中华物理医学与康复杂志
中華物理醫學與康複雜誌
중화물리의학여강복잡지
CHINESE JOURNAL OF PHYSICAL MEDICINE AND REHABILITATION
2013年
3期
161-166
,共6页
脊髓压迫性损伤%脱髓鞘病变%电针%超微结构%髓鞘碱性蛋白
脊髓壓迫性損傷%脫髓鞘病變%電針%超微結構%髓鞘堿性蛋白
척수압박성손상%탈수초병변%전침%초미결구%수초감성단백
Compression injury%Spinal cord%Demyelination%Electro-acupuncture%Ultrastructure%Inhibitor of DNA binding 2%Myelin basic protein
目的 观察电针对大鼠脊髓压迫性损伤(CSCI)后DNA结合抑制物2(Id2)和髓鞘碱性蛋白(MBP)的表达变化,探讨髓鞘再生的机制.方法 将54只SD大鼠分为对照组和治疗组,每组27只,再根据取材时间点的不同各分为3d、7d和14 d三个亚组,每个亚组9只大鼠.治疗组采用自行设计的大鼠脊髓压通器制作脊髓压通性损伤模型,针刺脊髓损伤节段局部夹脊穴、双侧足三里和双侧太溪穴,并于双侧足三里和太溪穴进行电针刺激(连续波,输出频率为2 Hz,电压1.5V,30 min).对照组只造模,不治疗.2组大鼠均于对应时间点取材,运用电镜观察脊髓损伤节段髓鞘的超微结构;采用免疫印迹技术和免疫荧光双标从分子水平检测Id2及MBP的表达变化.结果 CSCI后,荧光双标结果显示,造模后第3天,对照组大鼠的Id2免疫阳性少突胶质细胞数目为(20±2)个/高倍镜视野,并在造模后第14天下降至(16±1)个/高倍镜视野,组内各时间点Id2免疫阳性少突胶质细胞数目比较,差异无统计学意义(P＞0.05).造模后第3天,治疗组大鼠Id2免疫阳性少突胶质细胞为(13±1)个/高倍镜视野,造模后第14天降至(7±1)个/高倍镜视野,组内各时间点Id2免疫阳性少突胶质细胞数目比较,差异有统计学意义(P＜0.05),并与同时间点的对照组比较,差异亦有统计学意义(P＜0.05).Western结果显示,造模后第3天,对照组和治疗组Id2蛋白表达分别为(1.12±0.12)和(0.67±0.01),造模后第14天,对照组和治疗组Id2蛋白表达分别下降至(0.86±0.02)和(0.25±0.01),与组内造模后第3天比较,差异均有统计学意义(P＜0.05),2组相同时间点Id2蛋白表达组间比较,差异有统计学意义(P＜0.05).造模后第3天,对照组和治疗组的MBP蛋白表达分别为(0.44±0.02)和(0.67±0.04),造模后第14天,对照组和治疗组MBP蛋白表达均分别上调至(0.95±0.04)和(1.74±0.09),与组内造模后第3天比较,差异均有统计学意义(P＜0.05),2组相同时间点的MBP蛋白表达比较,差异有统计学意义(P＜0.05).结论 电针刺激可调节Id2的表达从而负向调控MBP的表达.
目的 觀察電針對大鼠脊髓壓迫性損傷(CSCI)後DNA結閤抑製物2(Id2)和髓鞘堿性蛋白(MBP)的錶達變化,探討髓鞘再生的機製.方法 將54隻SD大鼠分為對照組和治療組,每組27隻,再根據取材時間點的不同各分為3d、7d和14 d三箇亞組,每箇亞組9隻大鼠.治療組採用自行設計的大鼠脊髓壓通器製作脊髓壓通性損傷模型,針刺脊髓損傷節段跼部夾脊穴、雙側足三裏和雙側太溪穴,併于雙側足三裏和太溪穴進行電針刺激(連續波,輸齣頻率為2 Hz,電壓1.5V,30 min).對照組隻造模,不治療.2組大鼠均于對應時間點取材,運用電鏡觀察脊髓損傷節段髓鞘的超微結構;採用免疫印跡技術和免疫熒光雙標從分子水平檢測Id2及MBP的錶達變化.結果 CSCI後,熒光雙標結果顯示,造模後第3天,對照組大鼠的Id2免疫暘性少突膠質細胞數目為(20±2)箇/高倍鏡視野,併在造模後第14天下降至(16±1)箇/高倍鏡視野,組內各時間點Id2免疫暘性少突膠質細胞數目比較,差異無統計學意義(P＞0.05).造模後第3天,治療組大鼠Id2免疫暘性少突膠質細胞為(13±1)箇/高倍鏡視野,造模後第14天降至(7±1)箇/高倍鏡視野,組內各時間點Id2免疫暘性少突膠質細胞數目比較,差異有統計學意義(P＜0.05),併與同時間點的對照組比較,差異亦有統計學意義(P＜0.05).Western結果顯示,造模後第3天,對照組和治療組Id2蛋白錶達分彆為(1.12±0.12)和(0.67±0.01),造模後第14天,對照組和治療組Id2蛋白錶達分彆下降至(0.86±0.02)和(0.25±0.01),與組內造模後第3天比較,差異均有統計學意義(P＜0.05),2組相同時間點Id2蛋白錶達組間比較,差異有統計學意義(P＜0.05).造模後第3天,對照組和治療組的MBP蛋白錶達分彆為(0.44±0.02)和(0.67±0.04),造模後第14天,對照組和治療組MBP蛋白錶達均分彆上調至(0.95±0.04)和(1.74±0.09),與組內造模後第3天比較,差異均有統計學意義(P＜0.05),2組相同時間點的MBP蛋白錶達比較,差異有統計學意義(P＜0.05).結論 電針刺激可調節Id2的錶達從而負嚮調控MBP的錶達.
목적 관찰전침대대서척수압박성손상(CSCI)후DNA결합억제물2(Id2)화수초감성단백(MBP)적표체변화,탐토수초재생적궤제.방법 장54지SD대서분위대조조화치료조,매조27지,재근거취재시간점적불동각분위3d、7d화14 d삼개아조,매개아조9지대서.치료조채용자행설계적대서척수압통기제작척수압통성손상모형,침자척수손상절단국부협척혈、쌍측족삼리화쌍측태계혈,병우쌍측족삼리화태계혈진행전침자격(련속파,수출빈솔위2 Hz,전압1.5V,30 min).대조조지조모,불치료.2조대서균우대응시간점취재,운용전경관찰척수손상절단수초적초미결구;채용면역인적기술화면역형광쌍표종분자수평검측Id2급MBP적표체변화.결과 CSCI후,형광쌍표결과현시,조모후제3천,대조조대서적Id2면역양성소돌효질세포수목위(20±2)개/고배경시야,병재조모후제14천하강지(16±1)개/고배경시야,조내각시간점Id2면역양성소돌효질세포수목비교,차이무통계학의의(P＞0.05).조모후제3천,치료조대서Id2면역양성소돌효질세포위(13±1)개/고배경시야,조모후제14천강지(7±1)개/고배경시야,조내각시간점Id2면역양성소돌효질세포수목비교,차이유통계학의의(P＜0.05),병여동시간점적대조조비교,차이역유통계학의의(P＜0.05).Western결과현시,조모후제3천,대조조화치료조Id2단백표체분별위(1.12±0.12)화(0.67±0.01),조모후제14천,대조조화치료조Id2단백표체분별하강지(0.86±0.02)화(0.25±0.01),여조내조모후제3천비교,차이균유통계학의의(P＜0.05),2조상동시간점Id2단백표체조간비교,차이유통계학의의(P＜0.05).조모후제3천,대조조화치료조적MBP단백표체분별위(0.44±0.02)화(0.67±0.04),조모후제14천,대조조화치료조MBP단백표체균분별상조지(0.95±0.04)화(1.74±0.09),여조내조모후제3천비교,차이균유통계학의의(P＜0.05),2조상동시간점적MBP단백표체비교,차이유통계학의의(P＜0.05).결론 전침자격가조절Id2적표체종이부향조공MBP적표체.
Objective To observe the effects of electro-acupuncture on the expression and inhibition of DNA binding protein 2 (Id2) and myelin basic protein (MBP),and to explore the mechanism of remyelinization after compressive spinal cord injury (CSCI) in rats.Methods Fifty-four SD rats were randomly divided into a control group and a treatment group,and each was further subdivided into 3 time point subgroups:3,7 and 14 days.There were 9 rats in each subgroup.The CSCI models were made with a self-designed method.The acupuncture points Jiaji (EX-B2),bilateral Zusanli (ST36) and Taixi (KI3) were selected for treatment.Electro-acupuncture (continuous wave,2 Hz,1.5 V)was applied to the bilateral Zusanli (ST36) and Taixi (KI3) points.The control group received the injury but no treatment.The changes in the ultrastucture of the nerve fibers＇ white matter were de-termined by transmission electron microscopy (TEM).The alterations in the expression of MBP and Id2 were observed by double labeled immunofluorescence and Western blotting on the 3rd,7th and 14th day after the injury.Results TEM showed that the myelin sheaths in the control group had degenerated,swollen,and even broken down after CSCI.Changes to the myelin sheaths in the treatment group were milder than those in the control group.The immunofluorescence results showed the amount of Id2-immunoreactive oligodendrocytes in the control group to be (20 ±2) on the 3rd day after CSCI,becoming (16 ± 1) on the 14th day.The differences among the 3 control subgroups were not statistically significant.The amount of Id2-immunoreactive oligodendrocytes in the treatment group was (13 ± 1) on the 3rd day,reaching a minimum the 14th day.The differences among the 3 treatment groups were statistically significant.The differences compared with the control group at the same time points were also statistically significant.Western blotting showed that the expression of Id2 in the contrast and treatment groups was (1.12 ±0.12) and (0.67 ±0.01) respectively on the 3rd day after CSCI,and both decreased with time.The expression of Id2 in both groups reached their minima ((0.86 ±0.02) and (0.25 ±0.01) respectively) on the 14th day.The difference between the treatment groups and the contrast group was statistically significant at each time point.The expression of MBP in the contrast and treatment groups at day 3 was (0.44 ± 0.02) and (0.67 ± 0.04) respectively,and these increased with time.The expression of MBP in both groups peaked at the 14th day (at (0.95 ± 0.04) and (1.74 ± 0.09) respectively).These differences were again statistically significant.Conclusion Electro-acupuncture can regulate the expression of Id2 and MBP after CSCI.The down-regulation of Id2 which controls MBP negatively and the up-regulation of MBP may contribute to remyelination in the injured spinal cord.
