国际遗传学杂志
國際遺傳學雜誌
국제유전학잡지
INTERNATIONAL JOURNAL OF GENETICS
2012年
3期
151-156
,共6页
庞海燕%孟毅%赵薇%刘国艺%温海霞
龐海燕%孟毅%趙薇%劉國藝%溫海霞
방해연%맹의%조미%류국예%온해하
大豆异黄酮%Genistein%围绝经期%卵巢%颗粒细胞%LRH-1
大豆異黃酮%Genistein%圍絕經期%卵巢%顆粒細胞%LRH-1
대두이황동%Genistein%위절경기%란소%과립세포%LRH-1
Soy Isoflavones (SI)%Genistein%Perimenopause%Ovary%Granulosa cell%Liver receptor homolog-1(LRH-1)
目的 观察大豆异黄酮(soy isoflavones,SI)及主要活性成分Genistein对初老大鼠卵巢及体外培养卵巢颗粒细胞中肝受体类似物-1(liver receptor homolog-1,LRH-1)基因表达的影响,初步探讨大豆异黄酮作用于卵巢的分子机制.方法 采用自然老化法建立围绝经期大鼠动物模型,分别给予低剂量(50mg/kg)、中剂量(158 mg/kg)、高剂量(500 mg/kg)的SI灌胃处理8 w.分别采用原位杂交和RT-PCR检测卵巢组织中LRH-1 mRNA的表达.采用大鼠孕马血清促性腺激素(pregnant mare serum gonadotrophin,PMSG)皮下注射法建立促卵泡发育模型,分离、培养颗粒细胞24 h后,给予大豆异黄酮主要活性成分Genistein (0、0.1、1、5、10、100 μmol/L)以及雌激素受体(estrogen receptor,ER)阻断剂ICI182、780(1μmol/L)继续处理细胞24 h,采用RT- PCR方法检测卵巢颗粒细胞中LRH-1 mRNA的表达情况.结果 各剂量SI处理组卵巢LRH-1 mRNA的表达(低剂量0.563±0.037,中剂量0.926±0.127和高剂量1.223±0.134),与围绝经期模型组(0.460±0.082)相比均显著增加(P＜0.05).1～10 μmol/L的Genistein作用卵巢颗粒细胞24 h,LRH-1 mRNA的表达(分别为0.844±0.042,0.879±0.056,0.882±0.079)与空白对照组(0.678±0.052)相比均增加,结果具有统计学意义(P＜0.05).ER阻断剂ICI182,780预处理细胞30 min后再给予Genistein继续处理细胞24 h后,发现1～10 μmol/L的Genistein上调LRH-1表达作用,并未受ER阻断剂的影响.结论 一定剂量的大豆异黄酮可明显上调衰老卵巢LRH-1 mRNA的表达,1～10 μmol/L的Genistein可上调体外培养大鼠卵巢颗粒细胞中LRH-1 mRNA的表达,可能并非通过经典ER介导的.
目的 觀察大豆異黃酮(soy isoflavones,SI)及主要活性成分Genistein對初老大鼠卵巢及體外培養卵巢顆粒細胞中肝受體類似物-1(liver receptor homolog-1,LRH-1)基因錶達的影響,初步探討大豆異黃酮作用于卵巢的分子機製.方法 採用自然老化法建立圍絕經期大鼠動物模型,分彆給予低劑量(50mg/kg)、中劑量(158 mg/kg)、高劑量(500 mg/kg)的SI灌胃處理8 w.分彆採用原位雜交和RT-PCR檢測卵巢組織中LRH-1 mRNA的錶達.採用大鼠孕馬血清促性腺激素(pregnant mare serum gonadotrophin,PMSG)皮下註射法建立促卵泡髮育模型,分離、培養顆粒細胞24 h後,給予大豆異黃酮主要活性成分Genistein (0、0.1、1、5、10、100 μmol/L)以及雌激素受體(estrogen receptor,ER)阻斷劑ICI182、780(1μmol/L)繼續處理細胞24 h,採用RT- PCR方法檢測卵巢顆粒細胞中LRH-1 mRNA的錶達情況.結果 各劑量SI處理組卵巢LRH-1 mRNA的錶達(低劑量0.563±0.037,中劑量0.926±0.127和高劑量1.223±0.134),與圍絕經期模型組(0.460±0.082)相比均顯著增加(P＜0.05).1～10 μmol/L的Genistein作用卵巢顆粒細胞24 h,LRH-1 mRNA的錶達(分彆為0.844±0.042,0.879±0.056,0.882±0.079)與空白對照組(0.678±0.052)相比均增加,結果具有統計學意義(P＜0.05).ER阻斷劑ICI182,780預處理細胞30 min後再給予Genistein繼續處理細胞24 h後,髮現1～10 μmol/L的Genistein上調LRH-1錶達作用,併未受ER阻斷劑的影響.結論 一定劑量的大豆異黃酮可明顯上調衰老卵巢LRH-1 mRNA的錶達,1～10 μmol/L的Genistein可上調體外培養大鼠卵巢顆粒細胞中LRH-1 mRNA的錶達,可能併非通過經典ER介導的.
목적 관찰대두이황동(soy isoflavones,SI)급주요활성성분Genistein대초로대서란소급체외배양란소과립세포중간수체유사물-1(liver receptor homolog-1,LRH-1)기인표체적영향,초보탐토대두이황동작용우란소적분자궤제.방법 채용자연노화법건립위절경기대서동물모형,분별급여저제량(50mg/kg)、중제량(158 mg/kg)、고제량(500 mg/kg)적SI관위처리8 w.분별채용원위잡교화RT-PCR검측란소조직중LRH-1 mRNA적표체.채용대서잉마혈청촉성선격소(pregnant mare serum gonadotrophin,PMSG)피하주사법건립촉란포발육모형,분리、배양과립세포24 h후,급여대두이황동주요활성성분Genistein (0、0.1、1、5、10、100 μmol/L)이급자격소수체(estrogen receptor,ER)조단제ICI182、780(1μmol/L)계속처리세포24 h,채용RT- PCR방법검측란소과립세포중LRH-1 mRNA적표체정황.결과 각제량SI처리조란소LRH-1 mRNA적표체(저제량0.563±0.037,중제량0.926±0.127화고제량1.223±0.134),여위절경기모형조(0.460±0.082)상비균현저증가(P＜0.05).1～10 μmol/L적Genistein작용란소과립세포24 h,LRH-1 mRNA적표체(분별위0.844±0.042,0.879±0.056,0.882±0.079)여공백대조조(0.678±0.052)상비균증가,결과구유통계학의의(P＜0.05).ER조단제ICI182,780예처리세포30 min후재급여Genistein계속처리세포24 h후,발현1～10 μmol/L적Genistein상조LRH-1표체작용,병미수ER조단제적영향.결론 일정제량적대두이황동가명현상조쇠로란소LRH-1 mRNA적표체,1～10 μmol/L적Genistein가상조체외배양대서란소과립세포중LRH-1 mRNA적표체,가능병비통과경전ER개도적.
Objective To investigate the effects of Soy isoflavones on the expression of liver receptor homolog-1 (LRH-1) gene within aging ovary of rats and ovarian granulosa cell cultured in vitro treated with Genistein,which is a major active component of SI.Methods The animal model of perimenopause rats was established by unforced aging,were treated by intragastric administration ( ig ) with low (50 mg/kg),moderate (158 mg/kg) and high (500 mg/kg) dose of SI for 8 weeks.Expression of LRH-1 mRNA were detected by in situ hybridization and RT-PCR,respectively.Female Wistar rats were injected subcutaneously with pregnant mare serum gonadotrophin (PMSG),ovarian granulosa cells were collected and incubated for 24h,and then granulosa cells were administered with genistein (0,0.1,1,5,10,100 μmol/L) and ICI182,780( 1 μmol/L) for 24 h.LRH-1 mRNA expression levels were assessed by RT-PCR.Results Compared with the control model group ( 0.460 + 0.082 ),LRH-1 mRNA in the aging ovary(low dose 0.563 ±0.037,moderate dose 0.926 +0.127,high dose 1.223 ±0.134),were ihcreased significantly( P ＜ 0.05 ).1 ～ 10 μmol/L Genistein could increase the expression of LRH-1mRNA ( low dose 0.844 ± 0.042.Moderate dose 0.879 ± 0.056,high dose 0.882 ± 0.079 ) in the rats granulosa cells treated for 24 h ( P ＜ 0.05 ).UP - regulation of the expression of LRH-1 mRNA by 1 ～ 10 μmol/L Genistein was not in fluenced by ICI182,780 (1 μ mol/L,pretreated for 30 min).Conclusion Soy Isoflavones could up-regulate the expression of LRH-1 mRNA in aging ovaries.1 ～ 10 μmol/L Genistein could up-regulate the expressions LRH-1 mRNA of granulosa cells in rat ovaries,which was probably not mediated by classical ER pathway.
