化瘀消癥颗粒对miR-210过表达与低氧状态下滋养细胞血管生成、侵袭的影响

Effects of Huayu Xiaozheng Granules on Angiogenesis and Invasion in Trophoblast Cells with Overexpression of MiR-210 and Under Hypoxia

【目的】观察化瘀消癥颗粒对miR-210过表达与低氧状态下滋养细胞血管生成、侵袭的影响。【方法】体外培养HTR-8/SVneo细胞,通过转染建立miR-210过表达模型,以CoCl2法建立化学低氧模型,将实验分为空白组、miR-210过表达组、miR-210过表达阴性组、miR-210过表达+中药组、低氧+中药组及中药组,采用Western Blot法检测各组HTR-8/SVneo细胞中缺氧诱导因子1α(HIF-1α)、血管生成因子(VEGF)、侵袭相关因子[基质金属蛋白酶9(MMP-9)、金属蛋白酶组织抑制剂1(TIMP-1)]蛋白的表达。【结果】与空白组比较,miR-210过表达组中HIF-1α、VEGF、MMP-9、TIMP-1蛋白表达水平及MMP-9/TIMP-1比值升高(P<0.05);与miR-210过表达组比较,miR-210过表达阴性组、miR-210过表达+中药组、低氧+中药组、中药组中HIF-1α、VEGF、MMP-9、TIMP-1蛋白表达水平及MMP-9/TIMP-1比值均下降(P<0.05),中药组下降作用尤其明显;与miR-210过表达+中药组比较,低氧+中药组、中药组中HIF-1α、VEGF、MMP-9、TIMP-1蛋白表达水平及MMP-9/TIMP-1比值下降(P<0.05);与低氧+中药组比较,中药组上述各指标均下降(P<0.05)。【结论】化瘀消癥颗粒对miR-210过表达及低氧状态下滋养细胞所增强的血管生成、侵袭能力具有负调控作用,这可能是其治疗输卵管妊娠的作用机制之一。

Objective To observe the effect of Huayu Xiaozheng Granules on angiogenesis and invasion in trophoblast cells with overexpression of miR-210 and under hypoxia.Methods HTR-8/SVneo cells were cultured in vitro,a miR-210 overexpression model was established by transfection,and a chemical hypoxia model was established by CoCl2 method.The blank group,miR-210 overexpression group,miR-210 overexpressionnegative group,miR-210 overexpression+Chinese medicine group,hypoxia+Chinese medicine group and Chinese medicine group in experiment were set up.Western Blot method was used to detect protein expressions of hypoxia-inducible factor 1α(HIF-1α),vascular endothelial growth factor(VEGF),and invasion-associated factors[matrix metalloproteinase 9(MMP-9)and tissue inhibitors of matrix metalloproteinase 1(TIMP-1)]in HTR-8/SVneo cells.Results Compared with the blank group,the protein expression levels of HIF-1α,VEGF,MMP-9,TIMP-1 and MMP-9/TIMP-1 ratio were increased in the miR-210 overexpression group(P<0.05);compared with the miR-210 overexpression group,the HIF-1α,VEGF,MMP-9,TIMP-1 protein expression levels and MMP-9/TIMP-1 ratio in the miR-210 overexpression-negative group,miR-210 overexpression+Chinese medicine group,hypoxia+Chinese medicine group and Chinese medicine group were all decreased(P<0.05),and the decrease effects in Chinese medicine group was especially obvious;compared with the miR-210 overexpression+Chinese medicine group,the protein expression levels of HIF-1α,VEGF,MMP-9,TIMP-1 and MMP-9/TIMP-1 ratio in hypoxia+Chinese medicine group and Chinese medicine group were all decreased(P<0.05);compared with the hypoxia+Chinese medicine group,the above indicators were decreased in the Chinese medicine group(P<0.05).Conclusion Huayu Xiaozheng Granules can negatively regulate the increased angiogenesis and invasion of trophoblast cells with miR-210 overexpression and under hypoxia,which may be one of the action mechanisms of its treatment for tubal pregnancy.