CAJ | 학술논문

目的探讨滋养细胞内质网超微结构变化及内质网应激特征性分子--内质网分子伴侣葡萄糖调节蛋白(GRP)78、94和内质网凋亡因子--半胱氨酸天冬氨酸蛋白酶12(caspase-12)mRNA及蛋白表达与子痫前期发病的关系.方法选择2008年7月-2010年1月在南京医科大学第一附属医院分娩的孕妇共65例,其中子痫前期孕妇30例(子痫前期组),健康孕妇35例为对照组.应用透射电镜扫描观察胎盘组织中滋养细胞内质网超微结构改变,逆转录(RT)PCR技术检测胎盘组织中GRP78、GRP94和caspase-12 mRNA的表达,蛋白印迹法检测胎盘组织中GRP78、GRP94和caspase-12蛋白的表达.结果 (1)对照组胎盘组织中滋养细胞内质网体积无明显增大,内质网无扩张也无肿胀变化;子痫前期组滋养细胞内质网水肿,数量减少,内质网体积增大,内质网扩张及液泡化,且内质网脱颗粒改变明显.(2)子痫前期组胎盘组织中GRP78 mRNA及蛋白表达水平分别为2.59±0.09及0.81±0.31,显著高于对照组的1.16±0.07及0.40±0.10,两组分别比较,差异均有统计学意义(P<0.01).(3)子痫前期组胎盘组织中GRP94 mRNA和蛋白表达水平分别为1.31±0.91及0.55±0.24,显著高于对照组的0.63±0.57及0.22±0.09,两组分别比较,差异均有统计学意义(P<0.01).(4)子痫前期组胎盘组织中caspase-12 mRNA和蛋白表达水平分别为4.03±0.65及1.56±0.17,显著高于对照组的1.85±0.85及0.91±0.69,两组分别比较,差异均有统计学意义(P<0.01).结论子痫前期孕妇的滋养细胞内质网有明显的扩张及肿胀性改变;胎盘组织中GRP78、GRP94以及caspase-12 mRNA及蛋白表达水平比健康孕妇有明显升高.提示,内质网应激介导的滋养细胞凋亡可能是子痫前期发病的又一重要机制.
목적 탐토자양세포내질망초미결구변화급내질망응격특정성분자--내질망분자반려포도당조절단백(GRP)78、94화내질망조망인자--반광안산천동안산단백매12(caspase-12)mRNA급단백표체여자간전기발병적관계.방법 선택2008년7월-2010년1월재남경의과대학제일부속의원분면적잉부공65례,기중자간전기잉부30례(자간전기조),건강잉부35례위대조조.응용투사전경소묘관찰태반조직중자양세포내질망초미결구개변,역전록(RT)PCR기술검측태반조직중GRP78、GRP94화caspase-12 mRNA적표체,단백인적법검측태반조직중GRP78、GRP94화caspase-12단백적표체.결과 (1)대조조태반조직중자양세포내질망체적무명현증대,내질망무확장야무종창변화;자간전기조자양세포내질망수종,수량감소,내질망체적증대,내질망확장급액포화,차내질망탈과립개변명현.(2)자간전기조태반조직중GRP78 mRNA급단백표체수평분별위2.59±0.09급0.81±0.31,현저고우대조조적1.16±0.07급0.40±0.10,량조분별비교,차이균유통계학의의(P<0.01).(3)자간전기조태반조직중GRP94 mRNA화단백표체수평분별위1.31±0.91급0.55±0.24,현저고우대조조적0.63±0.57급0.22±0.09,량조분별비교,차이균유통계학의의(P<0.01).(4)자간전기조태반조직중caspase-12 mRNA화단백표체수평분별위4.03±0.65급1.56±0.17,현저고우대조조적1.85±0.85급0.91±0.69,량조분별비교,차이균유통계학의의(P<0.01).결론 자간전기잉부적자양세포내질망유명현적확장급종창성개변;태반조직중GRP78、GRP94이급caspase-12 mRNA급단백표체수평비건강잉부유명현승고.제시,내질망응격개도적자양세포조망가능시자간전기발병적우일중요궤제.
Objective To evaluate the relationship between pathogenesis of preeclampsia (PE) and the ultrastructure change of the endoplasmic reticulum in trophocyte, mRNA and protein expression levels of endoplasmic reticulum molecular chaperone glucose-regulated protein 78 (GRP78), glucose-regulated protein 94 (GRP94), endoplasmic reticulum apoptosis factor cysteine protease protein 12 (caspase-12).Methods Sixty-five pregnant women who were hospitalized in the First Affiliated Hospital of Nanjing Medical University from July 2008 to January 2010, were selected as the subject. Thirty pregnancy women diagnosed with PE were divided into PE group and 35 normal pregnant women were used as control group.Electron Microscopy was used to measure ultrastructure change of the endoplasmic reticulum in placenta trophocyte. Reverse transcription(RT) PCR and western blot were used to investigute the expression levels of GRP78, GRP94, caspase-12 mRNA and protein in placenta. Results (1) In control group the volume of endoplasmic reticulum does not increase; no swelling and no expansion of endoplasmic reticulum was found.In PE group the edema number of endoplasmic reticulum was reduced; the volume of endoplasmic reticulum increased; expansion and vacuolation of cavity and degranulation of the endoplasmic reticulum was observed significantly. (2) The mRNA and protein expression levels of GRP78 in placenta of PE group (2.59 ± 0. 09 and 0. 81 ±0. 31) were significantly higher than those in placenta of control group (1. 16 ±0. 07 and 0. 40 ± 0. 10, P <0. 01). (3) The mRNA and protein expression levels of GRP94 in placenta of PE group (1.31 ± 0. 91 and 0. 55 ±0. 24) were significantly higher than those in placenta of control group (0. 63 ±0. 57 and 0. 22 ±0. 09, P < 0. 01). (4) The mRNA and protein expression levels of caspase-12 in placenta of PE group (4. 03 ± 0. 65 and 1.56 ± 0. 17) were significantly higher than those in placenta of control group (1.85 ± 0. 85 and 0. 91 ± 0. 69, P < 0. 01). Conclusion The obvious expansion of endoplasmic reticulum in trophocyte and the increased expression levels of GRP78, GRP94 and caspase-12 indicate that endoplasmic reticulum stress-mediated apoptosis may be involved in the pathophysiological processes of PE.