CAJ | 학술논문

目的动态观察汉防己甲索( tetrandrine,Tet)对大鼠急性脊髓损伤(acute spinal cord injury,ASCI)后神经细胞凋亡和相关基因表达的影响,为该病治疗研究提供实验依据. 方法将95只成年SD大鼠随机分为空白对照组、急性脊髓损伤组(ASCI组)、汉防己甲素(Tet)组和甲基强的松龙组(Mp)组.空白对照组5只大鼠,其他每组30只大鼠.除空白对照组外,所有动物均用改良Allen重物打击法制作脊髓损伤模型.于术后8h、1,3,7,14和21 d每组处死5只大鼠,观察损伤段脊髓的形态结构、细胞凋亡调控基因(Bcl -xl和Bcl -xs)的表达,运用流式细胞仪对标本进行凋亡细胞半定量分析. 结果 (1)组织学改变:除空白组无明显改变外,ASCI组组织水肿,神经元变性,部分坏死,可见大量炎性细胞浸润；Tet组损伤程度减轻；Mp组更轻,无明显坏死及炎性细胞浸润.(2)凋亡基因表达检测:除空白组外,余各组在相同时相点,Bcl -xs表达量ASCI组较多,Tet组明显减少,Mp组更少；而Bcl -xl表达量则反之,且各组在伤后7d达高峰.(3)凋亡细胞半定量分析:除空白对照组外,各组凋亡细胞数均在伤后7d达峰值,之后逐渐减少；同一时相点各组间凋亡细胞比较,Tet组明显少于ASCI组,而多于Mp组. 结论 ASCI后Tet对凋亡调控基因的表达有明显的积极干预作用,可显著减少神经细胞凋亡；应用Tet治疗ASCI应越早越好,7d内有效；与甲基强的松龙相比,Tet对ASCI的疗效较弱.
목적 동태관찰한방기갑색( tetrandrine,Tet)대대서급성척수손상(acute spinal cord injury,ASCI)후신경세포조망화상관기인표체적영향,위해병치료연구제공실험의거. 방법 장95지성년SD대서수궤분위공백대조조、급성척수손상조(ASCI조)、한방기갑소(Tet)조화갑기강적송룡조(Mp)조.공백대조조5지대서,기타매조30지대서.제공백대조조외,소유동물균용개량Allen중물타격법제작척수손상모형.우술후8h、1,3,7,14화21 d매조처사5지대서,관찰손상단척수적형태결구、세포조망조공기인(Bcl -xl화Bcl -xs)적표체,운용류식세포의대표본진행조망세포반정량분석. 결과 (1)조직학개변:제공백조무명현개변외,ASCI조조직수종,신경원변성,부분배사,가견대량염성세포침윤；Tet조손상정도감경；Mp조경경,무명현배사급염성세포침윤.(2)조망기인표체검측:제공백조외,여각조재상동시상점,Bcl -xs표체량ASCI조교다,Tet조명현감소,Mp조경소；이Bcl -xl표체량칙반지,차각조재상후7d체고봉.(3)조망세포반정량분석:제공백대조조외,각조조망세포수균재상후7d체봉치,지후축점감소；동일시상점각조간조망세포비교,Tet조명현소우ASCI조,이다우Mp조. 결론 ASCI후Tet대조망조공기인적표체유명현적적겁간예작용,가현저감소신경세포조망；응용Tet치료ASCI응월조월호,7d내유효；여갑기강적송룡상비,Tet대ASCI적료효교약.
Objective To analyze the effect of Tetrandrine (Tet) on neuronal apoptosis and reletted gene expression after acute spinal cord injury (ASCI) in SD rats so as to provide experimental basis for treatment of ASCI. Methods A total of 95 adult SD rats were.randomly divided into four groups,ie,control group ( n =5 ),ASCI group ( n =30),Tet treatment group ( Tet group,n =30) and methylprednisolone treatment group ( Mp group,n =30).The ASCI model was established with the aid of modified Allen method.The five rats from each group were sacrificed at postoperative 8 hours and 1,3,7,14,21 days respectively to observe the morphology of the injury spinal cord and expressions of cell apoptosis regulation genes ( Bcl-xl,Bcl-xs).The flow cytometry ( FCM ) was used to semiquantitatively analyze the apoptotic cells. Results Except for the control group,the other three groups had morphological changes.Tissue edema,neuronal degeneration with some necrosis and marked inflammatory cell infiltration were founded in the ASCI group.Tissue injury was alleviated in the Tet group and was relatively much milder in the Mp group,with no obvious necrosis or inflammatory cell infiltration.Except for the control group,the number of Bcl-xs positive cells at the same time point was the most in the ASCI group in comparison with the Tet and Mp groups.Meanwhile,the expression of Bcl-xl in all groups reached the peak at day 7 post-injury.The semiquantitative analysis of the apoptotic cells displayed that,except for the control group,the number of apoptotic neurons in other three groups reached the peak at day 7 postinjury and then declined gradually.At the same time point,the apoptotic neurons in the Tet group were less than that in the ASCI group,but more than that in the Mp group. Conclusions After ASCI,Tet plays an active role in intervening the expression of the apoptosis regulation gene and can evidently decrease neuronal apoptosis.Tet treatment can work within 7 days and should be given for ASCI as early as possible.However,Tet exerts weaker effect on ASCI in comparison with MP.