lncRNA GHET1通过Wnt/β-catenin信号通路对子痫前期滋养层细胞生物学行为的影响

目的 探讨子痫前期（PE）患者胎盘组织中长链非编码RNA胃癌高表达转录本1（lncRNA GHET1）的表达及其对滋养层细胞增殖、细胞周期进展和侵袭能力的影响,并阐明其作用机制。 方法 30名孕产妇分为正常孕产妇组（正常组）15例和PE孕产妇组（PE组）15例,HE染色观察2组研究对象胎盘组织病理形态表现。体外培养滋养层HTR-8细胞,转染过表达lncRNA GHET1及对照序列质粒,并将滋养层细胞分为对照组（常规培养）、GHET1组（转染过表达lncRNA GHET1质粒）和阴性对照组（转染阴性对照序列质粒）。实时荧光定量PCR（RT-qPCR）法检测2组研究对象胎盘组织和各组HTR-8细胞中lncRNA GHET1 mRNA表达水平,CCK-8法检测各组HTR-8细胞增殖率,流式细胞术检测各组不同细胞周期HTR-8细胞百分率,Transwell小室实验检测各组HTR-8细胞侵袭能力,Western blotting法检测各组HTR-8细胞中细胞性骨髓细胞瘤病病毒癌基因（c-Myc）、细胞周期素D1（cyclinD1）、上皮细胞-间充质转化（EMT）相关蛋白E-钙黏蛋白（E-cadherin）和波形蛋白（Vimentin）蛋白表达 水平及Wnt/β-catenin信号通路中磷酸化糖原合成酶激酶3β（p-GSK3β）/糖原合成酶激酶3β（GSK3β）比值和β-连环蛋白（β-catenin）蛋白表达水平。 结果 与正常组比较,PE组患者胎盘组织绒毛发育不良,数量减少,绒毛内及间质血管分布紊乱,血管壁出现纤维素样坏死,钙化区域及绒毛上合体结节增加。与正常组比较,PE组患者胎盘组织中lncRNA GHET1 mRNA表达水平明显降低（P<0.05）。与对照组比较,GHET1组HTR-8细胞中lncRNA GHET1 mRNA表达水平明显升高（P<0.05）,阴性对照组HTR-8细胞中lncRNA GHET1 mRNA表达水平差异无统计学意义（P>0.05）。与对照组比较,GHET1组HTR-8细胞增殖率、S期细胞百分率和侵袭细胞数明显升高（P<0.05）,阴性对照组上述指标差异无统计学意义（P>0.05）。与对照组比较,GHET1组HTR-8细胞中c-Myc、cyclinD1、Vimentin和β-catenin蛋白表达水平及p-GSK3β/GSK3β比值均明显升高（P<0.05）,E-cadherin蛋白表达水平明显降低（P<0.05）,阴性对照组上述指标差异无统计学意义（P>0.05）。 结论 过表达lncRNA GHET1可能通过Wnt/β-catenin信号通路促进滋养层细胞增殖、细胞周期进展和细胞侵袭,发挥改善PE进展的作用。     

孕早中期孕妇肠道菌群差异与子痫前期发病关系的研究

背景 子痫前期病情严重者可进展至子痫并威胁母婴生命安全,而肠道菌群构成变化可能参与子痫前期的发生、发展,但尚无明确证据。 目的 探讨孕早中期孕妇肠道菌群差异与子痫前期发病的关系。 方法 选取2019年1月至2021年1月桂林医学院第二附属医院产科招募且符合本研究纳入标准的孕妇455例。将妊娠20周后确诊为子痫前期的孕妇作为子痫前期组（n=32）,未发生子痫前期的孕妇作为非子痫前期组（n=423）。收集孕妇的临床资料,并分别留取孕早期（≤12+6周）、孕中期（13~27+6周）粪便标本进行肠道菌群生物信息学分析,分析其与子痫前期发病的关系。 结果 子痫前期组和非子痫前期组年龄、孕早期Shannon指数和Simpson指数比较,差异均有统计学意义（P<0.05）。多因素Logistic回归分析结果显示,年龄≥35周岁〔OR=1.894,95%CI（1.432,2.369）〕、孕早期Shannon指数下降〔OR=0.709,95%CI（0.465,0.921）〕、孕早期Simpson指数下降〔OR=0.612,95%CI（0.354,0.893）〕是孕妇发生子痫前期的独立危险因素（P<0.05）。孕早期Shannon指数预测孕妇子痫前期发病的受试者工作特征（ROC）曲线下面积为0.745〔95%CI（0.652,0.838）〕,截断值为6.255,灵敏度为76.58%,特异度为60.00%;孕早期Simpson指数预测孕妇子痫前期发病的ROC曲线下面积为0.724〔95%CI（0.623,0.826）〕,截断值为0.945,灵敏度为62.90%,特异度为60.61%。 结论 孕早期粪便肠道菌群Shannon指数、Simpson指数降低是孕妇发生子痫前期的独立危险因素,且对子痫前期发病具有早期预测价值。     

Hcy D-D UA水平联合检测在子痫前期诊断及重度子痫前期预测中的应用

目的 探讨同型半胱氨酸(Hcy)、D-二聚体(D-D)、尿酸(UA)血清水平联合检测在子痫前期(PE)诊断及重度PE预测中的应用价值。方法 回顾性分析2020年1月至2022年4月铜陵市妇幼保健院收治的229例孕妇的临床资料,参照《妇产科学》第八版中PE诊断标准将其分为PE组(n=148)与非PE组(n=81),根据PE病情严重程度将PE患者分为轻度PE组(n=85)与重度PE组(n=63)。记录并比较各组患者Hcy、D-D、UA水平,以《妇产科学》第八版中的PE、轻度PE、重度PE诊断标准为“金标准”绘制受试者工作特征(ROC)曲线,评价Hcy、D-D、UA水平联合检测在PE诊断及重度PE预测中的应用价值。结果 PE组孕妇收缩压、舒张压、平均动脉压、剖宫产史比例以及24 h尿蛋白均高于非PE组(P均<0.05)。PE组孕妇Hcy(12.85±1.4[JP2]3)μmol/L、D-D(2.39±0.42)μg/mL及UA(407.83±48.86)μmol/L均高于非PE组(9.51±1.14)μmol/L、(1.61±0.27)μg/mL、(321.43±41.67)μmol/L(P均<0.05)。Hcy+D-D+UA联合检测用于诊断PE的曲线下面积(AUC)为0.976,灵敏度为98.65%,特异度为97.53%,Hcy+D-D+UA联合检测PE诊断的效能优于Hcy、D-D、UA单独及任意两指标联合。重度PE组患者Hcy(13.98±1.52)μmol/L、D-D(2.12±0.35)μg/mL及UA(443.17±51.39)μmol/L均高于轻度PE组(12.01±1.37)μmol/L、(2.12±0.35)μg/mL、(381.64±46.98)μmol/L(P均<0.05)。PE患者Hcy水平分别与D-D、UA水平呈正相关(r=0.739、0.861,P均<0.05),D-D水平与UA水平呈正相关(r=0.708,P<0.05);而Hcy、D-D及UA水平分别与PE严重程度均呈正相关(r=0.654、0.813、0.776,P均<0.05)。Hcy+D-D+UA联合检测对重度PE预测的AUC为0.941,灵敏度为90.48%,特异度为95.29%,Hcy+D-D+UA联合检测较Hcy、D-D、UA单独及任意两指标联合用于重度PE预测的效能更优。结论 PE患者Hcy、D-D、UA水平均升高且相互呈正相关,而重度PE患者Hcy、D-D、UA水平均升高更为明显,Hcy+D-D+UA联合检测可提高PE诊断效能与重度PE预测效果,值得借鉴。     

长链脂肪酸氧化酶在妊娠中期子痫前期样母鼠和胎鼠组织中的表达

目的 探讨长链脂肪酸氧化酶LCHAD在妊娠中期子痫前期样改变母鼠和胎鼠组织中的蛋白表达及分布.方法 将C57BL/6J孕鼠于孕1id皮下注射一氧化氮合酶抑制剂建立子痫前期样模型,同时设立注射生理盐水的正常妊娠对照组,模型鉴定成功后,于孕14 d进行标本取材.采用免疫组化方法检测母鼠及胎鼠组织中LCHAD蛋白的分布和表达状况.结果 孕14 d实验组平均动脉压(115.6±3.3)mmHg(1 mmHg =0.133 kPa)和尿蛋白(97.0±4.3)μg·g体质量-1·24h-1较对照组平均动脉压(96.2±4.9)mmHg和尿蛋白(60.8 ±4.9) μg·g体质量-1·24 h-1显著升高(P＜0.05).在两组动物中均可见到母鼠肝、肾、心、胎盘组织中LCHAD蛋白呈阳性表达,实验组母体肝、胎盘LCHAD的IA值强度较对照组明显降低(P＜0.05).胎鼠的肝、肾、心、肺和神经视网膜组织中,两组均有LCHAD表达,但IA值差异无统计学意义(P＞0.05).结论 妊娠中期LCHAD蛋白在母鼠和胎鼠重要器官均广泛表达分布,在妊娠期LCHAD与母体能量代谢、胎盘的发育及胎儿生长发育有关;子痫前期样改变时母体肝脏和胎盘的LCHAD表达存在异常.     

AKR1C3对子痫前期大鼠肾损伤的影响

目的 探讨AKR1C3对子痫前期大鼠肾脏损伤的影响及其可能的作用机制.方法 通过腹腔注射亚硝基左旋精氨酸甲酯(L-NAME)建立大鼠子痫前期的模型,治疗组同时采用灌胃的方法口服格列苯脲.40只Wistar雌鼠随机分为子痫前期组(孕鼠+L-NAME)、治疗组(孕鼠+L-NAME+格列苯脲)、未孕组(L-NAME)、对照组(孕鼠+生理盐水),每组10只.通过测血压、24 h尿蛋白和透射电镜观察肾脏结构确认子痫前期大鼠造模成功,采用RT-PCR、免疫印迹检测大鼠肾脏AKR1C3蛋白及mRNA含量,Elisa方法检测超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)的浓度.结果 造模前各组大鼠的血压值差异无统计学意义;造模后子痫前期组血压明显高于对照组与治疗组[(143.83±9.62)比(120.83±4.31)及(129.43±14.41) mmHg,均P＜0.05].造模前各组大鼠的24 h尿蛋白含量无差异;造模后子痫前期组的24h尿蛋白含量明显高于对照组与治疗组.子痫前期组大鼠肾脏中AKR1 C3蛋白及mRNA表达水平明显低于对照组[(0.48±0.09)比(0.98 ±0.27)、(0.05 ±0.02)比(0.87±0.45),均P＜0.05].与子痫前期组相比,治疗组AKR1 C3蛋白及mRNA表达水平升高[(0.48±0.09)比(1.05±0.20)、(0.05±0.02)比(0.22 ±0.06),均P＜0.05].与对照组相比子痫前期组SOD、CAT、GSH-Px的浓度明显降低,MDA的浓度升高;与子痫前期组相比,治疗组SOD、CAT、GSH-Px的浓度升高,MDA的浓度降低.结论 AKR1C3在子痫前期大鼠肾脏中表达降低,其作用途径可能与氧化应激有关.格列苯脲对子痫前期的治疗有一定的疗效.     

Differences in Liver Injury and Trophoblastic Mitochondrial Damage in Different Preeclampsia-like Mouse Models

Background:

Preeclampsia is a multifactorial disease during pregnancy. Dysregulated lipid metabolism may be related to some preeclampsia. We investigated the relationship between triglycerides (TGs) and liver injury in different preeclampsia-like mouse models and their potential common pathways.

Methods:

Preeclampsia-like models (Nw-nitro-L-arginine-methyl ester [L-NAME], lipopolysaccharide [LPS], apolipoprotein C-III [Apo] transgnic mice + L-NAME, β2 glycoprotein I [βGPI]) were used in four experimental groups: L-NAME (LN), LPS, Apo-LN and βGPI, respectively, and controls received saline (LN-C, LPS-C, Apo-C, βGPI-C). The first three models were established in preimplantation (PI), early-, mid- and late-gestation (EG, MG and LG). βGPI and controls were injected before implantation. Mean arterial pressure (MAP), 24-hour urine protein, placental and fetal weight, serum TGs, total cholesterol (TC) and pathologic liver and trophocyte changes were assessed.

Results:

MAP and proteinuria were significantly increased in the experimental groups. Placenta and fetal weight in PI, EP and MP subgroups were significantly lower than LP. Serum TGs significantly increased in most groups but controls. TC was not different between experimental and control groups. Spotty hepatic cell necrosis was observed in PI, EG, MG in LN, Apo-LN and βGPI, but no morphologic changes were observed in the LPS group. Similar trophoblastic mitochondrial damage was observed in every experimental group.

Conclusions:

Earlier preeclampsia onset causes a higher MAP and urine protein level, and more severe placental and fetal damage. Preeclampsia-like models generated by varied means lead to different changes in lipid metabolism and associated with liver injury. Trophoblastic mitochondrial damage may be the common terminal pathway in different preeclampsia-like models.     

妊娠期高血压疾病对胎龄28~34周早产儿外周静脉血细胞计数的影响

目的 探讨母亲妊娠期高血压疾病（hypertensive disorders of pregnancy,HDP）对胎龄28~34周早产儿外周静脉血细胞计数的影响。 方法 选取2020年1~12月昆明医科大学第一附属医院儿科收治的母亲合并HDP的胎龄28~34周早产儿227例为研究组,另选取同期收治的母亲无HDP的胎龄28~34周早产儿227例为对照组。研究组根据母亲妊娠期血压分为妊娠期高血压亚组（75例）、轻度子痫前期亚组（81例）、重度子痫前期亚组（71例）;根据早产儿出生体重分为小于胎龄儿（small for gestational age,SGA）亚组（113例）及适于胎龄儿（appropriate for gestational age,AGA）亚组（114例）。比较研究组和对照组、研究组各亚组间早产儿生后第1天外周血细胞计数的差异。 结果 研究组患儿生后第1天外周静脉血白细胞（white blood cell,WBC）计数、中性粒细胞绝对计数（absolute neutrophil count,ANC）及血小板（platelet,PLT）计数均低于对照组（P<0.05）,白细胞减少症、中性粒细胞减少症发生率高于对照组（P<0.05）。亚组分析中,轻度子痫前期亚组、重度子痫前期亚组WBC计数、ANC、PLT计数均低于妊娠期高血压亚组（P<0.05）;SGA亚组WBC计数、ANC、PLT计数低于AGA亚组（P<0.05）。 结论 HDP可对早产儿外周静脉血细胞计数产生影响,这一影响在母亲子痫前期及SGA早产儿中更为显著。     

胎盘早剥25例临床分析

目的:探讨胎盘早剥的发病原因、胎盘早剥与妊高症及急性弥漫性血管内凝血(DIC)的关系。方法:对2000年1月-2004年7月25例胎盘早剥患者的临床资料进行回顾性分析。结果:(1)25例胎盘早剥患者的发病原因主要是妊高症、机械性因素(包括私人不恰当操作)、羊水过多、胎膜早破;(2)25例胎盘早剥患者中妊高症10例(40%),重度妊高症6例(24%);(3)25例胎盘早剥患者并发DIC6例(24%);(4)25例胎盘早剥患者中死胎5例(20%),新生儿死亡2例(8%),死产1例(4%),新生儿窒息5例(20%),孕产妇死亡1例(4%),次全子宫切除5例(20%),产后出血9例(36%)。6例并发DIC患者中死胎4例(66.7%),新生儿重度窒息1例(16.7%);产妇死亡1例(16.7%),次全子宫切除4例(66.7%)。结论:胎盘早剥是严重威胁孕产妇及胎儿生命的妊娠晚期并发症,尤其是重度妊高症诱发的胎盘早剥,易并发急性DIC,威胁母婴安全。     

Angiogenic imbalance and plasma lipid alterations in women with preeclampsia from a developing country

Background: An imbalance between anti-angiogenic factors (e.g. soluble vascular endothelial growth factor receptor-1 (s-FLT1) and soluble endoglin (s-Eng)) and pro-angiogenic factors (e.g. placental growth factor (PlGF)) as well as increased oxidized low-density lipoprotein (ox-LDL) concentrations have been associated with preeclampsia (PE). Risk factors associated with the development of PE, however, are known to be different between developed and developing countries. The aim of the study was to determine the levels of s-FLT1, s-Eng, PIGF, and ox-LDL in women with PE from a developing country. Methods: A multi-center case–control study was conducted. One hundred and forty three women with PE were matched by age and parity with 143 healthy pregnant women without cardiovascular or endocrine diseases. Before delivery, blood samples were taken and serum was stored until analysis. Results: Women with PE had lower concentrations of PIGF (p < 0.0001) and higher concentrations of s-Eng (p = 0.001) than healthy pregnant women. There were no differences between the groups regarding ox-LDL or s-FLT1. Women with early onset PE had higher s-FLT1 concentrations (p = 0.0004) and lower PIGF concentrations (p < 0.0001) than their healthy pregnant controls. Women with late onset PE had higher concentrations of s-Eng (p = 0.005). Women with severe PE had higher concentrations of s-Eng (p = 0.0008) and ox-LDL (p = 0.01), and lower concentrations of PIGF (p < 0.0001). Conclusions: Women with PE from a developing country demonstrated an angiogenic imbalance and an increased rate of LDL oxidation. Findings from this study support the theory that PE is a multifactorial disease, and understanding differences in these subpopulations may provide a better target to approach future therapies.     

Role of microparticles in recurrent miscarriages and other adverse pregnancies: a review

The multiple functions attributed to microparticles (MPs) include blood coagulation, inflammation, tumorigenesis, angiogenesis, immunomodulatory functions and intercellular cross talk. These have drawn considerable interest during the last few years. The prothrombotic nature of MPs has linked them with almost all groups of thrombotic disorders including recurrent miscarriage (RM) and other abnormal pregnancy outcomes. Two authors (SS and RP) conducted a search independently on the computerized databases MEDLINE and EMBASE using relevant key words. Contradictory reports were observed on the association of MPs with RM. While most of the reports showed increased prevalence of MPs, both platelet and endothelial cell derived, in RM, some did not show any association. Almost all the reports showed a strong association of MPs with preeclampsia (PE), while the association with other adverse pregnancy conditions was not very conclusive. It may be concluded that MPs by themselves may result in adverse conditions or that they may be additive factors to an already existing prothrombotic state due to acquired or genetic thrombophilia or some unknown thrombophilic condition, besides the pre-existing hypercoagulable status of pregnancy.