本文關(guān)鍵詞： 巴馬香豬 肌纖維類型 MyHC miR-208b miR-499 AMPK PGC-1α　出處：《廣西大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：MicroRNAs是一類進(jìn)化上非常保守的小的非編碼RNA,在骨骼肌發(fā)育中起到了至關(guān)重要的調(diào)控作用。本實(shí)驗(yàn)以巴馬香豬為對象,探究miR-208b和miR-499在骨骼肌肌纖維類型轉(zhuǎn)化中的作用,為揭示巴馬香豬優(yōu)良肉質(zhì)形成機(jī)制的研究奠定基礎(chǔ)。獲得的主要研究結(jié)果如下：1.miR-208b和miR-499在巴馬香豬背最長肌、股二頭肌、心臟中均有高豐度表達(dá),在其他組織中的表達(dá)豐度較低。對巴馬香豬不同發(fā)育時期的背最長肌進(jìn)行檢測,發(fā)現(xiàn)miR-208b和miR-499的表達(dá)量在胚胎期持續(xù)升高,出生后表達(dá)量下降。2.巴馬香豬背最長肌發(fā)育過程中,MyHC Ⅰ的表達(dá)量顯著性逐漸降低(P0.05),ⅡA的表達(dá)量變化不大(P0.05),ⅡX的表達(dá)量顯著性逐漸升高(P0.05),ⅡB的表達(dá)量在E90開始升高。胚胎期氧化型肌纖維標(biāo)志基因的mRNA所占的比例高于酵解型,表達(dá)比例在E40最高,隨后下降；成年時期氧化型肌纖維標(biāo)志基因的mRNA所占的比例低于酵解型。3.巴馬香豬胚胎期,AMPK的表達(dá)與MyHC Ⅰ的表達(dá)呈顯著正相關(guān)(r=0.9561,P0.05),PGC-1α的表達(dá)與MyHC ⅡB的表達(dá)呈顯著負(fù)相關(guān)(r=-0.9410,P0.05)。4.miR-208b和miR-499的表達(dá)量均隨著C2C12和小鼠肌肉衛(wèi)星細(xì)胞的分化顯著上升(P0.05)。5.C2C12和小鼠肌肉衛(wèi)星細(xì)胞過表達(dá)miR-208b后,肌纖維形態(tài)更細(xì)長；在分化中后期顯著上調(diào)MyHC Ⅰ的表達(dá)量(P0.05),顯著下調(diào)MyHC ⅡB的表達(dá)量(P0.05)；分化早中期標(biāo)志基因MyoG在C2C12及肌肉衛(wèi)星細(xì)胞分化過程中的表達(dá)量實(shí)驗(yàn)組和對照組無差異(P0.05);C2C12及肌肉衛(wèi)星細(xì)胞分化過程中,AMPK的表達(dá)量實(shí)驗(yàn)組和對照組均無顯著差異(P0.05)；肌肉衛(wèi)星細(xì)胞中PGC-1α的表達(dá)水平顯著上調(diào)(P0.05)。6.C2C12和小鼠肌肉衛(wèi)星細(xì)胞過表達(dá)miR-499后,肌纖維形態(tài)更細(xì)長；在分化中后期顯著上調(diào)MyHCI的表達(dá)(P0.05),顯著下調(diào)MyHC ⅡB的表達(dá)(P0.05)；MyoG基因的表達(dá)量實(shí)驗(yàn)組和對照組無差異(P0.05)；AMPK、PGC-1α的表達(dá)水平均顯著上調(diào)(P0.05)。綜上所述,miR-208b和miR-499對氧化型肌纖維的形成有促進(jìn)作用,對酵解型肌纖維有抑制作用,可能通過調(diào)控AMPK、PGC-1α起作用。
[Abstract]:MicroRNAs is a very conservative small non RNA encoding a class of evolutionarily, during skeletal muscle development plays a vital role. In this experiment, Bama miniature pig as the object, to explore miR-208b and miR-499 in skeletal muscle fiber type transformation in the role of the study laid the foundation for revealing the excellent meat quality of Bama miniature pig formation mechanism. The main results are as follows: 1.miR-208b and miR-499 in Bama miniature pig longissimus muscle, unit two biceps, expressed in high abundance in the heart, expressed in other tissues. The lower of Bama miniature pigs in different developmental stages of longissimus muscle were detected and found that expression of miR-208b and miR-499 in the embryo the duration of elevated expression of.2. decreased after the birth of Bama miniature pig longissimus muscle during development, the expression of MyHC I was significantly decreased (P0.05), expression of A II (P0.05), little expression of X II Significantly increased (P0.05), the expression of B II began to increase at E90. The embryonic oxidative muscle fiber marker gene mRNA proportion is higher than that of glycolysis, expression ratio in the E40 is the highest, and then decreased; adult oxidative muscle fiber marker gene mRNA is lower than the proportion of glycolytic.3. pig embryos, there was a positive correlation between the expression of AMPK and MyHC I (r=0.9561, P0.05), negatively related to expression of PGC-1 alpha and MyHC II B (r=-0.9410, P0.05) expression of.4.miR-208b and miR-499 increased with the differentiation of C2C12 and mouse muscle satellite cells increased significantly (P0.05.5.C2C12) and mouse muscle satellite cells after overexpression of miR-208b, muscle fiber shape is more slender; in the late differentiation significantly up-regulated expression of MyHC I (P0.05), the amount of expression was significantly down regulated MyHC II B (P0.05) in the early stage; differentiation marker gene MyoG in C2C1 The amount of the experimental group and the control group had no difference in the expression of 2 and muscle satellite cells during differentiation (P0.05); C2C12 and muscle satellite cell differentiation, there were no significant differences in the expression of AMPK in the experimental group and the control group (P0.05); the expression level of PGC-1 alpha in muscle satellite cells increased significantly (P0.05) and.6.C2C12 mouse muscle satellite cells after overexpression of miR-499, muscle fiber morphology is more slender; expression in the differentiation of late significantly up-regulated the expression of MyHCI (P0.05), MyHC II B expression was significantly reduced (P0.05); the expression of MyoG gene in the experimental group and control group had no difference (P0.05); AMPK, PGC-1 protein expression significantly increase (P0.05). To sum up, miR-208b and miR-499 can promote the formation of oxidized muscle fibers, the glycolytic muscle fibers are inhibited, by regulating AMPK, PGC-1 alpha plays a role.

【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S828

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相關(guān)期刊論文 前1條

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本文編號：1524983