本文選題：β-胍基丙酸　切入點：果蠅　出處：《華中科技大學》2015年博士論文　論文類型：學位論文

【摘要】：第一部分β-胍基丙酸對果蠅壽命的影響 目的：β一胍基丙酸(p-guanadinopropionic acid, β-GPA)是肌酸類似物,可以慢性激活AMPK,同時還可以增加線粒體酶的活性,改善線粒體功能。由于AMPK的慢性激活和線粒體功能的增強可以延緩衰老,延長壽命。因此,β-GPA極可能有抗衰老,延長壽命的作用。因此,本實驗研究β-GPA對果蠅壽命的影響。 方法：在果蠅培養(yǎng)基中添加濃度分別為300、900和2700mM的β-GPA,觀察果蠅壽命的變化；分別采用只含有瓊脂的培養(yǎng)基來模擬饑餓環(huán)境,采用含有3MH202的培養(yǎng)基模擬強氧化環(huán)境,觀察加入含有β-GPA的培養(yǎng)基飼養(yǎng)30天后,果蠅在這兩種環(huán)境中的存活情況；采用羥胺法測定SOD活性,采用硫代巴比安酸(TBA)比色法測定MDA的含量,觀察β-GPA對果蠅SOD活性和MDA含量的影響。 結(jié)果：(1)濃度為900和2700mM的β-GPA能夠顯著延長果蠅(雌性和雄性)壽命。900mM和2700mM的β-GPA分別將雌性果蠅壽命由60d延長至65d和68d(n=200,p0.001)；將雄性果蠅的壽命由55d延長至59d和60d(n=200,p0.001)。(2)濃度為900mM的β-GPA能夠顯著延長果蠅在饑餓環(huán)境下的存活時間。900mM的β-GPA將饑餓環(huán)境下雌性和雄性果蠅的壽命均從5d延長至7d(n=100,p0.001),增強了果蠅對應激的抵抗力。(3)濃度為900mM的β-GPA能夠顯著延長果蠅在強氧化環(huán)境下存活時間。900mM β-GPA將強氧化環(huán)境下雌性和雄性果蠅的壽命均從1d延長至2d(n=100,p0.001),并顯著增加果蠅SOD活性,降低MDA含量,增強果蠅的抗氧化能力。 結(jié)論：β-GPA濃度依賴性的延長果蠅的壽命,且增加果蠅對不同應激的抵抗能力,增強其抗氧化能力。 第二部分β-胍基丙酸延長果蠅壽命的機制 目的：腺苷酸活化蛋白激酶(adenosine5'-monophosphate (AMP)-activated protein kinase, AMPK)的激活可以延緩衰老,延長壽命。p-GPA可以慢性激活AMPK,且后者可以通過調(diào)節(jié)哺乳動物雷帕霉素靶蛋白(mammalian target of rapamycin, mTOR)和unc-51樣激酶(Unc-51like kinase1, ULK1/Atg1)的活性對自噬進行調(diào)控。最近研究發(fā)現(xiàn),自噬和衰老,壽命密切相關(guān)。但尚不清楚β-GPA是否通過激活AMPK來增強自噬,從而發(fā)揮延長壽命的作用。本實驗旨在對β-GPA延長果蠅壽命的分子機制進行探討。 方法：Western blotting實驗檢測β-GPA對果蠅中AMPK磷酸化和自噬相關(guān)指標Atg5和p62的表達；分別采用藥理學方法和RNA干擾技術(shù)降低果蠅中AMPK和自噬關(guān)鍵分子Atg1和Atg5的表達后,觀察β-GPA對果蠅壽命的影響；采用藥理學方法和分子生物學方法抑制AMPK后,β-GPA對自噬相關(guān)指標和與延長壽命作用密切相關(guān)蛋白的影響。 結(jié)果：(1)濃度為900和2700mM的β-GPA能夠顯著增加果蠅各個部分AMPK的活性。果蠅各部位AMPK的磷酸化水平分別為：頭部：對照：100±13.54,900mM β-GPA:143.36±12.2,2700mMβ-GPA:147.13±8.54(n=6, p0.05);胸部：對照：100±15.49,900mM β-GPA:155.14±17.53,2700mM β-GPA:155.78±14.59(n=6,p0.05)；腹部：對照：100±13.35,900mM β-GPA:152.91±15.8,2700mM P-GPA:152.4±16.14(n=6, p0.05)。(2)濃度為900的β-GPA能夠顯著增加果蠅自噬水平：下調(diào)Atg5,降低果蠅自噬水平,P-GPA處理后使雌性果蠅壽命由62d下降至51d(n=200,p0.001),雄性果蠅壽命由59d減少至51d(n=200,p0.001),說明下調(diào)Atg5水平取消了P-GPA延長壽命的作用。(3)降低果蠅AMPK活性,β-GPA誘導的自噬增加作用消失,并且AMPK-RNAi使雌性果蠅壽命由63d下降至51d(n=200,p0.001),雄性果蠅壽命由57d減少至51d(n=200,p0.001),表明給予AMPK-RNAi阻礙了β-GPA延長果蠅壽命的作用,提示β-GPA的這些作用依賴于AMPK。(4)Atg1參與調(diào)節(jié)β-GPA誘導的自噬增加和壽命延長。(5) β-GPA可以通過激活AMPK抑制mTORC1的活性。 結(jié)論：β-GPA延長果蠅壽命作用是依賴于AMPK-Atg1-autophagy信號通路。
[Abstract]:Part 1 Effect of beta guanidinopropionic acid on lifespan of Drosophila melanogaster
Objective: beta guanidinopropionic acid (p-guanadinopropionic acid, P -GPA) is creatine analogs can be chronic activation of AMPK, but also can increase the activity of mitochondrial enzymes and improve mitochondrial function. Due to the enhancement of AMPK and chronic activation of mitochondrial function can delay aging, prolong life expectancy. Therefore, beta -GPA probably anti-aging to extend the life of the role. Therefore, this research studied the effect of beta -GPA on the lifespan of Drosophila melanogaster.
Methods: in Drosophila medium concentration were 300900 and 2700mM beta -GPA, to observe the change of the life span of Drosophila; using medium to simulate the hunger environment containing only agar medium containing 3MH202, the simulation of strong oxidizing environment, to observe the beta -GPA culture medium after 30 days of feeding, Drosophila survival in these two environments; SOD activity was determined by hydroxylamine method, using thio BABIAN acid (TBA) was measured by MDA colorimetry, observe the effect of -GPA on Drosophila beta SOD activity and MDA content.
Results: (1) the concentration of 900 2700mM and beta -GPA could prolong the life of Drosophila melanogaster (male and female).900mM and 2700mM -GPA respectively in female Drosophila beta extended from 60d to 65D and 68D (n=200, p0.001); the male Drosophila lifespan was extended from 55D to 59D and 60d (n=200, p0.001). (2) the survival time of.900mM concentration of 900mM beta -GPA can significantly prolong Drosophila hunger in the environment of the beta -GPA hunger environment of female and male Drosophila life are extended from 5D to 7d (n=100, p0.001), enhanced resistance to stress in Drosophila. (3) the concentration of 900mM beta -GPA could significantly prolong the survival time of.900mM beta -GPA in strong oxidizing environment the strong oxidation environment of female and male Drosophila life are extended from 1D to 2D (n=100, p0.001), Drosophila and Drosophila significantly increased SOD activity, decreased the content of MDA, enhance the antioxidant ability of Drosophila melanogaster.
Conclusion: the concentration of beta -GPA can prolong the life of Drosophila melanogaster, and increase the resistance of Drosophila melanogaster to different stress and enhance its antioxidant capacity.
The second part mechanism bgpa extend the life of Drosophila
Objective: adenosine monophosphate activated protein kinase (adenosine5'-monophosphate (AMP) -activated protein kinase, AMPK) activation can delay aging, prolong the life of chronic.P-GPA can activate AMPK, and the latter can be adjusted by the mammalian target of rapamycin (mammalian target of rapamycin, mTOR) and UNC-51 (Unc-51like kinase1, ULK1/Atg1 like kinase) activity regulation of autophagy was. A recent study found that autophagy and aging is closely related to life. But it is not clear whether beta -GPA through activation of AMPK enhanced autophagy, which play a role. To extend the life of the molecular mechanism of this experiment is to extend the life of Drosophila beta on -GPA.
Methods: the expression of Western blotting beta -GPA detection experiment of Drosophila AMPK phosphorylation and autophagy related indicators of Atg5 and p62 respectively; expression by pharmacological method and RNA interference technology to reduce AMPK and autophagy in Drosophila key molecules Atg1 and Atg5, to observe the beta -GPA on Drosophila life influence; inhibition of AMPK by pharmacological and molecular methods biological methods, effects of beta -GPA related indicators on autophagy and closely related with the longevity protein.
Results: (1) the concentration of 900 2700mM and beta -GPA could significantly increase the activity of AMPK. Each part of the Drosophila flies in different parts of the phosphorylation level of AMPK are: head of control: 100 + 13.54900mM + 12.22700mM + -GPA:147.13 beta -GPA:143.36 beta 8.54 (n=6, P0.05); chest: control: 100 + 15.49900mM beta -GPA:155.14 + 17.532700mM + -GPA:155.78 beta 14.59 (n=6, P0.05); abdomen: control: 100 + 13.35900mM + 15.82700mM + P-GPA:152.4 -GPA:152.91 beta 16.14 (n=6, P0.05). (2) the concentration of 900 P -GPA can significantly increase the level of autophagy in Drosophila: Atg5, Drosophila reduced autophagy, after P-GPA treatment to female flies life decreased from 62D to 51D (n=200, p0.001), male Drosophila melanogaster decreased from 59D to 51D (n=200, p0.001), that lowered the level of Atg5 cancelled the life extension of the P-GPA effect. (3) reduce Drosophila AMPK activity, beta -GPA induced autophagy Increase the effect disappeared, and AMPK-RNAi make the female Drosophila life decreased from 63d to 51D (n=200, p0.001), male Drosophila melanogaster decreased from 57d to 51D (n=200, p0.001), suggested that AMPK-RNAi blocked beta -GPA extend the life of Drosophila, the role of beta -GPA depends on AMPK. (4) Atg1 is involved in the regulation of beta -GPA induced autophagy and increase life expectancy. (5) beta -GPA can activate AMPK to inhibit mTORC1 activity.
Conclusion: the effect of beta -GPA on prolonging the life span of Drosophila is dependent on the AMPK-Atg1-autophagy signaling pathway.

【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2015
【分類號】：R965

【相似文獻】

相關(guān)期刊論文 前10條

1 于英君,管宇,宋高臣;蜂膠對果蠅壽命的影響[J];中醫(yī)藥學報;2004年05期

2 王俊林,宋高臣;黃芪對果蠅壽命影響的實驗研究[J];中醫(yī)藥學報;2005年04期

3 王秀琴;黃淑峰;李宗蕓;;硫酸鈰對果蠅壽命及繁殖力的影響[J];環(huán)境與職業(yè)醫(yī)學;2007年06期

4 劉玉荷;常江;王陸一;楊美霞;韓秀雁;劉萬珍;;枸杞子對炓腹果蠅壽命影響研究[J];包頭醫(yī)學院學報;2009年02期

5 陳忠杰;樸香蘭;李文虎;吳可仲;藍高武;崔箭;;藏藥“佐塔”對果蠅壽命的影響[J];時珍國醫(yī)國藥;2011年02期

6 葉恩賜,艾裕和;老年學實驗用果蠅壽命指標的選擇[J];老年學雜志;1988年02期

7 竇肇華,胡衛(wèi)紅,張繼偉,尚保紅,樊榮;茶葉對果蠅壽命影響的實驗研究[J];老年學雜志;1988年06期

8 張青葉，叢月珠，胡聰;延壽益智茶對果蠅壽命的影響[J];中成藥;1994年11期

9 谷建云;;回春湯對果蠅壽命的影響[J];東方食療與保健;2007年05期

10 劉丙進;田偉強;張s，

本文編號：1585009