CAJ | 학술논문

目的：观察复方丹参滴丸(compound Danshen dripping pills，CDDP)和人脐血单个核细胞(human umbilical cord blood mononuclear cells，HUCBMCs)静脉移植联合复方丹参滴丸对家兔急性心肌梗死(acute myocardial infarction， AMI)的心肌组织炎症反应、心肌细胞凋亡、氧化应激指标及心功能的影响，并探讨其联合治疗对急性心肌梗死可能的保护机制。方法：对健康家兔结扎冠状动脉左前降支，制备AMI模型。建模成功者40只随机分为4组，每组10只。对照组：术后24 h生理盐水0.5 mL耳缘静脉注射，生理盐水5 mL/d灌胃；CDDP组：术后24 h经耳缘静脉注入生理盐水0.5 mL，CDDP 270 mg/d溶入生理盐水5 mL灌胃；单纯移植组：术后24 h经耳缘静脉注入含3×107绿色荧光蛋白(green lfuorescent protein，GFP)标记人脐血单个核细胞生理盐水0.5 mL，生理盐水5 mL/d灌胃；联合治疗组：术后24 h经耳缘静脉注入含3×107 GFP标记人脐血单个核细胞的生理盐水0.5 mL，CDDP 270 mg/d溶入生理盐水5 mL灌胃。每组随机5只家兔分别于移植术后1周、4周超声心动图检测家兔心功能指标左室射血分数(letf ventricular ejection fraction，LVEF)及左室短轴缩短率(letf ventricular fractional shorting，LVFS)；苏木精-伊红(HE)染色光镜观察心肌病理变化并在光镜下进行白细胞计数。氮蓝四唑(nitrotetrazolium blue chloride， NBT)法测定心肌组织浆液超氧化物歧化酶(superoxide dismutase，SOD)活性；硫代巴比妥酸比色法测心肌组织浆液丙二醛(malondialdehyde，MDA)含量。根据荧光显微镜检测到的GFP阳性细胞进行移植细胞计数。结果：1)与对照组比较，CDDP组、单纯移植组、联合治疗组术后1周、4周心功能指标LVFS及LVEF改善(均P<0.05)，其中联合治疗组心功能改善优于单纯移植组、CDDP组(均P<0.05)，而后两组组间比较差异无统计学意义；2)与对照组比较，CDDP组、单纯移植组、联合治疗组术后1周、4周心肌组织白细胞计数及心肌细胞凋亡指数均显著减低(均P<0.05)，且联合治疗组均显著低于单纯移植组、CDDP组(均P<0.05)，而后两组比较差异无统计学意义；3)与对照组比较，CDDP组、单纯移植组、联合治疗组术后4周心肌组织MDA水平均显著降低(均P<0.05)，SOD活性显著增高(均P<0.05)，且联合治疗组均优于单纯移植组、CDDP组(均P<0.05)，而后两组比较差异无统计学意义；4)联合治疗组、单纯移植组术后4周均可见GFP阳性细胞散在分布于梗死周边区域，且联合治疗组细胞计数多于单纯移植组(P<0.05)。结论：HUCBMCs静脉移植联合CDDP治疗AMI，可提高移植细胞在心肌组织内存活率，拮抗心肌细胞凋亡，并进一步改善心功能。其机制可能与联合治疗减低心肌组织局部炎症反应及拮抗氧化应激作用增强有关。目的：觀察複方丹參滴汍(compound Danshen dripping pills，CDDP)和人臍血單箇覈細胞(human umbilical cord blood mononuclear cells，HUCBMCs)靜脈移植聯閤複方丹參滴汍對傢兔急性心肌梗死(acute myocardial infarction， AMI)的心肌組織炎癥反應、心肌細胞凋亡、氧化應激指標及心功能的影響，併探討其聯閤治療對急性心肌梗死可能的保護機製。方法：對健康傢兔結扎冠狀動脈左前降支，製備AMI模型。建模成功者40隻隨機分為4組，每組10隻。對照組：術後24 h生理鹽水0.5 mL耳緣靜脈註射，生理鹽水5 mL/d灌胃；CDDP組：術後24 h經耳緣靜脈註入生理鹽水0.5 mL，CDDP 270 mg/d溶入生理鹽水5 mL灌胃；單純移植組：術後24 h經耳緣靜脈註入含3×107綠色熒光蛋白(green lfuorescent protein，GFP)標記人臍血單箇覈細胞生理鹽水0.5 mL，生理鹽水5 mL/d灌胃；聯閤治療組：術後24 h經耳緣靜脈註入含3×107 GFP標記人臍血單箇覈細胞的生理鹽水0.5 mL，CDDP 270 mg/d溶入生理鹽水5 mL灌胃。每組隨機5隻傢兔分彆于移植術後1週、4週超聲心動圖檢測傢兔心功能指標左室射血分數(letf ventricular ejection fraction，LVEF)及左室短軸縮短率(letf ventricular fractional shorting，LVFS)；囌木精-伊紅(HE)染色光鏡觀察心肌病理變化併在光鏡下進行白細胞計數。氮藍四唑(nitrotetrazolium blue chloride， NBT)法測定心肌組織漿液超氧化物歧化酶(superoxide dismutase，SOD)活性；硫代巴比妥痠比色法測心肌組織漿液丙二醛(malondialdehyde，MDA)含量。根據熒光顯微鏡檢測到的GFP暘性細胞進行移植細胞計數。結果：1)與對照組比較，CDDP組、單純移植組、聯閤治療組術後1週、4週心功能指標LVFS及LVEF改善(均P<0.05)，其中聯閤治療組心功能改善優于單純移植組、CDDP組(均P<0.05)，而後兩組組間比較差異無統計學意義；2)與對照組比較，CDDP組、單純移植組、聯閤治療組術後1週、4週心肌組織白細胞計數及心肌細胞凋亡指數均顯著減低(均P<0.05)，且聯閤治療組均顯著低于單純移植組、CDDP組(均P<0.05)，而後兩組比較差異無統計學意義；3)與對照組比較，CDDP組、單純移植組、聯閤治療組術後4週心肌組織MDA水平均顯著降低(均P<0.05)，SOD活性顯著增高(均P<0.05)，且聯閤治療組均優于單純移植組、CDDP組(均P<0.05)，而後兩組比較差異無統計學意義；4)聯閤治療組、單純移植組術後4週均可見GFP暘性細胞散在分佈于梗死週邊區域，且聯閤治療組細胞計數多于單純移植組(P<0.05)。結論：HUCBMCs靜脈移植聯閤CDDP治療AMI，可提高移植細胞在心肌組織內存活率，拮抗心肌細胞凋亡，併進一步改善心功能。其機製可能與聯閤治療減低心肌組織跼部炎癥反應及拮抗氧化應激作用增彊有關。
목적：관찰복방단삼적환(compound Danshen dripping pills，CDDP)화인제혈단개핵세포(human umbilical cord blood mononuclear cells，HUCBMCs)정맥이식연합복방단삼적환대가토급성심기경사(acute myocardial infarction， AMI)적심기조직염증반응、심기세포조망、양화응격지표급심공능적영향，병탐토기연합치료대급성심기경사가능적보호궤제。방법：대건강가토결찰관상동맥좌전강지，제비AMI모형。건모성공자40지수궤분위4조，매조10지。대조조：술후24 h생리염수0.5 mL이연정맥주사，생리염수5 mL/d관위；CDDP조：술후24 h경이연정맥주입생리염수0.5 mL，CDDP 270 mg/d용입생리염수5 mL관위；단순이식조：술후24 h경이연정맥주입함3×107록색형광단백(green lfuorescent protein，GFP)표기인제혈단개핵세포생리염수0.5 mL，생리염수5 mL/d관위；연합치료조：술후24 h경이연정맥주입함3×107 GFP표기인제혈단개핵세포적생리염수0.5 mL，CDDP 270 mg/d용입생리염수5 mL관위。매조수궤5지가토분별우이식술후1주、4주초성심동도검측가토심공능지표좌실사혈분수(letf ventricular ejection fraction，LVEF)급좌실단축축단솔(letf ventricular fractional shorting，LVFS)；소목정-이홍(HE)염색광경관찰심기병리변화병재광경하진행백세포계수。담람사서(nitrotetrazolium blue chloride， NBT)법측정심기조직장액초양화물기화매(superoxide dismutase，SOD)활성；류대파비타산비색법측심기조직장액병이철(malondialdehyde，MDA)함량。근거형광현미경검측도적GFP양성세포진행이식세포계수。결과：1)여대조조비교，CDDP조、단순이식조、연합치료조술후1주、4주심공능지표LVFS급LVEF개선(균P<0.05)，기중연합치료조심공능개선우우단순이식조、CDDP조(균P<0.05)，이후량조조간비교차이무통계학의의；2)여대조조비교，CDDP조、단순이식조、연합치료조술후1주、4주심기조직백세포계수급심기세포조망지수균현저감저(균P<0.05)，차연합치료조균현저저우단순이식조、CDDP조(균P<0.05)，이후량조비교차이무통계학의의；3)여대조조비교，CDDP조、단순이식조、연합치료조술후4주심기조직MDA수평균현저강저(균P<0.05)，SOD활성현저증고(균P<0.05)，차연합치료조균우우단순이식조、CDDP조(균P<0.05)，이후량조비교차이무통계학의의；4)연합치료조、단순이식조술후4주균가견GFP양성세포산재분포우경사주변구역，차연합치료조세포계수다우단순이식조(P<0.05)。결론：HUCBMCs정맥이식연합CDDP치료AMI，가제고이식세포재심기조직내존활솔，길항심기세포조망，병진일보개선심공능。기궤제가능여연합치료감저심기조직국부염증반응급길항양화응격작용증강유관。
Objective:To explore the effects of compound Danshen dripping pills (CDDP) and CDDP combined with transplantation of human umbilical cord blood cells (HUMNCs) on the inlfammatory response, oxidative stress, myocardial cell apoptosis and cardiac function, and also to investigate the possible mechanisms of the combined therapy in the acute myocardial infarction (AMI). <br> Methods:Rabbit model of AMI successfully established by ligation of the letf anterior coronary artery (LAD). Forty rabbits were randomly divided into 4 groups (n=10 per group):a control group, injected with 0.5 mL of saline in 24 h atfer AMI and then gavaged with 5 mL of saline daily;a CDDP group, injected with saline 0.5 mL atfer AMI and then gavaged with CDDP (270 mg/d) daily;a transplantation group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs [labeled with green fluorescent protein (GFP)] and then gavaged with 5 mL of saline daily;a combined group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs (labeled with GFP) and then gavaged with CDDP (270 mg/d) daily. Cardiac function index such as left ventricular fractional shorting (LVFS) and ejection fraction(LVEF) were measured by echocardiography;the pathological changes were observed by HE staining and the white blood cells in the myocardium were determined by light microscopy. hTe superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardium were detected by nitrotetrazolium blue chloride (NBT) and thiobarbituric acid colorimetric measurement respectively. hTe number of transplanted cells in the myocardium was examined by GFP positive cells counted with lfuorescence microscopy. <br> Results:1) Compared with the control group (at 1 or 4 week), LVEF and LVFS were signiifcant improved in the CDDP group, the transplantation group and the combined groups (all P<0.05), the improvement degree of cardiac function in the combined group was the most significance. There was no significant difference between the CDDP group and the transplantation group. 2) Compared with the control group (at 1 or 4 week), the number of white blood cell, myocardial cell apoptosis ratio were decreased signiifcantly in the CDDP group, the transplantation group and the combined groups (all P<0.05), this decrease in the combined group was the most signiifcance, and there was no significant difference between the CDDP group and the transplantation group. 3) Compared with control (at 4 week), the SOD activity was increased signiifcantly, and MDA content in myocardium was decreased in the CDDP group, this change in the combined group was the most signiifcance. 4) GFP-positive cells were found to be present in the peri-myocardial infarction area in the transplantation group and the combined group at 1, 4 weeks post-transplantation. hTe number of the GFP positive cells in the combined group was more than that in the transplantation group (P<0.05). <br> Conclusion:The intravenous transplantation of HUMNCs combined with the CDDP in the treatment of rabbits with AMI could increase the survival rate of transplanted cells and inhibit the myocardial cell apoptosis, therefore improve the heart function. hTe possible mechanism of the combined treatment may be involved in the inhibition of the inlfammatory response and oxidative stress in the myocardium following AMI.