Application of divalent metal lactate in the preparation of products for repairing nerve damage

Abstract

The present invention belongs to the field of biomedicine technology, and specifically relates to the use of divalent metal lactates in the preparation of products for preventing, improving, repairing, and treating nerve damage. Preferably, the divalent metal lactate comprises a combination of one or more of calcium lactate, magnesium lactate, zinc lactate, and ferrous lactate. The present invention provides a new treatment for repairing central and peripheral nerve damage, and has broad application prospects in the treatment and repair of central nervous system damage caused by trauma, or peripheral neuritis or peripheral nerve paresthesia caused by viruses, diabetes, chemotherapy drugs, and other causes.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the application of divalent metal lactate in the preparation of products for preventing, improving, repairing and treating nerve damage. Background Art

[0002] The central and peripheral nervous systems comprise the human nervous system, playing a leading role in maintaining homeostasis, the integrity and unity of the body, and its coordinated balance with the external environment. With the deepening of social modernization, traffic accidents and sports accidents are inevitably increasing. Furthermore, potential violent incidents and natural disasters can cause damage to the nervous system.

[0003] Spinal cord injury (SCI) is a severe central nervous system injury that can lead to impaired sensory and motor function, placing a significant burden on patients' families and society. SCI is a complex and delicate process, categorized as primary and secondary based on the pathological process of neuronal damage. The severity of SCI is primarily determined by the secondary injury phase, which is primarily due to significant changes in the spinal cord microenvironment. These changes include elevated reactive oxygen species (ROS) levels, impaired oxygen delivery by damaged blood vessels, leading to hypoxia, and increased inflammatory responses. Furthermore, the poor recovery capacity of the nerves themselves prevents spontaneous regeneration after SCI. In clinical practice, surgery or rehabilitation training have limited efficacy in patients with SCI and fail to effectively promote nerve regeneration and functional recovery. Due to the complex pathophysiology of SCI, existing single therapies have been unsatisfactory in the treatment of SCI. Currently, autologous nerve transplantation is considered the gold standard for treating neurological deficits, but limited donor nerve resources and functional deficits in the donor site significantly limit its clinical application. Therefore, there is a continuous and urgent clinical demand for drugs for the treatment of spinal cord injury that are safe, effective, inexpensive, and easy to administer.

[0004] Furthermore, the repair and restoration of peripheral nerve function, particularly long-distance defects, remain global challenges in clinical treatment. Autologous or allogeneic nerve transplantation is currently a commonly used clinical repair method for peripheral nerve injury, but it is plagued by limited donor resources, structural differences between donor and recipient nerves, loss of nerve function in the donor site, scarring, and immune rejection. Peripheral neuritis and peripheral nerve paresthesia are currently the most common peripheral nerve injuries in clinical practice. Peripheral neuritis, also known as peripheral neuritis or polyneuritis, is a distal limb nerve dysfunction caused by various etiologies. Peripheral neuritis can be caused by a variety of factors, including nutritional metabolism, drugs and poisoning, vasculitis, tumors, trauma, or mechanical compression. Its manifestations primarily include abnormal sensory, motor, and autonomic nervous system function within the innervation of the damaged nerve. Symptoms of peripheral neuritis include decreased sensation, pain, numbness or tingling, tingling, burning, and hyperesthesia. Peripheral neuritis can be multiple or single, symmetrical or asymmetrical, and is one of the most common diseases of the nervous system. When the cause of peripheral neuritis is known, treatment targeting the cause can alleviate symptoms and restore damaged nerves, but the effect is poor for peripheral neuritis with unknown causes or refractory primary diseases. Data show that about 40% of patients with peripheral neuritis will have sequelae, with a high disability rate, which seriously affects the patient's physical and mental health and quality of life. Currently, there are not many clinical treatments for peripheral neuritis, and they are mainly Western medicine. For example, in clinical practice, peripheral neuritis caused by various reasons can be treated with large doses of B vitamins, such as vitamin B1, B6, B 12 For patients with significant pain, analgesics and sedatives may be used. Inflammatory demyelinating lesions can be treated with adrenal corticosteroids, such as methylprednisone, dexamethasone, or hydrocortisone. However, long-term use of these medications often has significant toxic side effects, can easily lead to drug dependence, and rarely results in a complete cure.

[0005] The inventors found in previous studies that during the degradation of poly-L-lactic acid (PLLA), the molecular structure of PLLA is gradually destroyed and slowly hydrolyzed into lactic acid. Lactic acid can induce human fibroblasts to increase the production of collagen, resulting in an increase in collagen fibers in the dermis, resulting in a filling and repair effect. On this basis, the inventors have found that PLLA and lactic acid and its related lactate compounds have beneficial effects in repairing tissues such as cartilage, connective tissue, tendons, fascia, and nerves (see CN202210028046.9). The present invention is a continuation of previous work. The present invention has discovered new uses for divalent metal lactates in the preparation of products for preventing, improving, repairing, and treating nerve damage. Summary of the Invention

[0006] To overcome the shortcomings and deficiencies of the prior art, the present invention provides a novel use of divalent metal lactates. The inventors unexpectedly discovered during their research that divalent metal lactates can effectively prevent, improve, repair, and treat central and peripheral nerve damage. Based on this discovery, they conducted in-depth research and completed the present invention.

[0007] In order to achieve the purpose of the present invention, the present invention adopts the following technical solutions:

[0008] The present invention provides the use of divalent metal lactate in preparing products for preventing, improving, repairing and treating nerve damage, wherein the tissue in which the nerve damage occurs is selected from nerve tissue rich in collagen.

[0009] As an optional mode, in the above application, the nerve damage is central nerve damage or peripheral nerve damage, and the prevention, improvement, repair and treatment of nerve damage is nerve regeneration, increase in nerve number and / or nerve repair.

[0010] As an option, in the above application, the central nervous system injury includes spinal cord injury or encephalomyelitis, and the peripheral nerve injury includes peripheral nerve injury, peripheral neuritis or peripheral neuropathy.

[0011] As an optional mode, in the above application, the peripheral neuritis is selected from one or more of the following: facial neuritis, diabetic peripheral neuritis, viral peripheral neuritis, peripheral neuritis caused by chemotherapy drugs or Guillain-Barré syndrome.

[0012] As an option, in the above application, the symptoms of peripheral neuritis include decreased sensation, pain, numbness or tingling, tingling, burning, and hyperesthesia.

[0013] As an optional manner, in the above application, the divalent metal lactate includes one or more combinations of calcium lactate, magnesium lactate, zinc lactate, and ferrous lactate.

[0014] As an optional mode, in the above application, the divalent metal lactate is selected from magnesium lactate or a combination of zinc lactate and magnesium lactate.

[0015] Preferably, in the composition, the ratio of the magnesium lactate to the zinc lactate is 1:10-10:1 by weight.

[0016] More preferably, in the composition, the ratio of the magnesium lactate to the zinc lactate is 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1 or 10:1 by weight.

[0017] Most preferably, in the composition, the ratio of the magnesium lactate to the zinc lactate is 4:1, calculated by weight.

[0018] As an optional mode, in the above application, the product is selected from one or more of drugs, kits, and medical devices.

[0019] The concentration of the divalent metal lactate in the product is 10-80 mmol / L, more preferably, the concentration of the divalent metal lactate in the product is 20-60 mmol / L.

[0020] As an optional manner, in the above application, the medical device includes one or more combinations of medical tape, bandage, gauze, dressing, sponge, and medical suture.

[0021] As an optional mode, in the above application, in the medicine, the divalent metal lactate is the active ingredient, and the medicine further comprises a pharmaceutically acceptable carrier or excipient.

[0022] Preferably, in the medicament, the divalent metal lactate is the only active ingredient.

[0023] As an option, in the above application, the dosage form of the drug includes injection or external preparation.

[0024] Preferably, the external preparation comprises one or more combinations of ointments, creams, patches, sprays, solutions, and lotions.

[0025] Preferably, the administration of the product includes one or more of the following: intravenous injection, in situ injection, intramuscular injection, subcutaneous injection, oral administration or topical application.

[0026] In addition, those skilled in the art will appreciate that the mode of use of the preparation, as well as the dosage and volume of administration, are related to the age, physical condition and disease of the subject, and can be determined by a clinician based on the circumstances.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The present invention provides a new treatment for central and peripheral nerve damage that is safe, effective, inexpensive, and simple to administer. Specifically, the present invention has discovered the use of a divalent metal lactate comprising a combination of one or more of calcium lactate, magnesium lactate, zinc lactate, and ferrous lactate in the preparation of products for preventing, ameliorating, repairing, or treating nerve damage. Therefore, this product has broad application prospects in the treatment and repair of central nervous system damage caused by trauma, as well as peripheral neuritis or peripheral nerve paresthesia caused by viruses, diabetes, chemotherapy drugs, and other factors. DETAILED DESCRIPTION

[0029] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0030] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0031] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0032] In the following examples, all experiments were repeated three times, with at least three samples in each group. The experimental data were statistically analyzed using SPSS 21.0 software. Mean ± standard deviation (mean ± SD) was used for statistical description of measurement data, and analysis of variance was used for comparison of indicators between groups. P < 0.05 was considered statistically significant. When P < 0.05, a pairwise test was performed using the LSD-t test.

[0033] Example 1: Effects of divalent metal lactate on sciatic nerve injury model animals 1. Establishment of sciatic nerve injury model and experimental grouping

[0034] Forty 8-week-old Sprague-Dawley rats (male and female, 1:1 ratio) weighing 200-220 g were housed in a temperature-controlled room (22 ± 2°C). The animals were anesthetized with 3.0% (w / v) sodium pentobarbital (0.2 ml / 100 g) intraperitoneally. After complete anesthesia, the left buttocks were skin-prepared and the skin was disinfected with alcohol and iodine swabs. An oblique incision was then made, and the left sciatic nerve was exposed through blunt dissection in the gluteal intermuscular space. The sciatic nerve was clamped 0.5 cm from the inferior edge of the piriformis muscle three times (10 seconds each, with 10 seconds between each clamp). The muscle sutures adjacent to the clamp were marked with surgical sutures, and the muscle and skin were immediately sutured and disinfected again. After surgery, the animals were randomly divided into four groups, each containing 10 rats. The rats were housed individually and subsequently received intraperitoneal injections of drugs or saline daily for four weeks, depending on group. The experimental groups and treatments are shown in Table 1. The general condition of the rats, skin ulcers around the incision, and functional recovery of the limbs on the operated side were closely observed after surgery.

[0035] Table 1: Experimental groups and treatments

[0036]

[0037]

[0038] 2. Footprint Collection and Sciatic Nerve Index Scoring

[0039] A homemade rat gait recording chamber was constructed with a 90 cm long, 15 cm wide, and 20 cm high passage. A 15 cm wide continuous recording paper was placed at the bottom. At 1, 2, 3, and 4 weeks postoperatively, the soles of the rats' hind feet were painted with a colored, non-toxic dye and the rats were instructed to walk on the continuous recording paper, leaving bilateral footprints. Footprints from the experimental (E) and normal (N) lateral feet were selected, and the following variables were measured: ① print length (PL), the distance from the heel to the toe; ② toe spread (TS), the distance from the first toe to the fifth toe; and ③ intertoe distance (IT), the distance from the first toe to the fourth toe. The three variables were substituted into the following formula to calculate the sciatic nerve function index (SFI): an SFI = 0 indicates normal function, and an SFI = -100 indicates complete injury. The calculation formula is as follows: SFI = -38.3×(EPL-NPL) / NPL+109.5×(ETS-NTS) / NTS+13.3×(EIT-NIT) / NIT-8.8

[0040] At various time points after surgery (1, 2, 3, and 4 weeks), rat footprints were collected and the SFI was calculated to evaluate motor function and recovery in each group. The results are shown in Table 2. The results showed that at 2, 3, and 4 weeks after surgery, the SFI in the low-, medium-, and high-dose magnesium lactate groups were significantly superior to that in the saline-treated control group (Table 2, P < 0.01 for the medium-dose magnesium lactate group, P < 0.05 for the low- and high-dose magnesium lactate groups). This suggests that low-, medium-, and high-dose magnesium lactate have a certain therapeutic effect and can promote the recovery of sciatic nerve function. Furthermore, at 2, 3, and 4 weeks after surgery, the SFI in the medium-dose magnesium lactate group was significantly superior to that in the low- and medium-dose magnesium lactate groups (P < 0.05), indicating that medium-dose magnesium lactate has a better therapeutic effect on nerve injury and is more conducive to the recovery of nerve function.

[0041] Table 2: Sciatic nerve index of each group at different time points after surgery (n=10)

[0042]

[0043]

[0044] Note: a Refers to comparison with group A, P < 0.05; aa Compared with group A, P < 0.01; b Compared with group B and group D, P < 0.05.

[0045] 3. Real-time Quantitative PCR

[0046] 3.1 Extraction of total RNA

[0047] Remove the sciatic nerve, rinse it in DEPC-treated water, add 0.5 mL of Trizol, grind thoroughly, and centrifuge to remove the supernatant. Add 0.1 mL of chloroform, centrifuge to remove the supernatant. Add 0.8 mL of isopropanol, centrifuge to remove the supernatant. Add 1 mL of 75% ethanol, centrifuge to remove the supernatant. Add 20 μL of DEPC-treated water to dissolve the RNA. Determine the concentration and purity of the total RNA solution using a nucleic acid protein analyzer.

[0048] 3.2 Real-time quantitative PCR reaction

[0049] The expressions of Caspase-3 and GAP-43 in the sciatic nerve tissues of each group were detected by RT-qPCR.

[0050] Primer design

[0051] Caspase-3 upstream primer 5`-GAGCTTGGAACGGTACGATA-3`

[0052] Downstream primer 5`-CCGTACCAGAGCGAGATGAC-3`

[0053] GAP-43 upstream primer 5`-GTGTGTGAGCCTGTCCTCTC-3`

[0054] Downstream primer 5`-AAAACCGGGGTACAGTGCAA-3`

[0055] Caspase-3 plays a key role in apoptosis and is a potential drug target. Studies have shown that caspase-3 can severely inhibit DNA replication, transcription, and damage repair, ultimately leading to irreversible apoptosis. Therefore, reducing caspase-3 activation can effectively inhibit apoptosis. The results of real-time quantitative PCR for caspase-3 are shown in Table 3. The results showed no significant changes in caspase-3 mRNA expression in the saline control group at any time point. Compared with the saline control group at the same time point, caspase-3 mRNA expression in the low-, medium-, and high-dose magnesium lactate groups was significantly downregulated on days 1, 3, and 7 after modeling (P < 0.05). Furthermore, caspase-3 mRNA levels in the low-, medium-, and high-dose magnesium lactate groups gradually decreased over time within 7 days after modeling (P < 0.05). Compared with the low-dose group, caspase-3 mRNA expression in the medium- and high-dose magnesium lactate groups was significantly downregulated on days 1, 3, and 7 (P < 0.05). Compared with the high-dose group, the medium-dose magnesium lactate group showed significantly lower Caspase-3 mRNA expression at all time points (P < 0.05). Caspase-3 expression in rat neurons after sciatic nerve injury was significantly enhanced, and the number of apoptotic neurons also increased. Magnesium lactate injection can reduce Caspase-3 protein and mitigate apoptosis caused by nerve injury.

[0056] Table 3: Comparison of Caspase-3 mRNA expression at different time points after modeling and at different time points after surgery

[0057]

[0058] Note: Compared with group A, a P<0.05; compared with group B, b P<0.05; compared with group D, d P<0.05; compared with the first day of the same group on the third day, e P<0.05; compared with the 3rd day in the same group on the 7th day, f P<0.05.

[0059] GAP-43 is a membrane phosphoprotein widely present in neurons and a hallmark of neuronal regeneration. It plays a key role in neuronal regeneration by regulating the actin cytoskeleton. The results of real-time quantitative PCR for GAP-43 are shown in Table 4. The results showed no significant changes in GAP-43 mRNA expression in the saline control group. Compared with the same group at the same time, GAP-43 mRNA expression in the low-, medium-, and high-dose magnesium lactate groups was significantly increased on days 1, 3, and 7 after modeling (P < 0.05). Furthermore, within 7 days after modeling, GAP-43 mRNA levels in the low-, medium-, and high-dose magnesium lactate groups gradually increased over time, reaching a peak on day 7 (P < 0.05). Compared with the low-dose group, GAP-43 mRNA expression in the medium- and high-dose magnesium lactate groups was significantly increased on days 1, 3, and 7 (P < 0.05). Compared with the high-dose group, the expression of GAP-43 mRNA in the medium-dose magnesium lactate group was significantly increased at all time points (P < 0.05). Magnesium lactate can increase the expression of GAP-43, alleviate sciatic nerve injury in rats, and promote nerve regeneration.

[0060] Table 4: Comparison of GAP-43 mRNA expression at different time points after modeling and at different time points after surgery

[0061]

[0062] Note: Compared with group A, aP<0.05; compared with group B, b P<0.05; compared with group D, d P<0.05; compared with the first day of the same group on the third day, eP<0.05; compared with the third day of the same group on the seventh day, f P<0.05.

[0063] Example 2: Effects of divalent metal lactates on spinal cord injury model animals

[0064] 1. Establishment of Rat Spinal Cord Injury Model

[0065] Forty 8-week-old Sprague-Dawley rats (male and female, 1:1 ratio) weighing 200-220 g were housed in a temperature-controlled room (22 ± 2°C). The animals were anesthetized with 3.0% (w / v) sodium pentobarbital (0.2 mL / 100 g) intraperitoneally. After complete anesthesia, the animals were placed in a prone position and their skin was prepared and disinfected routinely. A longitudinal incision (2-3 cm) was made along the dorsal midline, starting at the twelfth thoracic vertebra (T12). This incision was then made from the outside inward until the subcutaneous fascia was reached. The dorsal lamina of T9-11 was precisely excised to expose the dura mater of the spinal cord. An aneurysm clip was applied to the exposed midsection of the spinal cord for 40 seconds to create a spinal cord injury (SCI) model. During the experimental period, the bladders of the SCI rats were squeezed daily to facilitate urination, both in the morning and evening. After urination, the perineum was cleaned and kept dry. Extremities soaked in urine were promptly washed and air-dried, and body positions were changed frequently. In addition, to prevent infection, gentamicin (2000 U / kg / day) was routinely injected intramuscularly 3 days after surgery. After surgery, the animals were randomly divided into four groups, each containing 10 rats. Rats were housed in separate cages and subsequently received intraperitoneal injections of drugs or saline once daily for four weeks, depending on group assignment. The experimental groups and treatments are shown in Table 5.

[0066] Table 5: Experimental groups and treatments

[0067]

[0068] 2. Motor system score (BBB score)

[0069] The BBB scoring method, developed by American researchers Basso et al., was used to assess limb function in rats after spinal cord injury. The method involved placing the animals in an open basin and gently tapping the basin wall to induce crawling. The animals' hip, knee, and ankle joints were observed for walking, driving, and coordination. The results are listed in Table 6.

[0070] As shown in Table 6, all rats in each group showed varying degrees of functional recovery after magnesium lactate treatment, with the medium-dose magnesium lactate group recovering the fastest. One day after surgery, there was no statistically significant difference in the BBB scores among the rats (P>0.05). The BBB scores of the groups gradually increased over time, and the differences between the groups gradually increased. By four weeks after surgery, the medium-dose magnesium lactate group had the highest score, followed by the high-dose magnesium lactate group, then the low-dose magnesium lactate group, and the saline-treated control group had the lowest score. The difference between the low- and high-dose magnesium lactate groups was not statistically significant (P=0.213), but the differences among the remaining groups were statistically significant (P<0.05).

[0071] Table 6: BBB scores before and after surgery in each group

[0072]

[0073] Note: a Refers to comparison with group A, P < 0.05; b Compared with group B, P < 0.05; c Compared with group C, P < 0.05; d Compared with group D, P < 0.05.

[0074] 3. Real-time Quantitative PCR

[0075] The injured spinal cord was cut into 0.5 cm sections centered on the injured area, and RT-PCR was performed to measure the gene expressions of brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), and neuron-specific enolase (NSE).

[0076] 3.1 Extraction of total RNA

[0077] Add 1 mL to a homogenizer tube. Place 100 mg of tissue in a homogenizer tube, add RNA extraction solution, grind thoroughly, and centrifuge to remove the supernatant. Add 250 μL of chloroform, centrifuge, and remove the supernatant. Add 0.8 mL of isopropanol, centrifuge, and discard the supernatant. Add 1 mL of 75% ethanol, centrifuge, and discard the supernatant. Add 15 μL of RNase-free water to dissolve the RNA. Determine the concentration and purity of the total RNA solution using a nucleic acid protein analyzer.

[0078] 3.2 Reverse transcription reaction

[0079] 3.3 Real-time quantitative PCR reaction

[0080] Primer design

[0081] BDNF upstream primer 5`-GTCAAGTGCCTTTGGAGCCT-3`

[0082] Downstream primer 5`-CATGGGATTGCACTTGGTCTC-3` GFAP upstream primer 5`-AGTCGGCGAGTTACCAGGAG-3`

[0083] Downstream primer 5`-TTAATGACCTCGCCATCCCG-3`NSE upstream primer 5`-TATCCTGGAGAACAGCGAAGC-3`

[0084] Downstream primer 5`-GACAAAGTCCTGGTAGAGTGCCC-3`

[0085] BDNF is the most widely distributed and abundantly expressed neurotrophic factor in the mammalian brain. It plays a crucial role in the normal growth, development, and plasticity of synapses and can inhibit apoptosis by activating downstream pathways. Table 7 shows that one week after surgery, BDNF expression in the saline-treated control group was statistically significantly different among the low-, medium-, and high-dose magnesium lactate groups (P < 0.05). There was no statistically significant difference in BDNF expression between the low- and high-dose magnesium lactate groups (P = 0.628). Four weeks after surgery, BDNF expression was statistically significant among all groups, with the exception of the saline-treated control group and the low-dose magnesium lactate group (P = 0.820). GFAP and NSE expression also showed statistically significant differences among the four groups at four weeks after surgery (P < 0.05). As an important neurotransmitter, BDNF plays a crucial role in regulating neuronal growth, differentiation, and synaptic plasticity by binding to specific receptors and activating downstream pathways. As can be seen from Table 7, divalent metal lactate can increase the expression of BDNF, and then reduce the apoptosis of nerve cells after spinal cord injury by activating the downstream anti-apoptotic pathway, and participate in the neural remodeling process.

[0086] Table 7: Relative expression of BDNF in the spinal cord of rats 1 week and 4 weeks after surgery

[0087]

[0088] Note: a refers to comparison with group A, P < 0.05; b Compared with group B, P < 0.05; d Compared with group D, P < 0.05.

[0089] GFAP expression is an important indicator of spinal cord neuronal growth, proliferation, and differentiation and is considered a therapeutic target for acute spinal cord injury. Table 8 shows that one week after surgery, the differences in GFAP expression among the low, medium, and high-dose magnesium lactate groups in the saline-treated control group were statistically significant (P < 0.05). GFAP expression was also statistically significant among the four groups at one week after surgery (P < 0.05). At four weeks after surgery, the differences in GFAP expression among the four groups were also statistically significant (P < 0.05). GFAP is a marker protein specific for astrocytes, which are the most numerous cells in the central nervous system and therefore have higher expression levels after surgery. Compared with the other groups, GFAP expression was significantly reduced in the medium-dose divalent lactate group, indicating that it inhibits astrocyte proliferation and, consequently, glial scar formation, a process that promotes axonal regeneration and neural remodeling.

[0090] Table 8: Relative expression of GFAP in the spinal cord of rats 1 week and 4 weeks after surgery

[0091]

[0092] Note: a Refers to comparison with group A, P < 0.05; b Compared with group B, P < 0.05; d Compared with group D, P < 0.05.

[0093] NSE is a key enzyme in neuronal respiration and is widely present in brain tissue and other peripheral nerve cells. NSE participates in glycolysis and is released in response to damage such as hypoxia, ischemia, and poisoning. It enters the bloodstream as the blood-brain barrier is disrupted. Table 9 shows that one week after surgery, NSE expression in the saline-treated control group was statistically significantly different among the low, medium, and high-dose magnesium lactate groups (P < 0.05). NSE expression in the low, medium, and high-dose magnesium lactate groups was not statistically significant one week after surgery (P > 0.05). Four weeks after surgery, the differences in NSE expression among the four groups were statistically significant (P < 0.05). Both the medium and high-dose divalent lactate groups increased NSE expression, with the medium-dose group showing a more significant effect. Because NSE participates in catalytic reactions during cellular glucose metabolism, increased NSE expression may improve cellular energy metabolism and promote the recovery of damaged cells.

[0094] Table 9: Relative expression of NSE in the spinal cord of rats 1 week and 4 weeks after surgery

[0095]

[0096] Note: a Refers to comparison with group A, P < 0.05; b Compared with group B, P < 0.05; d Compared with group D, P < 0.05.

[0097] Example 3: Effects of divalent metal lactates on autoimmune neuritis model animals

[0098] 1. Establishment of Autoimmune Neuroinflammation Model and Experimental Grouping

[0099] SD rats (male and female, 1:1 ratio), 8 weeks old, weighing 200-220 g, a total of 40, were placed in a temperature-controlled room (22 ± 2 ° C). 180-199 A mixture of 1 mg H37Ra, 100 μL of saline, and 100 μL of IFA was thoroughly emulsified and injected subcutaneously into the soles of both hind limbs (100 μL per sole) as the dosage for one rat. After successful modeling, rats in each group were gavaged with the corresponding drug or saline once daily for four consecutive weeks.

[0100] Table 10: Experimental groups and treatments

[0101]

[0102] 2. Behavioral Scoring

[0103] During the administration period, the rats were observed and behavioral scores were performed daily. 0 points indicated no clinical symptoms; 5 points indicated quadriplegia or death.

[0104] On the first day after oral administration of normal saline, some rats in the autoimmune neuritis group began to show varying degrees of neurological impairment, including lethargy, matted fur, and redness and swelling of both hind paws. By the fifth day, all rats showed varying degrees of autoimmune neuritis, including loss of muscle tone, tail dragging, and paralysis. On the eighth day, autoimmune neuritis symptoms peaked, with the highest neurological deficit score of 4.0±0.5. In the low-dose magnesium lactate group, some rats showed varying degrees of neurological impairment on the sixth day of administration. On the eleventh day, all rats showed varying degrees of autoimmune neuritis. On the fifteenth day, autoimmune neuritis symptoms peaked, with the highest neurological deficit score of 3.0±0.5. Subsequently, autoimmune neuritis symptoms gradually subsided, and by the 28th day, the neurological deficit score was between 0 and 1. On day 10 of administration, some rats in the medium-dose magnesium lactate group developed varying degrees of neurological impairment. On day 16, all rats developed varying degrees of autoimmune neuritis. On day 21, autoimmune neuritis symptoms peaked, with the highest neurological deficit score of 2.0±0.5. Subsequently, autoimmune neuritis symptoms gradually resolved, and by day 25, the neurological deficit score was 0. In the high-dose magnesium lactate group, on day 8 of administration, some rats developed varying degrees of neurological impairment. On day 12, all rats developed varying degrees of autoimmune neuritis. On day 16, autoimmune neuritis symptoms peaked, with the highest neurological deficit score of 2.5±0.5. Subsequently, autoimmune neuritis symptoms gradually resolved, and by day 33, the neurological deficit score was between 0 and 1. These results suggest that magnesium lactate has a therapeutic effect on neurological deficits caused by autoimmune neuritis, with the medium-dose magnesium lactate being the most effective.

[0105] 3. Detection of inflammatory factor levels

[0106] Existing theories believe that inflammatory factors are one of the factors that aggravate the symptoms of autoimmune neuritis. IL-17, a characteristic cytokine secreted by Th17 cells, participates in the occurrence of autoimmune diseases by inducing an inflammatory cascade reaction in target organs. Blood was collected from the retroorbital venous plexus of rats and the supernatant was obtained by centrifugation. The concentrations of IL-17 and IFN-γ in the serum were determined by microplate reader, and the results are listed in Table 11. Compared with the autoimmune neuritis group gavaged with normal saline, the levels of IL-17 and IFN-γ in the serum of rats gavaged with magnesium lactate were significantly reduced (P < 0.05). Compared with the low- and high-dose magnesium lactate groups, the levels of IL-17 and IFN-γ in the medium-dose magnesium lactate group were significantly reduced (P < 0.05). This shows that magnesium lactate can reduce the secretion of inflammatory cytokines caused by autoimmune neuritis, among which the effect of medium-dose magnesium lactate is the most significant.

[0107] Table 11: Comparison of IL-17 and IFN-γ levels in serum of rats in each group

[0108]

[0109] Note: a Refers to comparison with group A, P < 0.05; b Compared with group B, P < 0.05; d Compared with group D, P < 0.05.

[0110] In the above application, the results of other divalent metal lactates including calcium lactate, zinc lactate, ferrous lactate and various combinations are similar to those of the above magnesium lactate, and will not be repeated here.

[0111] Example 4: Effects of divalent metal lactates on autoimmune encephalomyelitis model animals

[0112] 1. Establishment of Autoimmune Encephalomyelitis Model and Experimental Grouping

[0113] 40 SD rats (male and female, 1:1 ratio), 8 weeks old, weighing 200-220 g, were placed in a temperature-controlled room (22 ± 2°C). 35-55 The peptide was diluted to 6 mg / mL in PBS and mixed with an equal volume of CFA. A 5 mL syringe was connected using a dual-channel connector and repeatedly pushed and pulled to mix to prepare MOG. 35-55 Oil-in-water antigen emulsion. Experimental rats were immunized with antigens and the antigen emulsion was injected subcutaneously at multiple points on the back, 4 points per rat, for a total of 0.2 mL of antigen emulsion (MOG 35-55 Peptide content was 600 μg per rat). PTX was administered intraperitoneally at 0 and 48 hours after antigen emulsion immunization to increase the incidence of the model. After successful modeling, rats in each group were gavaged with the corresponding drug or saline once daily for 4 weeks.

[0114] Table 12: Experimental groups and treatments

[0115]

[0116] 2. Neurological impairment score

[0117] During the administration period, the rats were observed daily and neurological deficits were scored: 0, no clinical symptoms; 5, moribund or dead.

[0118] On the fifth day after oral administration of normal saline, some rats in the autoimmune encephalomyelitis group began to show varying degrees of neurological impairment. By the ninth day, all rats showed varying degrees of autoimmune encephalomyelitis symptoms, including loss of muscle tone, tail dragging, and paralysis. On the 12th day, autoimmune encephalomyelitis symptoms peaked, with the highest neurological deficit score of 3.5±0.5. In the low-dose magnesium lactate group, on the seventh day, some rats showed varying degrees of neurological impairment. On the 12th day, all rats showed varying degrees of autoimmune encephalomyelitis symptoms. On the 16th day, autoimmune encephalomyelitis symptoms peaked, with the highest neurological deficit score of 2.5±0.5. Subsequently, the autoimmune encephalomyelitis symptoms gradually subsided, and by the 28th day, the neurological deficit score was 0-1. On day 12 of administration, some rats in the medium-dose magnesium lactate group developed varying degrees of neurological impairment. On day 18, all rats developed varying degrees of autoimmune encephalomyelitis symptoms. On day 23, autoimmune encephalomyelitis symptoms peaked, with the highest neurological impairment score of 1.5±0.5. Subsequently, autoimmune encephalomyelitis symptoms gradually subsided, and by day 26, the neurological impairment score was 0. In the high-dose magnesium lactate group, on day 10 of administration, some rats developed varying degrees of neurological impairment. On day 15, all rats developed varying degrees of autoimmune encephalomyelitis symptoms. On day 19, autoimmune encephalomyelitis symptoms peaked, with the highest neurological impairment score of 2.5±0.5. Subsequently, autoimmune encephalomyelitis symptoms gradually subsided, and by day 31, the neurological impairment score was between 0 and 1. Magnesium lactate plays a certain therapeutic role in the neurological deficit caused by autoimmune encephalomyelitis, among which the effect of medium-dose divalent lactate is the most significant.

[0119] 3. Detection of inflammatory factor levels

[0120] After the rats were sacrificed, spinal cord tissue was obtained and homogenized with tissue lysis buffer. The supernatant was centrifuged and the concentrations of TNF-α, IL-6, and IL-β were measured. The results are listed in Table 13. Compared with the autoimmune encephalomyelitis group gavaged with normal saline, the levels of TNF-α, IL-6, and IL-β in the spinal cord of rats gavaged with magnesium lactate were significantly reduced (P < 0.05). Compared with the low- and high-dose magnesium lactate groups, the TNF-α, IL-6, and IL-β levels in the medium-dose magnesium lactate group were significantly reduced (P < 0.05). This indicates that magnesium lactate can reduce the secretion of inflammatory cytokines caused by autoimmune encephalomyelitis, thereby reducing the damage to myelin tissue, among which the effect of medium-dose magnesium lactate was more significant.

[0121] Table 13: Comparison of TNF-α, IL-6, and IL-β levels in the spinal cord of rats in each group

[0122]

[0123] Note: a Refers to comparison with group A, P < 0.05; b Compared with group B, P < 0.05; d Compared with group D, P < 0.05.

[0124] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. Use of divalent metal lactate in the preparation of products for preventing, improving, repairing and treating nerve damage, characterized by: The tissue in which nerve damage occurs is selected from nerve tissue rich in collagen, the nerve damage is central nervous system damage or peripheral nervous system damage, the central nervous system damage is spinal cord damage or encephalomyelitis, the peripheral nervous system damage is peripheral nervous system damage, peripheral neuritis or peripheral neuropathy, and the divalent metal lactate is selected from a combination of one or more of calcium lactate, magnesium lactate, zinc lactate and ferrous lactate.

2. The use according to claim 1, characterized in that: The prevention, improvement, repair and treatment of nerve damage are nerve regeneration, increase in nerve number and / or nerve repair.

3. The use according to claim 1, characterized in that: The peripheral neuritis is selected from one or more of the following: facial neuritis, diabetic peripheral neuritis, viral peripheral neuritis, peripheral neuritis caused by chemotherapy drugs or Guillain-Barré syndrome, and the symptoms of the peripheral neuritis include hypoesthesia, pain, numbness or tingling, tingling, burning, and hyperesthesia.

4. The use according to claim 1, characterized in that: The divalent metal lactate is selected from magnesium lactate or a combination of zinc lactate and magnesium lactate.

5. The use according to claim 4, characterized in that: In parts by weight, in the composition, the ratio of the zinc lactate to the magnesium lactate is 1:10-10:

1.

6. The use according to claim 1, characterized in that The product is selected from one or more of a medicine, a test kit, and a medical device.

7. The use according to claim 1, characterized in that: The concentration of the divalent metal lactate in the product is 5-250 mmol / L.

8. The use according to claim 7, characterized in that: The concentration of the divalent metal lactate in the product is 30-150 mmol / L.

9. The use according to claim 6, characterized in that: The medical device includes one or more combinations of medical adhesive tape, bandage, gauze, dressing, sponge, and medical suture.

10. The use according to claim 6, characterized in that: In the medicine, divalent metal lactate is an active ingredient, and the medicine further comprises a pharmaceutically acceptable carrier or excipient.

11. The use according to claim 6, characterized in that: The dosage form of the drug includes injection or external preparation.

12. The use according to claim 11, characterized in that: The external preparation includes one or more combinations of ointments, creams, patches, sprays, solutions, and lotions.

13. The use according to claim 6, characterized in that: The administration of the product includes one or more of the following: intravenous injection, in situ injection, intramuscular injection, subcutaneous injection, oral administration or topical application.

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