| 王锋,徐静,高亚南,等.黄芩苷对子痫前期大鼠胎盘滋养细胞增殖侵袭的作用机制研究[J].浙江中医药大学学报,2021,45(12):1358-1364, 1373. |
| 黄芩苷对子痫前期大鼠胎盘滋养细胞增殖侵袭的作用机制研究 |
| Mechanism of Baicalin on Proliferation and Invasion of Placental Trophoblast in Preeclampsia Rats |
| DOI：10.16466/j.issn1005-5509.2021.12.017 |
| 中文关键词: 子痫前期 黄岑苷 胎盘滋养细胞 增殖 侵袭 微小RNA-30d 神经胶质细胞缺失同源物-1 |
| 英文关键词: preeclampsia baicalin placental trophoblasts proliferation invasion microRNA-30d glial cells missing homolog-1 |
| 基金项目:河南省医学科技攻关计划(2018021025) |
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| 中文摘要: |
| [目的] 探讨黄岑苷对子痫前期大鼠胎盘组织滋养细胞增殖侵袭的影响及相关的作用机制。[方法] 选择36只健康清洁级SD大鼠，其中雌性24只，雄性12只，按照2∶1比例安排雌雄合笼，24只雌鼠妊娠第13天建立子痫前期大鼠模型，成功20只。从建模成功雌鼠中随机取5只进行原代滋养细胞分离、培养及传代，倒置显微镜下观察滋养细胞形态，以免疫荧光染色法鉴定滋养细胞纯度，四甲基偶氮唑蓝(methyl thiazolyl tetrazolium，MTT)法检测吸光度(optical density，OD)值，评估胎盘滋养细胞增殖活性。将其余15只建模成功雌鼠分为对照组(以0.3 mL蒸馏水灌胃)、黄岑苷低剂量组(以25 μg·mL-1黄岑苷灌胃)、黄岑苷高剂量组(以100 μg·mL-1黄岑苷灌胃)，灌胃后取腹主动脉血；同时将原代滋养细胞分为A组、B组、C组，分别以对照组、黄岑苷低剂量组、黄岑苷高剂量组大鼠血清培养48 h，采用Transwell法检测各组滋养细胞侵袭能力，实时荧光定量聚合酶链式反应(Real-time fluorescence quantitative polymerase chain reaction， Real-time qPCR)法检测微小RNA-30d(microRNA-30d，miR-30d)、神经胶质细胞缺失同源物-1(glial cells missing homolog-1，GCM-1)mRNA表达水平，免疫印迹法检测GCM-1蛋白相对表达量。[结果] 胎盘滋养细胞经消化、传代后，倒置显微镜下观察呈上皮样细胞形态，多为长多边形，体积大，核大呈卵圆形，呈片状延展生长，贴壁后呈铺路石样；滋养细胞纯度(90.65±5.64)%。与空白组比较，20、40、60、80、100 μg·mL-1黄岑苷组OD值增高(P＜0.05)。与A组比较，B组、C组侵袭细胞数增多，miR-30 d表达水平降低，GCM-1 mRNA及蛋白相对表达量升高(P＜0.05)；与B组比较，C组侵袭细胞数增多，miR-30 d表达水平降低，GCM-1 mRNA及及蛋白相对表达量升高(P＜0.05)。[结论] 黄岑苷可提升子痫前期大鼠胎盘滋养细胞增殖、侵袭能力，作用机制可能与抑制miR-30d表达、上调GCM-1蛋白表达有关。 |
| 英文摘要: |
| [Objective] To investigate the effect of baicalin on the proliferation and invasion of placental trophoblast in preeclampsia rats and the related mechanism. [Methods] Thirty-six clean and healthy SD rats(24 females and 12 males) were selected， the ratio of female to male was 2∶1. The preeclampsia rat model was established on the 13th day of pregnancy in 24 female rats， and 20 rats were successful. Five female rats were randomly selected for primary trophoblast isolation， culture and passage. The morphology of trophoblast was observed under inverted microscope， the purity of trophoblast was identified by immunofluorescence staining， and the optical density(OD) of placental trophoblast cells was evaluated by methyl thiazolyl tetrazolium(MTT) method. The remained 15 successful female rats were divided into control group(0.3 mL distilled water)， low-dose group(25 μg·mL-1) and high-dose group(100 μg·mL-1) of baicalin by gavage. At the same time， the trophoblasts were divided into group A， group B and group C， which were cultured in serum of control group， low-dose group and high-dose group of baicalin respectively for 48 hours. Transwell method was used to detect the invasion ability of placental trophoblast in each group. The expression levels of microRNA-30d(miR-30d) and glial cells missing homolog-1(GCM-1) mRNA were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time qPCR)， and the relative expression levels of GCM-1 protein were detected by Western blot. [Results] After digestion and passage， the placental trophoblast cells showed epithelioid cell morphology under inverted microscope， mostly long polygon， large volume， large nucleus， oval shape， elongated growth， and paving stone shape after adherence. The purity of trophoblast was (90.65±5.64)%. Compared with blank group， OD value of 20， 40， 60， 80， 100 μg·mL-1 baicalin groups increased(P<0.05). Compared with group A， the number of invaded cells in groups B and C was increased， the expression level of miR-30d was decreased， and the expression level of GCM-1 mRNA and protein were increased(P<0.05). Compared with group B， the number of invaded cells in group C was increased， the expression level of miR-30d was decreased， and the expression level of GCM-1 and protein were increased(P<0.05). [Conclusion] Baicalin can enhance the proliferation and invasion of placental trophoblasts in preeclampsia rats， and the mechanism may be related to the inhibition of miR-30d expression and up-regulation of GCM-1 protein expression. |
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