超声联合一氧化氮微泡对大鼠心肌组织的生物学效应

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目的:本研究旨在探讨超声联合一氧化氮(NO)微泡的作用对大鼠心肌组织局部基质衍生因子-1(SDF-1)的影响,及其对心肌组织作用的安全性。方法:将32只SPF级SD雄性大鼠随机分为4组,每组8只。第1、2、3组分别经尾静脉输入1 ml的NO微泡、普通脂质微泡以及PBS,并采用频率为1 MHz、强度为1 W/cm2的超声辐照大鼠心前区,辐照时间为10 min;第4组为空白对照组。4 h后每组处死4只大鼠,留取心肌组织进行HE染色,观察局部的炎症反应情况;同时留取血清标本用于检测肌酸激酶(CK),乳酸脱氢酶(LDH)和肌钙蛋白I(TnI)的含量。1周后处死剩余大鼠,取心肌组织进行RT-PCR和Western blot分别检测SDF-1的基因及蛋白相对表达量。结果:辐照4 h后留取心肌组织行HE染色显示各组均可见少量炎性细胞浸润,组间没有明显差异;血清学指标检测显示,第1组与第2组血清CK和LDH高于第3组和第4组,且差异有统计学意义(P<0.05);第1组血清TnI低于第2组,但高于第3组和第4组,且差异有统计学意义(P<0.05);辐照1周后留取的心肌组织行RT-PCR和Western blot分析结果均显示第1组SDF-1相对表达量最多,各组间进行两两比较,差异均有统计学意义(P<0.05)。结论:超声联合NO微泡辐照大鼠心肌组织,与内含全氟显气体的普通脂质微泡一样,对心肌组织无明显破坏;但能使大鼠心肌组织SDF-1表达增加,其程度高于应用超声联合普通脂质微泡,因此,NO微泡应用于干细胞归巢将优于目前普通脂质微泡。 OBJECTIVE: To investigate the effect of ultrasound combined with nitric oxide (NO) microbubbles on myocardial stromal derived factor-1 (SDF-1) in rats and its safety on myocardial tissue. Methods: Thirty-two SD male SD rats were randomly divided into 4 groups with 8 rats in each group. Groups 1, 2 and 3 were injected with 1 ml of NO microvesicles, ordinary lipid microbubbles and PBS respectively through the tail vein, and irradiated with 1 MHz and 1 W / Irradiation time was 10 min; the fourth group was blank control group. After 4 h, 4 rats were sacrificed in each group. HE staining was performed on the myocardium to observe the local inflammatory response. Meanwhile, serum samples were collected for determination of creatine kinase (CK), lactate dehydrogenase (LDH) and muscle The content of calpain I (TnI). One week later, the remaining rats were sacrificed and the myocardial tissues were harvested for RT-PCR and Western blot to detect the relative gene and protein expression of SDF-1. Results: After 4 h of irradiation, myocardial tissue was taken and stained with hematoxylin and eosin (HE). The infiltration of inflammatory cells in each group was observed with no significant difference between the groups. Serological tests showed that serum CK and LDH in group 1 and group 2 were higher than (P <0.05). The serum TnI of group 1 was lower than that of group 2, but higher than that of group 3 and group 4 (P <0.05), and the difference was statistically significant (P <0.05). The results of RT-PCR and Western blot showed that the expression of SDF-1 in group 1 was the highest among all the groups, and the differences were statistically significant (P <0.05). CONCLUSIONS: Ultrasound combined with NO microbubbles irradiates myocardial tissue in rats, which is similar to ordinary lipid microbubbles containing perfluorinated gas, and has no obvious damage to myocardial tissue. However, SDF-1 expression is increased in myocardial tissue of rats The degree is higher than the application of ultrasound combined with ordinary lipid microbubbles, therefore, NO microbubbles applied to stem cell homing will be superior to the current ordinary lipid microbubbles.
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