山西医药杂志
山西醫藥雜誌
산서의약잡지
SHANXI MEDICAL JOURNAL
2015年
9期
977-981
,共5页
孙长英%张宏其%鲁世金%黄佳%尹新华%王昱翔%郭强%曾科峰
孫長英%張宏其%魯世金%黃佳%尹新華%王昱翔%郭彊%曾科峰
손장영%장굉기%로세금%황가%윤신화%왕욱상%곽강%증과봉
RNA干扰%软骨细胞%青春期%椎体%生长板
RNA榦擾%軟骨細胞%青春期%椎體%生長闆
RNA간우%연골세포%청춘기%추체%생장판
RNA interference%Chondrocytes%Puberty%Vertebra%Growth plate
目的:初步探索雌激素受体(ER)α和ERβ基因在青春期椎体生长板软骨细胞分化和增殖中相互作用,以及Smad4基因在ERα和ERβ调控软骨细胞Runx2和BM P2基因表达过程中的信号传递作用。方法设计ERα、ERβ和Smad4 mRNA打靶的RNAi慢病毒载体并包装成为慢病毒颗粒。以小鼠椎体生长板软骨细胞为对象,采用分层设计的方法进行分组,分别转染目的基因RNAi载体,研究Smad4基因在椎板软骨细胞中介导ERα和ERβ调控Runx2和BM P2基因的表达的作用。分析各组细胞生长曲线;检测各组细胞Runx2、BM P2表达的变化。结果于转染后第3、5、7、9天时,各组细胞吸光度值(A490)在多个组间比较差异有统计学意义( P<0.05)。各组间Runx2基因表达相对于空白对照组改变的倍数由高到低的顺序与细胞生长曲线基本相同。而BM P2基因表达由高到低的顺序与 Runx2不同,多个亚组间差异有统计学意义( P <0.05)。然而,无论ERαRNAi组还是ERβRNAi组中,Smad4RNAi亚组与Smad4正常亚组比较,BM P2基因表达均差异无统计学意义( P >0.05)。结论 ERα和ERβ均具有调控椎体生长板软骨细胞分化与增殖的作用,且ERα的调控作用大于ERβ。在ERα和ERβ均正常表达时,ERβ具有部分抑制ERα的功能;当ERα表达显著降低时,ERβ可以部分补偿ERα的功能。另外,ERβ通过Smad4信号传递增强软骨细胞Runx2基因表达,但还可能存在非Smad4信号传递通路。然而,ERα和ERβ对BM P2基因表达的调控并非通过Smad4信号传递。
目的:初步探索雌激素受體(ER)α和ERβ基因在青春期椎體生長闆軟骨細胞分化和增殖中相互作用,以及Smad4基因在ERα和ERβ調控軟骨細胞Runx2和BM P2基因錶達過程中的信號傳遞作用。方法設計ERα、ERβ和Smad4 mRNA打靶的RNAi慢病毒載體併包裝成為慢病毒顆粒。以小鼠椎體生長闆軟骨細胞為對象,採用分層設計的方法進行分組,分彆轉染目的基因RNAi載體,研究Smad4基因在椎闆軟骨細胞中介導ERα和ERβ調控Runx2和BM P2基因的錶達的作用。分析各組細胞生長麯線;檢測各組細胞Runx2、BM P2錶達的變化。結果于轉染後第3、5、7、9天時,各組細胞吸光度值(A490)在多箇組間比較差異有統計學意義( P<0.05)。各組間Runx2基因錶達相對于空白對照組改變的倍數由高到低的順序與細胞生長麯線基本相同。而BM P2基因錶達由高到低的順序與 Runx2不同,多箇亞組間差異有統計學意義( P <0.05)。然而,無論ERαRNAi組還是ERβRNAi組中,Smad4RNAi亞組與Smad4正常亞組比較,BM P2基因錶達均差異無統計學意義( P >0.05)。結論 ERα和ERβ均具有調控椎體生長闆軟骨細胞分化與增殖的作用,且ERα的調控作用大于ERβ。在ERα和ERβ均正常錶達時,ERβ具有部分抑製ERα的功能;噹ERα錶達顯著降低時,ERβ可以部分補償ERα的功能。另外,ERβ通過Smad4信號傳遞增彊軟骨細胞Runx2基因錶達,但還可能存在非Smad4信號傳遞通路。然而,ERα和ERβ對BM P2基因錶達的調控併非通過Smad4信號傳遞。
목적:초보탐색자격소수체(ER)α화ERβ기인재청춘기추체생장판연골세포분화화증식중상호작용,이급Smad4기인재ERα화ERβ조공연골세포Runx2화BM P2기인표체과정중적신호전체작용。방법설계ERα、ERβ화Smad4 mRNA타파적RNAi만병독재체병포장성위만병독과립。이소서추체생장판연골세포위대상,채용분층설계적방법진행분조,분별전염목적기인RNAi재체,연구Smad4기인재추판연골세포중개도ERα화ERβ조공Runx2화BM P2기인적표체적작용。분석각조세포생장곡선;검측각조세포Runx2、BM P2표체적변화。결과우전염후제3、5、7、9천시,각조세포흡광도치(A490)재다개조간비교차이유통계학의의( P<0.05)。각조간Runx2기인표체상대우공백대조조개변적배수유고도저적순서여세포생장곡선기본상동。이BM P2기인표체유고도저적순서여 Runx2불동,다개아조간차이유통계학의의( P <0.05)。연이,무론ERαRNAi조환시ERβRNAi조중,Smad4RNAi아조여Smad4정상아조비교,BM P2기인표체균차이무통계학의의( P >0.05)。결론 ERα화ERβ균구유조공추체생장판연골세포분화여증식적작용,차ERα적조공작용대우ERβ。재ERα화ERβ균정상표체시,ERβ구유부분억제ERα적공능;당ERα표체현저강저시,ERβ가이부분보상ERα적공능。령외,ERβ통과Smad4신호전체증강연골세포Runx2기인표체,단환가능존재비Smad4신호전체통로。연이,ERα화ERβ대BM P2기인표체적조공병비통과Smad4신호전체。
Objective To explore the interaction of the ERαand ERβin regulating mice adolescence verte‐bral epiphyseal plate chondrocyte differentiation and proliferation ,and the role of smad4 signaling transduction in ERαand ERβregulating the Runx2 and BMP2 expression.Methods The siRNA sequences targeting and silencing ERα,ERβand Smad4 mRNA were designed which were constructed into gene silencing lentiviral vector and pack‐aged as lentiviral particles .Mice vertebral growth plate chondrocytes are extracted ,grouped and were grouped with hierarchical design and were transfected target gene RNAi vectors to observe the smad4 signaling in ERαand ERβregulating Runx2 and BMP2 gene expression in chondrocytes .The cells growth curves were plotted and the Runx2 and BMP2 expression was detected in each group .Results The A value in 10 subgroups at 3th ,5th ,7th and 9th day after transfected were significantly different among groups ( P < 0 .05) .Descending order of the Runx2 gene expression in each subgroup was basically the same as A value .The descending order of BMP2 gene expression was not same as Runx2 .Whether in ERβRNAi group or in ERαRNAi group .There were no significant differences between Smad4RNAi subgroup and Smad4 normal subgroup( P >0 .05).Conclusion Both ERα and ERβcould regulate vertebral growth plate cartilage cell differentiation and proliferation .The role of regulation of ERαwas greater than ERβ.When the expression of ERα and ERβ were normal ,ERβ has partially inhibit ERαfunction .However ,when the expression of ERα significantly reduced ,ERβ can partially compensate for ERαfunction .ERβalso enhanced the chondrocyte differentiation and proliferation by partly reinforced Runx2 gene ex‐pression by Smad4 signaling ,but it may also by non‐Smad4 signaling pathway .However ,the regulation of ERαand ERβon BMP2 gene expression was not through Smad4 signaling pathway.