Use of DMX-5804 in the preparation of a drug for inhibiting budding of aurelia sp.

Abstract

The application belongs to the technical field of jellyfish prevention and control, and discloses application of DMX-5804 in preparation of a medicine for inhibiting budding reproduction of Aurelia sp.1. By inhibiting budding reproduction of A.coerulea hydrocladium, the outbreak of Aurelia sp.1 can be prevented and controlled, and the adverse effects on marine ecological environment and human coastal economic activities can be reduced. Through experimental verification, 1 μM DMX-5804 can reduce the number of hydrocladium buds without affecting the living state of A.coerulea hydrocladium, which indicates that the compound DMX-5804 can inhibit the asexual reproduction of Aurelia sp.1 without toxicity at this concentration. The application provides a new theory and method for controlling the number of jellyfish, preventing the outbreak of Aurelia sp.1, and studying the biological conservative cell proliferation strategy.

Description

Technical Field

[0001] This invention relates to the field of jellyfish control technology, and more specifically, to the application of DMX-5804 in the preparation of drugs that inhibit the budding and reproduction of moon jellyfish. Background Technology

[0002] Jellyfish outbreaks are considered a very serious ecological disaster and have attracted widespread social attention in recent years. Among them, the moon jellyfish (Aurelia coerulea) has shown a year-on-year increasing trend along the Yellow and Bohai Sea coasts of my country, becoming one of the major jellyfish populations causing disasters in the country. The moon jellyfish (Aurelia coerulea) belongs to the phylum Cnidaria and has a complex life cycle consisting of several stages, including alternating generations of planktonic medusa and sessile polyps, which reproduce sexually and asexually, respectively. The polyp stage is the asexual generation of the moon jellyfish, primarily expanding its population through asexual reproduction. Its asexual reproduction is divided into two different directions: budding and metamorphosis. On the one hand, budding allows for self-replication, increasing the population size of the polyp stage and providing a quantitative basis for moon jellyfish outbreaks; on the other hand, metamorphosis releases transverse schizonts and discoid bodies, ultimately producing a large number of adult jellyfish, causing moon jellyfish outbreaks. By inhibiting the budding reproduction of A. coerulea hydroids, outbreaks of moon jellyfish can be prevented from the early stages of the hydroid stage, reducing the adverse impact on the marine ecological environment and human coastal economic activities.

[0003] Previously, due to limitations in research methods and the lack of genetic information on the polyps of the moon jellyfish, research on the population control of moon jellyfish has been limited to the reproductive effects of environmental factors such as temperature, food, and salinity on jellyfish and their polyps. In recent years, some studies have focused on the molecular mechanisms of metamorphosis in jellyfish polyps and their intervention. However, inhibiting metamorphosis can only reduce the number of adult jellyfish to a certain extent. The population reserve of the benthic polyp stage before metamorphosis is the root cause problem that needs to be solved to control jellyfish outbreaks. However, there is currently a lack of methods to regulate the budding and proliferation of jellyfish polyps through intervention with key inhibitors. Summary of the Invention

[0004] In order to overcome the shortcomings of the prior art, the purpose of this invention is to provide the application of DMX-5804 in inhibiting the budding reproduction of moon jellyfish.

[0005] The objective of this invention is achieved through the following technical solution:

[0006] This invention first provides the application of DMX-5804 in inhibiting the budding reproduction of moon jellyfish.

[0007] This invention has been experimentally verified to show that after incubation for 14 days with artificial seawater reagents at concentrations of 1 μM and 10 μM DMX-5804, the DMX-5804-treated group can significantly reduce the budding rate and number of buds of hydroids at the same time, thereby reducing the number of jellyfish.

[0008] Therefore, this invention also provides the use of DMX-5804 in the preparation of a drug that inhibits the budding and reproduction of moon jellyfish.

[0009] This invention also provides the application of DMX-5804 in the preparation of functional products for preventing and controlling moon jellyfish outbreaks.

[0010] Therefore, preferably, in the above applications, the concentration of DMX-5804 is 1-10 μM.

[0011] In the above applications, the DMX-5804 has the following functions:

[0012] (1) Reduce the budding rate of moon jellyfish polyps, and / or,

[0013] (2) Reduce the number of buds emerging from the polyps of the moon jellyfish, and / or.

[0014] Therefore, the present invention also provides a method for inhibiting budding reproduction in moon jellyfish, comprising the following steps:

[0015] A. Preparation of inhibitor seawater solution: Prepare a DMX-5804 solution with a concentration of 1-10 μM using seawater;

[0016] B. Place the hydra bodies of the moon jellyfish in DMX-5804 solution and culture for 2-14 days.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] This invention discovered that compound DMX-5804 can inhibit the budding rate of moon jellyfish polyps. Experiments verified that incubating moon jellyfish polyps with 1 μM and 10 μM DMX-5804 reduced the number of buds. Results showed that from day 2 to day 14 of the experiment, the number of buds in the DMX-5804-treated group was less than that in the control group, providing a theoretical basis for the prevention and control of jellyfish outbreaks. Attached Figure Description

[0019] Figure 1 The left and right images show the changes in the number of buds emerging from the 1 μM DMX-5804 treatment group and its control group during the experimental period.

[0020] Figure 2The changes in the number of buds of hydra in the 1 μM Homosalate treatment group and its control group during the experimental period (left) and the changes in the number of buds of hydra in the 10 μM Homosalate treatment group and its control group (right);

[0021] Figure 3 This is a microscopic image of the budding and proliferation process of a representative moon jellyfish polyp during the experiment. Detailed Implementation

[0022] To better illustrate the purpose, technical solution, and advantages of this invention, the invention will be further described below with reference to specific drawings and embodiments. Unless otherwise specified, the experimental methods used in the embodiments are conventional methods, and the materials and reagents used are commercially available unless otherwise specified.

[0023] I. Experimental Materials

[0024] Experimental animals: Hydra of the moon jellyfish (cultured by mECC Golden Club), reagents: DMX-5804 (2mg, Taoshu Bio T5483), DMSO (Boguang Bio), DMX-5804 incubation solution was prepared using artificial seawater with a concentration of 28‰-32‰.

[0025] Instruments: Constant temperature incubator (Bangxi Instrument Technology Co., Ltd.), continuous zoom stereo microscope (Chongqing Aote Optical Instrument Co., Ltd.), pipette (Eppendorf) (10μl, 100μl, 200μl, 1000μl), electronic balance AL204 (precision) (Mettler-Toledo Instruments Ltd.), seawater salinity meter (AS-ST10 Aicevoos official flagship store).

[0026] II. Experimental Methods

[0027] 1. Prepare sterile artificial seawater: Prepare artificial seawater with a salinity of 28‰–32‰, sterilize it at high temperature, and then adjust the salinity again.

[0028] 2. Preparation of laboratory animals

[0029] Two weeks in advance, select healthy moon jellyfish polyps from the polyp rearing tank and inoculate them into new 12-well plates. Observe the inoculation status one week after inoculation, and feed them appropriately (to ensure the health of the polyps). Set up two groups, an experimental group and a control group, with one 12-well plate in each group, three polyps per well, for a total of 36 polyps. After they have fully attached, start feeding them regularly, once every four days, and change the seawater two hours after feeding.

[0030] 3. Prepare DMX-5804 seawater solution

[0031] (1) Prepare 10mM DMX-5804 stock solution: Dissolve 1mg DMX-5804 in 276.7017μl DMSO to prepare 10mM DMX-5804 stock solution.

[0032] (2) Prepare 10μM DMX-5804 seawater solution: Take 30μl of 10mM DMX-5804 stock solution and add it to 29.97ml of artificial seawater to obtain a total volume of 30ml of 10μM DMX-5804 seawater solution.

[0033] (3) Prepare 1μM DMX-5804 seawater solution for the experimental group: Take 3ml of 10μM DMX-5804 seawater solution and add it to 27ml of artificial seawater to obtain a total volume of 30ml of 1μM DMX-5804 seawater solution.

[0034] 4. Prepare Homosalate seawater solution

[0035] (1) Prepare 100mM Homosalate stock solution: Dissolve 1mg Homosalate in 381.1847μl DMSO to prepare 10mM Homosalate stock solution.

[0036] (2) Preparation of 10μM Homosalate seawater solution: Take 30μl of 10mM Homosalate solution and add it to 29.97ml of artificial seawater to obtain a total volume of 30ml of 100μM Homosalate seawater solution.

[0037] (3) Preparation of 1μM Homosalate seawater solution for the experimental group: Take 3ml of 10μM Homosalate solution and add it to 27ml of artificial seawater to obtain a total volume of 30ml of 1μM Homosalate seawater solution.

[0038] 5. Prepare a control group seawater solution.

[0039] (1) Prepare 0.01% DMSO seawater solution for 1 μM inhibitor control group: Add 3 μl DMSO to 29.997 ml artificial seawater to obtain a total volume of 30 ml of 0.01% DMSO seawater solution.

[0040] (2) Prepare 0.1% DMSO seawater solution for 10 μM inhibitor control group: Add 30 μl DMSO to 29.97 ml artificial seawater to obtain a total volume of 30 ml of 0.1% DMSO seawater solution.

[0041] 6. Incubation of hydroids with inhibitory seawater solution

[0042] The artificial seawater in the 12-well plates of polyps in the DMX-5804 treatment group was completely replaced with 1 μM DMX-5804 seawater solution and 10 μM DMX-5804 seawater solution, respectively. The artificial seawater in the 12-well plates of polyps in the Homosalate treatment group was completely replaced with 1 μM Homosalate seawater solution and 10 μM Homosalate seawater solution, respectively. The artificial seawater in the 12-well plates of polyps in the control group was completely replaced with the corresponding 0.01% DMSO seawater solution and 0.1% DMSO seawater solution. The polyps were cultured in a constant temperature incubator at 20℃, and the water was changed every 4 days.

[0043] 7. Observation of bud emergence

[0044] Label the groups and times on 12-well plates. Add the corresponding solution to each group at the same time (2 ml to each well). Incubate in a 20°C incubator. Observe every 24 hours from day 1 to day 7 and record the observations using a stereomicroscope. Observe whether the hydra sprouts, the time of sprouting, and the number of sprouts, and calculate the sprouting rate and other indicators.

[0045] III. Results Analysis

[0046] The experiment found that during the experimental period, the number of hydra buds in the 1μM DMX-5804 treatment group and the 10μM DMX-5804 treatment group was significantly lower than that in the control group. Figure 1 During the experiment, the hydroid tentacles were in a good extended state. Figure 3 The normal feeding behavior indicates that DMX-5804 does not affect the life state of the moon jellyfish polyps. This suggests that at this concentration, compound DMX-5804 can inhibit the asexual proliferation of moon jellyfish without producing toxicity. Conversely, the number of polyp budding in the 1 μM and 10 μM Mososalate treatment groups showed a change from initial decrease to increase, ultimately exceeding the number of budding in the control group. Figure 2 This indicates that, unlike the inhibitor's effect on the metamorphosis of the polyp, Homosalate has no inhibitory effect on the budding reproduction of the moon jellyfish polyp.

[0047] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. Application of DMX-5804 in inhibiting budding of Aequorea victoria.

2. Application of DMX-5804 in preparing a drug for inhibiting budding of Aequorea victoria.

3. Use according to claim 1 or 2, characterized in that, The concentration of DMX-5804 is 1-10 μM.

4. A method of inhibiting budding of Aurelia sp. characterized by, The method comprises the following steps: A. Preparation of an inhibitor seawater solution: DMX-5804 solution with a concentration of 1-10 μM is prepared with seawater; B. Culturing Aequorea victoria hydrocladium in the DMX-5804 solution for 2-14 days.

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