丹参注射液联合脂肪干细胞移植改善脑梗死模型大鼠的神经功能

Salvia miltiorrhiza injection combined with adipose-derived stem cell transplantation improves neurologic function of cerebral infarction rats

背景:研究发现,丹参注射液、脂肪干细胞分别对脑梗死大鼠具有明显的神经保护作用。目的:观察丹参注射液联合脂肪干细胞移植对脑梗死大鼠脑组织及血清中环氧合酶2、β-淀粉样蛋白水平的影响。方法:选取脑梗死模型大鼠63只,随机分为3组:脑梗死组、脂肪干细胞组、联合组,每组21只。造模后24 h,脑梗死组于尾静脉注射30μL PBS,脂肪干细胞组尾静脉注射30μL脂肪干细胞悬液(含有3×10~6 L^(-1)个脂肪干细胞),联合组经尾静脉注射30μL脂肪干细胞悬液(含有3×10~6 L^(-1)个脂肪干细胞)+腹腔注射丹参注射液0.5 g/(kg·d),细胞注射1次,丹参注射液连续注射7 d。治疗后1,2,3,4周进行神经功能学缺损评分;治疗后3周,采用ELISA法检测血清环氧合酶2及β-淀粉样蛋白水平,TTC染色观察脑梗死范围,RT-PCR和蛋白免疫印迹技术检测脑组织环氧合酶2及β-淀粉样蛋白的mRNA和蛋白表达。结果与结论:①脂肪干细胞组、联合组神经功能缺损评分明显低于脑梗死组(P<0.05),联合组又明显低于脂肪干细胞移植组(P<0.05);②与脑梗死组比较,脂肪干细胞组、联合组血清环氧合酶2及β-淀粉样蛋白水平显著降低(P<0.05);联合组明显低于脂肪干细胞组(P<0.05);③联合组梗死体积小于脂肪干细胞组(P<0.05),脂肪干细胞组小于脑梗死组(P<0.05);④与脑梗死组比较,脂肪干细胞组、联合组脑组织中环氧合酶2及β-淀粉样蛋白mRNA和蛋白表达明显下调(P<0.05),联合组降低更为显著(P<0.05);⑤结果表明,丹参注射液联合脂肪干细胞移植可改善脑梗死大鼠的神经功能,可能是通过降低脑组织及血清中环氧合酶2及β-淀粉样蛋白发挥作用。

BACKGROUND:Both salvia miltiorrhiza injection and adipose-derived stem cells have evident neuroprotection againstcerebral infarction in rats.OBJECTIVE:To observe the effect of salvia miltiorrhiza injection combined with adipose-derived stem celltransplantation on cerebral infarction in rats and the levels of cyclooxygenase-2(COX-2)and beta-amyloid protein(Aβ)in the serum and brain tissues.METHODS:A rat model of cerebral infarction was successfully established in63rats,and then the model rats wererandomly divided into three groups(n=21per group):model group(cerebral infarction group with30μL of PBS via thetail vein),stem cell group(transplantation of adipose-derived stem cells,30μL,3×106/L,via the tail vein),salviamiltiorrhiza injection+adipose-derived stem cell transplantation(combined group with transplantation of adipose-derivedstem cells,30μL,3×106/L,via the tail vein and intraperitoneal injection of salvia miltiorrhiza injection,0.5g/(kg?d)forconsecutive7days).The modified neurological severity scores of each group were evaluated before and at1,2,3,and4weeks after treatment.At3weeks after treatment,MTT assay was used to detect the serum levels of COX-2and Aβ;2,3,5-triphenyltetrazolium chloride staining was performed to detect infarction size;and RT-PCR and western blot assaywere used to detect the mRNA and protein levels of COX-2and Aβin the brain tissues.RESULTS AND CONCLUSION:The modified neurological severity scores in the three groups were ranked as follows:combined group<stem cell group<model group,and there were significant differences between groups(P<0.05).Compared with the model group,the levels of COX-2and Aβin the serum and brain tissues at protein and mRNA levelswere significantly lower in the stem cell group and combined group;compared with the stem cell group(P<0.05),thelevels of COX-2and Aβwere significantly lower in the combined group(P<0.05).The infarct size was smallest in thecombined group,followed by the stem cell group,and biggest in the model group(P<0.05).To conclude,salviamiltiorrhiza injection combined with adipose-derived stem cell transplantation can improve the neurological function ofrats after cerebral infarction,probably through reducing the levels of COX-2and Aβin the rat brain and serum.