Anti-aging Effect of Ethanol Extract of Velvet Antler on Caenorhabditis elegans
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摘要: 以秀丽隐杆线虫为模式生物，探究鹿茸乙醇提取物对秀丽隐杆线虫抗衰老的作用。通过寿命、紫外应激、热应激、运动、产卵等实验检测鹿茸乙醇提取物对线虫衰老的影响；通过检测鹿茸乙醇提取物对野生型线虫和突变体线虫抗氧化应激能力的实验，初步探究其衰老作用及机理。结果表明，鹿茸乙醇提取物为2.5 mg/mL时能延长线虫寿命，最长寿命为28 d；具有抗紫外和热应激的能力，线虫平均寿命分别为2.81和3.28 d，优于对照组；能提高线虫运动和产卵能力；氧化应激条件下能显著（P<0.05）延长野生型线虫生存时间；加药组与对照组的clk-1突变体线虫在氧化应激条件下生存时间无明显差异，表明其作用机理可能为激活线粒体信号通路，上调抗氧化基因gst-4、gst-7和skn-1及热抗性基因hsp16.2的转录水平。因此，鹿茸乙醇提取物对线虫具有抗衰老的作用。Abstract: Taking Caenorhabditis elegans as model organism, the anti-aging effect of ethanol extract of velvet antler on Caenorhabditis elegans was studied. The effects of ethanol extract of velvet antler on the aging of C. elegans were tested by lifespan, UV stress, heat stress, exercise and oviposition. The anti-oxidative stress of wild-type nematodes and mutant nematodes were detected by ethanol extract of velvet antler. The results showed that the ethanol extract of velvet antler at 2.5 mg/mL could prolong the lifespan of nematodes, the longest lifespan was 28 days.The ethanol extract of velvet antler could resist UV and heat stress, the average lifespan of nematodes was 2.81 and 3.28 days, respectively, which was better than that of the control group.The ethanol extract of velvet antler could improve the locomotion and oviposition of nematodes.Under oxidative stress, the survival time of wild-type nematode was significantly prolonged ( P <0.05). The survival time of clk-1 mutant nematode in the treatment group and the control group was not significantly different under oxidative stress, indicating that the mechanism of action might be activation of mitochondrial signaling pathway and the transcription levels of anti-oxidation genes gst-4, gst-7 and skn-1 and heat resistance hsp16.2 were up-regulated. Therefore, ethanol extract of velvet antler has anti-aging effect on nematode.
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Key words:
- Caenorhabditis elegans /
- velvet antler ethanol extract /
- anti-aging /
- antioxidant
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图 1 鹿茸乙醇提取物对线虫寿命的影响
Figure 1. Effect of ethanol extract of velvet antler on life span of nematode
图 2 鹿茸乙醇提取物对线虫紫外应激和热应激的影响
注：A：VAEE对野生型N2线虫的紫外应激实验；B：VAEE对野生型N2线虫的热应激实验。
Figure 2. Effects of ethanol extract of velvet antler on ultraviolet and heat stress of nematode
图 3 鹿茸乙醇提取物对线虫运动能力和生殖能力的影响
注：A：VAEE对野生型N2线虫的运动实验；B：VAEE对野生型N2线虫的产卵实验；“ns”无显著性差异，“*”表示P<0.05，差异具有统计学意义；“**”表示P<0.01，存在显著性差异；“***”表示P<0.001，存在极显著差异；“****”表示P<0.0001，存在极极显著差异。
Figure 3. Effects of ethanol extract of velvet antler on locomotor and reproductive ability of nematode
图 4 鹿茸乙醇提取物对线虫氧化应激的影响
注：A：各浓度VAEE对野生型N2线虫的氧化应激实验；B：VAEE对daf-2突变体线虫的氧化应激实验；C：VAEE对eat-2突变体线虫的氧化应激实验；D：VAEE对clk-1突变体线虫的氧化应激实验。
Figure 4. Effect of ethanol extract of velvet antler on oxidative stress of nematode
图 5 鹿茸乙醇提取物对线虫基因转录水平的影响
注：“*”表示P<0.05，差异具有统计学意义；“**”表示P<0.01，存在显著性差异。
Figure 5. Effects of ethanol extract of velvet antler on gene transcription level of nematode
表 1 鹿茸乙醇提取物对线虫平均寿命的影响
Table 1. Effect of ethanol extract of velvet antler on the average life of nematode
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表 2 紫外应激、热应激下鹿茸乙醇提取物对线虫平均寿命的影响
Table 2. Effects of ethanol extract of velvet antler under ultraviolet and heat stress on the average life span of nematode
实验 类型 平均寿命 ± SEM（d） 总死亡数/总数 紫外应激实验 N2 Vehicle 1.84 ± 0.0980 90/90 N2 VAEE 1 mg/mL 2.08 ± 0.1033ns 90/90 N2 VAEE 2.5 mg/mL 2.81 ± 0.1350**** 90/90 N2 VAEE 5 mg/mL 2.46 ± 0.1249**** 90/90 热应激实验 N2 Vehicle 1.90 ± 0.1007 90/90 N2 VAEE1 mg/mL 2.41 ± 0.1276*** 90/90 N2 VAEE 2.5 mg/mL 3.28 ± 0.1406**** 90/90 N2 VAs 5 mg/mL 2.83 ± 0.1408**** 90/90
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表 3 鹿茸乙醇提取物对线虫在氧化应激下平均寿命的影响
Table 3. Effects of ethanol extract of velvet antler on the average life spans of C.elegans under oxidative stress
类型 平均寿命 (h) 总死亡数/总数 N2 Vehicle 3.03 ± 0.15 90/90 N2 VAEE 1 mg/mL 3.63 ± 0.19* 90/90 N2 VAEE 2.5 mg/mL 4.58 ± 0.24**** 90/90 N2 VAEE 5 mg/mL 4.69 ± 0.25**** 90/90 N2 Vehicle 3.68 ± 0.14 90/90 eat-2(ad1116) Vehicle 6.03 ± 0.24**** 90/90 eat-2(ad1116) VAEE 7.37 ± 0.28**** 90/90 N2 Vehicle 3.69 ± 0.15 90/90 daf-2(e1368) Vehicle 6.59 ± 0.26**** 90/90 daf-2(e1368) VAEE 8.13 ± 0.37*** 90/90 N2 Vehicle 5.01 ± 0.16 150/150 clk-1(mq130) Vehicle 4.21 ± 0.10**** 150/150 clk-1(mq130) VAEE 4.19 ± 0.11ns 150/150 注：野生型N2线虫氧化应激中的P值是与对照组相比，eat-2、daf-2、clk-1氧化应激中的P值是突变体线虫对照组与野生型对照组比，突变体线虫加药组与突变体线虫对照组比。
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