全凭静脉麻醉老龄大鼠海马tau蛋白磷酸化水平与认知功能的关系

目的 探讨咪唑安定、芬太尼、丙泊酚联用对老龄大鼠认知功能的影响以及这种影响是否与海马tau蛋白磷酸化水平有关.方法 雄性18月龄SD大鼠40只,随机分成全麻组和对照组,每组20只.每组分别于处理后第1和第7天各断头处死10只,HE染色光镜下观察海马神经元结构,免疫组化染色法检测海马tall蛋白Thr231和Ser404位点磷酸化蛋白的表达水平.其中后处死大鼠于处理后第1周行水迷宫实验.结果 处理后第1天,全麻组大鼠寻找平台的时间明显长于对照组(P＜0.01),但第2～7天与对照组比较差异无统计学意义.处理后第1天,全麻组海马tau蛋白Thr231位点的磷酸化水平高于对照组(P＜0.05).第7天,两组Thr231、Ser404位点的磷酸化水平差异无统计学意义.光镜下各组大鼠海马神经元结构未见明显异常.结论 咪唑安定、芬太尼、丙泊酚联用可引起老龄大鼠麻醉后第1天出现短暂的认知功能减退,海马tau蛋白Thr231位点的过度磷酸化可能与其有关.     

氯胺酮对SD幼鼠海马神经元细胞凋亡和NR2B蛋白的影响

目的观察氯胺酮对海马神经元细胞凋亡和N-甲基-D-天(门)冬氨酸受体2B亚基(NR2B)蛋白的影响。方法45只新生第7天SD大鼠随机均分为腹腔注射氯胺酮25 mg/kg组(K1组)、50 mg/kg组(K2组)和生理盐水50 ml/kg组(K0组)。24 h后每组取5只幼鼠处死,用TUNEL法检测海马神经元细胞凋亡,免疫组化法检测NR2B蛋白表达,剩余30只动物喂养至42 d,用Mor-ris水迷宫评价大鼠成年后的空间学习记忆能力。结果在腹腔注射24 h后,海马神经元细胞凋亡数明显增加(K1组9.5±4.2,K2组23.4±7.6,K0组5.3±1.7);NR2B蛋白免疫组化染色切片灰度值K1组为182.36±4.17,K2组为179.11±4.28,K0组为198.25±3.38;但是成年后在Morris水迷宫中的学习记忆能力并没有明显变化。结论单次注射氯胺酮24 h后,诱发新生第7天SD大鼠显著增强的海马神经元细胞凋亡,上调NR2B的表达,但对于成年后的空间学习记忆能力无明显影响。     

氯胺酮对新生大鼠海马CREB磷酸化水平的影响

目的 探讨氯胺酮对新生大鼠海马环磷腺苷反应元件结合蛋白(CREB)磷酸化水平的影响.方法 SD大鼠75只,日龄7 d,雌雄不拘,随机分为对照组(C组)、氯胺酮10 mg/kg组(K1组)和氯胺酮20 mg/kg组(K2组),每组25只.K1组和K2组分别皮下注射氯胺酮10、20 mg/kg,C组注射等容量生理盐水,每间隔90 min注射1次,共注射7次.于首次注射后24 h时断头取脑,分离海马,采用TUNEL法检测凋亡神经元,计算凋亡指数;免疫荧光双标法检测p-CREB表达水平;RT-PCR法半定量检测CREB下游基因BDNF mRNA及Bcl-2 mRNA表达水平;于首次注射后6周时采用Morris水迷宫实验测定各组大鼠认知功能.结果 与C组相比,K1组和K2组大鼠海马神经元凋亡指数升高,p-CREB、BDNF mRNA及Bcl-2 mRNA表达下调(P<0.05);与K1组相比,K2组大鼠海马神经元凋亡指数升高,p-CREB、BDNF mRNA及Bcl-2 mRNA表达下调(P<0.05);与C组和K1组相比,K2组逃避潜伏期延长(P<0.05).结论 氯胺酮10、20 mg/kg均可诱导发育期大鼠海马神经元凋亡,而氯胺酮20 mg/kg可导致大鼠发育成熟后认知功能降低,可能与其抑制CREB磷酸化后BDNF及Bcl-2表达下调,导致神经元凋亡,影响大鼠神经系统发育有关.     

氯胺酮对新生大鼠海马CREB磷酸化水平的影响

目的 探讨氯胺酮对新生大鼠海马环磷腺苷反应元件结合蛋白(CREB)磷酸化水平的影响.方法 SD大鼠75只,日龄7 d,雌雄不拘,随机分为对照组(C组)、氯胺酮10 mg/kg组(K1组)和氯胺酮20 mg/kg组(K2组),每组25只.K1组和K2组分别皮下注射氯胺酮10、20 mg/kg,C组注射等容量生理盐水,每间隔90 min注射1次,共注射7次.于首次注射后24 h时断头取脑,分离海马,采用TUNEL法检测凋亡神经元,计算凋亡指数;免疫荧光双标法检测p-CREB表达水平;RT-PCR法半定量检测CREB下游基因BDNF mRNA及Bcl-2 mRNA表达水平;于首次注射后6周时采用Morris水迷宫实验测定各组大鼠认知功能.结果 与C组相比,K1组和K2组大鼠海马神经元凋亡指数升高,p-CREB、BDNF mRNA及Bcl-2 mRNA表达下调(P<0.05);与K1组相比,K2组大鼠海马神经元凋亡指数升高,p-CREB、BDNF mRNA及Bcl-2 mRNA表达下调(P<0.05);与C组和K1组相比,K2组逃避潜伏期延长(P<0.05).结论 氯胺酮10、20 mg/kg均可诱导发育期大鼠海马神经元凋亡,而氯胺酮20 mg/kg可导致大鼠发育成熟后认知功能降低,可能与其抑制CREB磷酸化后BDNF及Bcl-2表达下调,导致神经元凋亡,影响大鼠神经系统发育有关.     

反复注射不同剂量氯胺酮对幼年小鼠认知功能的影响

目的 探讨反复注射不同剂量氯胺酮对幼年小鼠认知功能的影响.方法 21 d龄小鼠50只,体重10～15 g,随机分为5组(n=10),正常对照组(c组);生理盐水组(N组)腹腔注射生理盐水0.1 ml,1次/d,连续7 d;不同剂量氯胺酮组(K1组、K2组、K3组)分别腹腔注射氯胺酮25、50、100 mg/kg,1次/d,连续7 d.于给药结束后1 d时测定学习功能,于给药结束后2 d时测定记忆功能.记忆功能测定结束后每组处死4只小鼠,采用免疫组化法测定海马脑源性神经营养因子(BDNF)表达;处死6只小鼠,采用免疫印记法测定海马BDNF蛋白表达.结果 与C组比较,N组和K1组学习记忆功能减退,BDNF蛋白表达下调(P＜0.05或0.01),K2组和K3组差异无统计学意义(P＞0.05);N组和K1组学习记忆功能和BDNF蛋白表达差异无统计学意义(P＞0.05).结论 50、100 mg/kg(连续7 d反复给药)氯胺酮对幼年小鼠认知功能无影响,25 mg/kg(连续7 d反复给药)氯胺酮可降低认知功能.     

乌司他丁对氯胺酮麻醉后幼鼠认知功能及Tau、Aβ蛋白的影响

目的探讨乌司他丁预给药对氯胺酮麻醉后幼鼠认知功能的影响。方法健康雄性SPF级C57BL/6小鼠36只,21日龄,体重20~30 g,采用随机数字表法,将其分为3组(n=12):空白对照组(C组)、氯胺酮组(K组)、乌司他丁预先给药组(S组)。K组和S组腹腔注射氯胺酮30 mg/kg、每隔30 min重复注射一次,3次/d,连续7 d;S组于每天第1次注射氯胺酮前30 min腹腔注射乌司他丁50000 U/kg。末次给药结束后24 h立即行行为学测试,行为学测试完毕后立即处死大鼠,取海马组织,以western blot法测定海马tau蛋白、Aβ淀粉肽样蛋白的表达。结果与C组比较,K组逃避潜伏期延长,原平台停留时间,穿越平台次数减少,海马海马tau蛋白、Aβ淀粉肽样蛋白的表达含量升高(P0.05),S组上述指标差异无统计学意义(P0.05)。与K组比较,S组逃避潜伏期明显缩短,原平台停留时间延长,穿越原平台次数增多,海马tau蛋白、Aβ淀粉肽样蛋白的表达降低(P0.05)。结论乌司他丁预先给药可改善氯胺酮慢性暴露所致幼鼠认知功能障碍,其机制可能与抑制海马海马tau蛋白、Aβ淀粉肽样蛋白的表达有关。     

氯胺酮对大鼠海马神经元CREB磷酸化水平的影响

目的 探讨氯胺酮对大鼠海马神经元cAMP反应元件结合蛋白(CREB)磷酸化水平的影响.方法 原代培养1 d龄SD大鼠海马神经元5 d,随机分为对照组(C组)和氯胺酮组(K组),每组6孔.K组在终浓度为1 000 μmol/L氯胺酮的Neurobasel培养液中孵育3 h,C组不做任何处理.采用流式细胞仪Aanexin V-PI共染法检测凋亡神经元,计算神经元凋亡率;采用免疫组化法测定神经元Ser133位点磷酸化的CREB(pCREB)表达水平;采用RT-PCR法半定量测定CREB下游基因脑源性生长因子(BDNF)mRNA和抑凋亡基因Bcl-2 ndlNA的表达.结果 与C组相比,K组神经元凋亡率升高,pcREB、BDNFmRNA及Bcl-2 mRNA的表达均下调(P＜0.01).结论 氯胺酮可能通过抑制CREB磷酸化,下调BDNF及Bcl-2表达,诱导新生大鼠海马神经元凋亡.     

氯胺酮对新生大鼠海马CREB磷酸化水平的影响

目的 探讨氯胺酮对新生大鼠海马环磷腺苷反应元件结合蛋白(CREB)磷酸化水平的影响.方法 SD大鼠75只,日龄7 d,雌雄不拘,随机分为对照组(C组)、氯胺酮10 mg/kg组(K1组)和氯胺酮20 mg/kg组(K2组),每组25只.K1组和K2组分别皮下注射氯胺酮10、20 mg/kg,C组注射等容量生理盐水,每间隔90 min注射1次,共注射7次.于首次注射后24 h时断头取脑,分离海马,采用TUNEL法检测凋亡神经元,计算凋亡指数;免疫荧光双标法检测p-CREB表达水平;RT-PCR法半定量检测CREB下游基因BDNF mRNA及Bcl-2 mRNA表达水平;于首次注射后6周时采用Morris水迷宫实验测定各组大鼠认知功能.结果 与C组相比,K1组和K2组大鼠海马神经元凋亡指数升高,p-CREB、BDNF mRNA及Bcl-2 mRNA表达下调(P<0.05);与K1组相比,K2组大鼠海马神经元凋亡指数升高,p-CREB、BDNF mRNA及Bcl-2 mRNA表达下调(P<0.05);与C组和K1组相比,K2组逃避潜伏期延长(P<0.05).结论 氯胺酮10、20 mg/kg均可诱导发育期大鼠海马神经元凋亡,而氯胺酮20 mg/kg可导致大鼠发育成熟后认知功能降低,可能与其抑制CREB磷酸化后BDNF及Bcl-2表达下调,导致神经元凋亡,影响大鼠神经系统发育有关.     

氯胺酮麻醉及脾切除术对幼年大鼠学习记忆的影响

目的观察氯胺酮麻醉及脾切除术对幼年大鼠学习记忆的影响。方法健康幼年雄性SD大鼠30只,15日龄,体重29~31g,随机分为三组:生理盐水组(S组)、氯胺酮组(K组)、氯胺酮+脾切除组(KS组),三组大鼠腹腔分别给予等容量生理盐水0.3 ml、氯胺酮100 mg/kg(10mg/ml)、氯胺酮100mg/kg(10mg/ml),KS组大鼠氯胺酮麻醉后行脾切除。2周后三组大鼠进行Morris水迷宫实验,记录逃避潜伏期、Ⅱ象限穿越次数及Ⅱ象限游泳时间。水迷宫实验结束后杀死大鼠,采用ELISA法检测大鼠海马组织谷氨酸(Glu)和γ-氨基丁酸(GABA)含量。结果与S组比较,K组前3d和KS组前4d逃避潜伏期明显延长(P0.01),K组和KS组Ⅱ象限穿越次数明显减少,Ⅱ象限游泳时间明显缩短(P0.05);与K组比较,KS组逃避潜伏期明显延长,Ⅱ象限穿越次数明显减少,Ⅱ象限游泳时间明显缩短(P0.01)。与S组比较,K组和KS组Glu含量明显降低,Glu/GABA明显减小(P0.05),KS组GABA含量明显升高(P0.05);与K组比较,KS组Glu含量明显降低,Glu/GABA明显减小(P0.05)。结论氯胺酮麻醉及脾切除均能损伤幼年大鼠的学习记忆功能,且其记忆功能受损与海马区Glu含量及Glu/GABA下调有关。氯胺酮对幼年大鼠学习能力的损害只是暂时的,脾切除能延长并进一步损害幼年大鼠的学习能力。     

腹腔重复注射小剂量氯胺酮术后镇痛对趾部切口术大鼠认知功能的影响

目的观察腹腔重复注射小剂量氯胺酮术后镇痛对趾部切口术大鼠空间学习及记忆的影响。方法雄性3月龄SD大鼠72只,随机分为3组,对照组、模型组和氯胺酮组,每组又随机分为2个亚组(n=12),对照1周组(C1组)、模型1周组(M1组)、氯胺酮1周组(K1组)、对照3周组(C3组)、模型3周组(M3组)和氯胺酮3周组(K3组)。模型组和氯胺酮组按Brennan法制作趾部切口模型,对照组不作趾部切口;自趾部切口即日起,氯胺酮组腹腔注射氯胺酮10mg/kg,对照组和模型组腹腔注射生理盐水0.1ml,1次/d,连续7d。C1、M1、K1组于趾部切口术后1周后进行Morris水迷宫行为测试,C3、M3、K3组于趾部切口术后3周后进行水迷宫行为测试,4次/d,连续6d。大鼠的潜伏期和游泳距离由Morris水迷宫图像自动监视系统采集和分析。测试结束后,取大鼠海马组织作病理学观察。结果(1)水迷宫测试结果测试第1-6天大鼠的潜伏期和游泳距离:K1组均明显长于C1组和M1组(P<0.05或0.01),M1组与C1组比较差异无统计学意义;K3组均明显长于C3组和M3组(P<0.05或0.01),M3组与C3组比较差异无统计学意义。(2)病理学结果光镜下各组大鼠海马神经元结构未见明显异常。透射电镜下K1组和K3组大鼠海马神经元可见线粒体肿胀、溶酶体增多、粗面内质网扩张、出现空泡样改变。结论腹腔重复注射小剂量     

杭州健康女性定量骨超声测定原发性骨质疏松

目的 评价杭州健康女性骨超声速度(SOS)值随增龄减少和骨质疏松患病率，建立杭州地区女性骨超声速度值参考数据库。方法 定量超声法测定1208例杭州地区健康女性桡骨远端(RAD)，第3指骨近节(PLX)，第V跖骨(MTR)和胫骨中段(TIB)的超声速度值。结果 RAD、PLX、MTR和TIBSOS峰值(Peak of SOS)均出现在40-45岁，TJB的SOS峰值出现在35—40岁，此后随年龄增长而下降。绝经后妇女在绝经后早期和晚期各有1个SOS快速减少期，前见于桡骨近端，平均年减少率为2．4％，后见于胫骨中段，平均年减少率为1．8％。各部位骨SOS累积减少率随年龄增长而增加，到85岁4部位累积减少为13％-18％。60岁以后骨质疏松性症(OP)检出率为45％-70％，OP检出率以桡骨远端最高，60-70岁平均为67％，第3指骨近端次之约50％，胫骨中段最低为36％；75岁以后分别为70％，65％和45％。结论 全身各部位骨超声速度值到达峰值的年龄不同，峰值也各有差异。绝经后妇女骨超声速度值随年龄增加减少较快，应予激素和补钙治疗，桡骨远端为本地区SOS检测和OP检出的敏感部位。     

Importance of echocardiography in prognosis of surgical outcomes in cholecystitis in elderly patients

The authors propose to use more often echocardiography (EchoCG) in examination of elderly (over 60 years) of age patients with cholecystitis that permits to increase surgical activity to 92.4%. Left ventricular ejection fraction is the most informative. When this fraction is lower than 45% surgery must be recommended on vital indications only. EchoCG was used in 155 patients with cholecystitis, 131 of them were operated. 2 (1.52%) patients died due to acute cardio-vascular insufficiency and pulmonary artery thromboembolism.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

Objective To evaluate the role of gliocyte in the spinal cord in the development of bone cancer pain (BCP) in mice. Methods Forty male C3H/He mice aged 8-10 weeks weighing 18-22 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation (group S) , group II BCP, group Ⅲ PBS and group IV minocyline (group M) . In group BCP, PBS and M, bone cancer pain was produced by injection of NCTC2472 fibrosarcoma cell suspension (2 x 105 cells) 10 μl into medullary cavity of calcaneus bone, while in group S, PBS solution 10 μl was injected instead of cancer cell suspension. In group PBS and M, PBS 5 μl and minocyline 5 μl (dissolved to 0.2 mmol/L in PBS)_were given IT immediately before cancer cell inoculation once a day for 11 consecutive days respectively. Mechanical pain threshold was measured at 1 d before cancer cell inoculation, and at 0, 3, 5, 7, 9 and 11d after cancer cell inoculation. Cold pain threshold was measured at 3, 7, 9 and 11d after cancer cell inoculation. The animals were killed after measurement of pain threshold and L4-6, segment of spinal cord was removed for determination of GFAP and CD11b expression by Western blot. Results Compared with group S, mechanical pain threshold was significantly increased at 3-11 d after cancer cell inoculation in group BCP and PBS, and at 3 and S d after cancer cell inoculation in group M, and cold pain threshold was significantly increased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was up-regulated in group BCP, PBS and M ( P < 0.05) . Compared with group BCP, mechanical pain threshold was significantly decreased at 3-11 d after cancer cell inoculation, cold pain threshold was significantly decreased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was down-regulated in group M ( P <0.05) . ConclusionThe activiton of gliocyte in the spinal cord is involved in the development of bone cancer pian in mice.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

Objective To evaluate the role of gliocyte in the spinal cord in the development of bone cancer pain (BCP) in mice. Methods Forty male C3H/He mice aged 8-10 weeks weighing 18-22 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation (group S) , group II BCP, group Ⅲ PBS and group IV minocyline (group M) . In group BCP, PBS and M, bone cancer pain was produced by injection of NCTC2472 fibrosarcoma cell suspension (2 x 105 cells) 10 μl into medullary cavity of calcaneus bone, while in group S, PBS solution 10 μl was injected instead of cancer cell suspension. In group PBS and M, PBS 5 μl and minocyline 5 μl (dissolved to 0.2 mmol/L in PBS)_were given IT immediately before cancer cell inoculation once a day for 11 consecutive days respectively. Mechanical pain threshold was measured at 1 d before cancer cell inoculation, and at 0, 3, 5, 7, 9 and 11d after cancer cell inoculation. Cold pain threshold was measured at 3, 7, 9 and 11d after cancer cell inoculation. The animals were killed after measurement of pain threshold and L4-6, segment of spinal cord was removed for determination of GFAP and CD11b expression by Western blot. Results Compared with group S, mechanical pain threshold was significantly increased at 3-11 d after cancer cell inoculation in group BCP and PBS, and at 3 and S d after cancer cell inoculation in group M, and cold pain threshold was significantly increased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was up-regulated in group BCP, PBS and M ( P < 0.05) . Compared with group BCP, mechanical pain threshold was significantly decreased at 3-11 d after cancer cell inoculation, cold pain threshold was significantly decreased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was down-regulated in group M ( P <0.05) . ConclusionThe activiton of gliocyte in the spinal cord is involved in the development of bone cancer pian in mice.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

Objective To evaluate the role of gliocyte in the spinal cord in the development of bone cancer pain (BCP) in mice. Methods Forty male C3H/He mice aged 8-10 weeks weighing 18-22 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation (group S) , group II BCP, group Ⅲ PBS and group IV minocyline (group M) . In group BCP, PBS and M, bone cancer pain was produced by injection of NCTC2472 fibrosarcoma cell suspension (2 x 105 cells) 10 μl into medullary cavity of calcaneus bone, while in group S, PBS solution 10 μl was injected instead of cancer cell suspension. In group PBS and M, PBS 5 μl and minocyline 5 μl (dissolved to 0.2 mmol/L in PBS)_were given IT immediately before cancer cell inoculation once a day for 11 consecutive days respectively. Mechanical pain threshold was measured at 1 d before cancer cell inoculation, and at 0, 3, 5, 7, 9 and 11d after cancer cell inoculation. Cold pain threshold was measured at 3, 7, 9 and 11d after cancer cell inoculation. The animals were killed after measurement of pain threshold and L4-6, segment of spinal cord was removed for determination of GFAP and CD11b expression by Western blot. Results Compared with group S, mechanical pain threshold was significantly increased at 3-11 d after cancer cell inoculation in group BCP and PBS, and at 3 and S d after cancer cell inoculation in group M, and cold pain threshold was significantly increased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was up-regulated in group BCP, PBS and M ( P < 0.05) . Compared with group BCP, mechanical pain threshold was significantly decreased at 3-11 d after cancer cell inoculation, cold pain threshold was significantly decreased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was down-regulated in group M ( P <0.05) . ConclusionThe activiton of gliocyte in the spinal cord is involved in the development of bone cancer pian in mice.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

Objective To evaluate the role of gliocyte in the spinal cord in the development of bone cancer pain (BCP) in mice. Methods Forty male C3H/He mice aged 8-10 weeks weighing 18-22 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation (group S) , group II BCP, group Ⅲ PBS and group IV minocyline (group M) . In group BCP, PBS and M, bone cancer pain was produced by injection of NCTC2472 fibrosarcoma cell suspension (2 x 105 cells) 10 μl into medullary cavity of calcaneus bone, while in group S, PBS solution 10 μl was injected instead of cancer cell suspension. In group PBS and M, PBS 5 μl and minocyline 5 μl (dissolved to 0.2 mmol/L in PBS)_were given IT immediately before cancer cell inoculation once a day for 11 consecutive days respectively. Mechanical pain threshold was measured at 1 d before cancer cell inoculation, and at 0, 3, 5, 7, 9 and 11d after cancer cell inoculation. Cold pain threshold was measured at 3, 7, 9 and 11d after cancer cell inoculation. The animals were killed after measurement of pain threshold and L4-6, segment of spinal cord was removed for determination of GFAP and CD11b expression by Western blot. Results Compared with group S, mechanical pain threshold was significantly increased at 3-11 d after cancer cell inoculation in group BCP and PBS, and at 3 and S d after cancer cell inoculation in group M, and cold pain threshold was significantly increased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was up-regulated in group BCP, PBS and M ( P < 0.05) . Compared with group BCP, mechanical pain threshold was significantly decreased at 3-11 d after cancer cell inoculation, cold pain threshold was significantly decreased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was down-regulated in group M ( P <0.05) . ConclusionThe activiton of gliocyte in the spinal cord is involved in the development of bone cancer pian in mice.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

目的 评价脊髓胶质细胞在小鼠骨癌痛形成中的作用.方法 健康雄性C3H/He小鼠40只,周龄8～10周,体重18～22 g,随机分为4组(n=10):假手术组(S组)、骨癌痛组(B组)、PBS组(P组)和米诺环素组(M组).S组跟骨骨髓腔内注射PBS 10 μl;余3组跟骨骨髓腔内注射含2×105个骨纤维肉瘤细胞的PBS 10 μl制备骨癌痛模型,于造模前即刻开始PBS组鞘内注射PBS 5μl,M组鞘内注射米诺环素(用PBS溶解为0.2 mmol/L)5μl,1次/d,连续11 d.于造模前1 d、造模后即刻、3、5、7、9、11 d时测定机械痛阈;于造模后3、7、9、11 d机械痛阈测定结束后测定冷痛阈.痛阈测定结束后处死小鼠,取脊髓组织,测定神经胶质纤维酸性蛋白(GFAP)和CD11b的表达水平.结果 与S组比较,B组和P组造模后3-11 d时、M组造模后3、5 d时机械痛阈升高,B组、P组和M组造模后7～11 d时冷痛阈升高,脊髓CD11b和GFAP表达上调(P<0.05).与B组比较,M组造模后3-11 d时机械痛阈降低,造模后7-11 d时冷痛阈降低,脊髓CD11b和GFAP表达下调(P<0.05).结论 脊髓胶质细胞(星形胶质细胞和小胶质细胞)的激活参与了小鼠骨癌痛的形成.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

Objective To evaluate the role of gliocyte in the spinal cord in the development of bone cancer pain (BCP) in mice. Methods Forty male C3H/He mice aged 8-10 weeks weighing 18-22 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation (group S) , group II BCP, group Ⅲ PBS and group IV minocyline (group M) . In group BCP, PBS and M, bone cancer pain was produced by injection of NCTC2472 fibrosarcoma cell suspension (2 x 105 cells) 10 μl into medullary cavity of calcaneus bone, while in group S, PBS solution 10 μl was injected instead of cancer cell suspension. In group PBS and M, PBS 5 μl and minocyline 5 μl (dissolved to 0.2 mmol/L in PBS)_were given IT immediately before cancer cell inoculation once a day for 11 consecutive days respectively. Mechanical pain threshold was measured at 1 d before cancer cell inoculation, and at 0, 3, 5, 7, 9 and 11d after cancer cell inoculation. Cold pain threshold was measured at 3, 7, 9 and 11d after cancer cell inoculation. The animals were killed after measurement of pain threshold and L4-6, segment of spinal cord was removed for determination of GFAP and CD11b expression by Western blot. Results Compared with group S, mechanical pain threshold was significantly increased at 3-11 d after cancer cell inoculation in group BCP and PBS, and at 3 and S d after cancer cell inoculation in group M, and cold pain threshold was significantly increased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was up-regulated in group BCP, PBS and M ( P < 0.05) . Compared with group BCP, mechanical pain threshold was significantly decreased at 3-11 d after cancer cell inoculation, cold pain threshold was significantly decreased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was down-regulated in group M ( P <0.05) . ConclusionThe activiton of gliocyte in the spinal cord is involved in the development of bone cancer pian in mice.     

脊髓胶质细胞在小鼠骨癌痛形成中的作用

Objective To evaluate the role of gliocyte in the spinal cord in the development of bone cancer pain (BCP) in mice. Methods Forty male C3H/He mice aged 8-10 weeks weighing 18-22 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation (group S) , group II BCP, group Ⅲ PBS and group IV minocyline (group M) . In group BCP, PBS and M, bone cancer pain was produced by injection of NCTC2472 fibrosarcoma cell suspension (2 x 105 cells) 10 μl into medullary cavity of calcaneus bone, while in group S, PBS solution 10 μl was injected instead of cancer cell suspension. In group PBS and M, PBS 5 μl and minocyline 5 μl (dissolved to 0.2 mmol/L in PBS)_were given IT immediately before cancer cell inoculation once a day for 11 consecutive days respectively. Mechanical pain threshold was measured at 1 d before cancer cell inoculation, and at 0, 3, 5, 7, 9 and 11d after cancer cell inoculation. Cold pain threshold was measured at 3, 7, 9 and 11d after cancer cell inoculation. The animals were killed after measurement of pain threshold and L4-6, segment of spinal cord was removed for determination of GFAP and CD11b expression by Western blot. Results Compared with group S, mechanical pain threshold was significantly increased at 3-11 d after cancer cell inoculation in group BCP and PBS, and at 3 and S d after cancer cell inoculation in group M, and cold pain threshold was significantly increased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was up-regulated in group BCP, PBS and M ( P < 0.05) . Compared with group BCP, mechanical pain threshold was significantly decreased at 3-11 d after cancer cell inoculation, cold pain threshold was significantly decreased at 7-11 d after cancer cell inoculation, and expression of CD11b and GFAP was down-regulated in group M ( P <0.05) . ConclusionThe activiton of gliocyte in the spinal cord is involved in the development of bone cancer pian in mice.