国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2015年
3期
193-197,217
,共6页
目的 通过鞘内注射不同剂量驱动蛋白17(kinesin superfamily 17,KIF17)反义寡核苷酸,研究其在骨癌痛小鼠疼痛发生中的作用. 方法 55只雄性C3H/HeJ小鼠,4周～6周龄,体重20 g～25 g.用随机数字表法分为骨癌组(A组,41只)和假手术组(S组,14只).A组小鼠右侧股骨骨髓腔内注射20μl含约2×105个纤维肉瘤细胞(NCTC2472)的最小必须培养基(α-minimal essence medium,α-MEM)以制备骨癌痛模型,S组小鼠右侧股骨骨髓腔内注射等体积不含肿瘤细胞的α-MEM.各组小鼠于接种前(base)及接种后第4、7、10、14天进行痛行为学测试,包括自发抬足次数(the number of spontaneous flinches,NSF)和机械缩足反射阈值(paw withdrawal mechanical threshold,PWMT).每组每个时间点各取1只小鼠断头处死,取L3-5脊髓,用免疫印迹法(Western blot)测定KIF17蛋白含量.在接种后第14天,再将剩余A组小鼠按随机数字表法分为KIF17反义寡核苷酸2.5 μg组(A1组)、KIF17反义寡核苷酸5.0 μg组(A2组)、KIF17反义寡核苷酸10.0 μg组(A3组)以及对照组(AC组),每组9只.A1组～A3组小鼠分别鞘内给予相应剂量KIF17反义寡核苷酸,S组和AC组小鼠鞘内给予生理盐水,注药容积均为5μl.给药结束后2、12、24、36、48 h每个时间点取1只小鼠处死,再行痛行为学测试和脊髓水平KIF17蛋白含量. 结果 与S组[(2.15±0.49)次、(1.80±0.39)g]和基础值[(2.08±0.34)次、(1.88±0.35)g]比较,接种后第14天,A组小鼠NSF显著增加[(11.95±0.78)次],PWMT显著降低[(0.34±0.11)g](P＜0.05);同时A组小鼠脊髓水平KIF17表达增多;与AC组和给药前比较,A1组小鼠在给药后2、12h和24 h,NSF[2 h(4.80±0.40)次]减少,PWMT[2 h(0.95±0.33)g]升高(P＜0.05),A2组和A3组小鼠在给药后2、12、24 h和36 h,NSF[2 h(2.35±0.26)、(2.53±0.21)次]减少,PWMT[2 h(1.70±0.32)、(1.73±0.40)g]升高(P＜0.05),且与A1组比较,A2组和A3组小鼠痛行为学的改善更明显,A2组与A3组各时间点小鼠NSF和PWMT比较,差异无统计学意义(P＞0.05);给药后2h,A1组～A3组小鼠脊髓水平KIF17表达减少. 结论 骨癌痛小鼠脊髓水平KIF17表达上调,鞘内给予KIF17反义寡核苷酸能有效改善骨癌痛小鼠的痛行为学,其镇痛效果和作用时间与剂量有关.
目的 通過鞘內註射不同劑量驅動蛋白17(kinesin superfamily 17,KIF17)反義寡覈苷痠,研究其在骨癌痛小鼠疼痛髮生中的作用. 方法 55隻雄性C3H/HeJ小鼠,4週～6週齡,體重20 g～25 g.用隨機數字錶法分為骨癌組(A組,41隻)和假手術組(S組,14隻).A組小鼠右側股骨骨髓腔內註射20μl含約2×105箇纖維肉瘤細胞(NCTC2472)的最小必鬚培養基(α-minimal essence medium,α-MEM)以製備骨癌痛模型,S組小鼠右側股骨骨髓腔內註射等體積不含腫瘤細胞的α-MEM.各組小鼠于接種前(base)及接種後第4、7、10、14天進行痛行為學測試,包括自髮抬足次數(the number of spontaneous flinches,NSF)和機械縮足反射閾值(paw withdrawal mechanical threshold,PWMT).每組每箇時間點各取1隻小鼠斷頭處死,取L3-5脊髓,用免疫印跡法(Western blot)測定KIF17蛋白含量.在接種後第14天,再將剩餘A組小鼠按隨機數字錶法分為KIF17反義寡覈苷痠2.5 μg組(A1組)、KIF17反義寡覈苷痠5.0 μg組(A2組)、KIF17反義寡覈苷痠10.0 μg組(A3組)以及對照組(AC組),每組9隻.A1組～A3組小鼠分彆鞘內給予相應劑量KIF17反義寡覈苷痠,S組和AC組小鼠鞘內給予生理鹽水,註藥容積均為5μl.給藥結束後2、12、24、36、48 h每箇時間點取1隻小鼠處死,再行痛行為學測試和脊髓水平KIF17蛋白含量. 結果 與S組[(2.15±0.49)次、(1.80±0.39)g]和基礎值[(2.08±0.34)次、(1.88±0.35)g]比較,接種後第14天,A組小鼠NSF顯著增加[(11.95±0.78)次],PWMT顯著降低[(0.34±0.11)g](P＜0.05);同時A組小鼠脊髓水平KIF17錶達增多;與AC組和給藥前比較,A1組小鼠在給藥後2、12h和24 h,NSF[2 h(4.80±0.40)次]減少,PWMT[2 h(0.95±0.33)g]升高(P＜0.05),A2組和A3組小鼠在給藥後2、12、24 h和36 h,NSF[2 h(2.35±0.26)、(2.53±0.21)次]減少,PWMT[2 h(1.70±0.32)、(1.73±0.40)g]升高(P＜0.05),且與A1組比較,A2組和A3組小鼠痛行為學的改善更明顯,A2組與A3組各時間點小鼠NSF和PWMT比較,差異無統計學意義(P＞0.05);給藥後2h,A1組～A3組小鼠脊髓水平KIF17錶達減少. 結論 骨癌痛小鼠脊髓水平KIF17錶達上調,鞘內給予KIF17反義寡覈苷痠能有效改善骨癌痛小鼠的痛行為學,其鎮痛效果和作用時間與劑量有關.
목적 통과초내주사불동제량구동단백17(kinesin superfamily 17,KIF17)반의과핵감산,연구기재골암통소서동통발생중적작용. 방법 55지웅성C3H/HeJ소서,4주～6주령,체중20 g～25 g.용수궤수자표법분위골암조(A조,41지)화가수술조(S조,14지).A조소서우측고골골수강내주사20μl함약2×105개섬유육류세포(NCTC2472)적최소필수배양기(α-minimal essence medium,α-MEM)이제비골암통모형,S조소서우측고골골수강내주사등체적불함종류세포적α-MEM.각조소서우접충전(base)급접충후제4、7、10、14천진행통행위학측시,포괄자발태족차수(the number of spontaneous flinches,NSF)화궤계축족반사역치(paw withdrawal mechanical threshold,PWMT).매조매개시간점각취1지소서단두처사,취L3-5척수,용면역인적법(Western blot)측정KIF17단백함량.재접충후제14천,재장잉여A조소서안수궤수자표법분위KIF17반의과핵감산2.5 μg조(A1조)、KIF17반의과핵감산5.0 μg조(A2조)、KIF17반의과핵감산10.0 μg조(A3조)이급대조조(AC조),매조9지.A1조～A3조소서분별초내급여상응제량KIF17반의과핵감산,S조화AC조소서초내급여생리염수,주약용적균위5μl.급약결속후2、12、24、36、48 h매개시간점취1지소서처사,재행통행위학측시화척수수평KIF17단백함량. 결과 여S조[(2.15±0.49)차、(1.80±0.39)g]화기출치[(2.08±0.34)차、(1.88±0.35)g]비교,접충후제14천,A조소서NSF현저증가[(11.95±0.78)차],PWMT현저강저[(0.34±0.11)g](P＜0.05);동시A조소서척수수평KIF17표체증다;여AC조화급약전비교,A1조소서재급약후2、12h화24 h,NSF[2 h(4.80±0.40)차]감소,PWMT[2 h(0.95±0.33)g]승고(P＜0.05),A2조화A3조소서재급약후2、12、24 h화36 h,NSF[2 h(2.35±0.26)、(2.53±0.21)차]감소,PWMT[2 h(1.70±0.32)、(1.73±0.40)g]승고(P＜0.05),차여A1조비교,A2조화A3조소서통행위학적개선경명현,A2조여A3조각시간점소서NSF화PWMT비교,차이무통계학의의(P＞0.05);급약후2h,A1조～A3조소서척수수평KIF17표체감소. 결론 골암통소서척수수평KIF17표체상조,초내급여KIF17반의과핵감산능유효개선골암통소서적통행위학,기진통효과화작용시간여제량유관.
Objective To investigate the effect of different doses of kinesin superfamily protein 17 (KIF17) antisense oligodeoxy nucleotide on a mouse model of bone cancer pain.Methods Fifty-five C3H/HeJ mice,4-6 weeks of age,weighting 20 g～25 g,were randomly divided into two groups:bone cancer pain group (group A,n=41) and sham operation group (group S,n=14).Group A was induced by implantation of 20 μl α-minimal essence medium (α-MEM) which containing 2×105 osteosarcoma (NCTC 2472) cells into the intramedullary space of the right femur.And 20 μl α-MEM with no cancer cell was injected instead to group S.The number of spontaneous flinches (NSF) and the paw withdrawal mechanical threshold (PWMT) were measured at the day before inoculation (base) and the day 4,7,10 and 14 after inoculation.According to the corresponding time points,ten mice were sacrificed,and the lumbar segment of spinal cord(L3-5) was removed for determination the expression of KIF17 using Western blot.At the day 14 after inoculation,the mice of group A were randomly divided into four groups,KIF17 antisense oligodeoxy nucleotide 2.5,5.0,10.0 μg group (group A1-group A3) and the control group (group Ac).Each group had nine mice.group A1-group A3 were treated by intrathecal injection the corresponding dose of KIF17 antisense oligodeoxy nucleotide group S and group AC was treated by saline.The volume of all drugs was 5 μl.The pain behaviors were measured at 2,12,24,36 h and 48 h after administration and determinated the expression of KIF17.Results Compared with group S [(2.15±0.49),(1.80±0.39) g] and the base[(2.08±0.34),(1.88±0.35) g],the NSF[(11.95±0.78)] was significantly increased and PWMT[(0.34±0.11) g] was decreased at 14 d after inoculation in the bone caner pain groups(P＜0.05),the expression of KIF17 were increased,compared with group AC,the NSF [2 h (4.80±0.40)] was decreased and the PWMT [2 h (0.95±0.33) g] was increased at 2,12,24 h after administration in group A1 (P＜0.05),and the NSF[2 h(2.35±0.26),(2.53±0.21)] was decreased and the PWMT[2 h(1.70±0.32),(1.73±0.40) g] was increased at 2,12,24,36 h after administration in group A2-group A3,group A2 compared with group A3,the NSF and PWMT were no statistically significant difference (P＞0.05),after intrathecal administration,the expression of KIF17 was reduced in group A1-group A3 (P＜0.05).Conclusions Intrathecal KIF17 antisense oligodeoxy nucleotide in the mice of bone cancer pain improve the pain behaviors,and the analgesic effect is time-and dose-related.
