缺氧诱导基因-1α和gax基因共转染对移植静脉内膜重塑态的效应与分子机制

目的 探讨缺氧诱导基因(HIF-1α)和同源盒基因(gax)过表达对移植静脉内膜过度增生的影响及其机制.方法 自体静脉移植术大鼠28只均分为基因转染组和非基因转染组.于核酸、蛋白、细胞超微结构及细胞形态层次分析移植静脉中HIF-1α和gax基因表达与血管平滑肌细胞(VSMC)表型、中膜厚度等指标.结果 基因转染组与非基因转染组比较:HIF-1α和gax的mRNA及其蛋白表达增强(P＜0.05);增殖型VSMC和增殖细胞核抗原(PCNA)阳性细胞减少(P＜0.05);TUNEL阳性细胞增多(P＜0.05);内皮修复较显著,较多肌-内皮连接结构形成;内膜过度增生程度有所减弱.结论 HIF-1α和gax基因联合转染对移植静脉内膜过度增生具有一定的抑制作用.     

重组腺相关病毒介导HIF-1α基因对移植静脉再内皮化的影响

目的　探讨缺氧诱导因子 1α(HIF 1α)过表达对大鼠移植静脉再内皮化的影响。方法　56只大鼠,随机分为实验组和对照组,均行自体右颈内静脉移植至同侧颈总动脉手术,实验组移植静脉经HIF 1α重组腺病毒工作液预处理。分别于术后7 d或14 d切取移植静脉和血清。检测和观察组织标本中HIF 1αmRNA、血管内皮细胞生长因子(VEGF)、E 选择素(E selectin)的表达及内皮细胞再内皮化的情况。结果　实验组与对照组比较,HIF 1αmRNA表达增强(P<0 01);HIF 1α和VEGF蛋白表达明显增加(P<0 01);血清E selectin含量明显降低(P<0 05)。内皮细胞增殖和肌内皮连接重建显著。结论　移植静脉重塑早期HIF 1α过表达可促进受损血管内皮细胞(VEC)结构和功能的重建,加速再内皮化进程。     

STAT3信号通路与血管重塑中平滑肌细胞表型转化及增殖关系

目的 观察静脉桥再狭窄模型中血管平滑肌细胞(VSMC)表型转化和增殖活性的变化,探讨信号转导子和激活子3蛋白(STAT3)的表达与VSMC表型转化及增殖活性的关系.方法 建立猪静脉桥再狭窄模型,采用血管病理形态学、免疫组织化学和免疫印迹(Western blot)方法,观察术后7、14、30 d血管蓖塑及血管壁中增殖细胞核抗原(PCNA)、平滑肌ot肌动蛋白(SM-α actin)和STAT3表达变化及其相关性.结果 (1)术后7 d新牛内膜形成逐渐增厚,于术后30 d达最大;重塑指数和外弹力板围绕面积(EELA)术后7 d稍有增大,其后不断减小,术后14～30 d明显减小(P<0.05).(2)免疫组织化学和Western blot测定STAT3蛋白显示,术后7 d中膜VSMC中阳性表达明显,术后14 d中膜VSMC和内膜VSMC中阳性表达均增加达高峰;术后30 d中膜VSMC中有较少部分阳性表达,内膜VSMC中阳性表达较14 d减少.血管中膜中STAT3和PCNA的蛋白呈显著性正相关(r=0.726,P<0.05).结论 血管平滑肌细胞的表型转化和增殖活性改变对内膜增生和血管重塑起着重要作用,STAT3信号通路与血管重塑中VSMC的增殖高度相关,参与并促进了VSMC的表型转化和增殖.     

脂质体与腺病毒转染EDRz抑制自体移植静脉内膜增生的比较研究

目的探讨以脂质体和腺病毒为载体,早期生长反应基因-1 DNA酶(Egr-1 DNA enzyme,EDRz)局部外用对自体移植静脉血管平滑肌(VSMC)增殖和内膜增生的抑制作用。方法大鼠建立自体静脉移植模型后随机分为脂质体组、腺病毒组和对照组,两实验组分别用脂质体-EDRz和腺病毒-EDRz涂抹移植静脉行在体转染。于术后1,2,6,24 h及3,7,14,28 d取材,荧光显微镜检测移植静脉的转染情况;采用原位杂交方法检测Egr-1 mRNA的表达;用免疫组织化学方法检测Egr-1蛋白表达,同时进行组织形态学观察。结果术后1 h,EDRz主要位于移植静脉的外膜、中膜和部分内皮细胞。脂质体组荧光灰度值为70.3±13.5,腺病毒组荧光灰度值为60.5±11.2。术后2～24 h,EDRz主要位于移植静脉的中膜;移植7 d,EDRz主要位于移植静脉的内膜。移植早期Egr-1蛋白表达主要位于中膜的血管平滑肌细胞(VSMC)和部分单核细胞、内皮细胞;移植后期未检测到Egr-1 DNA酶的表达。移植后期在中膜和新生内膜的VSMC均未检测到Egr-1蛋白的表达。移植2 h阳性表达率,脂质体组为(15.3±4.2)%,腺病...     

反义c-fos核酸对自体移植静脉内膜增生的影响

目的:利用反义核酸技术和显微外科技术,探讨反义c-fos核酸对自体移植静脉内膜增生的影响.方法:选择20只新西兰家免,等分为实验组和对照组,均行自体颈外静脉、颈总动脉移植手术,实验组移植静脉周围和血管吻合口周围应用反义c-fos核酸凝胶涂布;对照组仅行凝胶涂布.术后2周取出移植血管,分别行病理学、免疫组织化学检测移植血管内膜厚度,内膜平滑肌细胞数及PCNA阳性表达情况.结果:实验组移植血管内膜厚度、管腔狭窄度及VSMC数均较对照组减少,PCNA阳性表达情况亦较对照组明显减少.结论:反义c-fos核酸可有效的抑制移植静脉内膜的增生,是一种比较有发展前途的防治移植静脉内膜增生的基因疗法.     

RhoA/SRF信号通路介导Nelin诱导人血管平滑肌细胞表型转化

目的 探讨Nelin基因对人血管平滑肌细胞(vascular smooth muscle cell,VSMC)表型转化的调控机制.方法 应用过表达慢病毒载体[Nelin-VSMC]及干扰慢病毒载体[LV-Nelin-SiRNA-VSMC]制备稳定转染的VSMC细胞模型,通过荧光定量PCR以及蛋白质印迹分析(Western blotanalysis,WB)等技术手段,观察Nelin蛋白过表达及表达抑制对人VSMC表型转化的影响及其调控机制.结果 Nelin-VSMC组细胞呈收缩表型,细胞细长,成“谷-峰”样生长,Nelin mRNA、Nelin蛋白和平滑肌分化标志蛋白平滑肌α-肌动蛋白(SMoα-actin)表达显著增加,同时SMα-actin相关调控因子-血清反应因子(SRF)入核转位,RhoA总蛋白表达上调,经Rho激酶特异性抑制剂Y-27632作用后,SRF在细胞核中表达显著减少,同时SMα-actin表达下调;LV-Nelin-SiRNA-VSMC组细胞呈合成表型,细胞体积变大,细胞极性消失,生长状态无序,Nelin mRNA、Nelin蛋白和SMα-actin蛋白表达显著降低,同时伴有SRF出核转位及RhoA总蛋白表达下调.结论 在体外培养的人VSMC中,Nelin依次激活SMα-actin蛋白相关调控因子RhoA和SRF引起SMα-actin表达增加,促进VSMC向收缩表型转换.     

缺氧诱导因子-1α与同源盒基因共转染在移植静脉重塑态的表达

目的 探讨缺氧诱导因子(HIF-1α)和同源盒基因(gax)共转染在移植静脉组织中的表达状态.方法 Wistar大鼠16只随机分为实验组和对照组,每组8只,均行自体静脉移植术,实验组移植静脉行缺氧诱导因子(HIF-1α)和同源盒基因(gax)共转染,对照组则未行基因转染,于术后14 d取出移植的静脉,分别采用逆转录-聚...     

反义雷帕霉素靶蛋白基因局部转染对移植静脉内膜增生的影响

目的:观察以纳米粒子为载体的反义雷帕霉素靶蛋白(mTOR)基因局部转染对移植静脉内膜增生的影响。方法:应用聚乳酸聚乙醇酸共聚物(PLGA)和聚乙烯醇(PVA)包载mTOR基因,制备纳米级粒子混合物。检测其包埋率、体外释放情况及粒子大小。建立自体静脉移植模型,随机分成转基因组、空载体组、对照组。转基因组移植静脉转染以纳米粒子为载体的反义mTOR基因,空载体组单纯转染纳米粒子包载的空载体,对照组不予特殊处理。分别于术后3d、7d、14d、28d取材,常规HE、Verhoeff染色,RT-PCR、Westernblot检测mTOR基因的mRNA及蛋白的变化,TUNEL法观察血管平滑肌细胞(VSMC)凋亡的动态变化。结果:转基因组内膜中mTOR基因的mRNA及蛋白产物表达较其他两组明显减少(P<0.05);转基因组内膜增生厚度7d、14d、28d较其他组明显减少(P<0.01);转基因组凋亡细胞较其他组明显增高(P<0.05)。结论:纳米粒子可以作为转基因载体,反义mTOR基因的表达能够有效抑制自体移植静脉内膜的增生,促进VSMC的凋亡。     

联合转染eNOS基因反义ET核酸对自体移植静脉内膜增生的影响

目的;探讨联合转染eNOS基因和反义ET核酸对自体移植静脉内膜增生的影响。方法：制作20只自体颈静脉腹主动脉移植Wistar大鼠模型,实验组,对照组各10只,实验组移植血管行腺病毒介导的eNOS溶液浸泡和反义ET核酸凝胶涂布,对照组仅行空载腺病毒溶液浸泡和凝胶兴布。术后2周取出移植血管,利用病理学,免疫组织化学,RT－PCR方法检测移植血管内膜厚度,管腔狭窄度,内膜VSMC数及PCNA阳性表达,血管ETmRNA,eNOSmRNA表达情况。结果：实验组移植血管内膜厚度,管腔狭窄及VSMC数均较对照组减小或减少,PCNA阳性表达及ETmRNA表达较对照组减少,而eNOSmRNA表达则明显增加。结论;联合转染NOS基因和反义ET核酸可有效地抑制移植静脉内膜的增生,是一种有效地防治移植静脉再狭窄的基因疗法。     

平滑肌22α启动子/增强子靶向调控哺乳动物雷帕霉素靶蛋白信号通路对移植血管平滑肌细胞增殖的影响

目的 采用血管平滑肌细胞(SM)特异性SM22α启动子/平滑肌肌球重链蛋白(SMHC)增强子,构建靶向大鼠哺乳动物雷帕霉素靶蛋白(mTOR)基因的干扰RNA真核表达载体,研究其对血管平滑肌细胞(VSMC)增殖及移植血管新生内膜增生的影响.方法 构建大鼠特异性SM22α真核表达载体SM22α-p/e-mTOR-短发卡RNA(shRNA),建立大鼠颈静脉-动脉移植模型,在血管吻合完成后通过Pluronic F-127质粒缓释系统对血管内平滑肌增殖进行局部RNA干扰.实验分5组,每组20只SD大鼠.A组:对照组(25％ Pluronic F-127组);B组:SM22α组(SM22α-p/e-mTOR-shRNA);C组:巨细胞病毒(CMV)组(CMV-p/e-mTOR-shRNA);D组:阴性对照组(空质粒pGenesil-10);E组:阳性对照组;其中,B组和C组统称实验组.根据实验分组的不同,将含有50 μg shRNA质粒,或50 μg渥曼青霉素(wortmannin)凝胶,或单纯200μl 25％ Pluronic F-127凝胶均匀涂抹在移植静脉周围,分别于术后1、3、7、14 d获取移植血管.苏木素-伊红(HE)染色观察新生内膜厚度;免疫组织化学检测平滑肌肌动蛋白(α-SM-actin)、增殖细胞核抗原(PCNA)、磷酸化-mTOR (Ser2448),原位末端转移酶标记技术(TUNEL)检测细胞凋亡,评估mTOR信号通路的变化以及平滑肌细胞的增殖.结果 术后7d各组新生内膜厚度差异有统计学意义(P＜0.05);术后14 d,A组新生内膜较术后1d增厚12.4倍;B、C、D、E组分别增厚9.7、4.8、7.6、2.6倍.术后14d,各实验组和阳性对照组移植静脉α-SM-actin表达阳性面积均明显低于对照组(P＜0.05),PCNA阳性表达面积均低于对照组(P＜0.05),磷酸化-mTOR(Ser2448)阳性表达面积均低于对照组(P＜0.05);实验组、对照组、阴性对照组术后移植静脉凋亡细胞阳性面积比较差异无统计学意义(P＞0.05),阳性对照组凋亡面积明显高于其他组(P＜0.05).结论 SM22α-p/e-mTOR-shRNA可通过抑制血管平滑肌细胞mTOR信号通路而抑制血管平滑肌细胞增殖,预防或延缓移植静脉狭窄.     

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.