Application of nanobubble hydrogen water in prevention and treatment of noise-induced hearing loss and composition

Abstract

The invention discloses application of nanobubble hydrogen water in prevention and treatment of noise-induced hearing loss and a composition. The nano-bubble hydrogen water is hydrogen-saturated nano-bubble water and is used as a medicine component for preventing and treating noise-induced hearing loss. The bubble number density of the nano-bubble hydrogen water is 1.4-3E8 / ml, and the bubble diameter is 30-200nm. More specifically, the nanobubble hydrogen water is an oral preparation. The dosage of the nano-bubble hydrogen water is 10ml / day. The nano-bubble hydrogen water has good protection and treatment effects on noise-induced hearing impairment; the nano-bubble hydrogen water is non-toxic to a human body and almost has no side effect. The application of the nano-bubble hydrogen water provides a new path for prevention and treatment of noise-induced hearing loss.

Description

Technical Field

[0001] The present invention relates to the technical field of prevention and treatment of hearing loss, and more specifically, to an application of nano-bubble hydrogen water in the prevention and treatment of noise-induced hearing loss and a composition. Background Art

[0002] Noise-induced hearing loss (NIHL) is a progressive sensorineural deafness mainly caused by the damage of inner ear hair cells due to exposure to damaging noise environments. With the development of modern society, noise has become an inevitable potential hazard in daily life. The World Health Organization report points out that due to excessive noise damage, 1.1 billion young people globally are at risk of hearing impairment. Therefore, we need to conduct in-depth research on NIHL intervention measures.

[0003] The damage mechanism of noise to the inner ear is complex and diverse, among which the main ones are hair cell apoptosis, free radical damage, and cochlear immune inflammatory response. Currently, the most effective method to prevent NIHL is to physically attenuate the noise intensity entering the inner ear. Clinically, current research mainly focuses on enhancing the antioxidant capacity and anti-inflammatory response ability of tissues. Considering the efficacy and safety, there are currently no drugs or compounds approved by the FDA for its treatment.

[0004] Some studies have shown that hydrogen can indirectly improve the antioxidant capacity of the body by antioxidant and reducing apoptosis, and has a certain therapeutic effect on various diseases and injuries. However, in animal experiments, the stability of orally administered or intraperitoneally injected hydrogen-saturated water is poor, and its therapeutic effect on noise-induced hearing loss is very small. Therefore, its generalization and replicability are poor, and it cannot become a potential drug for preventing or treating noise-induced hearing loss. There is an urgent need to develop a highly efficient and safe compound or composition for the prevention and treatment of noise-induced hearing loss. Summary of the Invention

[0005] In view of the above problems, the present invention provides an application of nano-bubble hydrogen water in the prevention and treatment of noise-induced hearing loss and a composition, aiming to provide a highly efficient and safe drug ingredient for the prevention and treatment of noise-induced hearing loss.

[0006] To achieve the above object, on the one hand, the present invention provides an application of nano-bubble hydrogen water in the prevention and treatment of noise-induced hearing loss, characterized in that: the nano-bubble hydrogen water is used as a drug ingredient for the prevention and treatment of noise-induced hearing loss, and its bubble number density is 1.4 - 3E8 bubbles / ml, and the bubble diameter is 30 - 200 nm.

[0007] Further, the nano-bubble hydrogen water is an oral preparation.

[0008] Further, the dosage of the nano-bubble hydrogen water is 10 ml / day.

[0009] Furthermore, the nano-bubble hydrogen water is hydrogen-saturated nano-bubble hydrogen water with a hydrogen concentration of 1.5 ppm.

[0010] On the other hand, the present invention provides a composition containing nano-bubble hydrogen water for preventing and treating noise-induced hearing loss, characterized in that the number density of bubbles in the nano-bubble hydrogen water is 1.4 - 3E8 bubbles / ml, and the bubble diameter is 30 - 200 nm.

[0011] Furthermore, the composition is an oral preparation.

[0012] Furthermore, the dosage of nano-bubble hydrogen water in the composition is 10 ml / day.

[0013] Furthermore, the nano-bubble hydrogen water in the composition is hydrogen-saturated nano-bubble hydrogen water with a hydrogen concentration of 1.5 ppm.

[0014] Compared with the prior art, the above invention has the following advantages or beneficial effects:

[0015] The present invention discloses that nano-bubble hydrogen water has extremely excellent effects on the prevention and treatment of noise-induced hearing loss compared with ordinary hydrogen water. Experiments have confirmed that compared with the control group, the auditory brainstem response (ABR) threshold of the model mice given nano-bubble hydrogen water recovered faster and more significantly, indicating that nano-bubble hydrogen water can make the threshold recover faster after noise exposure and treat noise-induced hearing loss; compared with the control group, the amplitude of wave I of the auditory brainstem response of the model mice given nano-bubble hydrogen water was lower and the latency was prolonged, indicating that nano-bubble hydrogen water has a protective effect on noise-induced injury; the experimental results show that hydrogen in nano-bubble hydrogen water may penetrate the cochlear barrier and reach the organ of Corti in the cochlea, acting more directly on the lesion site and having a protective effect on cochlear hair cells and synaptic structures, thus achieving unexpected therapeutic effects; in addition, the bubbles in nano-bubble hydrogen water have very special properties, with a negative charge on the surface, having a good interfacial effect, having strong penetrability in tissues, and showing significant biological effects. Nano-bubble hydrogen water is non-toxic to the human body and has almost no side effects. These results of the present invention will provide a new path for the prevention and treatment of noise-induced hearing loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] By reading the following detailed description of the non-limiting embodiments with reference to the accompanying drawings, the present invention and its features and advantages will become more obvious.

[0017] Figure 1 This is the change of ABR threshold of mice in the control group I and H2 group before and after noise exposure in an embodiment of the present invention;

[0018] Figure 2 Changes in ABR thresholds of mice in control group II and H2 group before and after noise exposure in an embodiment of the present invention;

[0019] Figure 3 Changes in ABR wave I amplitudes of mice in control group I and H2 group after noise exposure in an embodiment of the present invention;

[0020] Figure 4 Changes in latency of mice in control group I and H2 group after noise exposure in an embodiment of the present invention;

[0021] Figure 5 Changes in the number of hair cells of mice in control group I and H2 group after noise exposure in an embodiment of the present invention;

[0022] Figure 6 Changes in the number of synapses of mice in control group I and H2 group after noise exposure in an embodiment of the present invention. Detailed implementation manners

[0023] The technical solutions in the embodiments will be described clearly and completely below. Obviously, the following described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments.

[0024] The experimental materials, experimental instruments, and detection instruments involved in the following embodiments are all commercially available, and the usage methods of the instruments or the detection methods adopted are the existing technologies that can be retrieved.

[0025] The present invention relates to an application of nano-bubble hydrogen water in preventing and treating noise-induced hearing loss and a composition containing nano-bubble hydrogen water for preventing and treating noise-induced hearing loss. The nano-bubble hydrogen water, as a drug component for preventing and treating noise-induced hearing loss, has a bubble number density of 1.4 - 3E8 bubbles / ml and a bubble diameter of 30 - 200 nm.

[0026] Example 1

[0027] 1. Experimental materials:

[0028] 1.1. Reagents: Hydrogen-saturated nano-bubble water (nano-bubble hydrogen water, with a bubble number density of 1.4 - 3E8 bubbles / ml and a bubble diameter of 30 - 200 nm), stored at 4°C. Ordinary hydrogen-saturated water, stored at 4°C. Pure water, stored in a cool place.

[0029] 1.2 Animals: A number of 6-week-old male CBA / CaJ mice were selected, excluding middle ear and inner ear diseases, and auditory brainstem response (ABR) audiometry was performed to screen mice with normal ABR thresholds as experimental subjects. The experimental animals were maintained at a room temperature of 24°C, provided with sufficient water and food, with a 12-hour light-dark cycle, and the ambient noise was less than 40 dB SPL.

[0030] 2. Experimental methods:

[0031] 2.1 Animal grouping and treatment: The experimental animals were randomly divided into 3 groups, including: Control Group I, in which the control mice were orally administered the same volume of pure water as the experimental mice. Control Group II, in which the control mice were orally administered the same volume of ordinary hydrogen-saturated water as the experimental mice. Experimental Group (H2 group), in which the mice were orally administered nano-bubble hydrogen water at a dose of 10 ml / day 7 days before and after noise exposure. The nano-bubble hydrogen water was preferably nano-bubble hydrogen water saturated with hydrogen, with a hydrogen concentration of 1.5 ppm.

[0032] 2.2 Noise exposure of experimental animals: All experimental mice were placed in a unique noise exposure device, and the mice could move freely in the device space. The noise signal was generated by a real-time signal processor (TDT system). The sound of the speaker was calibrated before each noise exposure, and the sound intensity change at each part in the device during exposure was <1 dB. During noise exposure, a noise with a frequency of 8 - 16 kHz and a sound intensity of 98 dB SPL was used for exposure, and the exposure time was 2 hours.

[0033] 3. Detection:

[0034] 3.1 Auditory brainstem response (ABR)

[0035] The baseline value and the changes in hearing thresholds after noise exposure were measured on the day before noise exposure and on days 1, 3, 7, and 14 after noise exposure. Short pure tones with frequencies of 4, 8, 16, 22.6, and 32 kHz were used as stimulus sounds by the TDT system, and the hearing thresholds were recorded. When approaching the threshold, the detection was repeated more than twice to compare whether the waveforms were consistent to determine the final threshold. Each index was displayed, recorded, calculated, and stored in real time by the software attached to the test instrument.

[0036] 3.2 Recording the amplitude and latency of wave I

[0037] Based on the measured ABR, the amplitude and latency of wave I at 16 and 22.6 kHz on day 1, the amplitude and latency of wave I at 16 kHz on day 3, and the amplitude and latency of wave I at 22.6 kHz on day 14 were obtained.

[0038] 3.3. Immunofluorescent staining of the cochlear basilar membrane to count the number of outer hair cells and the number of ribbon synapses of inner hair cells

[0039] The experimental mice were sacrificed and the cochleae were quickly removed and fixed thoroughly with 4% paraformaldehyde. The basilar membrane was dissected, and the tectorial membrane and vestibular membrane were removed. Immunofluorescent staining of the hair cell cytoskeletal protein (MyosinVIIa) and the proteins related to the ribbon synapses of inner hair cells (Ctbp2, GluA2) was performed. A Zeiss LSM 880 laser confocal microscope was used to perform layer scanning on the specimens (Z-step 0.38 μm), and the number of ribbon synapses in each inner hair cell was counted.

[0040] The experimental data were expressed in the form of Mean±SD, and one-way analysis of variance was performed.

[0041] 4. Experimental results

[0042] 4.1. Effects of nano-bubble hydrogen water on the auditory function of mice after noise exposure

[0043] As Figure 1 and Figure 2 shown, first, by detecting the changes in ABR of mice in control group I, control group II, and H2 group before and after noise exposure, the efficacy of nano-bubble hydrogen water against noise-induced injury was reflected. The results showed that the hearing thresholds of mice in control group I, control group II, and H2 group increased on the 1st day after noise exposure and recovered on the 3rd, 7th, and 14th days, indicating that we successfully established an animal model of noise-induced hearing loss. On the 3rd, 7th, and 14th days, the ABR thresholds of the H2 group (corresponding to TTS+H2 in the figure) at sound frequencies of 16, 22.6, and 32 kHz were significantly restored compared with those of control group I (corresponding to TTS in the figure) and control group II (corresponding to TTS+ordinary hydrogen water in the figure), indicating that nano-bubble hydrogen water can make the thresholds recover faster after noise exposure and can protect against noise-induced hearing loss.

[0044] The amplitude of ABR wave I mainly reflects the excitability of cochlear afferent nerve fibers. After the hearing thresholds recovered, the amplitudes and latencies of wave I at 16 and 22.6 kHz on the 1st day after noise exposure of the experimental mice were analyzed respectively. See Figure 3 and Figure 4 , the amplitudes and latencies of wave I at 16 KHz on the 3rd day and 22.6 KHz on the 14th day. The results showed that the amplitude of ABR wave I in the control group mice was lower than that in the experimental group, and the latency was prolonged, indicating that nano-bubble hydrogen water has a protective effect against noise-induced injury.

[0045] Figure 5 and Figure 6The immunofluorescence staining of the cochlear basilar membrane was used to count the number of outer hair cells (OHC loss rate) and the number of ribbon synapses per inner hair cell (synapses per IHC) in control group I and the experimental group before and after noise exposure. After noise exposure, the number of outer hair cells and ribbon synapses in the basal turn of the cochlea in control group I decreased, but the loss of ribbon synapses in the experimental group was significantly less than that in the control group.

[0046] From the test results of the above examples and application examples, it can be seen that ordinary hydrogen-saturated water has no obvious therapeutic effect on noise-induced hearing loss. Nano-bubble hydrogen water has good protective and therapeutic effects on noise-induced hearing loss due to its optimized number of nano-bubbles and good interfacial effects. Hydrogen in nano-bubble hydrogen water may penetrate the cochlear barrier and reach the Organ of Corti in the cochlea, acting more directly on the lesion site, thus achieving unexpected therapeutic effects. In addition, the bubbles in nano-bubble hydrogen water have very special properties, are negatively charged on the surface, have good interfacial effects, and have strong penetrability in tissues, showing significant biological effects.

[0047] In summary, the present invention discloses an application of nano-bubble hydrogen water in the prevention and treatment of noise-induced hearing loss and a composition. The nano-bubble hydrogen water is hydrogen-saturated nano-bubble water and is used as a pharmaceutical ingredient for preventing and treating noise-induced hearing loss. The number density of the bubbles in the nano-bubble hydrogen water is 1.4 - 3E8 bubbles / ml, and the bubble diameter is 30 - 200 nm. More specifically, the nano-bubble hydrogen water is an oral preparation. The dosage of the nano-bubble hydrogen water is 10 ml / day. The nano-bubble hydrogen water in the present invention has good protective and therapeutic effects on noise-induced hearing loss; the nano-bubble hydrogen water is non-toxic to the human body and has almost no side effects. The application of nano-bubble hydrogen water will provide a new way for the prevention and treatment of noise-induced hearing loss.

[0048] Those skilled in the art should understand that those skilled in the art can achieve variation examples by combining the prior art and the above examples, which will not be elaborated here. Such variation examples do not affect the essence of the present invention and will not be elaborated here.

[0049] The preferred embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the above specific embodiments, and the devices and structures not described in detail should be understood to be implemented in a common manner in the art; any person skilled in the art can make many possible changes and modifications to the technical solution of the present invention, or modify it into an equivalent embodiment with equivalent changes, without departing from the scope of the technical solution of the present invention, which does not affect the essence of the present invention. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the protection of the technical solution of the present invention.

Claims

1. Application of nano-bubble hydrogen water in preventing and treating noise-induced hearing loss, characterized in that: The nano-bubble hydrogen water, as a pharmaceutical ingredient for preventing and treating noise-induced hearing loss, has a bubble number density of 1.4 - 3E8 bubbles / ml and a bubble diameter of 30 - 200 nm.

2. Use of a nano-bubble hydrogen water in the prevention and treatment of noise-induced hearing loss according to claim 1, characterized in that, The nano-bubble hydrogen water is an oral preparation.

3. Use of a nano-bubble hydrogen water according to claim 1 or 2 in the prevention and treatment of noise-induced hearing loss, characterized in that, The dosage of the nano-bubble hydrogen water is 10 ml / day.

4. Use of a nano-bubble hydrogen water in the prevention and treatment of noise-induced hearing loss according to claim 1 or 2, characterized in that, The nano-bubble hydrogen water is hydrogen-saturated nano-bubble hydrogen water with a hydrogen concentration of 1.5 ppm.

5. A composition containing nano-bubble hydrogen water for preventing and treating noise-induced hearing loss, characterized in that, The nano-bubble hydrogen water has a bubble number density of 1.4 - 3E8 bubbles / ml and a bubble diameter of 30 - 200 nm.

6. A composition containing nano-bubble hydrogen water for preventing and treating noise-induced hearing loss according to claim 5, characterized in that, The composition is an oral preparation.

7. A composition containing nano-bubble hydrogen water for preventing and treating noise-induced hearing loss according to claim 5 or 6, characterized in that, The dosage of the nano-bubble hydrogen water in the composition is 10 ml / day.

8. A composition containing nano-bubble hydrogen water for preventing and treating noise-induced hearing loss according to claim 5 or 6, characterized in that The nano-bubble hydrogen water in the composition is hydrogen-saturated nano-bubble hydrogen water with a hydrogen concentration of 1.5 ppm.

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