CN115634235A - Application of molecular hydrogen in preparation of medicine for preventing or relieving jellyfish sting - Google Patents
Application of molecular hydrogen in preparation of medicine for preventing or relieving jellyfish sting Download PDFInfo
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- CN115634235A CN115634235A CN202210788030.8A CN202210788030A CN115634235A CN 115634235 A CN115634235 A CN 115634235A CN 202210788030 A CN202210788030 A CN 202210788030A CN 115634235 A CN115634235 A CN 115634235A
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Abstract
The invention relates to the technical field of medicines, and provides application of molecular hydrogen in preparation of a medicine for preventing or relieving jellyfish sting. Experiments prove that the molecular hydrogen mixed administration and the advanced intraperitoneal administration both effectively inhibit the vola swelling of mice caused by Tentacle Extract (TE), and in addition, the molecular hydrogen can promote the cell proliferation and improve the LD of TE acting on cells 50 Provides a new theoretical basis for preventing or relieving poisoning symptoms such as skin inflammatory swelling and cell injury caused by jellyfish sting by molecular hydrogen. In addition, the molecular hydrogen has the anti-inflammatory and antioxidant effects with safety and no side effect, and the hydrogen-rich water, the hydrogen absorber, the derivative products thereof and the like are clinically approved, so the new indication of the molecular hydrogen provided by the invention can quickly realize clinical transformation. Since molecular hydrogen has great potential in the application of injury protection caused by jellyfish sting, the invention also aims to prevent or relieve systemic jellyfish stingThe toxic reaction and cell damage provide a new clinical treatment means.
Description
Technical Field
The invention belongs to the field of biological medicine, provides new application of molecular hydrogen or a medical hydrogen-rich medium, and particularly relates to application of the molecular hydrogen or the hydrogen-rich medium in preparation of a medicine for preventing or relieving jellyfish sting.
Background
Jellyfish is one of the most harmful marine organisms to human beings, the number of jellyfish stings is remarkably increased in coastal areas due to the frequent outbreak of the jellyfish, and more than 1.5 hundred million people are stinged by the jellyfish estimated in the world every year. It has been reported that jellyfish stings account for 30% to 80% of biological stings in coastal areas of china, and that between 1983 and 2019, there are over 2,000 clinical cases of jellyfish stings in china, of which at least 13 cause death. In australia, nearly 10,000 people are jellyfish-wounded each year, and the number of jellyfish-wounded people has sharply increased in japan in recent years. The toxic jellyfish can cause serious injury to a human body, wherein local skin symptoms comprise erythema, swelling, ulcer and necrosis accompanied by obvious pain and pruritus, the clinical sign is jellyfish dermatitis, and severe skin inflammatory reaction brings huge pain to a patient; systemic toxic symptoms include fever, vomiting, arrhythmia, acute pulmonary edema, and even organ failure, which in severe cases result in death of the patient. In addition, studies have shown that the jellyfish toxin has various toxicological effects such as hemolytic activity, muscle toxicity, neurotoxicity and cytotoxicity, but the jellyfish sting diagnosis and treatment is difficult due to complex and various toxin components and unclear toxicity mechanism.
Hydrogen, a hydrophobic, nonpolar molecule, has a solubility in water (0.8 mM at 20 ℃) lower than that of oxygen (1.34 mM at 20 ℃), and has been considered to be a physiologically inert and non-functional gas in the past. However, ohsawa et al, 2007 reported that hydrogen molecules could be a therapeutic antioxidant by selectively reducing intracellular toxic oxygen radicals, after which hydrogen gas was of interest as a "medical gas". Molecular hydrogen can selectively neutralize toxic radicals such as hydroxyl radical, nitrite anion, etc., without interfering with physiologically active oxygen, and thus is defined as a selective antioxidant. To date, the medical properties of molecular hydrogen have been explored for animal disease models and human diseases, where the mechanism of action of hydrogen is mainly related to antioxidant stress, as well as anti-inflammatory and anti-apoptotic effects in different organ systems. Hydrogen can protect the body from oxidative and inflammatory effects by reducing the amount of active oxygen that is cytotoxic, thereby preventing tissue damage. Due to the insecurity caused by long-term hydrogen inhalation, the hydrogen-rich water and the hydrogen-rich water are produced as derivatives and substitutes, and the hydrogen-rich water also show the same treatment and damage-resistant effects as the hydrogen in clinical tests and animal damage models. Research shows that the hydrogen-rich water has obvious protective effect on obstructive jaundice and chronic ethanol-induced liver injury; drinking hydrogen-rich water can be used for treating metabolic syndromes such as atherosclerosis, chemotherapy nephrotoxicity, brain injury, and insulin resistance; the hydrogen-rich water has redox balance effect in atopic dermatitis and other ultraviolet-mediated skin injury models, and can effectively inhibit endoplasmic reticulum stress, etc.
Aiming at inflammatory injury in vivo and in cells caused by jellyfish toxin and whether the antagonistic effect of molecular hydrogen is effective or not, no relevant research report is found at present, and the method adopts molecular hydrogen to research the antagonistic effect so as to find a treatment method for jellyfish sting and the wider application of molecular hydrogen.
Disclosure of Invention
The invention aims to provide a new medical application of molecular hydrogen.
In a first aspect of the invention, the application of molecular hydrogen in preparing a medicament for preventing or relieving jellyfish sting is provided. In a second aspect, there is provided the use of a hydrogen-rich medium in the manufacture of a medicament for preventing or alleviating jellyfish stings.
Preferably, the hydrogen-rich medium refers to medical hydrogen-rich normal saline
Preferably, the agent for preventing or alleviating jellyfish sting is an agent for alleviating inflammatory swelling of skin or tissue damage caused by jellyfish sting, or an agent for increasing the proliferation rate of Raw264.7 cells and increasing LD of jellyfish toxin acting on cells 50 The medicament of (1).
The medicine for reducing the inflammatory swelling of skin or tissue injury caused by jellyfish sting is a medicine for improving the edema of epidermis acanthosis, liquefaction and degeneration of basal layer, telangiectasis of dermal layer, separation and fracture of collagen fiber and infiltration of a large number of inflammatory cells caused by TE.
Preferably, the jellyfish sting prevention or alleviation medicine of the present invention is molecular hydrogen as the only active ingredient or is a pharmaceutical composition containing molecular hydrogen. The form of the pharmaceutical composition is preferably injection, oral liquid and the like.
Through mouse experiments, the inflammatory swelling degree of the foot palm of the mouse can be remarkably reduced by mixed injection or pre-injection of hydrogen-rich water and jellyfish TE. Pathological examination shows that the molecular hydrogen group is effective in improving the symptoms of edema of epidermis acanthosphere and basal layer, telangiectasis of dermis layer, separation and breakage of collagen fiber and infiltration of inflammatory cells caused by TE.
Cell experiments prove that the hydrogen can obviously improve the proliferation rate of Raw264.7 cells and improve LD of the aequorin acting on the cells 50 。
The method is suitable for the situations that sea workers, naval soldiers, seaside visitors and sea survivors suffer jellyfish sting, can be used for preventing or relieving jellyfish sting, recommends drinking hydrogen-rich water in advance or inhaling hydrogen and continuously injecting the hydrogen-rich water or inhaling the hydrogen immediately when the jellyfish sting occurs, and the administration mode is not limited to oral administration, injection and the like.
Action and Effect of the invention
Through experimental verification, the molecular hydrogen is simultaneously administrated and extractedThe administration of the compound to the anterior abdominal cavity effectively inhibits the vola swelling of mice caused by TE, and molecular hydrogen can promote cell proliferation and improve LD of TE acting on cells 50 Provides a new theoretical basis for preventing or relieving poisoning symptoms such as skin inflammatory swelling and cell injury caused by jellyfish sting by molecular hydrogen.
In addition, the molecular hydrogen has the anti-inflammatory and antioxidant effects with safety and no side effect, and the hydrogen-rich water, the hydrogen absorption machine and derivative products thereof and the like are clinically approved, so the new indication of the molecular hydrogen provided by the invention can quickly realize clinical transformation. The molecular hydrogen has great potential in the application of injury protection caused by jellyfish sting, and the invention also provides a new clinical treatment means for preventing or relieving systemic toxic reaction and cell injury caused by jellyfish sting.
Drawings
FIG. 1 is a graph showing the toxicity evaluation of molecular hydrogen against TE at the animal level. Wherein A is the mouse intracutaneous injection of TE and the mixed injection of hydrogen-rich water-TE (TE + H) 2 Mix), intraperitoneal injection of hydrogen-rich water-intradermal injection of TE (TE + H) 2 IP), intracutaneous injection of PBS footpad morphogram; b is the intradermal injection of TE and the mixed injection of hydrogen-rich water-TE (TE + H) to mice 2 Mix), intraperitoneal injection of hydrogen-rich water-intradermal injection of TE (TE + H) 2 IP), intracutaneous injection of PBS paw thickness profile; c is the intradermal injection of TE and the mixed injection of hydrogen-rich water-TE (TE + H) to mice 2 Mix), intraperitoneal injection of hydrogen-rich water-intradermal injection of TE (TE + H) 2 IP), percent change in intradermally injected PBS footpad swelling,
*，TE+H 2 the Mix group is significantly different from the TE group; #, TETE + H 2 The difference between the IP group and the TE group is remarkable; c: control, n =6,. Sup./# @&，P<0.05；**/##/&&，P<0.01。
FIG. 2 shows the intradermal injection of TE and the mixed injection of hydrogen-rich water-TE (TE + H) into mice 2 Mix), intraperitoneal injection of hydrogen-rich water-intradermal injection of TE (TE + H) 2 IP), histopathological sections of the paw injected intradermally with PBS at three time points of 12h, 48h, n =4, magnification 400 ×, scale bar =200px.
Fig. 3 is an evaluation of the toxicity of TE at cellular level for molecular hydrogen antagonism, where a and B are both the cytotoxicity of TE in hydrogen incubator and general cell incubator, n =6, P <0.05; * P <0.01.
Detailed Description
The present invention will be described in detail below with reference to examples and the accompanying drawings. The following examples should not be construed as limiting the scope of the invention.
1. Experimental Material
Preparing hydrogen-rich water: and filling hydrogen into the NS until saturation after water molecule electrolysis is carried out by utilizing a Shenzhenjian ryen hydrogen-rich cup, wherein the hydrogen content is about 1.5ppm.
The device for protecting cells by hydrogen adopts a hydrogen incubator provided by Shanghai Nanobibarbus.
2. Experimental methods
1. Animal model experiment
ICR mice were selected, and 7-week-old male mice were tested by intradermal injection of the left paw: injecting 1 XPBS into right sole as control group; jellyfish TE is used as an experimental group, and diluted by 1 × PBS to ensure that the final concentration of TE is 400 μ g/mL; molecular Hydrogen group, TE was diluted with 1.5ppm hydrogen-rich water to a final TE concentration of 400. Mu.g/mL as TE + H 2 Mix group; another way of molecular hydrogen adopts the prior intraperitoneal injection of 1mL of 1.5ppm hydrogen-rich water for 0.5H as TE + H 2 In the IP group, TE was injected into the sole of the foot at a final concentration of 400. Mu.g/mL.
The injection volumes of the sole of the experimental group, the molecular hydrogen group and the control group were 50. Mu.L, and the thickness of the sole was measured and photographed at 0h,0.5h,1h,2h,4h,12h,24h,36h and 48h after injection.
1 XPBS, TE + H were selected respectively 2 Mix and TE + H 2 IP groups 12h and 48h were examined for pathological sections of the soles.
2. Cell assay
Raw264.7 cells with good growth are selected for cell experiments. A bottle of Raw264.7 cells was taken out, and according to the cell concentration, 8,000 cells/70. Mu.L of cell suspension was prepared by diluting with a cell culture medium, incubated after adding the corresponding reagent to a 96-well plate according to the conditions shown in Table 1 below, and the light absorption value (OD 450) at 450nm was measured with a microplate reader, wherein the cell suspension was replaced with the cell culture medium in the blank group and TE was replaced with 1 XPBS in the control group.
TABLE 1 TE sample application table for Raw264.7 cytotoxicity determination
One 96-well plate is placed in a common cell incubator, and the other 96-well plate is placed in a hydrogen incubator.
3. Analysis of results
Experiments show that in the molecular hydrogen group, hydrogen-rich water is injected intraperitoneally-TE (TE + H) is injected intradermally 2 IP) and hydrogen-rich water-TE mixture (TE + H) 2 Mix) both hydrogen administration modes of injecting the paw effectively reduce the swelling percentage of the mouse paw: the abdominal cavity group can remarkably inhibit TE-induced sole swelling within 6-24 h (P in 6h and 24 h)<0.01; at a time of 12h, P<0.05 And the mixed group can obviously inhibit the sole swelling caused by TE within 6 to 12 hours (P when the time is 6 hours)<0.01; at a time of 12h, P<0.05 FIGS. 1A to C. The results show that the molecular hydrogen effectively inhibits inflammatory swelling of the soles of the mice caused by TE, and the anti-inflammatory and antioxidant treatment can be a potential effective method for treating jellyfish sting.
The pathological examination shows that the molecular hydrogen group effectively improves the conditions of intercellular edema of epidermis acanthosis, liquefaction and denaturation of basal layer, telangiectasis of dermis layer, light dyeing and separation and fracture of collagen fiber and infiltration of inflammatory cells of dermis shallow middle layer caused by TE. Compared with the TE group, the intraperitoneal injection of hydrogen-rich water effectively reduces the generation of edema in 24 hours and effectively reduces the infiltration of inflammatory cells in 48 hours; while the hydrogen-rich water-TE mixed injection group has no obvious improvement at 24 hours, and effectively weakens the infiltration of inflammatory cells at 48 hours (figure 2). The results show that the antioxidant can obviously inhibit the inflammatory and edema-causing activity caused by TE and improve the degree of tissue damage.
It was found by experiment that TE has dose-dependent killing power on Raw264.7 cells, and the higher the concentration of TE, the lower the survival rate of Raw264.7 cells. When the cell activity is calculated by taking the cells of the control group in the common incubator as the control, the hydrogen incubator has different concentrationsThe activity of TE group cells and control group cells (0-12 mu g/mL) in the TE group cells is obviously higher than that in a common incubator (when TE is 2 mu g/mL and 12 mu g/mL, P is<0.05; TE 0. Mu.g/mL, 1.2. Mu.g/mL, and 6. Mu.g/mL, P<0.01 Demonstrated that molecular hydrogen itself can promote cell proliferation (fig. 3A); in addition, the results of the control groups in the respective incubators showed the LD of the cells in the hydrogen incubator 50 6.12 mu g/mL of the total volume of the culture solution is higher than the LD in a common incubator 50 4.96. Mu.g/mL, which is probably due to the antioxidant effect of molecular hydrogen, the cells were attacked less by active oxygen and thus were more viable than those in the ordinary incubator (FIG. 3B). The results indicate that molecular hydrogen can promote cell proliferation and increase LD of TE acting on cells 50 。
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 given by way of illustration of the principles of the present invention, and that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (7)
1. Application of molecular hydrogen in preparing medicine for preventing or relieving jellyfish sting.
2. Application of hydrogen-rich medium in preparing medicine for preventing or relieving jellyfish sting is provided.
3. Use according to claim 1 or 2, characterized in that:
wherein, the hydrogen-rich medium refers to hydrogen-rich normal saline.
4. Use according to claim 1 or 2, characterized in that:
wherein the jellyfish sting preventing or relieving medicine is used for relieving inflammatory swelling of skin or tissue injury caused by jellyfish stingOr increasing proliferation rate of Raw264.7 cells and increasing LD of aequorin acting on cells 50 The medicament of (1).
5. Use according to claim 4, characterized in that:
the medicine for reducing the inflammatory swelling of the skin or tissue damage caused by jellyfish sting is a medicine for improving the edema of an epidermal acantho layer, liquefaction and degeneration of a basal layer, telangiectasis of a dermal layer, separation and fracture of collagen fibers and infiltration of a large number of inflammatory cells caused by TE.
6. Use according to claim 1 or 2, characterized in that:
wherein the jellyfish sting prevention or alleviation medicine is a medicine composition containing molecular hydrogen as the only active ingredient.
7. Use according to claim 6, characterized in that:
wherein, the pharmaceutical composition is an injection or an oral liquid.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111700906A (en) * | 2020-05-20 | 2020-09-25 | 山东第一医科大学(山东省医学科学院) | Application of hydrogen molecule and construction method of mouse model for relieving inflammatory bowel disease |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111700906A (en) * | 2020-05-20 | 2020-09-25 | 山东第一医科大学(山东省医学科学院) | Application of hydrogen molecule and construction method of mouse model for relieving inflammatory bowel disease |
Non-Patent Citations (2)
| Title |
|---|
| 揣云海等: "氢气生物学及其医学应用", 生物物理学报, vol. 28, no. 9, pages 705 - 718 * |
| 肖良等: "水母毒素研究与转化医学", 国际药学研究杂志, vol. 38, no. 6, pages 439 * |
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