汉黄芩素调控Akt/mTOR通路对流产大鼠滋养层细胞凋亡及自噬的影响  

Influences of wogonin on the apoptosis and autophagy of trophoblast cells in aborted rats by regulating Akt/mTOR pathway

作　　者：王静[1] 李芬霞[1] 郑艳丽 WANG Jing;LI Fenxia;ZHENG Yanli(Department of Obstetrics and Gynecology,Second Affiliated Hospital of Xi’an Medical University,Xi’an,Shaanxi 710038,China)

机构地区：[1]西安医学院第二附属医院产一科,陕西西安710038

出　　处：《中国优生与遗传杂志》2023年第11期2206-2211,共6页Chinese Journal of Birth Health & Heredity

摘　　要：目的探讨汉黄芩素(Wog)对流产大鼠滋养层细胞凋亡及自噬的影响以及可能的机制。方法将正常妊娠大鼠在妊娠第7天时灌胃8.3mg/kg米非司酮和0.1 mg/kg米索前列醇以构建流产大鼠模型,取出正常妊娠大鼠及流产大鼠胎盘,并分离出绒毛膜滋养层细胞,分别命名为对照组、模型组。将模型组滋养层细胞随机分为Wog低剂量组(Wog-L组)、Wog中剂量组(Wog-M组)、Wog高剂量组(Wog-H组)、Wog-H+MK2206(Akt抑制剂)组,流式细胞术检测细胞凋亡;透射电镜观察滋养层细胞中自噬小体的形成;免疫荧光检测细胞自噬标志蛋白微管相关蛋白1轻链3(LC3)的表达;Westernblot检测滋养层细胞中Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved-caspase-3)、Beclin1、磷酸化蛋白激酶B(p-Akt)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)蛋白表达。结果与对照组比较,模型组滋养层细胞凋亡率、LC3阳性率、Bax、cleaved-caspase-3、Beclin1蛋白表达及自噬小体数量升高,p-Akt/Akt、p-mTOR/mTOR表达降低(P<0.05);与模型组比较,Wog-L组、Wog-M组、Wog-H组滋养层细胞凋亡率、LC3阳性率、Bax、cleaved-caspase-3、Beclin1蛋白表达及自噬小体数量降低,p-Akt/Akt、p-mTOR/mTOR表达升高,且呈剂量依赖性(P<0.05);MK2206减弱了高剂量Wog对流产大鼠滋养层细胞凋亡及自噬的抑制作用。结论Wog可能通过激活Akt/mTOR通路抑制流产大鼠滋养层细胞凋亡及自噬。Objective To investigate the effect of wogonin(Wog)on apoptosis and autophagy of trophoblast cells in aborted rats and its possible mechanism.Methods On the seventh day of pregnancy,normal pregnant rats were given 8.3 mg/kg mifepristone and 0.1 mg/kg misoprostol by gavage to establish an abortion rat model.The placentas of normal pregnant rats and aborted rats were taken out,and chorionic trophoblast cells were isolated,which were named as control group and model group,respectively.The trophoblast cells in model group were randomly divided into Wog low-dose group(Wog-L group),Wog middle-dose group(Wog-M group),and Wog high-dose group(Wog-H group),and Wog-H+MK2206(Akt inhibitor)group.Flow cytometry was used to detect apoptosis.The formation of autophagosomes in trophoblast cells was observed by transmission electron microscope.The expression of autophagy marker protein microtubule-associated protein 1 light chain 3(LC3)was detected by immunofluorescence.Western blot was used to detect the protein expression of Bcl-2-associated X protein(Bax),activated cysteine-containing caspase-3(cleaved-caspase-3),Beclin1,phosphorylated protein kinase B(p-Akt),phosphorylated mammalian target of rapamycin(p-mTOR)protein expression in trophoblast cells.Results Compared with the control group,the model group had higher trophoblast cell apoptosis rate,LC3 positive rate,Bax,cleaved-caspase-3,Beclin1 protein expression and the number of autophagosomes increased,and p-Akt/Akt and p-mTOR/mTOR expression decreased(P<0.05).Compared with model group,Wog-L group,Wog-M group,Wog-H group had decreased trophoblast cell apoptosis rate,LC3 positive rate,Bax,cleaved-caspase-3,Beclin1 protein expression and the number of autophagosomes,and increased expression of p-Akt/Akt and p-mTOR/mTOR,in a dose-dependent manner(P<0.05).MK2206 attenuated the inhibitory effects of high-dose Wog on apoptosis and autophagy of trophoblast cells in aborted rats.Conclusion Wog may inhibit apoptosis and autophagy of trophoblast cells in aborted rats by activating Akt/

关 键 词：汉黄芩素 蛋白激酶B/哺乳动物雷帕霉素蛋白通路 滋养层细胞 自噬 凋亡 

分 类 号：R285.5[医药卫生—中药学]