背景:脑内肿瘤坏死因子α可通过诱导脑缺血再灌注时内皮细胞黏附分子的表达促进炎症反应及血栓形成.目的:观察头穴百会透曲鬓对缺血性脑损伤的良性调节,以及对脑内肿瘤坏死因子α过度表达的抑制作用.设计:随机对照实验.单位:青岛大学医学院附属医院急诊神经科.材料:实验于2002-08/2003-08在青岛大学医学院脑血管病研究所完成.清洁级雌性SD大鼠15只,体质量230～270 g,随机分为假手术组、针刺组和对照组,5只/组.干预:①针刺组和对照组建立局灶性脑缺血再灌注模型,假手术组手术步骤同对照组,但不阻塞大脑中动脉.针刺组在缺血后10min给予针刺治疗,在大鼠头顶部相当于人体百会、风府穴常规剪毛消毒,用0.35 mmx40 mm毫针在皮下百会穴处左前下方相当于人体曲鬓穴处平刺,进针深度约0.8 sm.另于风府穴直刺一针,深度约0.5 cm.连接全能脉冲电疗仪,百会穴接阳极,风府穴接阴极,电针频率7Hz,强度6mA,时间30min.②脑片制备及肿瘤坏死因子α原位杂交检测:各组大鼠于缺血2 h再灌注12 h后深麻醉,左心室插管经升主动脉快速灌注生理盐水200 mL,再持续灌注质量浓度40 g/L多聚甲醛300mL,断头取脑,相同固定液后固定1 h.自视交叉处开始向后取冠状切片,片厚约7μm.肿瘤坏死因子α阳性细胞胞浆着色呈棕黄色,阴性对照细胞不着色.应用VIDAS-21图像分析系统进行图像分析处理,结果用吸光度表示.③缺血脑组织病理学观察:常规苏木精-伊红染色,光镜下观察针刺组和对照组病变区病理变化.主要观察指标:①各组脑缺血再灌注后脑皮层及纹状体内肿瘤坏死因子α的表达.②针刺组和对照组缺血脑组织病理变化.结果:15只大鼠全部进入结果分析.①脑缺血再灌注大鼠脑皮质及纹状体内肿瘤坏死因子α的表达:与假手术组比较,对照组和针刺组表达明显增强(0.302±0.04,0.320±0.02;0.466±0.08,0.423±0.02;0.367±0.03,0.362±0.02;P＜0.05);与对照组比较,针刺组表达明显降低(P＜0.05).②缺血脑组织病理变化:对照组脑组织变性坏死程度较重,神经元变性坏死,胶质细胞呈空泡样变性,血管内有明显的白细胞聚集及附壁现象,血管周围亦有明显白细胞浸润;针刺组仅有少量神经元变性坏死及胶质细胞空泡样变性,血管内白细胞聚集减少.结论:脑缺血再灌注后脑皮层及纹状体内肿瘤坏死因子α水平升高,头曲鬓穴处平刺可明显抑制其表达,从而有效阻止炎性细胞因子过量表达及其生物学作用的发挥,减少脑缺血后的炎性损伤.
揹景:腦內腫瘤壞死因子α可通過誘導腦缺血再灌註時內皮細胞黏附分子的錶達促進炎癥反應及血栓形成.目的:觀察頭穴百會透麯鬢對缺血性腦損傷的良性調節,以及對腦內腫瘤壞死因子α過度錶達的抑製作用.設計:隨機對照實驗.單位:青島大學醫學院附屬醫院急診神經科.材料:實驗于2002-08/2003-08在青島大學醫學院腦血管病研究所完成.清潔級雌性SD大鼠15隻,體質量230～270 g,隨機分為假手術組、針刺組和對照組,5隻/組.榦預:①針刺組和對照組建立跼竈性腦缺血再灌註模型,假手術組手術步驟同對照組,但不阻塞大腦中動脈.針刺組在缺血後10min給予針刺治療,在大鼠頭頂部相噹于人體百會、風府穴常規剪毛消毒,用0.35 mmx40 mm毫針在皮下百會穴處左前下方相噹于人體麯鬢穴處平刺,進針深度約0.8 sm.另于風府穴直刺一針,深度約0.5 cm.連接全能脈遲電療儀,百會穴接暘極,風府穴接陰極,電針頻率7Hz,彊度6mA,時間30min.②腦片製備及腫瘤壞死因子α原位雜交檢測:各組大鼠于缺血2 h再灌註12 h後深痳醉,左心室插管經升主動脈快速灌註生理鹽水200 mL,再持續灌註質量濃度40 g/L多聚甲醛300mL,斷頭取腦,相同固定液後固定1 h.自視交扠處開始嚮後取冠狀切片,片厚約7μm.腫瘤壞死因子α暘性細胞胞漿著色呈棕黃色,陰性對照細胞不著色.應用VIDAS-21圖像分析繫統進行圖像分析處理,結果用吸光度錶示.③缺血腦組織病理學觀察:常規囌木精-伊紅染色,光鏡下觀察針刺組和對照組病變區病理變化.主要觀察指標:①各組腦缺血再灌註後腦皮層及紋狀體內腫瘤壞死因子α的錶達.②針刺組和對照組缺血腦組織病理變化.結果:15隻大鼠全部進入結果分析.①腦缺血再灌註大鼠腦皮質及紋狀體內腫瘤壞死因子α的錶達:與假手術組比較,對照組和針刺組錶達明顯增彊(0.302±0.04,0.320±0.02;0.466±0.08,0.423±0.02;0.367±0.03,0.362±0.02;P＜0.05);與對照組比較,針刺組錶達明顯降低(P＜0.05).②缺血腦組織病理變化:對照組腦組織變性壞死程度較重,神經元變性壞死,膠質細胞呈空泡樣變性,血管內有明顯的白細胞聚集及附壁現象,血管週圍亦有明顯白細胞浸潤;針刺組僅有少量神經元變性壞死及膠質細胞空泡樣變性,血管內白細胞聚集減少.結論:腦缺血再灌註後腦皮層及紋狀體內腫瘤壞死因子α水平升高,頭麯鬢穴處平刺可明顯抑製其錶達,從而有效阻止炎性細胞因子過量錶達及其生物學作用的髮揮,減少腦缺血後的炎性損傷.
배경:뇌내종류배사인자α가통과유도뇌결혈재관주시내피세포점부분자적표체촉진염증반응급혈전형성.목적:관찰두혈백회투곡빈대결혈성뇌손상적량성조절,이급대뇌내종류배사인자α과도표체적억제작용.설계:수궤대조실험.단위:청도대학의학원부속의원급진신경과.재료:실험우2002-08/2003-08재청도대학의학원뇌혈관병연구소완성.청길급자성SD대서15지,체질량230～270 g,수궤분위가수술조、침자조화대조조,5지/조.간예:①침자조화대조조건립국조성뇌결혈재관주모형,가수술조수술보취동대조조,단불조새대뇌중동맥.침자조재결혈후10min급여침자치료,재대서두정부상당우인체백회、풍부혈상규전모소독,용0.35 mmx40 mm호침재피하백회혈처좌전하방상당우인체곡빈혈처평자,진침심도약0.8 sm.령우풍부혈직자일침,심도약0.5 cm.련접전능맥충전료의,백회혈접양겁,풍부혈접음겁,전침빈솔7Hz,강도6mA,시간30min.②뇌편제비급종류배사인자α원위잡교검측:각조대서우결혈2 h재관주12 h후심마취,좌심실삽관경승주동맥쾌속관주생리염수200 mL,재지속관주질량농도40 g/L다취갑철300mL,단두취뇌,상동고정액후고정1 h.자시교차처개시향후취관상절편,편후약7μm.종류배사인자α양성세포포장착색정종황색,음성대조세포불착색.응용VIDAS-21도상분석계통진행도상분석처리,결과용흡광도표시.③결혈뇌조직병이학관찰:상규소목정-이홍염색,광경하관찰침자조화대조조병변구병리변화.주요관찰지표:①각조뇌결혈재관주후뇌피층급문상체내종류배사인자α적표체.②침자조화대조조결혈뇌조직병리변화.결과:15지대서전부진입결과분석.①뇌결혈재관주대서뇌피질급문상체내종류배사인자α적표체:여가수술조비교,대조조화침자조표체명현증강(0.302±0.04,0.320±0.02;0.466±0.08,0.423±0.02;0.367±0.03,0.362±0.02;P＜0.05);여대조조비교,침자조표체명현강저(P＜0.05).②결혈뇌조직병리변화:대조조뇌조직변성배사정도교중,신경원변성배사,효질세포정공포양변성,혈관내유명현적백세포취집급부벽현상,혈관주위역유명현백세포침윤;침자조부유소량신경원변성배사급효질세포공포양변성,혈관내백세포취집감소.결론:뇌결혈재관주후뇌피층급문상체내종류배사인자α수평승고,두곡빈혈처평자가명현억제기표체,종이유효조지염성세포인자과량표체급기생물학작용적발휘,감소뇌결혈후적염성손상.
BACKGROUND: Tumor necrosis factor-alpha (TNF-α) in the brain can promote inflammative reaction and thrombosis through inducing the expression of endothelioal adhesion molecule during cerebral ischemic reperfusion.OBJECTIVE: To investigate the good regulation of scalp point through point acupucture with Baihui (GV-20) to Qubin (GB-7) to ischemic cerebral injury, and to inhibition of TNF-α over-expression within the brain as well.DESIGN: A randomized controlled trial.SETTING: Emergency Neurological Department of the Hospital Affiliated to Medical College of Qingdao University.MATERIALS: The experiment was completed from August 2002 to August 2003 at the Institute of Cerebrovascular Disease in Medical College of Qingdao University. A total of 15 healthy female SD rats of clean grade, weighing 230-270 g, provided by Shanghai Experimental Animal Center of Chinese Academy of Sciences, were at random divided as sham operation group,acupuncture group and control group, with 5 rats in each group.for rats in acupuncture group and control groups, the procedur for rats in sham operation group was similar to those in control group, but without obstruction of the middle cerebra artery. After ten minutes ischemia, the rats in acupuncture group were given acupuncture treatment. The hairs in parts correspondent to Baihui (GV-20) and Fengfu (GV-16) points in human were cut down and conventionally sterilized, then a filiform needle in 0.35 mmx40 mm was used to point to point acupucture correspondent to Baihui and Qubin in human to the depth of 0.8 cm. Another needle was used to perpendicularly insert at Fengfu (GV-16) point to the depth of 0.5 cm. A pulse electric machine was applied for 30 minutes, with the positive electrode connecting to Baihui (GV-20) and negative electrode to the Fengfu GV-16), the frequensection and in situ hybridization assay of TNF-α: All rats of every group,after treatment of 2 hours ischemia and 12 hours reperfusion, were deeply anesthetized, quickly given 200 mL normal saline through the catheterized ascending aorta, and continually given 300 mL paraformaldehyde of 40 g/L concentration; then they were decapitated for collecting the brain, the brain was fixed for 1 hour with the same solidification solution. The coronal sec-tion about 7 μm was prepared starting from the part of optic chiasm towards the back. The cytoplasm of TNF-α positive cell was stained brownish yellow, and that of TNF-α negative control cell was not stained. VIDAS-21 image analysing system was used to analyse the image, the result was extissue: After conventional staining with hematoxylin-eosin, the pathological changes of focal areas in both acupuncture group and control group were observed under light microscope.ischemic cerebral tissues in acupuncture group and control group.TNF-α in the cortex and striate body of brain in rats with ischemic reperfusion: Compared with sham operation group, the expressions in acupuncture group and control group were significantly increased (0.302±0.04,0.320±0.02; 0.466±0.08, 0.423±0.02; 0.367±0.03, 0.362±0.02; P ＜ 0.05);compared with acupuncture group, the expression in control group was sigbral tissues: In control group the extent of degeneration and necrosis was more serious, the neuron was degenerative necrosis, the neuroglia cell was in vacuolar degeneration, there was obvious phenomenon of leukocytic aggregation and margination within the vessel, and also leukocytic infiltration around the vessels. In acupuncture group there were only a small nub of neurons being in degenerative degeneration and neuroglia cells in vacuolar degeneration, and less leukocytic aggregation within the vessel.CONCLUSION: The level of TNF-α in the cortex and striate body of brain was increased after ischemic reperfusion. Scalp point through point acupuncture with Baihui (GV-20) to Qubin (GB-7) could markedly inhibit its expression, so that the therapy could effectively check over-expression of inflammatory mediators and their biological effects, hence reducing the inflammatory injury after cerebral ischemia.
