Application of protocatechuic acid in preparation of fish nonyl phenol toxicity resistant preparation
By adding procatechic acid to water or feed, the reproductive toxicity of nonylphenol to fish is solved, especially gonad damage in male fish, effectively recovering from nonylphenol-induced damage, and providing treatment of endocrine treatment An effective method for interfering substance toxicity.
Patent Information
- Application Number
- CN202411580421.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-07
- Publication Date
- 2025-01-14
AI Technical Summary
The prior art is difficult to effectively solve the reproductive toxicity of nonylphenol to fish, especially gonad damage in male fish, and the purity and content of existing resistant materials are low, so the dosage and efficacy cannot be effectively evaluated.
Procatechic acid is used as a method for preparing fish anti-nonylphenol toxic preparations. By adding 50 μg/L of procatechic acid to water or feed, it is used to prevent or treat fish gonadal damage induced by nonylphenol.
Protocatechic acid significantly restored fish gonadal damage induced by nonylphenol and restored its reproductive activity, providing an effective solution to control the toxicity of endocrine disruptors, and filling the gap in restoring the toxic effects of pollutants on animals.
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Figure CN119302938A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of drugs for treating toxicity of biological interfering substances, and particularly relates to application of protocatechuic acid in preparing a fish anti-nonylphenol toxicity preparation. Background Art
[0002] Nonylphenol (NP) is a degradation product of nonylphenol polyoxyethylene ether, both of which are mainly used in surfactants and modified phenolic resins, and exist in large quantities in human daily life. Nonylphenol is a typical endocrine disruptor and an artificial compound of exogenous estrogen, which has a strong estrogenic effect. Low doses can cause damage to male fish, but almost no significant harm to female fish. Studies in recent decades have found that nonylphenol is widely distributed and exists in large quantities in waters, posing many serious hazards to aquatic organisms, freshwater, estuarine and marine ecosystems. At the same time, the substance can also be bioaccumulated through the food chain, ultimately posing a huge threat to human health and survival.
[0003] At present, a large number of studies have focused on its "three-hazard" effects (teratogenicity, carcinogenicity, mutagenicity), acute and chronic toxicity, genotoxicity, bioaccumulation and amplification, DNA methylation, etc. on organisms. There is still a lack of research on how to restore the reproductive toxicity of fish induced by nonylphenol. Limited data show that Tang et al. used Lycium barbarum polysaccharides to mitigate the reproductive damage of zebrafish induced by nonylphenol (Tang et al., 2017); Wang et al. also found that Rheum mycelium polysaccharide can mitigate the toxic effects of nonylphenol (Wang et al., 2022). However, both of the above substances contain polysaccharides, their composition is relatively complex, and the specific active substances are not very clear; and the purity and content are low, and the relationship between dose and efficacy cannot be effectively evaluated. Secondly, both substances have a mitigating effect, and there is no report of recovery, and they may cause secondary pollution to the water body.
[0004] Protocatechuic acid (PCA) is a natural organic compound found in many higher plants. Most studies have found that it has typical antibacterial effects and protective effects on the heart and cardiovascular system. There are currently no reports on its effects related to nonylphenol. Summary of the invention
[0005] In view of the above-mentioned technical problems, the present invention provides the use of protocatechuic acid in the preparation of a fish anti-nonylphenol toxicity preparation to solve the problem of the existing nonylphenol accumulation in water bodies causing reproductive toxicity to fish.
[0006] The invention provides application of protocatechuic acid in preparing a fish anti-nonylphenol toxicity preparation.
[0007] Furthermore, the fish anti-nonylphenol toxicity preparation is used to prevent or treat the toxicity induced by nonylphenol to fish.
[0008] Furthermore, the toxicity induced by the nonylphenol to fish manifests as reproductive damage.
[0009] Furthermore, the reproductive damage is gonadal damage of male fish.
[0010] Furthermore, the fish anti-nonylphenol toxicity preparation is a medicine or feed.
[0011] Furthermore, the protocatechuic acid is used by adding it into water.
[0012] Furthermore, the concentration of protocatechuic acid in the water was 50 μg / L.
[0013] Furthermore, the protocatechuic acid is used by adding it into medicine or feed.
[0014] Furthermore, the fish is zebrafish.
[0015] According to the application of protocatechuic acid provided by the present invention in the preparation of fish anti-nonylphenol toxicity preparations, nonylphenol induced male gonadal damage in fish, and through specific experiments it was found that protocatechuic acid had obvious therapeutic effects on nonylphenol toxicity in fish, and could significantly restore the gonadal damage induced by nonylphenol in fish, and restore its reproductive activity. This discovery provides positive guidance for the program of controlling the toxicity of endocrine disruptors to animals, and fills the gap in restoring the toxic effects of pollutants on animals. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0017] Figure 1 Schematic diagram of the PCA recovery protocol for zebrafish toxic injury induced by nonylphenol.
[0018] Figure 2 Histological sections showing protocatechuic acid recovery of nonylphenol-induced gonadal damage in male zebrafish.
[0019] Figure 3 Figure 3. Cell-free area analysis of protocatechuic acid restoring nonylphenol-induced gonadal damage in male zebrafish.
[0020] Figure 4 This is a correlation diagram of gene expression levels among samples under different treatment modes.
[0021] Figure 5 The following is the Venn diagram of differentially expressed genes in each group of experiments.
[0022] Figure 6 The figure shows the clustering heat map of differentially expressed genes in each group of experiments.
[0023] Figure 7 This is a transcriptome DEG enrichment analysis diagram (A: GO terms; B: KEGG pathway). DETAILED DESCRIPTION
[0024] The invention discloses application of protocatechuic acid in preparing a fish anti-nonylphenol toxicity preparation.
[0025] The following will be combined with the accompanying drawings in the present invention to clearly and completely describe the technical solutions in the present invention. Obviously, what is described is only a part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0026]
[0027] The present invention provides a method for recovering the toxicity induced by nonylphenol in fish. Zebrafish is used as a model organism, nonylphenol is used as a target pollutant for exposure experiment, and after 15 days, the fish are put into water without nonylphenol and water containing protocatechuic acid for recovery experiment respectively. The test scheme is as follows: Figure 1 As shown, the grey part is the exposure period, the white part is the unexposed period, and the orange part is the PCA recovery period.
[0028] (1) Preliminary preparation
[0029] 700 fertilized eggs were selected for incubation, and the incubation time was 3-4 days. Dead eggs and fry were removed during the incubation process. On the 5th day, paramecium was fed, and every 100 zebrafish fry were cultured in a fish tank containing 5L of aerated oxygenated water, and divided into 7 fish tanks for feeding.
[0030] Culture conditions: The water temperature is 26±1℃, and red worms are fed as bait, once in the morning, noon and evening each day. The best feeding amount is when the fish can finish eating the worms within 10 minutes.
[0031] (2) Experimental and evaluation procedures
[0032] Starting from the 20th day, 40 zebrafish were placed in each glass tank for breeding, including 3 blank control groups and 6 nonylphenol-treated groups.
[0033] The nonylphenol treatment group was exposed to 150 μg / L nonylphenol for 15 days. After the exposure, the zebrafish that had differentiated into females and those that had not differentiated into females were removed, leaving only male zebrafish, with no less than 15 male zebrafish in each fish tank. During the exposure process, half of the nonylphenol exposure solution was replaced every day to maintain its concentration.
[0034] After the exposure, the 6 groups in the nonylphenol treatment group were randomly divided into 3 groups and transferred to aerated clean water for recovery. No substances were added to this group, and they were fed normally, with half of the water changed every two days. The other 3 groups were transferred to aerated water containing protocatechuic acid, where the protocatechuic acid content was 50μg / L, and they were fed normally, with half of the water changed every day. The recovery period was 25 days.
[0035] After the recovery period, the zebrafish gonads were dissected for pathological tissue sections to study gonadal damage. Six zebrafish were randomly selected from each group and placed in cryopreservation tubes, frozen in liquid nitrogen, and sent to Beijing Novogene Technology Co., Ltd. for transcriptome sequencing.
[0036] (3) Experimental results
[0037] 3.1 Analysis of tissue sections of protocatechuic acid restoring nonylphenol-induced gonadal damage in male zebrafish
[0038] Figure 2 The figure shows the tissue sections of protocatechuic acid restoring the nonylphenol-induced gonadal damage in male zebrafish. Figure 2 -D, E, and F have fewer sperm numbers and densities, fewer spermatogonia, and larger intercellular spaces. Figure 2 -G,H,I), the sperm count and density were effectively restored, and the number of spermatogonia was significantly increased.
[0039] Comparing the cell-free areas in the gonads of different treatment groups, the analysis showed that the cell-free area in the water recovery group was 37.27±2.24%, while that in the blank control group and the protocatechuic acid recovery group was 12.75±4.22% and 16.32±1.29%, respectively, which was significantly lower than that in the water recovery group, and there was no significant difference between the control group and the protocatechuic acid group. Figure 3 .
[0040] 3.2 Transcriptomic analysis of protocatechuic acid restoring nonylphenol-induced gonadal damage in male zebrafish
[0041] Figure 4 is the correlation of gene expression levels between samples under different treatment modes. Figure 4 It can be seen that the R 2<0.5, indicating that the correlation is poor. However, the correlation between the protocatechuic acid recovery group and the blank control group is high, indicating that the difference in gene expression levels between the protocatechuic acid recovery group and the blank control group is small.
[0042] from Figure 5 It can be seen that there are 18,892 genes with inconsistent regulation between the PCA and NP groups, while there are only 149 genes with inconsistent regulation between the PCA and blank control groups, indicating that the results of the PCA and blank control groups after NP treatment are relatively similar, which further indicates that protocatechuic acid has a more obvious effect in restoring nonylphenol-induced damage in male zebrafish.
[0043] from Figure 6 It can be seen that the up- and down-regulated genes of the blank control group and the PCA group are basically the same, while the up- and down-regulated genes of the NP and clean water recovery groups have significant differences with the blank control group and the PCA group. Among them, there are 19079 genes with significant differences between the PCA group and the NP group (p<0.01), and there are 336 genes with significant differences between the PCA group and the blank control group (p<0.01). The results show that the differential genes between the PCA group and the blank control group are far less than those between the PCA group and the NP group, indicating that PCA has played a role in recovery.
[0044] GO analysis ( Figure 7 -A(a)(b)(c)), the comparison between PCA and NP groups revealed that RNA synthesis, carbohydrate synthesis, and enzyme activities involved in related pathways were up- and down-regulated to varying degrees; while the comparison between PCA and the blank control group revealed that the up- and down-regulation was concentrated between oxidoreductase activity and growth factors, indicating that the NP group had affected the regulation of basic vital signs, while the PCA group had improved this. From the KEGG pathway analysis ( Figure 7 -B(a)(b)(c)), the comparison between PCA and NP groups revealed that RNA metabolism, sugar and fat synthesis, and cell synthesis pathways were downregulated, while the comparison between PCA and blank control groups revealed changes in the regulatory pathways of some enzymes, which further indicated that PCA had restored the damage caused by NP.
[0045] The above contents are further detailed descriptions of the present invention in combination with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For ordinary technicians in the technical field to which the present invention belongs, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.
Claims
1. Application of protocatechuic acid in the preparation of fish anti-nonylphenol toxicity preparations.
2. The use according to claim 1, characterized in that: The fish anti-nonylphenol toxicity preparation is used for preventing or treating the toxicity induced by nonylphenol to fish.
3. The use according to claim 2, characterized in that: The nonylphenol-induced toxicity in fish manifests itself in reproductive damage.
4. The use according to claim 3, characterized in that: The reproductive damage refers to gonad damage of male fish.
5. The use according to claim 1, characterized in that: The fish anti-nonylphenol toxicity preparation is a medicine or feed.
6. The use according to claim 1, characterized in that: The protocatechuic acid is used by adding it into water.
7. The use according to claim 6, characterized in that: The concentration of protocatechuic acid in water is 50μg / L.
8. The use according to claim 1, characterized in that: The protocatechuic acid is used by adding it into medicine or feed.
9. The use according to claim 1, characterized in that: The fish is zebrafish.