利用秀丽线虫评价神经传递抑制肽的祛皱功效研究

Evaluation of anti-wrinkle effect of neurotransmission inhibitory peptides by Caenorhabditis elegans

随着化妆品行业迈进高科技发展时期,人们更加注重皮肤的保养,特别是对皮肤纹理平整和皱纹淡化的需求剧增,然而抗皱类化妆品或原料的功效评估存在周期较长、费用较高且无法实现高通量检测的问题。因此,建立可行、有效的替代检测模型实现化妆品原料祛皱功效的快速评价对化妆品开发具有重要意义。作者以秀丽线虫为模式生物,研究几种神经传递抑制类化妆品祛皱多肽对其运动行为和神经递质传导相关酶活性的影响,从而实现化妆品原料祛皱功效的定性定量检测。结果表明,红蝎素、蛇毒肽、芋螺肽以及红蝎素EX(红蝎素和芋螺肽的复配肽)均有一定的阻断神经与肌肉间神经冲动、抵御兴奋刺激条件下线虫神经与肌肉间神经冲动以及显著抑制线虫的运动行为的能力。此外,神经递质传导相关酶活性的数据表明上述神经抑制类多肽是通过刺激钙调神经磷酸酶CaN过表达同时抑制乙酰胆碱酯酶AchE的活力,从而阻断神经与肌肉间神经递质的传递,抑制神经冲动,使肌肉达到放松进而平整皮肤表面、达到祛皱的效果。

With the cosmetics industry entering the high-tech development period,people pay more attention to the maintenance of the skin,especially the sharp increase in the demand for skin texture smoothing and wrinkle desalinization.However,the efficacy evaluation of anti-wrinkle cosmetics or raw materials has the problems of long cycle,high cost and inability to achieve high-throughput detection.Therefore,it is of great significance for product development to establish a feasible and effective alternative detection model,in order to achieve the rapid evaluation of the wrinkle-removing efficacy of cosmetic raw materials.In this study,Caenorhabditis elegans(C.elegans)was used as the model organism to test the inhibitory effects of several kinds of cosmetic peptides on neuromuscular synapses.The movement behaviors of the treated C.elegans were observed and two main enzymes function in neuromuscular junctions,the Acetylcholinesterase(AChE)and the Calcineurin(CaN),were also tested.The results show that,after treatment with all tested peptides(including scorpion venom,snake venom peptide,conotoxin and the compound peptide by scorpion venom and conotoxin),the movement behavior of C.elegans(head swinging and body bending)is inhibited to a certain extent(9%-33%).And the tail muscles can not contract normally,showing the movement state of trailing and crawling.With the intervention of neostigmine methylsulfate,the body bending amplitude of the C.elegans after exercise injury recovers little,and the movement behavior of C.elegans is inhibited about 5%-17%.These data indicate that all of the tested peptides show strong inhibitory activities on normal synapse formation between motor neuron and muscle,with remarkably inhibiting the locomotor behavior of C.elegans.In addition,we further find that these peptides stimulate the overexpression of CaN while inhibiting the enzyme activity of AchE,thereby blocking the functions of neuromuscular junction,which leads to muscles relax and achieves the antiwrinkle effects.Various types of neurosuppressive peptides have different targets and different action mechanisms for exerting anti-wrinkle activity.Compared with the anti-wrinkle polypeptide with a single target,the polypeptide raw material compounded by the red scorpion element and the cono peptide can exert more excellent anti-wrinkle activity.Furthermore,by testing the movement recovery ability of C.elegans,the head swing frequency of C.elegans recovers to the level of the blank group(P>0.05),which indicates that these neuroprotective peptides have no neurotoxicity to C.elegans,and are immediate and safe.In conclusion,the methods described in this study can provide scientific reference for the development and efficacy verification of anti-wrinkle ingredients for skin care products.