探讨微小RNA-383(miR-383)靶向调控可溶性血管内皮生长因子受体1(sFlt-1)介导的磷酸肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路，在子痫前期(PE)发生中的作用机制。

选择2022年1月至12月在海南西部中心医院产科住院分娩的58例PE孕妇作为研究对象，将其分别纳入研究组1(n＝29，重度PE孕妇)与研究组2(n＝29，轻度PE孕妇)，选择同期在同一家医院住院分娩的29例健康孕妇作为对照，纳入对照组1。选择12只无特殊病原体(SPF)级健康妊娠5 d的小鼠为研究对象，分别纳入模型＋转染组1[n＝3，采用尾静脉注射磷脂酰丝氨酸(PS)/磷脂酰胆碱(PC)乳化微团成功构建小鼠PE病理模型后，再用miR-383 mimics转染复合物处理]，模型＋转染组2(n＝3，采用尾静脉注射PS/PC乳化微团成功构建小鼠PE病理模型后，再用NC agomir转染复合物处理)，模型组(n＝3，采用尾静脉注射PS/PC乳化微团成功构建小鼠PE病理模型后，再用生理盐水处理)，对照组2(n＝3，仅采用尾静脉注射生理盐水处理措施)。采用酶联免疫吸附试验(ELISA)方法检测3组孕妇、4组孕鼠的血清sFlt-1蛋白水平，采用实时荧光定量逆转录聚合酶链反应(RT-qPCR)方法测定3组孕妇、4组孕鼠胎盘组织miR-383和sFlt-1 mRNA相对表达水平。采用Western blot法测定4组孕鼠胎盘组织PI3K、磷酸化PI3K(p-PI3K)、Akt、磷酸化Akt(p-Akt)蛋白相对表达水平。采用单因素方差分析分别对3组孕妇、4组孕鼠上述检查结果进行统计学比较，进一步分别两两比较，均采用Student-Newman-Keuls q检验。本研究经过海南西部中心医院伦理委员会审批(审批文号：LLKY-2023-10)，并与所有孕妇签署临床研究知情同意书。3组孕妇年龄、初产妇与经产妇构成比、民族等一般临床资料比较，差异均无统计学意义(P>0.05)。

①3组孕妇分娩时收缩压、舒张压、分娩孕龄、24 h尿蛋白水平分别比较，差异均有统计学意义(F＝214.94、226.74、9.40、449.76，P<0.001)。②研究组1、2与对照组1孕妇的血清sFlt-1蛋白水平分别为(2 941.3±490.1)、(2 189.5±455.3)、(1 409.1±318.1)ng/L，3组总体比较，差异有统计学意义(F＝93.08，P<0.001)。进一步对其进行两两比较的结果显示，孕妇血清sFlt-1蛋白水平依次为研究组1>研究组2>对照组1，并且差异均有统计学意义(均为P<0.001)。③3组孕妇胎盘组织miR-383和sFlt-1 mRNA相对表达水平分别总体比较，差异均有统计学意义(P<0.001)。进一步分别对其进行两两比较结果显示，miR-383相对表达水平依次为研究组1<研究组2<对照组1，sFlt-1 mRNA相对表达水平依次为研究组1>研究组2>对照组1，并且差异亦均有统计学意义(均为P<0.05)。④ 4组孕鼠的血清sFlt-1蛋白表达水平总体比较，差异有统计学意义(F＝136.49，P<0.001)。进一步对其进行两两比较的结果显示，模型＋转染组1孕鼠血清sFlt-1蛋白表达水平低于模型＋转染组2和模型组，对照组2低于其余3组，并且差异均有统计学意义(均为P<0.001)。⑤4组孕鼠胎盘组织中miR-383和sFlt-1 mRNA相对表达水平分别总体比较，差异均有统计学意义(均为P<0.001)。进一步对其分别进行两两比较的结果显示，模型＋转染组1孕鼠miR-383相对表达水平高于其余3组，对照组2高于模型＋转染组2和模型组，并且差异均有统计学意义(均为P<0.001)。在sFlt-1 mRNA相对表达水平方面，模型＋转染组1孕鼠明显低于模型＋转染组2和模型组，对照组2孕鼠低于其余3组，并且差异均有统计学意义(均为P<0.001)。⑥4组孕鼠胎盘组织PI3K、p-PI3K、Akt、p-Akt蛋白相对表达水平分别总体比较，差异均有统计学意义(P<0.001)。进一步对其进行两两比较的结果显示，模型＋转染组1这4种蛋白水平均显著高于模型＋转染组2和模型组，对照组2均高于其余3组，并且差异均有统计学意义(均为P<0.05)。

miR-383与sFlt-1在孕鼠与孕妇PE发病中，均发挥着重要作用，miR-383表达降低，导致sFlt-1表达上调，进而介导PI3K/Akt信号通路下调，参与孕鼠与孕妇的PE发生、发展。

To investigate the mechanism of microRNA-383 (miR-383) in regulating soluble fms-like tyrosine kinase-1 (sFlt-1)-mediated phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in the development of preeclampsia (PE).

A total of 58 pregnant women with PE who delivered at the Department of Obstetrics of Hainan Western Central Hospital from January to December 2022 were selected as the study subjects. They were included into the severe PE group (n=29) and mild PE group (n=29) according to the severity of PE. Twenty-nine healthy pregnant women who were hospitalized and delivered in the same hospital during the same period were enrolled as controls and assigned to control group 1. In parallel, 12 specific pathogen-free (SPF) healthy female pregnant Kunming mice on day 5 of gestation were selected as the study subjects and randomly included in the following groups: model + transfection group 1 [n=3, mice were first treated with tail vein injection of phosphatidylserine (PS)/phosphatidylcholine (PC) emulsion microdroplets to establish a PE-like pathological model, and then administered with the miR-383 mimics transfection complex], model + transfection group 2 (n=3, mice were first treated with tail vein injection of PS/PC emulsion microdroplets to establish a PE-like pathological model, and then administered with the NC agomir mimics transfection complex), model group (n=3, mice were first treated with tail vein injection of PS/PC emulsion microdroplets to establish a PE-like pathological model, and then administered with saline), and control group 2 (n=3, treated with tail vein injection of saline). Serum sFlt-1 protein levels in the three groups of pregnant women and four groups of pregnant mice were measured by enzyme-linked immunosorbent assay (ELISA). The relative expression levels of miR-383 and sFlt-1 mRNA in placental tissues from the three groups of pregnant women and four groups of pregnant mice were determined by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). The relative protein expression levels of PI3K, phosphorylated PI3K (p-PI3K), Akt, and phosphorylated Akt (p-Akt) in placental tissues of four groups of pregnant mice were measured by Western blot. One-way ANOVA method was used to statistically compare the above examination results of the three groups of pregnant women and the four groups of pregnant mice. Further pairwise comparisons were performed using the Student-Newman-Keuls q test. This study was approved by the Ethics Committee of Hainan Western Central Hospital (Approval No. LLKY-2023-10), and written informed consents were obtained from all pregnant women. There were no statistically significant differences in general clinical data, including maternal age, proportion of primiparas and multiparas, and ethnicity, among the three groups of pregnant women (all P>0.05).

① There were statistically significant differences among the three groups of pregnant women in systolic blood pressure, diastolic blood pressure, gestational age at delivery, and 24-hour urinary protein levels (F=214.94, 226.74, 9.40, 449.76; all P<0.001). ② Serum sFlt-1 protein levels in the pregnant women of severe PE group, mild PE group, and control group 1 were (2 941.3±490.1), (2 189.5±455.3), and (1 409.1±318.1) ng/L, respectively, with a statistically significant difference among the three groups (F=93.08, P<0.001). Further pairwise comparisons revealed that serum sFlt-1 protein levels were highest in the severe PE group, followed by the mild PE group and the control group 1, and all the differences were statistically significance (all P<0.001). ③ The relative expression levels of miR-383 and sFlt-1 mRNA in placental tissues of the three groups of pregnant women were compared overall, respectively, and both the differences were statistically significant (P<0.001). Further pairwise comparisons showed that the relative expression levels of miR-383 were lowest in the severe PE group, followed by the mild PE group and the control group 1, while the relative expression levels of sFlt-1 mRNA were highest in the severe PE group, followed by the mild PE group and the control group 1, and all the differences were statistically significant (all P<0.05). ④ Overall comparison of serum sFlt-1 protein levels among the four groups of pregnant mice revealed a statistically significant difference (F=136.49, P<0.001). Further pairwise comparisons showed that the model + transfection group 1 had had significantly lower serum sFlt-1 protein level than the model + transfection group 2 and model group, control group 2 had significantly lower serum sFlt-1 protein levels than the other three groups, and with all differences being statistically significant (all P<0.001). ⑤ Overall comparisons of relative expression levels of placental miR-383 and sFlt-1 mRNA among the four groups of pregnant mice showed statistically significant differences (both P<0.001). Further pairwise comparisons revealed that, for miR-383 expression, model + transfection group 1 had significantly higher levels than the other 3 groups, and the control group 2 had higher level than the model + transfection group 2 and the model group, and all the differences were statistically significant (all P<0.001). For sFlt-1 mRNA expression, the model + transfection group 1 had significantly lower level than the model + transfection group 2 and the model group, and the control group 2 had lower level than the other three groups, and all the differences were statistically significant (all P<0.001). ⑥ Overall comparisons of relative expression levels of placental PI3K, p-PI3K, Akt, and p-Akt protein among the four groups of pregnant mice showed statistically significant differences (all P<0.001). Further pairwise comparisons demonstrated that the model + transfection group 1 had significantly higher levels of these four proteins than the model + transfection group 2 and the model group, the control group 2 had higher levels of these four proteins than the other three groups, and all the differences were statistically significant (all P<0.05).

miR-383 and sFlt-1 play important roles in the pathogenesis of PE in pregnant mice and women. Downregulation of miR-383 leads to upregulation of sFlt-1, which in turn mediates the downregulation of the PI3K/Akt signaling pathway, thereby contributing to the onset and progression of PE in pregnant mice and women.