目的 研究棉子糖抗衰老活性并探討其作用機(jī)制。方法 棉子糖(150、200、250μmol·L^-1)加到大腸桿菌中孵育秀麗隱桿線蟲后,顯微鏡下觀察線蟲身體彎曲頻率、頭尾擺動(dòng)頻率、體長,觀察熱應(yīng)激刺激下線蟲的存活時(shí)間;通過脂褐素?zé)晒鈴?qiáng)度檢測評價(jià)棉子糖對線蟲抗衰老能力的影響;采用DAF-16核定位測定和超氧化物歧化酶-3(SOD-3)表達(dá)測定研究其抗衰老的相關(guān)機(jī)制。結(jié)果 與對照組比較,棉子糖3個(gè)濃度均使線蟲的身體彎曲頻率加快(P＜0.001);200μmol·L^-1棉子糖使線蟲頭尾擺動(dòng)頻率加快(P＜0.01);體長測試結(jié)果顯示棉子糖3個(gè)濃度均對線蟲生長發(fā)育無毒害作用。熱應(yīng)激測試中,與對照組比較,棉子糖3個(gè)濃度均顯著延長線蟲在熱應(yīng)激條件下的存活時(shí)間(P＜0.05)。脂褐素測試中,與對照組比較,棉子糖3個(gè)濃度均使線蟲體內(nèi)脂褐素含量顯著降低(P＜0.01)。DAF-16核定位測定中,棉子糖3個(gè)濃度均促進(jìn)DAF-16蛋白入核表達(dá)。SOD-3表達(dá)測定中,與對照組比較,棉子糖250μmol·L^-1濃度組的SOD-3蛋白表達(dá)水平顯著上升(P＜0.05),與DAF-16核定位測定一并說明棉子糖抗衰老活性依賴ⅡS分子信號通路。結(jié)論 棉子糖能夠促進(jìn)線蟲的活動(dòng)能力,對其生長發(fā)育無毒害性,并能延緩線蟲的衰老速度,促進(jìn)DAF-16蛋白入核表達(dá),提高SOD-3蛋白表達(dá)水平,其抗衰老機(jī)制依賴ⅡS分子信號通路。

Objective To study the anti-aging activity of raffinose and to explore its anti-aging mechanism.Methods Raffinose(150,200, and 250 μmol·L^-1) was added to Escherichia coli to incubate Caenorhabditis elegans, the body bending frequency, head and tail swinging frequency, and body length of the nematodes were observed under a microscope, and the survival time of the Caenorhabditis elegans under heat stress stimulation was observed. The effect of raffinose on the anti-aging ability of nematode was evaluated by lipofuscin, and the anti-aging mechanism was determined by DAF-16 nuclear localization and SOD-3 expression.Results In the body bending test, the three concentration groups of raffinose accelerated the body bending frequency of Caenorhabditis elegans campared with control group(P＜0.001). In the head-tail swing test, the raffinose of 200 μmol·L^-1 concentration group accelerated the head-tail swing frequency of nematodes campared with control group(P＜0.01). In the body length test, the three concentration groups of raffinose had no toxic effect on the growth and development of nematode. In the heat stress test, the three concentration groups of raffinose prolonged the survival time of Caenorhabditis elegans under heat stress(P＜0.05). In the lipofuscin test, the three concentration groups of raffinose decreased the content of lipofuscin in Caenorhabditis elegans. In DAF-16 nuclear localization assay, raffinose at three concentrations promoted the expression of DAF-16 protein into the nucleus. In the determination of SOD-3 expression, the level of SOD-3 protein expression increased in raffinose 250 μmol·L^-1 group,which together with DAF-16 nuclear localization assay showed that raffinose's anti-aging activity depended on ⅡS molecular signal pathway.Conclusion Raffinose can promote the activity of Caenorhabditis elegans, has no toxicity to its growth and development,and can delay the senescence of Caenorhabditis elegans, promote the expression of DAF-16 protein into nucleus and increase the expression level of SOD-3 protein. The anti-aging mechanism of raffinose depends on ⅡS molecular signal pathway.

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