中华危重病急救医学
中華危重病急救醫學
중화위중병급구의학
Chinese Critical Care Medicine
2014年
8期
589-593
,共5页
李涛%李金河%李浩波%夏正远%史晓勇%李玄英%刘友坦
李濤%李金河%李浩波%夏正遠%史曉勇%李玄英%劉友坦
리도%리금하%리호파%하정원%사효용%리현영%류우탄
高渗氯化钠羟乙基淀粉%蛛网膜下腔出血%脑血管痉挛%凋亡
高滲氯化鈉羥乙基澱粉%蛛網膜下腔齣血%腦血管痙攣%凋亡
고삼록화납간을기정분%주망막하강출혈%뇌혈관경련%조망
Hypertonic sodium chloride hydroxyethyl starch solution%Subarachnoid hemorrhage%Cerebral vasospasm%Apoptosis
目的 探讨高渗氯化钠羟乙基淀粉溶液(HSH)对蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)的影响及其相关机制.方法 按随机数字表法将24只雄性SD大鼠分为4组,每组6只.利用枕大池二次注血法建立SAH-CVS模型.模型组和HSH治疗组于制模后每日分别经尾静脉注射生理盐水或HSH 8 mL/kg;假手术组进行SAH手术但仅在枕大池内注射1.5 mL/kg生理盐水并每日给予生理盐水8 mL/kg治疗;正常组不予任何处理.各组大鼠均于7d后处死,苏木素-伊红(HE)染色分析检测基底动脉血管壁厚度及管腔面积.分离血管平滑肌细胞(VSMC),采用流式细胞仪检测细胞凋亡率;荧光法检测细胞天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)活性;蛋白质免疫印迹试验(Western Blot)检测细胞Bax及Bcl-2蛋白表达;H2DCFDA荧光探针检测细胞活性氧(ROS)水平.结果 与正常组比较,模型组基底动脉血管壁厚度明显增加(μm:27.72±1.94比18.30±1.11,P＜0.05),管腔面积明显减小(μm2:26 115±1 991比55 080±2 091,P＜0.05),细胞凋亡率显著增加[(35.05±5.54)％比(5.93±1.53)％,P＜0.05].与模型组比较,HSH治疗组基底动脉血管壁厚度明显减小(μm:22.55±1.50比27.72±1.94,P＜ 0.05),管腔面积明显增加(μm2:48 115±2 460比26 115±1 991,P＜0.05),细胞凋亡率明显下降[(16.54±5.94)％比(35.05±5.54)％,P＜0.05].模型组细胞caspase-3活性、ROS水平、Bax和Bcl-2表达分别为正常组的(188.40±19.35)％、(163.50±17.02)％、(208.71±26.04)％和(44.52±9.61)％,与正常组比较差异均有统计学意义(均P＜0.05);HSH治疗组caspase-3活性、ROS水平、Bax和Bcl-2表达分别为正常组的(135.05±19.52)％、(119.44±11.50)％、(139.20±18.04)％和(85.35±13.12)％,与模型组比较均明显改善(均P＜0.05).假手术组各指标与正常组比较差异均无统计学意义.结论 HSH可抑制SAH后CVS程度,改善基底动脉血管壁增厚及管腔狭窄,其机制可能与抑制VSMC凋亡有关.
目的 探討高滲氯化鈉羥乙基澱粉溶液(HSH)對蛛網膜下腔齣血(SAH)後腦血管痙攣(CVS)的影響及其相關機製.方法 按隨機數字錶法將24隻雄性SD大鼠分為4組,每組6隻.利用枕大池二次註血法建立SAH-CVS模型.模型組和HSH治療組于製模後每日分彆經尾靜脈註射生理鹽水或HSH 8 mL/kg;假手術組進行SAH手術但僅在枕大池內註射1.5 mL/kg生理鹽水併每日給予生理鹽水8 mL/kg治療;正常組不予任何處理.各組大鼠均于7d後處死,囌木素-伊紅(HE)染色分析檢測基底動脈血管壁厚度及管腔麵積.分離血管平滑肌細胞(VSMC),採用流式細胞儀檢測細胞凋亡率;熒光法檢測細胞天鼕氨痠特異性半胱氨痠蛋白酶3(caspase-3)活性;蛋白質免疫印跡試驗(Western Blot)檢測細胞Bax及Bcl-2蛋白錶達;H2DCFDA熒光探針檢測細胞活性氧(ROS)水平.結果 與正常組比較,模型組基底動脈血管壁厚度明顯增加(μm:27.72±1.94比18.30±1.11,P＜0.05),管腔麵積明顯減小(μm2:26 115±1 991比55 080±2 091,P＜0.05),細胞凋亡率顯著增加[(35.05±5.54)％比(5.93±1.53)％,P＜0.05].與模型組比較,HSH治療組基底動脈血管壁厚度明顯減小(μm:22.55±1.50比27.72±1.94,P＜ 0.05),管腔麵積明顯增加(μm2:48 115±2 460比26 115±1 991,P＜0.05),細胞凋亡率明顯下降[(16.54±5.94)％比(35.05±5.54)％,P＜0.05].模型組細胞caspase-3活性、ROS水平、Bax和Bcl-2錶達分彆為正常組的(188.40±19.35)％、(163.50±17.02)％、(208.71±26.04)％和(44.52±9.61)％,與正常組比較差異均有統計學意義(均P＜0.05);HSH治療組caspase-3活性、ROS水平、Bax和Bcl-2錶達分彆為正常組的(135.05±19.52)％、(119.44±11.50)％、(139.20±18.04)％和(85.35±13.12)％,與模型組比較均明顯改善(均P＜0.05).假手術組各指標與正常組比較差異均無統計學意義.結論 HSH可抑製SAH後CVS程度,改善基底動脈血管壁增厚及管腔狹窄,其機製可能與抑製VSMC凋亡有關.
목적 탐토고삼록화납간을기정분용액(HSH)대주망막하강출혈(SAH)후뇌혈관경련(CVS)적영향급기상관궤제.방법 안수궤수자표법장24지웅성SD대서분위4조,매조6지.이용침대지이차주혈법건립SAH-CVS모형.모형조화HSH치료조우제모후매일분별경미정맥주사생리염수혹HSH 8 mL/kg;가수술조진행SAH수술단부재침대지내주사1.5 mL/kg생리염수병매일급여생리염수8 mL/kg치료;정상조불여임하처리.각조대서균우7d후처사,소목소-이홍(HE)염색분석검측기저동맥혈관벽후도급관강면적.분리혈관평활기세포(VSMC),채용류식세포의검측세포조망솔;형광법검측세포천동안산특이성반광안산단백매3(caspase-3)활성;단백질면역인적시험(Western Blot)검측세포Bax급Bcl-2단백표체;H2DCFDA형광탐침검측세포활성양(ROS)수평.결과 여정상조비교,모형조기저동맥혈관벽후도명현증가(μm:27.72±1.94비18.30±1.11,P＜0.05),관강면적명현감소(μm2:26 115±1 991비55 080±2 091,P＜0.05),세포조망솔현저증가[(35.05±5.54)％비(5.93±1.53)％,P＜0.05].여모형조비교,HSH치료조기저동맥혈관벽후도명현감소(μm:22.55±1.50비27.72±1.94,P＜ 0.05),관강면적명현증가(μm2:48 115±2 460비26 115±1 991,P＜0.05),세포조망솔명현하강[(16.54±5.94)％비(35.05±5.54)％,P＜0.05].모형조세포caspase-3활성、ROS수평、Bax화Bcl-2표체분별위정상조적(188.40±19.35)％、(163.50±17.02)％、(208.71±26.04)％화(44.52±9.61)％,여정상조비교차이균유통계학의의(균P＜0.05);HSH치료조caspase-3활성、ROS수평、Bax화Bcl-2표체분별위정상조적(135.05±19.52)％、(119.44±11.50)％、(139.20±18.04)％화(85.35±13.12)％,여모형조비교균명현개선(균P＜0.05).가수술조각지표여정상조비교차이균무통계학의의.결론 HSH가억제SAH후CVS정도,개선기저동맥혈관벽증후급관강협착,기궤제가능여억제VSMC조망유관.
Objective To investigate the protective effect and potential mechanisms of hypertonic sodium chloride hydroxyethyl starch solution (HSH) against the cerebral vasospasm (CVS) following subarachnoid hemorrhage (SAH).Methods Twenty-four male Sprague-Dawley (SD) rats were randomly assigned to four groups according to the random number table,with 6 rats in each group.The SAH-CVS model was reproduced by injection of the blood twice through the cisterna magna.Rats in both model and HSH treatment groups received 8 mL/kg normal saline (NS) or HSH treatment everyday via caudal vein.Rats in sham group were injected with 1.5 mL/kg NS into cisterna magna followed by 8 mL/kg NS treatment.Rats in normal group received no treatment.Rats were sacrificed to harvest basilar artery after 7 days.The thickness of vessel wall and lumen area were measured using hematoxylin-eosin (HE) staining.The rate of apoptosis of vascular smooth muscle cell (VSMC) was assessed using flow cytometry.Caspase-3 activity was measured by a fluorometric assay.The expressions of Bax and Bcl-2 were determined by Western Blot.Intracellular reactive oxygen species (ROS) was detected by H2DCFDA.Results Compared with normal group,increased thickness of vessel wall (μm:27.72 ± 1.94 vs.18.30 ± 1.10,P＜0.05),decreased lumen area (μm2:26 115 ± 1 991 vs.55 080 ± 2 091,P＜0.05),and elevation of rate of apoptosis of VSMCs [(35.05 ± 5.54) ％ vs.(5.93 ± 1.53) ％,P＜ 0.05] were found in model group.Compared with model group,decreased thickness of vessel wall (μm:22.55 ± 1.50 vs.27.72 ± 1.94,P＜0.05),increase of lumen area (μm2:48 115 ±2 460 vs.26 115 ± 1 991,P＜0.05),and depressed rate of apoptosis of VSMCs [(16.54 ± 5.94) ％ vs.(35.05 ± 5.54) ％,P＜ 0.05] were found in HSH treatment group.Caspase-3 activity,intracellular ROS level,Bax and Bcl-2 expressions in model group were (188.40 ± 19.35)％,(163.50 ± 17.02)％,(208.71 ± 26.04)％ and (44.52 ± 9.61) ％ of those of normal group,and the differences of these parameters between model and normal groups were statistically significant (all P＜0.05).Caspase-3 activity,intracellular ROS level,Bax and Bcl-2 expressions in HSH treatment group were (135.05 ± 19.52)％,(119.44 ± 11.50)％,(139.20 ± 18.04)％ and (85.35 ± 13.12)％ of those of normal group,respectively,and the differences of these parameters between HSH treatment and model groups were statistically significant (all P＜0.05).The differences of all measurements between sham and normal groups were not statistically significant.Conclusion The current results demonstrate that HSH attenuates the SAH-induced CVS,alleviates thickness of vessel wall,and increases lumen area via inhibition of VSMCs apoptosis.
