脑深部电刺激双侧伏核对吗啡成瘾大鼠复吸行为的影响

Objective To investigate the influence on the behavior of withdrawal and relapse after deep brain stimulation of bilateral nucleus accumbens in morphine-dependent rats. Methods The rats with a strong unconditioned preference were discarded in preconditioning test, the selected rats were distributed into five groups randomly. After operation,morphine hydrochloride was injected subcutaneously into SD rats for 12 days (once every day,initial 5 mg/kg,increasing by 5 mg/kg per time,stable in 20 mg/kg ). A modified electrical circuit was used to procedure the DBS,the parameter was 130 Hz,150 A,60 s,l h/d,14 d. CPP test was used to exam the effect of DBS. A minor morphine dose (3 mg/kg) was injected to induce the behavior of relapse, and CPP was tested again after 24 h. Two-way ANOVA was performed on the data with Bonferroni posttest. Result ①After CPP training,CPP score of group morphine, morphine + sham and morphine + DBS was ( 155. 87 ± 20. 45 ) s, (107.33 ± 18.10)s,(135.45 ±22.09)s,and had significant difference with group of control( ( -70.34 ± 15.40) s)(t = 9.45,P＜0.01; t = 6.94,P＜0.01;t = 8.04,P＜0.01).②After 7 days' DBS,the CPP score in group of morphine + DBS reduced significantly compared to group of morphine( t = 4.21, P＜0.01) and morphine + sham( t=1.10, P＜0.05).0n the 14th day,there was more pronounced reduction ( t = 5. 15, P＜0.01; t = 3.92, P＜ 0.01). ③ 24 hours after the minor morphine dose was injected,the CPP score in morphine + DBS didn't increase significantly, and had significant difference with group of morphine ( t = 4.04, P＜0.01) and morphine + sham ( t= 4. 13, P＜0.01). Conclusion DBS bilateral nucleus accumbens in morphine-dependent rats can interfere the behavior of morphine-induced CPP and relapse.     

脑深部电刺激伏核对吗啡成瘾依赖大鼠学习记忆的影响

目的 探讨脑深部电刺激(DBS)伏核对吗啡成瘾依赖大鼠学习记忆的影响.方法 通过DBS电极刺激吗啡成瘾依赖大鼠双侧伏核,于吗啡注射第10天和刺激结束后第1天,进行Morris水迷宫实验比较假刺激组、DBS组、生理盐水组(NS组)大鼠学习记忆能力的变化.结果 刺激前3组间的平均潜伏期总体比较差异有显著性(F=7.49,P=0.0077),吗啡依赖组大鼠的平均潜伏期明显长于NS组(P<0.01);刺激后3组间的平均潜伏期总体比较差异有显著性(P=7.76,P=0.0069),刺激后DBS组大鼠的平均潜伏期缩短,与假刺激组相比差异具有显著性(P=0.014),较NS组差异无显著性(P=0.43);假刺激组与NS组相比差异具有显著性(P=0.0056).刺激前NS组(51.94±3.06)与吗啡依赖组穿越Ⅱ象限时间差异有显著性(P=0.001);刺激后NS组[(48.53±2.29)s]较假刺激组[(20.74±2.13)s]差异有显著性(P=0.001),较DBS组[(38.34±1.68)s]差异无显著性(P=0.062).结论 脑深部电刺激伏核改善了已被吗啡损害的大鼠的学习记忆功能.     

异丙酚对新生大鼠空间学习记忆功能及内源性神经干细胞增殖的影响

目的 探讨不同剂量的异丙酚对新生大鼠空间学习记忆功能发育及对内源性神经干细胞增殖的影响.方法 5窝新生SD大鼠按照随机区组化分为对照组(C组)、3种不同剂量异丙酚组(P10组、P50组、P50D组),每组15只.P10、P50组分别单次皮下注射异丙酚10mg/kg、50mg/kg;P50D组先后2次皮下注射异丙酚(50mg/kg);C组注射等容量的脂肪乳剂.3d后用BrdU法检测海马齿状回神经元的增殖;使用水迷宫检测出生后28d的幼鼠空间学习记忆能力.结果 免疫组化的检测中,P10组齿状回单位面积含BrdU阳性细胞的数量[(1225±154)个/mm2,P＜0.05 ]明显多于Con组[(840±76)个/mm2],而P50D组[(225±66)个/mm2,P＜0.05]明显减少.水迷宫试验中,P50D组空间探索的潜伏期[(42.68±6.18)s,P＜0.05]明显长于Con组[(15.12±3.43)s],P50D组目标象限停留时间[(32.18±5.38)s,P＜0.05]明显少于Con组[(55.66±8.57)s],而P50组、P10组与Con组的差异均无统计学意义.结论 大剂量异丙酚可以抑制新生大鼠的空间学习、记忆能力发育,并可抑制内源性神经干细胞增殖;小剂量的异丙酚反而可以促进内源性神经干细胞增殖.

Abstract：

Objective To investigate the effects of propofol on the development of spatial learning and memory and neuron proliferation of neonatal rats at different doses. Methods 60 neonatal rats were divided into four groups among per litter by using a randomized block design. Three different doses of propofol group were induced with propofol 10 mg/kg( group P10) ,50 mg/kg( group P50) or 50 mg/kg twice( group P50D) by subcutaneous injection respectively. Neuron proliferation at dentate gyrus was detected by using BrdU marker 3 days later.Morris water maze test was carried out on postnatal day 28. Escape latency,time in probe quadrant were recorded.Results Compared to the control group,neuron marked with BrdU at dentate gyrus in group P50D was significantly decreased( (840±76) vs (225 ±66), P＜0.05) ,group P10 was significantly increased( (840 ±76) vs ( 1225± 154), P＜0.05). Compared to the control group,latency of group P50D was significantly increased( ( 15.12 ±3.43 ) s vs (42.68 ± 6. 18 ) s, P ＜ 0. 05 ), time in probe quadrant of group P50D were significantly decreased ( ( 55.66 ± 8.57 ) s vs (32. 18 ± 5. 38 ) s, P＜ 0. 05 ). Compared to the control group, there was no significant difference between group P50 and group P10. Conclusion Propofol given to seven-day-old rats with 50 mg/kg twice by subcutaneous injection suppresses neuron proliferation and impairs development of memory and learning in neonatal rats,but propofol given with 10 mg/kg once promotes neuron proliferation.     

ENDOGENOUS HEME OXYGENASE／CARBON MONOXIDE SYSTEM MEDIATES LIPOPOLYSACCHARIDE- INDUCED INTUSSUSCEPTION IN RATS

Objectives. To investigate the role of endogenous heme oxygenase (HO)/carbon monoxide (CO) system in regulating the process of intussusception (IN) induced by administration of lipopolysaccharide (LPS) in rats.Methods. IN model of rats were induced by lipopolysaccharide. HO activity was determined by the amount of bilirubin formation which was measured with a double-beam spectrophotometer, and HbCO formation was measured by CO-oximeter.Results. The results showed that LPS (10mg/kg) caused IN in up to 40% of the rats at 6h after treatment of LPS. The incidence of IN were significantly increased by 50% (P＜0.05) and by 83.2%(P＜0.01) in HO substrate(heme-L-lysinate)-treated rats and in exogenous CO-treated rats, respectively; but it was significantly decreased by 41.8%(P＜0.05) after administration of ZnDPBG, an inhibitor of heme oxygenase (HO) activity. Furthermore, LPS increased HO activity, HbCO formation cGMP content within colic smooth muscle and the plasma level of cGMP, and these parameters were significantly elevated by 62.6%(P＜0.01), 40.0%(P＜0.01), 49.3%(P＜0.05) and 38.9%(P＜0.05), respectively, compared with LPS-non-IN rats.Conclusion. It is suggested that endogenous HO/CO system plays an important role in the process of IN induced by LPS, and inhibition of HO activity may decrease the formation of IN.     

噻萘普汀与碳酸锂对慢性应激抑郁模型大鼠海马pCREB表达的影响

目的 研究噻萘普汀与碳酸锂对慢性应激抑郁模型大鼠海马磷酸化Camp反应元件结合蛋白(Pcreb)表达的影响.方法 将大鼠随机排列法分为抑郁模型组、噻萘普汀组、碳酸锂组和对照组.模型组、噻萘普汀组和碳酸锂组给予21 d的应激刺激,此期间对照组正常饲养,刺激期间噻萘普汀组每天灌胃噻萘普汀(50mg/kg),碳酸锂组每天灌胄碳酸锂(60mg/kg),模型组和对照组每天灌胃等体积的生理盐水.行为学检测应用open-field法和液体消耗实验.采用Western-blotting法检测各组大鼠海马Pcreb的表达情况.结果 应激后模型组水平穿越格数[(23.2±23.0)格]、竖立次数[(8.1±7.2)次]、修饰次数[(3.6±3.5)次]、糖水消耗百分比[(55.4±11.7)%]均显著低于对照组[分别为(46.0±18.9)格、(20.3±11.3)次、(8.4±2.7)次、(68.5±8.2)%;均P＜0.01].应激后噻萘普汀组水平穿越格数[(28.1±23.0)格]、竖立次数[(12.1±9.4)次]和修饰次数[(5.5±3.2)次]低于对照组(P＜0.05),与模型组差异无显著性;糖水消耗百分比[(62.7±10.6)%]与对照组差异无显著性,但高于模型组(P＜0.05).应激后碳酸锂组水平穿越格数和糖水消耗百分比低于对照组(P＜0.05),竖立次数和修饰次数低于对照组(P＜0.01),各项数值均高于模型组,但差异无显著性(P＜0.05).在Western-blotting法检测中,模型组大鼠海马Pcreb的表达水平显著低于对照组(P＜0.01);噻萘普汀组大鼠海马Pcreb的表达水平与对照组差异无显著性(P＞0.05),但显著高于模型组(P＜0.01);碳酸锂组大鼠海马Pcreb的表达水平显著低于对照组(P＜0.01).高于模型组(P＜0.01).结论 噻萘普汀可以逆转慢性应激抑郁模型大鼠海马中Pcreb表达的降低;碳酸锂可以部分逆转慢性应激抑郁模型大鼠海马中Pcreb表达的降低.

Abstract：

Objective To research the effects of tianeptine and lithium on expression of pCREB in hippocampus of chronic stress depression rats. Methods All the experimental rats were divided by random into : Group of depression,Group of tianeptine,Group of lithium and Group of control. The rats of Group of depression, Group of tianeptine and Group of lithium were applied stress for 21 days,and meanwhile Group of control had no stress. The rats of Group of tianeptine were fed with tianeptine (50 mg/kg) , Group of lithium were fed with lithium (60 mg/kg) , while another groups were fed with normal sodium of the same volume. The ethology examination was performed by using method of open-field and experiment of fluid consumption. The expression of pCREB was detected by Western-blotting method. Results After the chronic stress,the horizontal crossing numbers,the erection times,the modification times and the percentage of sacchar-consumption of the rats of Group of depression were 23.2±23.0;8. 1 ±7.2; 3.6 ±3.5 and (55.4 ±11.7)% respectively, which were less than Group of control (46.0±18.9;20.3±11.3;8.4±2.7 and (68.5 ±8.2)% ; P＜0.01). The horizontal crossing numbers(28. 1 ±23.0) ,the erection times(12. 1 ± 9.4) and the modification times(5.5 ±3.2) of Group of tianeptine are less than those of Group of control (P ＜ 0. 05), but no significant difference compared with Group of depression; the percentage of sacchar-consumption(62.7 ± 10.6) % ,Group of tianeptine was more than Group of depression (P＜ 0.05 ) , but no obvious difference with Group of control. The horizontal crossing numbers, the erection times, the modification times and the percentage of sacchar-consumption of Group of lithium were less than those of Group of control (P ＜ 0.05), more than those of Group of depression but no significant difference (P ＞ 0.05). In Westernblotting method,the level of pCREB in the hippocampus of Group of depression was less than that of Group of control (P＜ 0.01); that of Group of tianeptine was more than that of Group of depression (P ＜ 0.01) but no obvious difference with Group of control; that of Group of lithium was less than that of Group of control (P＜0. 01) and more than Group of depression (P＜0.01). Conclusion Tianeptine could reverse the reduction of expression of pCREB in hippocampus of chronic stress depression rats and lithium partly did it.     

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.     

电磁辐射对大鼠海马NMDA受体NR1亚单位蛋白及其mRNA表达的影响

目的 探讨2000 μW/cm2电磁辐射对大鼠海马N-甲基-D-门冬氨酸(NMDA)受体NR1亚单位蛋白及其mRNA水平表达的影响,揭示电磁辐射对大鼠学习记忆功能的损伤机制.方法 实验分为空白对照组,假辐射组,1 h/d、2h/d、3 h/d辐射组.将辐射组大鼠固定体位,头部接受功率密度为2000μW/cm2的近场辐射,连续辐射30d.通过Morris水迷宫检测大鼠的空间学习记忆能力,采用免疫组化法和Western-Blot法检测大鼠海马组织NR1蛋白表达的变化,RT-PCR法检测大鼠海马组织NR1 mRNA表达的变化.结果 各辐射组大鼠在水迷宫检测第4天寻找安全平台的逃避潜伏期分别为1 h/d[(12.29±1.36)s]、2 h/d[(17.99±2.25)s]、3 h/d[(24.66±5.56)s],均明显长于空白对照组[(8.8±1.66)s](P＜0.05);1 h/d、2 h/d和3 h/d辐射组大鼠海马神经元均排列紊乱,NR1阳性细胞比率明显低于空白对照组,海马组织NR1蛋白[分别为(0.122±0.026)、(0.102±0.023)、(0.060±0.009)]及其mRNA[分别为(0.46±0.07)、(0.35±0.05)、(0.12±0.02)]表达水平较空白对照组[(10.70±0.11)、(0.68±0.11)]均明显降低(P＜0.05).而假辐射组大鼠各项指标与空白对照组相比均无显著差异(P＞0.05).结论 2000μW/cm2电磁辐射可导致大鼠学习记忆功能下降,其机制可能与大鼠海马组织NR1蛋白及其mRNA的表达降低有关.

Abstract：

Objective To evaluate the effects of electromagnetic irradiation of 2000 μW/cm2 exposure on mRNA and protein expression levels of immunoreactive protein and mRNA of NMDAR1 in rats hippocampal,and to explore the impaired mechanism of electromagnetic irradiation on learning and memory.Methods Rats were randomly divided into normal control group, sham-radiated group, and 1 h/d, 2 h/d, and 3 h/d radiation groups.The rats in the radiation groups were fixed and recieved microwave exposure of 2000 μW/cm2, then their learning and memory abilities were tested by Morris water maze experiment, the change of NR1 protein in hippocampal neurons of each group of rats was measured with immunohistochmistry and western blot techniques, and the expression of NR1 mRNA in hippocampus was determined by RT-PCR.Results In the water maze test,compared with the normal control group (8.8 ± 1.66 ), the escape latency of three radiated groups rats ( 1 h/d ( 12.29 ±1.36) s,2 h/d ( 17.99 ±2.25) s,and 3 h/d (24.66 ±5.56) s) were significantly longer (P＜0.05).In the radiation group,the hippocampal neurons of rats showed evident reduction in the ratio of NR1 positive cells,irregular,and arrayed in disorder.Moreover,compared with the normal control group ( (0.70 ±0.11 ), (0.68 ±0.11 ) ) ,the expession of NR1 protein ( 1 h/d (0.122 ±0.026) ,2 h/d (0.102 ±0.023) ,and 3 h/d (0.060 ± 0.009) ) and its mRNA ( 1 h/d (0.46 ±0.07) ,2 h/d (0.35 ±0.05) ,and 3 h/d (0.12 ±0.02) ) in hippocampal neurons was significantly decreased (P＜0.05).Among the indicators, there was no significant difference between sham-radiated group and normal control group.Conclusions Electromagnetic irradiation of 2000 μW/cm2 exposure can impair the learning and memory abilities of rats possibly through a mechanism correlated with the lower expression of NR1 protein and its mRNA in hippocampus.     

苯并芘对断乳大鼠学习记忆能力的影响及机制研究

目的 探讨不同剂量苯并(a)芘[B(a)P]对断乳大鼠学习记忆能力的影响及其机制.方法 将40只SD断乳大鼠(28d)随机分为5组,空白对照组、溶剂对照组、3个染毒组(浓度分别为5,10和20mg/kg体质量),隔日腹腔染毒30d.染毒结束后用Morris水迷宫试验观察,免疫组化法测定海马组织中N-甲基-D天门冬氨酸受体NR2B亚基(NMDAR2B)和脑源性生长因子(BDNF)含量.结果 Morris水迷宫试验结果显示,随B(a)P染毒剂量的增加,逃避潜伏期呈递减趋势,高剂量组[(62.78 ±47.25)s]与对照组[(40.60 4-38.79)s]相比,差异有统计学意义(P<0.01);跨台次数随染毒剂量的增加逐渐减少,高剂量组[(4.33 ± 2.08)次]与对照组[(11.25 ± 2.63)次]相比.差异有统计学意义(P＜0.05);随染毒剂量增加,海马区NMDAR2B灰度值呈递减趋势,高剂量组(150.38 ± 15.34)与对照组(162.23±6.56)相比,差异有统计学意义(P＜0.05);BDNF灰度值除低剂量组外呈递减趋势,高剂量组(141.83 ±13.37)与对照组(163.13±8.09)、低剂量组(164.56±9.10)相比均差异有统计学意义(P＜0.05).结论 亚急性B(a)P暴露可降低断乳大鼠空间学习记忆能力,其机制可能与B(a)P影响海马NMDAR2B和BDNF的表达有关.

Abstract：

Objective To investigate the changes and mechanism of learning and memory in rats by different doses of benzo (a) pyrene (B(a)P). Methods Forty weaned rats (28 days) were randomly divided into control group (NS), solvent group ( vegetable oil) and three B (a) P dosage groups (the doses were 5,10 and 20 mg / kg body weight respectively ). And all rats were administrated intraperitoneally every other day to one month. The capability of learning and memory in rats were measured by Morris water maze test, and the brain-derived neurotrophic factor ( BDNF) and NMDAR2B content in hippocampus were tested by immunohistochemistry. Results In training of Morris water maze,the average escape latency was extended gradually with increasing dose, and there was a statistically significant difference between high-dose group((62. 78 ±47. 25 )s) and the control group((40.60±38.79)s)(P＜ 0.01). Compared with the control group(11.25 ±2.63), the number of crossplatform of high-dose group(4.33 ±2.08) was statistically reduced (P＜0.05). B(a)P at 10 and 20 mg/kg decreased NMDAR2B and BDNF expression in hippocampus of rats in immunohistochemistry. The level of NMDAR2B was (162.23 ±6.56) in the high-dose group and (150.38 ± 15.34) in the control group(P＜0.05);the expression level of BDNF was (163. 13 ± 8.09) in the high-dose group and (141.83 ± 13.37) in the control group(P＜ 0.05). Conclusion Subacute B(a)P exposure can reduce spatial learning and memory in weaning rats, it may be related to decreased levels of NMDAR2B and BDNF in hippocampus.     

兔原发性甲状旁腺机能亢进甲状旁腺细胞增生与凋亡的相关性

目的 探讨兔甲状旁腺细胞增殖与凋亡在原发性甲状旁腺机能亢进症(PHPT)发病机制中的作用.方法 健康成年中国白兔80只,随机分成两组,对照组40只以正常饮食(Ca:P,1:0.7)喂养,实验组40只以高磷饮食(Ca:P,1:7)喂养诱发原发性甲状旁腺机能亢进动物模型.在第3、4、5、6个月动物死亡后,分别对实验组和对照组动物行甲状旁腺细胞计数,同时采用免疫组织化学法观察腺体增殖细胞核抗原(PCNA)和Bcl-2的表达及采用DNA片段末端标记作细胞凋亡的定量检测,并与对照组正常腺体作对照研究.结果 PHPT组腺体细胞计数(个/高倍视野)是正常对照组的1.61倍(分别为673±151与418±25,t=12.112,P＜0 01);PHPT组凋亡指数明显高于对照组(分别为200.2±125.6与11.0±3.0,t=-10 193,P＜0.01);PHPT组腺体细胞PCNA阳性率(50.5‰±11.6‰)显著高于对照组(26 7‰±2.8‰)(t=-13 120,P＜0 01),Bcl-2表达(460‰±190‰)显著高于对照组(67‰±4‰)(t=-14.120,P＜0.01);PCNA和Bcl-2表达与腺体细胞凋亡指数均呈正相关(r值分别为0.861和0.871,P＜0.05).结论 甲状旁腺细胞增生与凋亡失衡可能是导致PHPT发生的主要原因.

Abstract：

Objective To evaluate the effect of proliferation and apoptosis of parathyroid cell in rabbits with primary hyperparathyroidism (PHPT) . Methods A total of 80 adult Chinese rabbits were randomly divided into two groups (n=40 each) . The control group was fed with a normal diet (Ca: P,1:0.7) while the experimental group a high phosphate diet (Ga: P, 1: 7) for3-,4- ,5-, or 6-month intervals to establish the animal model of PHPT. The parathyroid was totally removed for pathological examination after all rabbits were sacrificed. The thyroparathyroid complex was removed en bloc, fixed in neutral formalin and prepared for histological examination. The number of parathyroid cell in PHPT was calculated. Proliferation was determined by immunohistochemistry of proliferation cell nuclear antigen (PCNA) while apoptosis assessed by in situ dUTP biotin nick-end labeling (TUNEL). Results The number of parathyroid cell was 1.61 times in PHPT than that in the normal control (673±151, 418 ±25, t=-12.112, P＜0.01).Apoptotic index (AI) increased significantly more in PHPT than that in normal control (200.2‰±125.6‰, 11.0‰±3.0‰, t=-10.193, P＜0.01). The rate of PCNA positive-cell increased significantly more in PHPT than that in control (50.5%＞±11.6‰, 26.7‰±2.8‰,t=-13.120, P＜0.05).So did Bcl-2 (460‰±190‰, 67‰±4‰, t=-14.120,P＜0.05). There was a positive correlation between AI and PCNA (r=0.861, P＜0.05). It was the same as between AI and Bcl-2 (r=0.871, P＜0.05). The value of bone mineral density decreased significantly more in PHPT than that in normal control (152±34,189±12, t=9.236, P＜0.05). Conclusion PHPT may be mainly induced by an excessive proliferation of parathyroid cells and an acceleration of apoptotic process.     

糖皮质激素受体通过细胞外信号调节激酶途径参与吗啡耐受机制

目的 探讨糖皮质激素受体参与吗啡耐受形成的机制及细胞外信号调节激酶(ERK)信号途径在其中的作用.方法 将40只健康雄性SD大鼠行鞘内置管后随机分为4组(n=10),盐水对照组(C组)鞘内注射生理盐水lOμl;慢性吗啡耐受组(M组)鞘内注射吗啡10μg;糖皮质激素受体拮抗剂组(MR组)鞘内注射吗啡10μg,再注射糖皮质激素受体拮抗剂RU38486 2μg;糖皮质激素受体激动剂组(MD组)鞘内注射吗啡10μg,再注射糖皮质激素受体激动剂地塞米松4μg,鞘内注射每种药物均为2次/d,连续7 d.采用甩尾实验评价大鼠热痛觉,于给药后第8天取出腰膨大处脊髓分别进行Western印迹检测μ阿片受体(MOR)、糖皮质激素受体(GR)、磷酸化ERK(pERK)蛋白表达及TUNEL染色.结果 地塞米松和RU38486分别对吗啡耐受诱导的热痛敏具有促进和抑制作用.与M组(凋亡细胞数5.4±1.1,蛋白表达MOR 37±5,GR 20±6,pERK1 39±4,pERK2 41±5)比较,MR组凋亡细胞数(3.2±0.4)显著少(P<0.01);MD组(16.0±1.6)显著多(P<0.01);MR组MOR(28±5)、GR(12±6)、pERK1(33±4)、pERK2(34±5)蛋白表达均下调(均P<0.01);MD组MOR(55±6)、GR(28±9)、pERKl(49±6)、pERK2(59±5)蛋白表达均上调(均P<0.01).结论 糖皮质激素受体的活性可影响吗啡诱导的脊髓神经凋亡及吗啡耐受中脊髓pERK的表达.

Abstract：

Objective To investigate the mechanism of glucocorticoid receptors(GR)participating in morphine tolerance development via the extracellular signal-regulated kinase(ERK)signal pathway.Methods Forty healthy male SD rats were implanted with intrathecal catheters and then randomized into 4 groups:Group C received an intrathecal injection of 10 μl saline,Group M 10μg morphine,Group MR 10μg morphine followed by 2μg GR antagonist RU38486 and Group MD 10μg morphine followed by 4μg GR agonist dexamethasone(DEX)respectively.Each intrathecal drug was administered twice daily for 7 days.Tail-flick test was employed to evaluate the thermal hyperalgesia.After tail-flick test at Day 8,the lumbar intumescentias were isolated by Western blot to examine the protein expressions of Mu opioid receptor(MOR).GR and pERK.And terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling(TUNEL)stain was performed.Results DEX and RU38486 enhanced and inhibited morphine induced hyperalgesia to heat respectively.Compared with Group M(count of spinal dorsal horn apoptotic cells 5.4±1.1,protein expression:MOR 37±5,GR 20±6,pERK1 39±4,pERK2 41±5),apoptotic cells decreased in Group MR(3.2±0.4)(P<0.01)and increased in Group MD (16.0±1.6)(P<0.01);the protein expressions of MOR(28±5),GR(12±6),pERK1(33±4)and pERK2(34±5)were down-regulated in Group MR(P<0.01)while up-regulated in Group MD(55±6,28±9,49±6,59±5)(P<0.01).Conclusion The activity of glucocorticoid receptors affects the morphine-induced spinal neural apoptosis and the expression of spinal pERK in morphine tolerance.     

盐酸戊乙奎醚对吗啡依赖大鼠戒断症状及位置偏爱的影响

目的 探讨盐酸戊乙奎醚(PHC)对吗啡依赖大鼠戒断症状及条件性位置偏爱效应的影响.方法 (1)48只雄性SD大鼠,随机分为吗啡依赖组(MOR组),纳洛酮促戒断组(NAL组),PHC治疗组(PHCl,2,3组),对照组(NS组),每组8只.剂量递增法连续5 d皮下注射吗啡(10～50mg/kg,每日2次)及纳洛酮催促戒断(5 mg/kg),建立吗啡躯体依赖大鼠模型.实验第6天上午,纳络酮催促戒断前30min腹腔注射不同剂量的PHC(0.5,1.0,1.5 mg/kg).观察各组大鼠在20 min内的体质量丧失情况及戒断症状.(2)40只雄性SD大鼠,随机分为吗啡诱导组(MOR组),PHC治疗组(PHC1,2,3组),对照组(NS组),每组8只.连续7d交替皮下注射吗啡(10mg/kg,每天1次)或生理盐水,诱导大鼠的吗啡位置偏爱效应.实验第8天停用吗啡,NS组与MOR组腹腔注射等体积的生理盐水;PHC治疗组则分别腹腔注射PHC 0.5,1.0,1.5 mg/kg.各组大鼠行CPP测试.结果 (1)PHC治疗组能明显缓解吗啡依赖大鼠的催促戒断症状,其体质量丧失[(8.53±1.20)g、(7.36±1.06)g、(5.40±1.79)g,(12.63±2.22)g,F=83.16,P＜0.01]和戒断症状评分[(25.36±3.11)分、(21.38±3.50)分、(17.06±1.78)分,(31.69±2.76)分,F=256.56,P＜0.01)]明显低于NAL组,且呈剂量依赖性.(2)PHC治疗组的灰区停留时间与MOR组比较显著缩短[(529±83)s、(460±107)s、(418±97)s,(643±111)s,F=13.22,P＜0.01],且呈剂量依赖性.结论 PHC急性治疗能剂量依赖的抑制吗啡依赖大鼠戒断症状和条件性位置偏爱的表达.     

青龄大鼠和成龄大鼠在吗啡条件性位置偏爱和行为敏感化效应上的差异

目的 考察青龄大鼠在吗啡奖赏效应和行为激活效应上是否比成龄大鼠更加敏感.方法 使用条件化位置偏爱程序检验吗啡奖赏效应和行为敏感化效应,青龄雄性大鼠(出生后35d)和成龄雄性大鼠(出生后67d)每天接受盐水或吗啡(3mg/kg)注射,进行6d的位置条件化训练,同时记录条件化训练期间大鼠在伴药箱的水平运动量,训练结束24h后对条件性位置偏爱效应进行检测.结果 青龄大鼠没有形成吗啡条件性位置偏爱效应[盐水组(20.89±31.14)s,吗啡组(90.75±27.91)s,P=0.15];成龄大鼠形成了吗啡条件性位置偏爱效应[盐水组(31.5±41.24)s,吗啡组(266.13±32.26)s,P<0.01];成龄大鼠的吗啡条件性位置偏爱效应显著高于青龄大鼠(p<0.01).青龄吗啡组大鼠d3～d6的水平运动量均显著高于dl(P<0.05或P<0.01),成龄吗啡组大鼠d2～d6的水平运动量均显著高于d1(P<0.05或P<0.01);对吗啡给药大鼠进行二因素重复测量方差分析表明:年龄主效应不显著[F_(1,14)=0.33,JP=0.57],年龄×处理交互作用不显著[F_(5,70)=0.85,P=0.52].结论 青龄大鼠对吗啡的奖赏效应不如成龄大鼠敏感,但与成龄大鼠有着类似的行为敏感化效应.     

内侧前额叶皮层神经元可塑性改变在吗啡奖赏记忆形成中的作用

目的 观察吗啡诱导的大鼠内侧前额叶皮质（mPFC）神经元可塑性改变对吗啡奖赏记忆形成的影响.方法 40只SD大鼠分为生理盐水对照组和吗啡组,分别腹腔注射生理盐水（2 ml/kg）和盐酸吗啡（10 mg/kg）,注射后0、2、4、8h断头取脑,Western Blot分析mPFC区Arc/Arg 3.1蛋白变化;另取大鼠60只,分别腹腔注射0、5、10或20 mg/kg吗啡,Western Blot（n=5）分析mPFC区Arc/Arg 3.1蛋白变化;免疫组化法（n=5）检测mPFC区Arc/Arg 3.1阳性细胞数量变化;Golgi-cox改良法（n=5）检测mPFC区神经元棘突数量变化;再取大鼠40只,经8天生理盐水、吗啡交替训练建立条件性位置偏爱（CPP）模型,吗啡模型组大鼠在每次吗啡注射前15 min给予mPFC区注射Arc/Arg 3.1基因的反义寡核苷酸（AS,n=10）及其对照（CS,n=10）,观察其对吗啡CPP评分的影响.结果 与生理盐水对照组相比,10 mg/kg吗啡注射后2 h mPFC内Arc/Arg 3.1蛋白水平、Arc/Arg 3.1阳性细胞数、棘突数量[（1.01±0.04） vs （1.58±0.18）,P＜0.01;（42.80±7.63） vs （74.47±8.02）,P＜0.01;（17.27±5.64） vs （39.47±7.56）,P＜0.01]均显著增加;与对照组相比,5、10或20 mg/kg吗啡注射后2h均可诱导mPFC的Arc/Arg 3.1蛋白水平显著增加,无剂量依赖效应;对于吗啡CPP模型组大鼠,与mPFC区注射CS相比（0.74±0.02）,AS显著降低吗啡CPP的评分（0.51±0.01）,差异具有统计学意义（P＜0.01）.结论 单次吗啡注射可以诱导大鼠mPFC区Arc/Arg 3.1蛋白表达增加并伴有神经元可塑性的增强,增加的Arc/Arg 3.1蛋白介导了吗啡奖赏记忆的形成.     

肾上腺切除对强迫游泳大鼠吗啡成瘾行为的影响

为探索应激和肾上腺切除在药物成瘾行为中的作用机制,将４０只雄性Ｗｉｓｔａｒ大鼠随机分为肾上腺切除组、糖皮质激素Ⅰ组（肾上腺切除＋氢化考的松２０ｍｇ／ｋｇ）、糖皮质激素Ⅱ组（肾上腺切除＋氢化考的松４０ｍｇ／ｋｇ）及生理盐水对照组,每组各１０只,观察肾上腺切除及给予糖皮质激素对强迫游泳大鼠条件性位置偏爱形成的影响。结果：①肾上腺切除组动物在药物搭配侧箱体中停留的时间与在对侧箱体中停留时间相比无明显差异     

尿酸对6-羟基多巴胺致大鼠黑质纹状体系统毒性的影响

目的 评价尿酸对6-羟基多巴胺致大鼠黑质纹状体系统多巴胺能神经元毒性的影响.方法 30只雄性SD大鼠,分为生理盐水组(10只)、100 mg/kg尿酸组(5只)、200 mg/kg尿酸组(10只)和250 mg/kg尿酸组(5只).每天2次(间隔2 h),每次腹腔注射生理盐水或尿酸5 d,于第6天第1次腹腔注射生理盐水或尿酸后,在立体定向仪指导下,右侧纹状体内两点注射6-羟基多巴胺.此后继续给予大鼠腹腔注射生理盐水或尿酸5 d.于注射6-羟基多巴胺后第3、4周分别进行自主活动计数,安非他明旋转实验,前肢功能测定.注射6-羟基多巴胺后第5周进行纹状体多巴胺和高香草酸水平测定.结果 200 mg/kg尿酸组大鼠自主活动计数[(14±4)次/2 min]明显高于生理盐水组[(4±5)次/2 min,P＜0.01];安非他明诱导的旋转实验100 mg/kg尿酸组(11.2 ±4.2)和200 mg/kg尿酸组每分钟旋转次数(10.8±7.5)显著低于生理盐水组(19.3 ±5.2,P＜0.01).200 mg/kg尿酸组前肢功能测定5 s内移动90 cm的前患肢步数(9.89±3.41)明显高于生理盐水组(4.36±3.72,P＜0.01);200 mg/kg尿酸组毁损侧纹状体多巴胺(0.29±0.19)、高香草酸水平(1.22±0.5)显著高于生理盐水组多巴胺(0.05 ±0.03,P＜0.01)、高香草酸水平(0.24±0.13,P＜0.05).结论 适当提高体内的尿酸水平能减轻6-羟基多巴胺对SD大鼠黑质纹状体系统多巴胺神经元的毒性作用.     

青风藤及青藤碱对吗啡依赖小鼠位置偏爱效应和脑内组胺水平的影响

目的探讨中药青风藤及其有效成分青藤碱对吗啡诱导的小鼠条件性位置偏爱(CPP)及脑内组胺(HA)水平的影响。方法采用位置偏爱箱法,连续皮下注射吗啡(9mg/kg·b.w.)6d,引起小鼠产生显著的条件性位置偏爱效应。实验分空白对照组、吗啡模型组(9mg/kg·b.w.)、青风藤醇提液组(10g/kg·b.w.)、青藤碱组(60mg/kg·b.w.)、苯海拉明组(30mg/kg·b.w.)、CP48/80组(5mg/kg·b.w.)和L-组氨酸组(750mg/kg·b.w.),后5个给药组分别在位置偏爱训练的第4天开始给药,连续用药3d。脑内组胺含量采用荧光分光光度法测定。本实验同时检测青风藤、青藤碱、苯海拉明、CP48/80及L-组氨酸对小鼠的奖赏效应或厌恶效应。结果吗啡模型组小鼠在伴药箱中停留的时间明显延长,小鼠脑内HA水平显著升高(P<0.01)。青风藤或青藤碱连续用药可显著抑制吗啡引起的小鼠位置偏爱的形成,降低脑内的HA含量。青风藤、青藤碱及L-组氮酸对正常小鼠脑内组脑水平有升高作用(P<0.01),但3药本身并不使小鼠产生奖赏或厌恶效应。CP48/80能使正常及吗啡依赖小鼠脑内组胺含量明显减少(P<0.01),但该药对CPP无明显影响。结论吗啡诱导的小鼠位置偏爱效应与脑内HA水平升高、中枢组胺能神经系统激活有关。青风藤及青藤碱能消除吗啡诱导的小鼠条件性位置偏爱的形成,对脑内组胺水平的改变具有调节作用。     

Clobenpropit及组氨酸对大鼠吗啡诱导的条件位置偏爱重燃的作用

目的:研究组胺H3受体拮抗剂clobenpropit及组胺前体物质组氨酸对小剂量吗啡诱导的大鼠条件位置偏爱重燃的作用.方法:在吗啡诱导的大鼠条件位置偏爱表达、消退后,利用小剂量吗啡诱发复吸,然后评价clobenpropit及组氨酸对其的作用.clobenpropit干预组:在测试前15 min,大鼠腹腔内注射吗啡(1 mg/kg),合并脑室给予clobenpropit(2、5、10 μg/rat);组氨酸干预组:在给予吗啡(1 mg/kg)前1 h,腹腔内注射组氨酸(100、200、500 mg/kg);分别观察15 min测试时间大鼠各个阶段,各个处理组在伴药盒停留的时间.结果:脑室内注射clobenpropit(5、10 μg/rat)和腹腔内注射组氨酸(100、200、500 mg/kg),都可以显著抑制小剂量吗啡诱发的大鼠条件位置偏爱的重燃.结论:clobenpropit及组氨酸均能剂量依赖性地抑制小剂量吗啡诱导的大鼠条件位置偏爱的重燃,在一定程度上抑制吗啡的复吸过程,表明内源性组胺具有抑制吗啡复吸的作用.     

吗啡依赖大鼠海马CA1区突触界面结构变化的易感性差异

目的 通过研究高、低吗啡条件性位置偏爱(conditioned place preference,CPP)大鼠海马CA1区突触界面结构参数的变化,为吗啡依赖易感性差异提供形态学依据.方法 将雄性SD大鼠随机分为实验组(130只)和生理盐水对照组(30只).实验组按剂量递增法腹腔注射吗啡建立吗啡依赖模型.分别对2组大鼠进行CPP训练和测评,根据测评值将实验组大鼠再次分为高、中、低偏爱组,中偏爱组淘汰.于末次吗啡注射后3h、戒断后3d和14d将高、低偏爱组和对照组各选取8只大鼠处死,取海马CAI区按照标准程序制成电镜样本,电镜观察摄像,图像分析软件分析测量突触界面结构参数.结果 ①预测试时3组大鼠CPP值之间的差异无显著性(F=0.78,P=0.47);处理因素终止后3h、3d、14d 3组大鼠CPP值差异有极显著性(P<0.01);两两比较显示高偏爱组CPP值高于低偏爱组,差异有极显著性(P<0.01).②在3h和3d时,3组大鼠PSD厚度(突触后致密物质厚度)、突触间隙宽度差异有极显著性(P=0.01～0.03),并且高偏爱组的PSD厚度[(15.20±3.65)nm]小于低偏爱组[(17.63±6.61)nm],差异具有显著性(P<0.05);高偏爱组间隙宽度[(5.77±2.08)nm]大于低偏爱组[(4.92±1.65)nm],差异具有显著性(P<0.05);在14d时,3组大鼠PSD厚度差异有极显著性(P=0.00),并且高偏爱组的PSD厚度[(16.22±4.93)nm]小于低偏爱组[(18.42±3.78)nm],差异具有显著性(P<0.01).结论 高偏爱组大鼠海马CA1区突触间隙宽度的测量值大于低偏爱组,PSD厚度的测量值小于低偏爱组,上述变化可能是吗啡依赖易感性差异的突触界面结构基础.     

精神分裂症模型鼠腹腔中的中性粒细胞趋化运动的动态可视化分析

目的：应用细胞动态可视化系统观察并分析正常鼠和精神分裂症模型鼠腹腔中的中性粒细胞趋化运动能力及其差异。方法：18只健康昆明小鼠随机分为对照组（6只）、0.3 mg/kg 5-甲基二氢二苯并环庚烯亚氨马来酸（dizocilpine maleate,MK-801）处理组（6只）、0.6 mg/kg MK 801处理组（6只）,分别提取中性粒细胞,在显微镜下观察形态并计数。将每组细胞分成两份进行趋化实验,分别为趋化剂处理组和趋化剂未处理组,即对照组 1、0.3 mg/kg MK-801处理组1、0.6 mg/kg MK-801处理组 1和对照组 2、0.3 mg/kg MK-801处理组 2、0.6 mg/kg MK-801处理组 2。使用NIS-Elements软件记录细胞动态迁移过程。使用TAXIScan Analyzer 2软件每组细胞随机选取30个细胞（即n=30）分析细胞迁移的轨迹、速度和距离,并采用配对检验和单因素方差分析进行统计学分析。结果：对照组、0.3 mg/kg MK-801处理组、0.6 mg/kg MK-801处理组的中性粒细胞的数量分别是（1.00±0.03）×104/mL、（0.05±0.02）×104/mL、（0.32±0.01）×104/mL,差异均有统计学意义（P<0.05）。在趋化剂的作用下,对照组1、0.3 mg/kg MK-801处理组1、0.6 mg/kg MK-801处理组1的中性粒细胞定向迁移能力分别是（0.85±0.11） radian、（1.00±0.11） radian、（0.96±0.10） radian,差异均有统计学意义（P<0.05）;中性粒细胞迁移速度分别是（0.09±0.02） μm/s、（0.12±0.01） μm/s、（0.14±0.01） μm/s,差异均有统计学意义（P<0.05）;中性粒细胞迁移距离分别是（94.26±0.02） μm、（134.61±0.01） μm、（156.19±0.01） μm,差异均有统计学意义（P<0.05）。结论：与对照组腹腔中的中性粒细胞比较,精神分裂症模型鼠腹腔中的中性粒细胞具有更强的趋化运动能力。