<0.01)]、空腹血糖[(6.07±2.09)mmol/L,(5.96±1.53)mmol/L比(5.67±1.92)mmol/L,P=0.021]和肌酐[(84.9±21.7)μmol/L,(90.2±30.9)μmol/L比(82.2±25.8)μmol/L]水平高(P均<0.05).与正常体重的患者相比,在调整了其他因素后,超重组的全因死亡风险(HR 0.769,95%CI 0.442～1.338)和MACCE(HR 0.998,95% CI 0.754～1.322)并未增加,而肥胖组全因死亡(HR 0.285,95%CI 0.104～0.777)和MACCE(HR 0.596,95% CI 0.401～0.885)风险降低.BMI对心性死亡无显著影响.结论 在进行血管重建的冠心病心衰患者中,尽管超重和肥胖者相对体重正常者有更多的危险因素,但是接受血管重建治疗后的预后不比体重正常者差.     

胆囊收缩素对豚鼠结肠平滑肌及其细胞膜L-型钙电流和膜电位的影响

目的 探讨胆囊收缩素8肽(CCK-8S)对豚鼠近端结肠平滑肌及细胞膜L-型钙电流和膜电位的电生理作用及其机制.方法 (1)采用RM6240多道生理信号采集处理系统记录CCK-8S对豚鼠近端结肠平滑肌肌条收缩的影响;(2)检测Fluo-2/AM标记的近端结肠平滑肌细胞膜内钙离子浓度([Ca2+]i)变化;(3)用EPC-10膜片钳放大器测量全细胞模式下CCK-8S对近端结肠平滑肌细胞膜静息电位(RP)、动作电位(AP)和L-型钙通道电流(ICa-L)的影响.结果 CCK-8S(10-1mol/L)作用后:(1)豚鼠近端结肠平滑肌肌条收缩幅值和频率分别为对照组的(149±12)%和(132±13)%(均P＜0.05);(2)近端结肠平滑肌细胞膜的[Ca2+]i为对照组的(738±24)%(P＜0.05);(3)RP、AP峰值及AP快速复极时间分别为对照组的(52±9)%、(140±4)%和(61±13)%(均P＜0.05),该效应可被预先加入的CCK1受体拮抗剂地伐西匹和(或)L-型钙通道阻断剂尼非地平阻断(n=8,均P＜0.05),而预先向灌流液中加入CCK2受体拮抗剂CI 988后,CCK-8S对RP、AP峰值及AP快速复极时间的作用仍存在;(4)从-60 mV去极化至+10 mV,ICa-L为对照组的(138±7)%(P＜0.05),但分别可被相应拮抗剂抑制;当向电极内液加入肝素(10-6 mol/L)或向细胞外液加入staurosporine(10-6 mol/L)后,CCK-8S对Ica-L均没有影响(2.9%±2.6%和1.9%±2.3%,均P＞0.05).结论 CCK-8S通过CCK1受体促进近端结肠平滑肌自发性收缩频率和强度;其机制与激活1,4,5-三磷酸肌醇介导的蛋白激酶C途径进而促进L-型钙通道开放有关.

Abstract：

Objective To investigate the effects and mechanism of cholecystokinin octapeptide (CCK-8S) on the contractile activity of smooth muscles,L-type calcium current and membrane potentials of proximal colon myocytes in guinea pig.Methods ( 1 ) Strips of proximal colon were obtained from adult guinea pigs.The contraction of these stripes was measured by a RM6240 multi-channel physiological signal system.(2) Suspension of single smooth muscle cells (SMCs) were obtained from proximal colon and isolated by enzymatic digestion.The effect of CCK-8S on intracellular calcium concentration ( [Ca2+] i) of SMCs was examined by fura-2-1oaded miscrofluorimetric measurement.(3) Resting potential ( RP),action potential (AP) and L-type calcium current (ICa-L ) were recorded by patch-clamp technique.Results ( 1 )The contractile amplitude and frequency of muscle stripes enhanced by CCK-8S ( 10 -7 mol/L) were ( 149 ±12)% and (132 ± 13 )% respectively of those of control group (all P ＜ 0.05 ).They were significantly attenuated by pretreating strips with CCK1 receptor antagonist devazepide ( 10-7 mol/L),L-type calcium channel blocker nifedipine ( 10 -5 mol/L),Ca2+ -ATPase inhibitor TG (thapsigargin) ( 10-5 mol/L) and BA (boric acid) (10-5 mol/L) respectively.(2) [Ca2+]i of SMCs intensified by CCK-8S was (738 ±24)% of that of control group.And it was inhibited by pretreating SMCs with devazepide(all P ＜0.05).(3) After the superfusion of CCK-8S,RP depolarized to (52 ±9)%,the exogenously stimulated peak values of AP rose to (140±4)% and fast repolarization time of AP decreased to (61 ± 13)% (all P ＜0.05).They were significantly inhibited when these cells were pretreated with devazepide and/or nifedipine (n = 8,P ＜0.05 for each group) whereas CI 988 had little effect.(4) The CCK-8S-evoked ICa-L of SMCs at the voltage of + 10 mV was boosted to ( 138 ± 7 )%.Such an effect was suppressed by a pretreatment with nifedipine,devazepide,TG and BA respectively.In the presence of an inhibitor of inositol 1 ,4,5-trisphosphate (IP3 ) receptors,heparin (10-6 mol/L) and an protein kinase C (PKC) inhibitor,saurosporine (10-6 mol/L),CCK-8S did not significantly intensify ICa-L (all P ＞ 0.05 ).Conclusion CCK-8S promotes proximal colon contraction by CCK1 receptors through the activation of IP3-mediated PKC pathway.     

大麻素WIN55,212-2对体外培养牛眼小梁细胞MMP-3、MMP-9及TIMP-1表达的影响

目的研究大麻素WIN55,212-2对体外培养的牛眼小梁细胞MMP-3、MMP-9及TIMP-1表达的影响,从而探讨其降眼压机制。方法采用组织块培养法对牛眼小梁细胞进行原代及传代培养,应用免疫组化方法（NSE,Ⅷ因子相关抗原染色）鉴定细胞,应用透射电镜对细胞进行形态学及生长特性观察;对传3代的小梁细胞分别施加含大麻素WIN55,212-2终浓度为0（对照组）、1、10、20、40μmol/L的培养液,48h后行MMP-3和MMP-9免疫组化SP染色,结果进行计算机图像分析并进行统计学检验。提取上清液用ELASA法检测随浓度的不同TIMP-1量的变化。结果体外培养的牛眼小梁细胞表达MMP-3及MMP-9,大麻素WIN55,212-2可促进MMP-3及MMP-9的表达,并抑制TIMP-1的表达。结论一定剂量的大麻素可以促进牛眼小梁细胞MMP-3及MMP-9的表达（P〈0.05）,并抑制TIMP-1的表达（P〈0.01）,从而达到降低眼压的目的。     

美沙酮维持治疗者中枢神经递质功能的变化

目的 检测美沙酮维持治疗（methadone maintenance treatment,MMT）患者脑神经递质功能及脑功能指数,了解其大脑功能的变化特点.方法 应用脑涨落图仪（EFG）采集58例MMT患者及44例健康成年人的脑电功率信号,分析MMT患者脑神经递质变化的特点及各神经递质与脑功能状态的相关性.结果 （1）与对照组相比,MMT组的神经递质实测功率普遍降低,其中γ-氨基丁酸（GABA）[（17.73±3.54）μV2 vs.（121.48±44.64） μV2,P＜0.01）、谷氨酸（Glu）[（42.18±12.84） μV2 vs.（105.31±34.95） μV2,P＜0.05]均差异有统计学意义.（2） MMT组的GABA[（17.10±51.72）μV2 vs.（78.67±10.93） μV2,P＜0.001]、Glu[（30.48±21.61） μV2 vs.（69.23±42.26） μV2,P＜0.001]相对功率降低,差异有统计学意义;而五羟色胺（5-HT）[（297.18±31.54）μV2 vs.（280.18±31.54） μV2,P＜0.01]、乙酰胆碱（Ach）[（235.08±37.72） μV2 vs.（217.23±40.60） μV2,P＜0.05]、去甲肾上腺素（NE）[（164.11±33.05） μV2 vs （146.39±30.80） μV2,P＜0.01]、多巴胺（DA）[（98.87±22.48） μV2 vs.（91.49±21.04） μV2,P＜0.05]则升高,均差异有统计学意义;（3） MMT组的全脑总功率低于对照组[（1012.01±195.09） μV2 vs.（1775.94±458.99） μV2,P＜0.01],差异有统计学意义;兴奋抑制指数（2.19±1.46 vs.0.99±0.47,P＜0.001）及相对熵[（89.45±9.71）％ vs.（75.48±9.97）％,P＜0.01]高于对照组,均差异有统计学意义;（4）MMT组各种神经递质功率与全脑总功率均具有正相关,均差异有统计学意义（P＜0.001）,Glu与兴奋抑制指数（r=0.264,P＜0.05）、NE与血管舒缩指数则有正相关,均差异有统计学意义（r=0.269,P＜0.05）,5-HT（r=-0.276,P＜0.05）和DA（r=-0.375,P＜0.01）分别与相对熵存在负相关,均差异有统计学意义.结论 MMT患者的大脑功能显著降低,神经递质的兴奋性和抑制性明显失衡.     

卡通表情自动加工的失匹配负波研究

目的 采用事件相关电位(Event-related potential,ERP)技术探讨人脑对卡通表情自动加工的脑电生理机制.方法 受试者为16名健康人(男7,女9).要求受试者观看卡通表情图片并计数靶刺激绿色图片的呈现次数,而忽略非靶刺激红色图片(包括作为标准刺激的中性表情及作为偏差刺激的情绪性表情).记录32导脑电,对受试者的视觉失匹配负波(Mismatch negativity,MMN)的波幅进行分析.结果 (1)MMN1:波幅分另为(-0.570±0.076)μV(TP7/TP8)、(-0.840±0.119)μV(A1/A2)、(-1.199±0.105)μV(T5/T6)、(-1.184±0.102)μV(O1/O2).波幅的电极位置主效应显著(F(3,45)=8.340,P＜0.05).方向与半球存在交互效应显著(F(1,15)=11.977,P＜0.05).(2)MMN2:波幅分别为(-0.469±0.126)μV(TP7/TP8)、(-1.014±0.255)μV(A1/A2)、(-1.071±0.182)μV(T5/T6)、(-0.915±0.178)μV(O1/O2).方向主效应差异有显著性(F(1,15)=7.232,P＜0.05),且方向与表情交互效应显著(F(1,15)=6.458,P＜0.05),方向与半球交互效应显著(F(1,15)=11.907,P＜0.05).结论 人脑对正、负性卡通表情均可进行自动加工,而负性表情自动加工较正性表情更强;表情信息自动加工存在右侧偏侧化效应.     

海马神经干细胞体外诱导分化过程中延迟整流钾通道的电生理特性

目的 研究大鼠海马源神经干细胞(NSC)分化前及其体外诱导分化的神经元样细胞在不同发育阶段延迟整流钾电流(IDR)的电生理特性.方法 利用无血清培养、单细胞克隆技术,体外培养SD大鼠海马组织源NSC.应用膜片钳技术全细胞模式记录IDR的电生理特性.结果 IDR的电流密度在NSC分化前和体外分化(DIV)0～6 d分别为45 pA/pF±4 pA/pF和56 pA/pF±10 pA/pF(+50 mV,标本数=9),而在DIV＞6 d的发育过程中保持稳定;IDR的半数最大激活膜电位(V1/2)在NSC分化前和DIV 0～6 d分别为9 mV±3 mV和12 mV±3 mV(标本数=9,P＜0.05),激活曲线右移,斜率参数K值无明显改变,但IDR激活特性在DIV＞6 d的发育过程中无明显改变;IDR的失活特性NSC分化前后及不同发育阶段的神经元样细胞中均无改变.结论 NSC分化/发育过程中,IDR的特性改变均发生在DIV 0～6 d,提示IDR通道在神经发育过程中发挥作用,且发育初始阶段对于细胞功能的成熟尤为重要.     

Effects of Chinese herbs on multiple ion channels in isolated ventricular myocytes

Background Shensong Yangxin (SSYX) is one of the compound recipe of Chinese materia medica. This study was conducted to investigate the effects of SSYX on sodium current (I(Na)), L-type calcium current (I(Ca,L)), transient outward potassium current (I(to)), delayed rectifier current (I(K)), and inward rectifier potassium currents (I(K1)) in isolated ventricular myocytes. Methods Whole cell patch-clamp technique was used to study ion channel currents in enzymatically isolated guinea pig or rat ventricular myocytes. Results SSYX decreased peak I(Na) by (44.84±7.65)% from 27.21±5.35 to 14.88±2.75 pA/pF (n=5, P&lt;0.05). The medicine significantly inhibited the I(Ca,L). At concentrations of 0.25, 0.50, and 1.00 g/100 ml, the peak I(Ca,L) was reduced by (19.22±1.10)%, (44.82±6.50)% and (50.69±5.64)%, respectively (n=5, all P&lt;0.05). SSYX lifted the I-V curve of both I(Na) and I(Ca,L) without changing the threshold, peak and reversal potentials. At the concentration of 0.5%, the drug blocked the transient component of I(to) by 50.60% at membrane voltage of 60 mV and negatively shifted the inactive curve and delayed the recovery from channel inactivation. The tail current density of I(K) was decreased by (30.77±1.11)% (n=5, P&lt;0.05) at membrane voltage of 50 mV after exposure to the medicine and the time-dependent activity of I(K) was also inhibited. Similar to the effect on I(K), the SSYX inhibited I(K1) by 33.10% at the test potential of –100 mV with little effect on reversal potential and the rectification property. Conclusions The experiments revealed that SSYX could block multiple ion channels such as I(Na) I(Ca,L), I(k), I(to) and I(K1), which may change the action potential duration and contribute to some of its antiarrhythmic effects.     

Aβ1-42诱导下大鼠海马细胞胰岛素受体表达的变化

目的 检测大鼠海马细胞在Aβ1-42诱导下胰岛素受体表达水平的变化,从而在受体水平探讨中枢胰岛素信号紊乩及阿尔茨海默病发病的分子机制.方法 体外培养原代海马神经细胞,经不同浓度Aβ1-42处理,流式细胞术检测细胞凋亡,实时定量PCR和Western印迹法检测胰岛素受体的表达.结果 原代培养的细胞在第7天时发育成熟,鉴定为海马神经细胞;经Aβ1-42(0～150μmol/L)处理后,30 μmol/L以上处理组的早期凋亡率(32.4%,36.1%,51.0%,53.6%)较对照组(13.4%)呈浓度依赖型增高.PCR结果显示,30 μmol/L(2.56±0.19)和60 μmol/L(3.44±0.23)处理组的胰岛素受体水平较对照组(设为1)明显增高(P＜0.01),100 μmol/L组(0.74±0.15)则较对照组降低(P＜0.01).Western结果与PCR结果趋势相同,30和60 μmol/L处理组的蛋白水平为1.27±0.13,1.82±0.10(P＜0.01),100 μmol/L组为0.82±0.08(P＜0.05).结论 Aβ1-42诱导大鼠海马细胞胰岛素受体的表达发生改变,致使其出现功能缺陷,提示可能为中枢胰岛素抵抗的原因.     

细胞外高K^＋浓度对HCN（1，2）通道电生理特性的影响

目的：研究细胞外高K^＋浓度对异源转染的中国仓鼠卵巢（CHO）细胞上的超极化激活的环核苷酸门控阳离子通道（HCN）1,2电生理特性的影响．方法：将HCN1,HCN2质粒DNA转染至CHO细胞,在两种不同细胞外K^＋浓度条件下用全细胞膜片钳记录细胞膜上HCN1,HCN2通道电流．结果：细胞外高K^＋浓度时,HCN1（pA／pF,112．30±21．50 vs42．33±15．89;-140mV,n=6,P〈0．05）,HCN2（pA／pF,130．57±17．70 vs 26．98±4．62;-140mV,n=8,P〈0．05）通道的电流密度增加,加快了HCN1（激活时间常数：ms,176．59±8．23 vs 306．12±45．66;-80rnV,P〈0．05）,HCN2（激活时间常数：ms,385．24±31．58 vs 763．55±106．17;-80mV,P〈0．05）通道的激活,减小了HCN2通道的半时最大激活电压[mv,-（110．05±1．73）vs -（126．09±2．85）,P〈0．05]和倾斜因子（11．70±0．46 vs 15．87±1．17,P〈0．05）,但对HCN1无影响．结论：细胞外高K^＋浓度对细胞膜上的HCN1和HCN2通道的电生理特性具有明显影响．     

5一羟色胺对人胎盘滋养细胞L-型钙通道的影响

目的探讨5一羟色胺对胎盘滋养细胞钙通道的影响。方法将体外培养的胎盘滋养细胞分为对照组和低（1．01xmol／L）、高（10．0μmol／L）浓度5一羟色胺组。采用全细胞膜片钳实验方法,在待测胎盘滋养细胞外液中分别加入Nimodpine（10μmol／L）、Flunarizne（10μmol／L）和替曲朵辛（Trxl01．Lmol／L）,进行电流检测;同样的实验条件和方法,在待测胎盘滋养细胞外液中分别加入5-TH（1μmol／L和101xmol／L）,测试电流值;在细胞外液中分别加入5-TH（μmol／L）和Nimodpine（10μmol／L）、5-TH（10μmol／L）和Nimodpine（10μmol／L）后再检测电流。结果胎盘滋养细胞L-型钙通道开放在一60--50mV之间,最大峰值电流在一40～30mV为82．31±6．46（pA）。加入Nimodpine（10μmol／L）。可阻断此电流大部分成份。Flunarizne（10Ixmol／L）和’兀X（10肛m0ⅣL）不能抑制该电流成份。加入5-TH（1μmol/L和10μmol／L）峰值电流明分别为104．77±10．84（pA）和117．16±9．67（pA）,与对照组相比有显著差异（P〈0．05）;加入5-TH（1μmo]／L）和Nimodpine（101xmol／L）、5一TH（10μmol／L）和Nimodpine（10μmol／L）后;峰值电流分别为85．35±6．54（pA）、89．42±7．62（pA）,与对照组相比无显著差异（P〉0．05）。结论5-TH可激动胎盘滋养细胞L-型钙通道,促使钙电流值增加。