CN111759837A - Application of mother chrysanthemum lactone in preparing anti-stress medicine or life prolonging medicine for caenorhabditis elegans - Google Patents
Application of mother chrysanthemum lactone in preparing anti-stress medicine or life prolonging medicine for caenorhabditis elegans Download PDFInfo
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
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Abstract
The invention relates to application of wild chrysanthemum lactone in preparing an anti-stress medicament or a life prolonging medicament for caenorhabditis elegans, belonging to the technical field of traditional Chinese medicines. The invention also discloses the application of the wild chrysanthemum lactone in preparing the caenorhabditis elegans anti-stress medicament or the life prolonging medicament, the human anti-stress food or the medicament additive; in addition, a kit for researching the stress resistance or the life prolonging of caenorhabditis elegans is also disclosed, and a positive compound in the kit is wild chrysanthemum lactone. The invention discloses the average survival rate and median life of nematodes under stress conditions, the maximum life is reduced: in the experiment, the wild chrysanthemum lactone with the concentration of 50 mu M can prolong the life of the caenorhabditis elegans and promote the caenorhabditis elegans to resist various stresses. The invention explores new application of the active ingredient wild chrysanthemum lactone in the wild chrysanthemum extract and lays a foundation for developing novel health-care food or medicine in the future.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to application of wild chrysanthemum lactone in preparation of an anti-stress medicament or a life prolonging medicament for caenorhabditis elegans and application of wild chrysanthemum lactone in preparation of an anti-stress food or a medicament additive for human.
Background
The England scientist Sydney Brenne uses caenorhabditis elegans as a mechanism for researching apoptosis genetic regulation in 1965, the caenorhabditis elegans has the advantages of short life cycle, rapid development, easy culture, easy observation, easy storage, easy operation, high evolution conservation, vigorous fertility, clear and stable genetic background, complete genome sequence, simple operation, aging mechanism similar to mammals and the like, and the caenorhabditis elegans becomes an important model organism for researching molecular genetics, developmental biology, neuroscience and cell biology and researching the influence of compounds on life. The application of the caenorhabditis elegans has important theoretical and clinical significance for research and development of novel medicaments for delaying senescence. Most importantly, due to the similarity of nematodes and human genomes, many senescence-associated signaling pathways found in nematodes have been validated in humans, and senescence-and longevity-associated interventions using nematode models have also been validated in mammals, including humans, including metformin, resveratrol, and the like. Generally, the death of an individual is closely related to the environment.
The wild chrysanthemum lactone is one of active monomer molecules in the secondary metabolite of the wild chrysanthemum and has high content, however, no related research on caenorhabditis elegans by adopting the wild chrysanthemum lactone is found at present.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provides the application of the wild chrysanthemum lactone in preparing the caenorhabditis elegans anti-stress medicine or the life prolonging medicine and the application of the wild chrysanthemum lactone in preparing the human anti-stress food or the medicine additive.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the invention provides application of wild chrysanthemum lactone in preparing anti-stress medicaments or life prolonging medicaments of caenorhabditis elegans.
Further, preferably, the stress includes heat stress and oxidative stress.
The invention provides a kit for researching the stress resistance or the life prolonging of caenorhabditis elegans in a second aspect, which comprises a positive compound, wherein the positive compound is wild chrysanthemum lactone.
The third aspect of the invention provides the application of the wild chrysanthemum lactone in preparing human anti-stress drugs or life prolonging drugs.
The fourth aspect of the invention provides the application of the wild chrysanthemum lactone in preparing human anti-stress food or medicine additives.
The invention takes the wild chrysanthemum lactone as an active ingredient, and adopts the model organism caenorhabditis elegans to observe the influence of the wild chrysanthemum lactone on the life of the caenorhabditis elegans. The 50 mu M wild chrysanthemum lactone (wild chrysanthemum lactone) can resist liquid heat stress at 35 ℃, the average survival rate of a 50 mu M wild chrysanthemum lactone medicine adding drying and pretreatment group is improved by 13%, and the wild chrysanthemum lactone can resist 0.003% H to a certain extent2O25mM paraquat and 240 muM juglone. The influence of the wild chrysanthemum lactone on the stress resistance of the nematode can also indicate that the wild chrysanthemum lactone can delay the aging process.
The wild chrysanthemum lactone can be extracted by self or can be a commercially available product, and the wild chrysanthemum lactone is not limited in this respect, and is purchased from Shanghai leaf Biotech Co.
Compared with the prior art, the invention has the beneficial effects that:
1. the study of the invention reveals that the average survival rate and the median life of the nematodes under the stress condition are reduced: in the experiment, the wild chrysanthemum lactone with the concentration of 50 mu M can prolong the life of the caenorhabditis elegans and promote the caenorhabditis elegans to resist various stresses.
2. The invention explores new application of the active ingredient wild chrysanthemum lactone in the wild chrysanthemum extract and lays a foundation for developing novel health-care food or medicine in the future.
Drawings
FIG. 1 is a graph showing the survival rate of the group administered with different doses of wild chrysanthemum lactone at 20 ℃ and 25 ℃ in example 1 of the present invention; wherein a is 25 ℃ and b is 20 ℃;
FIG. 2 is a survival curve chart of an influence experiment of 50 μ M wild chrysanthemum lactone on the healthy life of nematodes, a pharyngeal pump movement rate test and a body movement test in embodiment 1 of the invention; wherein, a is a pharyngeal pump movement rate test, and b is a body movement test;
FIG. 3 is an effect of 50 μ M of mother chrysanthemum lactone on nematode resistance to heat stress; wherein a is a solid heat stress experiment at 35 ℃, and b is a liquid heat stress experiment at 35 ℃;
FIG. 4 is an effect of 50 μ M of mother chrysanthemum lactone on nematode resistance to oxidative stress; wherein a is H2O2Oxidative stress; b is the oxidizing stress of paraquat; c is juglone oxidative stress;
FIG. 5 is an effect of 50 μ M of wild chrysanthemum lactone on nematode resistance to bacterial infection; wherein, a is after synchronizing, culturing with normal Escherichia coli OP50 to adult five days, and then adding gram-negative bacterium PA14 for intervention to carry out life test; b is after the synchronization, culturing the strain by using normal Escherichia coli OP50 to a period of L4, and directly performing a life test by using a gram-negative bacterium PA14 mixed with 50 mu M wild chrysanthemum lactone.
Detailed Description
The present invention will be described in further detail with reference to examples.
It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The materials or equipment used are not indicated by manufacturers, and all are conventional products available by purchase.
Example 1: pharmacodynamic experiment of wild chrysanthemum lactone for prolonging life of nematode
1. Material
1.1 Experimental materials and reagents
The C.elegans strain and E.coli OP50 used in the assay were purchased from Caenorhabditis Genetics Center (CGC);
c.elegans strain: wild-type nematode N2;
wild chrysanthemum lactone: purchased from Shanghai leaf Biotech, Inc.;
juglone, paraquat was purchased from sigma aldrich trade ltd, and the remaining organic and inorganic agents were purchased from beijing bo mingsai biotechnology ltd.
Standard nematode NGM medium: adding NaCl 3g, peptone 2.5g, and agar 17g per liter of culture medium, adding water to a constant volume of 1L, sterilizing, adding 1ml of 5mg/ml cholesterol solution, and 1ml of 1mol/L MgSO 2425ml of 1mol/L PK buffer (pH = 6);
s-basal culture solution: 5.9g NaCl, 50ml 1M potassium phosphate buffer, adding water to a constant volume of 1L, sterilizing, cooling, and adding 1ml 5mg/ml cholesterol solution;
s-complete culture solution: 977ml S-basal culture, 10ml 1M potassium citrate buffer, 10ml Tracemetals solution, 3ml 1M NaOH, 3ml 1mol/L MgSO 24；
The Bleach lysate was prepared from 4ml of deionized water, 1.5ml of 5mol/L NaOH, and 2ml of NaClO.
PK buffer (pH = 6): 119g KH2PO4，21.5g K2HPO4Adding water to constant volume of 1L, and sterilizing at 121 deg.C for 20 min.
LB liquid medium: adding 10g of NaCl, 10g of peptone and 5g of yeast powder, adding water to a constant volume of 1L, and sterilizing at 121 ℃ for 20 min.
1.2 instruments
A constant temperature incubator (Shanghai New Miao medical instruments manufacturing Co., Ltd.), a body type microscope (Karl. Zeiss Co., Ltd.), and an autoclave (Shanghai Shenan medical instruments Co., Ltd.).
2. Method of producing a composite material
2.1 Experimental groups
The wild type caenorhabditis elegans N2 was selected to eat normal food (live E.coil OP 50) and life-span determination experiments were performed at 25 ℃ and 20 ℃ respectively. The control group is 0 muM wild chrysanthemum lactone (containing 2% DMSO solvent by mass), the intervention concentration of the wild chrysanthemum lactone in the administration group is 50 muM and 0 muM (each containing 2% DMSO solvent), and a 40 muM Metformin solution (Metformin) (containing 2% DMSO by mass) which is proved to prolong the service life of nematodes is used as a positive control of the experiment.
(1) Culture of OP50
One OP50 single colony was picked up and placed in 100ml LB liquid medium, and shake-cultured overnight at 37 ℃ in a shake incubator and 200rpm until OD600 reached 0.4-0.6, and stored in a refrigerator at 4 ℃ until use.
(2) Culture of caenorhabditis elegans
1) Caenorhabditis elegans was cultured in standard nematode NGM medium. The normal culture temperature is 20 ℃/25 ℃ (constant temperature incubator), and the culture humidity is controlled within 40% -60%. Generally, only one actively-active dauer-stage nematode is selected from different nematode strains and placed into a culture medium with a smooth surface after 200 mu l of OP50 bacterial liquid is dripped for overnight culture, amplification culture is carried out, and experiments are carried out after the caenorhabditis elegans is synchronously treated for 2-3 times after the caenorhabditis elegans are propagated to a certain number.
2) S-complete medium was used to culture C.elegans. The normal culture temperature is 20 ℃/25 ℃. The culture was performed as described above.
(3) Synchronization
Bleach lysis method: washing hermaphrodite in oviposition with M9 buffer solution from NGM culture medium, centrifuging in 15ml centrifuge tube at 1700rpm and 21 deg.C for 1-2min, and discarding supernatant. And (3) repeatedly washing the dish twice by using the same method, adding an equal volume of Bleach lysate, and quickly shaking for 3-5min until the liquid is clear and contains a large amount of worm eggs under a microscope. The lysis was stopped by adding a volume of M9 buffer, and after centrifugation, the supernatant was discarded, leaving a pellet. The pellet was washed with 2ml of M9 buffer and the procedure was repeated twice. The eggs were allowed to develop into L1 larvae by overnight incubation on a shaker at 20 ℃.
An egg laying method: and (3) selecting adults in the egg laying peak period to an NGM culture medium flat plate containing OP50 (bacterial liquid with the diameter of 6 cm and the diameter of 300 mu lOD600 of 0.6 is dripped into a culture dish), picking the adults out after 5 hours of egg laying in a biochemical incubator at 25 ℃, and continuously culturing in the incubator to obtain the nematodes in the same period.
(3) Observation of Life
Synchronized L4 nematodes were picked into groups of standard nematode NGM media, 90 per group, with 30 in each plate. Nematode survival days were counted from the moment of transfer, which was recorded as day 0 of the life test (day 0). The nematodes were observed daily and the number of nematode deaths recorded. Nematode death criteria: loss of motor ability and no response to multiple touch. The mean life span, median life span (median life span of each group of nematodes), and maximum life span (mean life span of the last 10 nematodes living in each group) were statistically compared.
(4) Drug addition
The drug is added directly into OP50 bacteria, that is, wild chrysanthemum lactone or metformin dissolved in DMSO solution is added into OP50 bacteria solution to make the final concentration be the final concentration shown in figure 1 a.
3. Analysis of results
Survival plots or histograms were plotted using GraphPad Prism 8. The data comparison of the life cycle curves was performed using a time series test (Long-rank test), and the other data comparisons were analyzed using a one-way ANOVA. The 25 ℃ experiment result shows that the Median life (media life span) of the control group (0 mu M wild chrysanthemum lactone) is 14 days, and the maximum life (Max life span) is 19 days. The median life of the dosing group (concentration of 50 μ M) was 15 days (50 μ M), and the maximum life was 22 days (50 μ M) in order. The median life of the positive control 40 μ M metformin was 16 days, with a maximum life of 20 days. Wherein 50 μ M of wild chrysanthemum lactone prolonged the average life span of nematodes by 16% (P < 0.01) (FIG. 1 a). The results show that at 25 ℃, 50 μ M of wild chrysanthemum lactone significantly prolongs the life of wild type caenorhabditis elegans, and the degree of the prolongation is close to that of the positive compound (metformin).
The 20 ℃ experiment result shows that the median life of the control group (0 mu M wild chrysanthemum lactone) is 17 days, and the maximum life is 21 days. The median life of the dosing groups (concentration of 50 μ M) was 19 days (50 μ M) in order, and the maximum life was 25 days (50 μ M) in order. The median life of the positive control 40 μ M metformin was 19 days, with a maximum life of 27 days. Wherein 50 μ M of wild chrysanthemum lactone prolonged the average life span of nematodes by 19% (P < 0.001) (FIG. 1 b). The result shows that at 20 ℃, the service life of wild type caenorhabditis elegans is remarkably prolonged by 50 mu M of wild chrysanthemum lactone, and the prolonging degree is lower than that of a positive compound metformin.
These results demonstrate that 50 μ M of wild chrysanthemum lactone significantly extended the life of caenorhabditis elegans at both nematode life test temperatures of 20 ℃ and 25 ℃. In conclusion, the 50 mu M wild chrysanthemum lactone obviously prolongs the life of caenorhabditis elegans, the prolonging action is close to that of the positive compound metformin, and the prolonging action is not influenced by temperature.
Example 2: experiment for influence of 50 mu M wild chrysanthemum lactone on health life of nematode
(1) Pharyngeal pumping rate test
The nematodes were picked for 8, 10 and 12 days of administration and the number of pharyngeal pumping swallows per nematode within 1min was recorded. The pharyngeal pumping rate experiment was performed in triplicate for each group of dosing and control groups, each for 10 nematodes.
Through pharyngeal pump movement experiments on the wild type N2 nematodes in the middle and old age period (8 days, 10 days and 12 days after imago), the mean pharyngeal pump movement rate of the caenorhabditis elegans in the control group is 82 times/min and the administration group is 110 times/min when 50 mu M wild chrysanthemum lactone intervenes in the wild type N2 at the end of L4 or the early stage of the imago for 8 days (the administration group is 110 times/min) ((P<0.001). At 10 days of intervention, the mean pharyngeal pumping rate of the caenorhabditis elegans in the control group is 41 times/min, and the mean pharyngeal pumping rate of the caenorhabditis elegans in the administration group is 72 times/min (P<0.001). The mean pharyngeal pumping movement rate of the caenorhabditis elegans in the control group is 13 times/mi at the time of 12 days of interventionn, the administration group is 27 times/min (P<0.05) (fig. 2 a). The 50 mu M wild chrysanthemum lactone-treated pharyngeal pump motor capacity of the caenorhabditis elegans is obviously higher than that of a control group, namely Handeiln obviously improves the pharyngeal pump motor function of the caenorhabditis elegans in the middle and old age period.
(2) Physical exercise testing
After 8 days of the administration treatment of the nematodes at the end of the L4 period or the early stage of the adult, the number of body swings of each nematode in the S-complete culture solution within 1min was recorded every two days. Each group of body movements was tested in triplicate, each paralleling approximately 10 worms.
Through body swing rate experiments of wild type N2 nematodes in the middle and old age period (8 days, 10 days and 12 days after imago), the average swing rate of the nematodes in the control group is 50 times/min and the average swing rate of the nematodes in the administration group is 74 times/min (P < 0.001) when 50 mu M wild chrysanthemum lactone intervenes in wild type N2 at the end of L4 or the initial period of the imago for 8 days. The mean swing rate of the nematode bodies in the control group is 22 times/min and the mean swing rate in the administration group is 39 times/min (P < 0.001) at 10 days of intervention. At 12 days of intervention, the mean rate of nematode body oscillation was 12 times/min in the control group and 28 times/min in the administered group (P < 0.01) (FIG. 2 b). The body swing capacity of the caenorhabditis elegans treated by the 50 mu M wild chrysanthemum lactone is obviously higher than that of a control group, namely the wild chrysanthemum lactone obviously improves the body movement function of the caenorhabditis elegans in the middle and old age period.
Example 3: influence of mother chrysanthemum lactone on resistance of nematode to heat stress
(1) Solid heat stress test at 35 ℃: after the nematodes cultured to the final stage of L4 or the initial stage of the adult are transferred to NGM culture media of the administration group and the control group for 5 days after synchronization, the NGM culture media are replaced by new NGM culture media and placed at 35 ℃ for 8 hours for culture, a heat stress experiment is started, the nematodes cultured to the final stage of L4 or the initial stage of the adult are transferred to the NGM culture media of the administration group and the control group for culture for 5 days after synchronization, and the nematodes are transferred to a 35 ℃ incubator for culture. Survival and mortality of nematodes were recorded every 2 hours. Each set of experiments was in triplicate, each with 40 worms in parallel.
(2) 35 ℃ liquid heat stress test: after synchronized post-culture until late L4 or early adult nematodes were transferred to NGM in the dosed and control groups for 5 days, 1mL of freshly prepared S-complete medium (containing dosed and control groups) was added to each well of a 12-well plate, 10 nematodes were placed in each well, and nematode mortality and survival were recorded every 2 hours. Each group was three replicates, each replicate 40 worms.
The results of the 35 ℃ solid heat stress experiment show that the median life of the control group caenorhabditis elegans is 29 hours, and the maximum life is 40 hours. The median life of 50 mu M wild chrysanthemum lactone administration group nematodes is 31 hours, and the maximum life is 40 hours. Under solid heat stress conditions, 50 μ M wild chrysanthemum lactone prolonged the median life of wild type N2 nematodes and significantly prolonged the overall life of the nematodes (P < 0.01) (fig. 3 a).
The liquid heat stress experiment result at 35 ℃ shows that the average survival rate of the nematodes in the control group is 34% and the average survival rate of the nematodes in the 50 mu M wild chrysanthemum lactone administration group is 47% after 8 hours of intervention. Under the liquid heat stress condition, the life of the wild type N2 nematode was prolonged by 50 μ M of wild chrysanthemum lactone (FIG. 3 b).
The experiment results show that 50 mu M of wild chrysanthemum lactone remarkably enhances the resistance of caenorhabditis elegans to heat stress no matter the NGM culture medium is used for carrying out the heat stress experiment at 35 ℃ or the liquid S-complete culture medium is used for carrying out the heat stress experiment at 35 ℃.
Example 4 Effect of mother Chrysanthemum lactone on combating antioxidant stress of nematodes
1. H2O2Oxidative stress
After the nematodes cultured to the late stage of L4 or the early stage of adult after synchronization were transferred to NGM medium of the administration group and the control group and cultured for 5 days, 1mL of freshly prepared H with a mass concentration of 0.003% was added to each of 12-well plates2O2The S-complete culture solution of (2) was placed in each well for 10 nematodes, and the number of dead nematodes and the number of alive nematodes were recorded every 2 hours. Each group was three replicates, each replicate 40 worms.
After 50 mu M of wild chrysanthemum lactone intervenes wild type N2 (adult day 0) nematodes for 5 days (t =0 is set at this time), the nematodes are placed in a solution containing 0.003% H2O2Subjecting to Oxidative Stress (OS) in the S-complex liquid medium of (1)And (6) testing. The results show that the median life span of the control nematodes was 18 hours and the maximum life span was 30 hours. The median life of 50 mu M wild chrysanthemum lactone administration group nematodes was 20 hours, and the maximum life was 34 hours, indicating 0.003% H2O2Under the induced oxidative stress, 50 mu M wild chrysanthemum lactone has certain influence on the median life and the maximum life of the wild type N2 nematodes (fig. 4 a).
2. Paraquat oxidative stress
After the nematodes cultured after synchronization to the late stage of L4 or the early stage of adult were transferred to NGM medium of the administered group and the control group for 5 days, they were transferred to NGM medium containing paraquat at a concentration of 5mM, and the number of dead and alive nematodes were recorded every 1 hour. Each group was three replicates, each replicate 40 worms.
After 50 μ M of chrysanthemumlactone intervenes in wild type N2 (adult day 0) nematodes 5 days later (at this time, t =0 was set), the nematodes were placed in NGM medium containing 5mM paraquat for oxidative stress experiments. The results show that the median life span of the control group nematodes was 6 days and the maximum life span was 9 days. The median life of 50 mu M wild chrysanthemum lactone administration group nematodes is 6 days, and the maximum life is 10 days. It was shown that 50. mu.M of wild chrysanthemum indicum lactone has a certain effect on the maximum life span of wild type N2 nematodes under oxidative stress caused by 5mM paraquat (FIG. 4 b).
3. Juglone oxidative stress
After the nematodes cultured to the late stage of L4 or the initial stage of adult after synchronization were transferred to NGM culture media of the administration group and the control group for 5 days, a 12-well plate was taken, 1ml of freshly prepared S basal buffer solution containing juglone at a concentration of 240 μ M was added to each well, 10 nematodes were placed in each well, the number of dead nematodes and the number of surviving nematodes were recorded every 2 hours, 3 nematodes in each group were paralleled, and 40 nematodes were paralleled.
After 50 mu M of wild chrysanthemum lactone intervenes wild type N2 (adult day 0) nematodes for 5 days (t =0 is set at the moment), the nematodes are placed in an S-complete culture solution containing 240 mu M juglone (juglone) for an oxidative stress experiment. The results show that the average survival rate of the nematodes in the control group after 6 hours of intervention is 27%, and the average survival rate of the nematodes in the 50 μ M wild chrysanthemum lactone administration group after 6 hours of intervention is 33% (fig. 4 c). The result shows that under oxidative stress caused by 240 mu M juglone, 50 mu M wild chrysanthemum lactone has a certain effect on the overall life of the wild type N2 nematode within 6 hours, but the effect is not significant.
In summary, the wild chrysanthemum lactone can resist 0.003% of H to a certain extent2O25mM paraquat and 240 muM juglone.
Example 5 Effect of wild Chrysanthemum lactone on nematode antibacterial experiments and activating immunity
1. Bacterial infection assay
The experiment takes two intervention modes: (1) after synchronization, the adult worms were cultured for five days with normal E.coli OP50, and then were subjected to a longevity test with the intervention of gram-negative bacteria PA 14. (2) After the synchronization, the strain is cultured by normal Escherichia coli OP50 to the L4 stage, and the life test is directly carried out by using a gram-negative bacterium PA14 mixed with 50 mu M wild chrysanthemum lactone.
Bacterial infection experiments employed two different intervention periods: 1) five days after normal medicated feeding, a life test of bacterial infection was performed with Pseudomonas aeruginosa (PA 14) intervention. 2) After the culture is finished at the end of the L4 period or at the early stage of imago, the pseudomonas aeruginosa PA14 is directly fed with drugs for carrying out the bacterial infection life test.
In the first experimental mode, the median life of the nematodes in the control group was found to be 18 hours. The maximum lifetime is 48 hours. The median life of 50 mu M wild chrysanthemum lactone administration group nematodes is 20 hours, and the maximum life is 48 hours. The wild chrysanthemum lactone extended the median life of PA14 bacteria infected nematodes, but did not significantly reduce mortality due to caenorhabditis elegans PA14 bacterial infection as a whole (fig. 5 a). In the second experimental mode, the median life span of the control group nematodes was found to be 4.5 days. The median lifetime was 10 days. The median life of 50 mu M wild chrysanthemum lactone administration group nematodes is 6 days, and the maximum life is 11 days. The wild chrysanthemum lactone prolongs the median life and the maximum life of the PA14 bacteria infected nematodes and reduces the mortality rate (P < 0.05) caused by the bacterial infection of the caenorhabditis elegans PA14 (FIG. 5 b).
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (5)
1. The application of the wild chrysanthemum lactone in preparing anti-stress medicaments or life prolonging medicaments for caenorhabditis elegans.
2. The use of wild chrysanthemum lactone according to claim 1 for the preparation of an anti-stress or life-prolonging drug for caenorhabditis elegans, characterized in that: the stress includes heat stress and oxidative stress.
3. A kit for researching the stress resistance or the life prolonging of caenorhabditis elegans is characterized by comprising a positive compound, wherein the positive compound is wild chrysanthemum lactone.
4. Application of wild chrysanthemum lactone in preparing anti-stress medicine or life prolonging medicine for human.
5. Application of wild chrysanthemum lactone in preparing human anti-stress food or medicine additive.
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