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作 者:殷荷 吴琪瑞 高丽娜 张玉月 王艳华 朱亚飞 吴凯[2] 张慧萍[3,4,5] 姜怡邓[2,3,4] YIN He;WU Qi-rui;GAO Li-na;ZHANG Yu-yue;WANG Yan-hua;ZHU Ya-fei;WU Kai;ZHANG Hui-ping;JIANG Yi-deng(School of Clinical Medical,Ningxia Medical University,Yinchuan 750000,Ningxia Hui Autonomous Region,China;School of Basic Medical,Ningxia Medical University,Yinchuan 750000,Ningxia Hui Autonomous Region,China;National Health Commission Key Laboratory of Metabolic Cardiovascular Diseases Research,Ningxia Medical University,Yinchuan 750000,Ningxia Hui Autonomous Region,China;Ningxia Key Laboratory of Vascular Injury and Repair Research,Ningxia Medical University,Yinchuan 750000,Ningxia Hui Autonomous Region,China;General Hospital of Ningxia Medical University,Yinchuan 750000,Ningxia Hui Autonomous Region,China)
机构地区:[1]宁夏医科大学临床医学院,宁夏回族自治区银川750000 [2]宁夏医科大学基础医学院,宁夏回族自治区银川750000 [3]宁夏医科大学国家卫生健康委代谢性心血管疾病研究重点实验室,宁夏回族自治区银川750000 [4]宁夏医科大学宁夏血管损伤与修复研究重点实验室,宁夏回族自治区银川750000 [5]宁夏医科大学总医院妇产科,宁夏回族自治区银川750000
出 处:《中国临床药理学杂志》2021年第16期2180-2183,共4页The Chinese Journal of Clinical Pharmacology
基 金:国家自然科学基金资助项目(81760270);宁夏回族自治区重点研发计划重大基金资助项目(2019BFG02004,2018BEG02004,2018BEG03026);宁夏自然科学基金重点基金资助项目(2020AAC02038,2020AAC03380)。
摘 要:目的探讨环状RNA-RBM39(circRBM39)在缺氧致胎盘滋养细胞凋亡中的作用及其分子机制。方法以缺口末端标记法检测子痫前期(PE)患者胎盘滋养细胞凋亡情况。体外培养人源胎盘滋养细胞株(HTR8-S/Vneo),将其分为3组:第1转染组(空白对照)、第2转染组(转染5μL小干扰RNA阴性对照)和第3转染组(转染5μL circRBM39小干扰RNA),浓度均为20μmol·L^(-1),于1%O_(2)缺氧培养箱中培养48 h。通过蛋白质印迹法检测胱天蛋白酶-3(Caspase-3)、胱天蛋白酶-9(Caspase-9)、B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)蛋白的表达。结果PE患者与健康妊娠孕妇比较,胎盘组织凋亡明显增加。第1转染组、第2转染组和第3转染组的Caspase-3蛋白表达量分别为0.49±0.10,0.53±0.07,0.19±0.04;Caspase-9蛋白表达量分别为0.58±0.09,0.61±0.07和0.36±0.05;Bcl-2蛋白表达量分别为1.67±0.18,1.79±0.17和2.78±0.25;Bax蛋白表达量分别为3.07±0.20,3.11±0.18和2.30±0.21。第3转染组与第1转染组和第2转染组比较,上述指标差异均有统计学意义(均P<0.05)。结论下调circRBM39可抑制缺氧诱导的胎盘滋养细胞凋亡。Objective To investigate the effect and molecular mechanism of circular RNA RBM39(circRBM39) on the apoptosis of placental trophoblast cells induced by hypoxia. Methods Placental tissue apoptosis of preeclampsia(PE) patients was used by TdT-mediated dUTP nick-end labeling. Human placental trophoblast cell lines(HTR8-S/Vneo) were cultured in vitro and divided into three groups: transfection-1 group(blank control),transfection-2 group(transfected with 5 μL small interfering RNA negative control) and transfection-3 group(transfected with 5 μL circBM39 small interfering RNA) were both 20 μmol·L-1 and cultured in 1 % O2 hypoxia incubator for 48 h. The protein expressions of Cysteinyl aspartate specific proteinase-3(Caspase-3),Cysteinyl aspartate specific proteinase-9(Caspase-9),B-cell lymphoma-2(Bcl-2) and Bcl-2 associated X protein(Bax) were detected by Western blot. Results Compared with healthy pregnant women, PE patients’ placental tissue apoptosis significantly increased. The expression levels of Caspase-3 protein in transfection-1 group,transfection-2 group and transfection-3 group were 0. 49 ± 0. 10,0. 53 ± 0. 07,0. 19 ± 0. 04;the expression levels of Caspase-9 protein in the three groups were 0. 58 ± 0. 09,0. 61 ± 0. 07,0. 36 ± 0. 05;the expression levels of Bcl-2 in the three groups were 1. 67 ± 0. 18,1. 79 ± 0. 17,2. 78 ± 0. 25;the expression levels of Bax protein in the three groups were 3. 07 ± 0. 20,3. 11 ± 0. 18,2. 30 ± 0. 21. Comparison between transfection-3 group and transfection-1 group,or transfection-2 group,the differences in the above indicators are statistically significant( all P < 0. 05). Conclusion Downregulation of circRBM39 can inhibit apoptosis of placental trophoblast cells induced by hypoxia.
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