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

doi:10.3969/j.issn.2095-4344.2017.29.014

Abstract

Abstract:

BACKGROUND: Both salvia miltiorrhiza injection and adipose-derived stem cells have evident neuroprotection against cerebral infarction in rats.
OBJECTIVE: To observe the effect of salvia miltiorrhiza injection combined with adipose-derived stem cell transplantation 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 in 63 rats, and then the model rats were randomly divided into three groups (n=21 per group): model group (cerebral infarction group with 30 μL of PBS via the tail vein), stem cell group (transplantation of adipose-derived stem cells, 30 μL, 3×10⁶/L, via the tail vein), salvia miltiorrhiza injection + adipose-derived stem cell transplantation(combined group with transplantation of adipose-derived stem cells, 30 μL, 3×10⁶/L, via the tail vein and intraperitoneal injection of salvia miltiorrhiza injection, 0.5g/(kg•d) for consecutive 7 days). The modified neurological severity scores of each group were evaluated before and at 1, 2, 3, and 4 weeks after treatment. At 3 weeks after treatment, MTT assay was used to detect the serum levels of COX-2 and Aβ; 2,3,5-triphenyltetrazolium chloride staining was performed to detect infarction size; and RT-PCR and western blot assay were used to detect the mRNA and protein levels of COX-2 and Aβ in 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-2 and Aβ in the serum and brain tissues at protein and mRNA levels were significantly lower in the stem cell group and combined group; compared with the stem cell group (P < 0.05), the levels of COX-2 and Aβ were significantly lower in the combined group (P < 0.05). The infarct size was smallest in the combined group, followed by the stem cell group, and biggest in the model group (P < 0.05). To conclude, salvia miltiorrhiza injection combined with adipose-derived stem cell transplantation can improve the neurological function of rats after cerebral infarction, probably through reducing the levels of COX-2 and Aβ in the rat brain and serum.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Brain Infarction, Adipose Tissue, Salvia miltiorrhiza, Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Yang Peng, Gao Da-wei. Salvia miltiorrhiza injection combined with adipose-derived stem cell transplantation improves neurologic function of cerebral infarction rats[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(29): 4673-4678.

Figures/Tables 2

2.1 脂肪干细胞的形态在显微镜下观察第3代脂肪干细胞生长旺盛，形态渐趋一致，趋向长梭形，以一点为中心向两头呈放射状。细胞呈复层生长，胞膜完整、细胞状态良好，细胞间联系紧密，见图1A。经PKH26标记的脂肪干细胞，荧光显微镜观察可见红色荧光，标记率达98%，见图1B。 2.2 神经功能学mNSS评分由表1可见，治疗前各组大鼠神经功能损伤评分差异无显著性意义(P > 0.05)；治疗1-4周后脂肪干细胞组的神经功能缺损评分显著低于脑梗死组，联合组又明显低于脂肪干细胞组(P < 0.05)。提示脂肪干细胞移植治疗能够改善脑梗死模型大鼠神经功能，而联合治疗方法对大鼠神经功能的改善更显著。 2.3 血清环氧合酶2及β-淀粉样蛋白水平由表2可见，与脑梗死组比较，脂肪干细胞组大鼠血清环氧合酶2及β-淀粉样蛋白水平显著降低(P < 0.05)；联合组大鼠血清环氧合酶2及β-淀粉样蛋白水平降低更明显(P < 0.05)。联合组血清环氧合酶2及β-淀粉样蛋白水平低于脂肪干细胞组(P < 0.05)。 2.4 脑梗死体积比较在治疗后3周，脑梗死组大鼠脑梗死体积为(245±24) mm3，脂肪干细胞组为(158±14) mm3，联合组为(104±12) mm3。联合组与脑梗死组、脂肪干细胞组相比，差异有显著性意义(P < 0.05)。 2.5 脑组织病理形态和PKH26阳性细胞数治疗3周后，脑梗死组可见梗死处脑组织被胶质细胞和胶质纤维充填，形成胶质瘢痕，局部脑萎缩，软化灶形成(图2A)。脂肪干细胞组移植部位出现典型的神经细胞样形态学改变，组织瘢痕和软化灶小于脑梗死组，而大于联合组(图2B)。联合组出现典型的神经细胞样形态学改变且软化灶消失，有少量瘢痕(图2C)。荧光显微镜观察损伤灶脑组织中的PKH26阳性细胞数：脑梗死组为(0±0.00)个/高倍视野，脂肪干细胞组为(34.32±5.58)个/高倍视野，联合组为(56.34±7.42)个/高倍视野，见图3。"

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