CN113425733A

CN113425733A - Medicine for treating acute myelitis and application thereof

Info

Publication number: CN113425733A
Application number: CN202110808768.1A
Authority: CN
Inventors: 陈晓莉; 张国平
Original assignee: Shaanxi Provincial Peoples Hospital
Current assignee: Shaanxi Provincial Peoples Hospital
Priority date: 2021-07-16
Filing date: 2021-07-16
Publication date: 2021-09-24

Abstract

The invention relates to a medicament for treating acute myelitis and application thereof, wherein the medicament composition takes ganglioside, vitamin B6 and vitamin C as active ingredients, and can provide nutrition support for patients and enhance organism immunity through scientific compatibility. Compared with the existing medicines for treating spinal cord injury diseases, the medicine has the advantages of definite curative effect, quick response, small side effect and the like.

Description

Medicine for treating acute myelitis and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a medicine for treating acute myelitis and application thereof.

Background

Acute myelitis (acutemyelitis) is a spinal disease which is possibly caused by congestion, edema and demyelination of nerve fibers caused by viral and bacterial infection, vaccine injection and other factors and mainly shows paralysis, sensory disorder and autonomic dysfunction below the level of acute onset myelopathy. The disease mostly occurs in young and strong years, and has no sex difference. The symptoms of upper respiratory tract infection are common days or 12 weeks before the disease, and cold catching, overstrain, trauma and the like are frequently the causes of the disease. In modern clinic, glucocorticoid is mainly used for treating the disease, and a better effect can be achieved by matching with gamma globulin intravenous drip. But the recovery of the nerve function of the patient is not ideal, and the related nerve function sequelae are easy to leave.

In the 60's of the 20 th century, scientists first demonstrated that the central nervous system of mammals has some characteristic of endogenous repair, with neuronal axons and major branches of the adult brain and spinal cord repaired after some degree of injury, and that the phenomenon of new neuronal birth in the adult avian, non-primate and human brain is called neural repair. The neurorestorative is an agent which promotes neuron repair by directly acting on neural stem cells in the brain of a human or an animal and has an effect of increasing neurons in the brain, and the neurorestorative can be used as a therapeutic agent for a neural disease caused by neurodegeneration or accompanying injury.

To date, there are many drugs for the treatment of spinal cord injury, but only methylprednisolone is used in large quantities with its effect of treating spinal cord injury being confirmed. Many other medicines are still in the test stage, based on the deepening of the basic research on the mechanism of spinal cord injury, new medicines or old medicines are continuously proved to be effective in treating experimental spinal cord injury, but due to various reasons, clinical verification is difficult to enter, so that a plurality of new medicines are difficult to widely apply to clinic, most medicines only act on a certain link of spinal cord injury, and the spinal cord injury relates to various injury links and mechanisms, so that the effects of the new medicines are limited, the characteristic treatment window period after the spinal cord injury also influences the medicine development, at present, the treatment of the spinal cord injury tends to combine the application of various medicines, and the curative effect is better than that of a single medicine method. Among studies of factors that promote nerve repair, the first nerve cell regulatory factor reported to elucidate its molecular structure exclusively was Nerve Growth Factor (NGF). Meanwhile, ganglioside and basic fibroblast growth factor (bFGF) are also intensively researched, and are respectively proved to have nerve growth promoting effects of different degrees. At present, the research and application of single factors are mainly used, but the factors have very limited effect on nerve repair, uncertain curative effect and high price, and are difficult to popularize and apply clinically. The progress of clinical research and application of nerve growth factors, substitutes and inducers of neural tube trophic factors and the like is slow at present abroad, and no reliable medicine for promoting nerve repair exists so far. Therefore, how to promote the repair of the damaged nerve and restore the function thereof has become an important research point.

Acute myelitis (acutemellitis) refers to a non-specific inflammatory lesion of the spinal cord, which is mostly followed by infection, often involving the gray substance of several segments of the cord and the surrounding meninges, and producing symptoms of transverse spinal cord injury as the thoracic cord is most vulnerable to invasion. The clinical manifestations are characterized by paralysis of the extremities below the level of the lesion, sensory disturbance of conduction tracts and dysuria. Early treatment, careful care and early rehabilitation training of acute myelitis are very important for prognosis. The location of the affected spinal segment is one of the main factors affecting the therapeutic effect. The spinal cord segments gradually develop upwards from the lower segments, and the treatment effect is poor. The current treatments mainly comprise: the first is immunosuppressive therapy, mainly corticosteroid hormones, immunoglobulins, and the like; the second is symptomatic treatment, mainly consisting of nerve nourishment, anti-infection and supportive therapy. However, the existing treatment method has great side effect and insignificant curative effect, thereby limiting the application of the treatment method.

Clinical observation of methylprednisolone combined with ganglioside for treating acute myelitis (Liyu et al, J. China's. Utility neurological diseases) discloses that the methylprednisolone treatment is combined with ganglioside (GM-1) to treat acute myelitis, and a better recovery level can be achieved.

The method and the experimental result show that the clinical treatment effect is obvious, the muscle strength and the urination condition are obviously improved, and the method is worthy of wide popularization and application.

Chinese granted patent CN102772407B discloses a pharmaceutical composition for promoting nerve injury repair and application thereof. Each unit of the pharmaceutical composition contains 0.5-8 g of L-ornithine, 1-5 g of aspartic acid, 3-10 g of arginine, 63-10 g of vitamin B, and the balance of auxiliary materials and/or other components. The pharmaceutical composition can remarkably promote the repair of spinal nerve function, can be used for preparing medicines for repairing nerve injury, and especially has certain curative effect on acute myelitis.

However, the currently used hormones (such as methylprednisolone) have serious side effects on human bodies, but the hormone-free drugs have certain difficulty in rapidly relieving acute myelitis. Therefore, the present invention aims to overcome the above-mentioned drawbacks of the prior art and provide a drug for treating acute myelitis and its use, which can rapidly relieve the symptoms of acute myelitis without using hormones.

Disclosure of Invention

Based on the background technology, the technical problem to be solved by the invention is to provide a medicine for treating acute myelitis and application thereof. In order to realize the purpose of the invention, the following technical scheme is adopted:

one aspect of the present invention relates to a medicament for the treatment of acute myelitis, which comprises a combination of ganglioside, vitamin B6 and vitamin C as active ingredients. The invention does not use hormone and other active ingredients, and can provide nutrition support for patients and enhance the immunity of organisms through scientific compatibility. Compared with the existing medicines for treating spinal cord injury diseases, the medicine has the advantages of definite curative effect, quick response, small side effect and the like.

In a preferred embodiment of the present invention, the medicament is an injectable formulation.

In another preferred embodiment of the present invention, the injection further comprises glucose.

In a preferred embodiment of the invention, the weight ratio of ganglioside, vitamin B6 and vitamin C is 5-10: 1-2: 1-2.

In a preferred embodiment of the invention, the ratio of ganglioside, vitamin B6 and vitamin C is 5-6: 0.8-1.2: 0.8-1.2; particularly preferably 5: 1: 1. this ratio contributes to a rapid onset of action of the drug.

The invention also relates to application of the medicine in preparation of medicines for treating acute myelitis.

In a preferred embodiment of the invention, said use comprises the restoration of motor capacity in patients with acute myelitis.

Advantageous effects

The pharmaceutical composition of the invention takes ganglioside, vitamin B6 and vitamin C as raw materials, and can provide nutrition support for patients and enhance the immunity of organisms through scientific compatibility. Compared with the existing medicines for treating spinal cord injury diseases, the medicine has the advantages of definite curative effect, quick response, small side effect and the like.

Detailed Description

In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Unless otherwise stated, all reagents involved in the embodiments of the present invention are commercially available products, and all the reagents can be purchased from commercial sources, and the molding and pharmaceutical experiments of the present invention are performed with reference to the chinese granted patent CN 102772407B.

Example 1:

(1) experimental drugs:

experimental group 1: adding 5g ganglioside, 1g vitamin B6 and 1g vitamin C into 100ml of 5% glucose solution to prepare injection;

experimental group 2: adding 5g ganglioside, 2g vitamin B6 and 1g vitamin C into 100ml of 5% glucose solution to prepare injection;

positive control group 1: adding 7g ganglioside into 100ml of 5% glucose solution to prepare injection;

positive control group 2: adding 5g ganglioside, 1g vitamin B1 and 1g vitamin C into 100ml of 5% glucose solution to prepare injection;

positive control group 3: adding 5g ganglioside, 1g vitamin B12 and 1g vitamin C into 100ml of 5% glucose solution to prepare injection;

negative control group: 5% glucose solution.

(2) The molding and administration method comprises the following steps:

60 clean-grade standard adult healthy Wistar rats with unlimited male and female parts, weight of 250-300 g, 1% sodium pentobarbital (50mg/kg) are used for intraperitoneal injection anesthesia, prone position fixation is performed, T9-T10 vertebral lamina spinous processes are exposed under the aseptic condition, the vertebral laminae are cut off, the dura mater is exposed, and a weight falling method (10g multiplied by 4cm) is used for manufacturing a spinal cord injury model. The successful making standard of the spinal cord injury model is spinal cord tissue edema, subdural congestion and hemorrhage after the strike, and the score of BBB (Basso-Beattie-Bresnahan) is less than or equal to 1 point after 24h of behavioral examination. Timely hemostasis and suture after operation, strengthening warm keeping, anti-infection and other nursing, massaging the bladder twice every day, and disinfecting the perineum part by iodophor until the bladder function is recovered. 60 successfully molded rats were randomly divided into 6 groups: the injection is administered intraperitoneally at a dose of 2mL/kg 1 time daily for 4 weeks.

(3) Inclined plate test:

the rats were placed on the modified Rivlin inclined plate with their heads facing left, and the final results were obtained by repeating 3 times with the maximum angle at which the animals could stay on the plate for 5 seconds without falling as the scoring criterion, with increasing angles from the horizontal position. Groups 6 were performed at 1d, 2d, 3d, 1w, 2w, 3w and 4w post-surgery, respectively.

(4) BBB motor score:

the method comprises the following steps of scoring the motor functions of 6 groups of rats by a double-blind method, placing animals on a flat, non-slip and light-sufficient table top to observe the movement conditions of each joint of hind limbs, scoring the motor functions of the animals, scoring the rats once before injury, scoring 1d, 2d, 3d, 1w, 2w, 3w and 4w respectively after model building, detecting for 3 times in total, taking the mean value, scoring mainly embodying the recovery of the motor functions, scoring the highest score (21 score) of normal rats, and scoring the most seriously injured rats by 0 score.

(5) Tissue material taking and section preparation:

after 3 time points (1, 48 and 72 hours) after operation, animals are anesthetized, about 200mL of heparin normal saline is perfused into the left ventricle until clear liquid flows out, then about 200mL of 4% paraformaldehyde is used for lavage and fixation, spinal cord tissue with the length of about 8mm taking injury as the center is taken, the spinal cord tissue is fixed by 4% paraformaldehyde for 24 hours, the spinal cord tissue is embedded by conventional paraffin, and is continuously sliced, wherein each sample slice is 25, the slice thickness is 5 mu m, and the spinal cord tissue is stained by conventional hematoxylin-eosin.

(6) Immunohistochemical staining:

NF expression was detected by Super Picture immunohistochemical staining. The NF-antibody working concentration is 1:400, and the instant generation 2 immunohistochemical broad spectrum KIT SuperPicTure (Mouse/Rabbit KIT) is dispensed by Zymed. DAB color development kit (DAB-0031). The staining result was observed under an optical microscope, and the immunopositive cells were colored in a tan color or a tan color. Selecting 10 different fields of vision for each section under a 400-time optical microscope, adopting a pathological image analyzer and Leica Qwin image analysis software, determining the same standard for the sections dyed in the same batch according to the gray level of dyeing, and calculating the average gray level value of the neurofilament protein (NF) according to the background color of the dyeing. The grey scale value can indirectly reflect the expression activity of the cell protein, the larger the grey scale value is, the lower the expression activity is, and the smaller the grey scale value is, the higher the expression activity is.

(7) Histological observation of spinal cord in each group of rats

After 72 hours of molding, pathological examination results show that the rat spinal cord tissue structure of the negative control group is disordered, more flaky bleeding and capsule cavities formed after hemorrhagic necrosis exist, the nerve cell swelling is obvious, a large number of inflammatory cells are gathered, some nerve cells cannot be identified, the gray white boundary is unclear, the central tube part disappears or deviates, and the edge of the spinal cord is adhered. In the positive control groups 1-3, the tissue structure of the spinal cord of the rat is fuzzy, a small amount of flaky bleeding and a capsule cavity formed after hemorrhagic necrosis exist, the swelling of nerve cells is obvious, a certain amount of inflammatory cells are gathered, a small amount of nerve cells cannot be identified, the grey white boundary is unclear, the central tube part disappears or deviates, and the edge of the spinal cord is adhered. The rat spinal cord tissue structures of the experimental group 1 and the experimental group 2 are generally clear, the bleeding and necrosis degrees are light, the formed capsule cavities are few, the nerve cell swelling is not obvious, the inflammatory cell infiltration quantity is small, the nerve cells are rich, the white matter area is edematous, and the edge of the spinal cord cortex area is unclear.

(8) Expression of NF

NF expression of spinal cord tissue neurons progressively increases with time after spinal cord injury, with motor neurons appearing first at the anterior horn of the spinal cord and later at the posterior horn of the spinal cord. Morphologically, both the soma and the nucleus of neurons are enlarged. The NF expression of the drug treatment groups 1 and 2 can be increased after 1h of injury, the high expression can still be seen after 96h, the NF expression water average of the drug composition treatment group at each time point is higher than that of a normal saline group, the NF gray value of the drug treatment groups 1 and 2 is compared with that of a negative control group and that of a positive control group 1-3, the difference is significant (P is less than 0.05), and the experimental result is shown in table 1.

TABLE 1 values of NF-staining positive cells in spinal cord surrounding injury zone at different times

(9) Behavioral changes in rats in each group

The BBB scores of the rats in the 6 groups before modeling are 21 scores, after modeling, the rats show double lower limb complete paralysis, all the rats show urine retention, spinal shock is expressed, the scores are lower in the first 3 days, but the spinal cord functions of the rats are gradually recovered along with the time extension, the muscle strength of hind limbs is continuously enhanced, the rats in the 6 groups 7 days after injury are all increased, and the drug treatment groups 1 and 2 have significant differences (P < 0.05) compared with a negative control group and a positive control group 1-3. The postoperative inclined plate test result shows that the maximum angle of the inclined plate is increased in 6 groups from 3 days after injury, and the drug treatment groups 1 and 2 have significant difference (P < 0.05) compared with the negative control group and the positive control group 1-3 (see table 2 and table 3). The results show that the injection of the drug treatment groups 1 and 2 can obviously promote the recovery of spinal nerve function after injury, and particularly, the injection of the drug treatment group 1 has more obvious treatment effect and more rapid effect.

Table 2 oblique plate test results of different time periods after each group of rats was injured

TABLE 3 BBB scoring results after injury in rats of each group

The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations of the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.

Claims

1. A medicine for treating acute myelitis contains combination of ganglioside, vitamin B6 and vitamin C as active ingredients.

2. The medicament of claim 1, which is an injectable formulation.

3. The medicament of claim 2, the injection further comprising glucose.

4. The medicament of claim 2, wherein the weight ratio of ganglioside, vitamin B6 and vitamin C is 5-10: 1-2: 1-2.

5. The medicament of claim 2, wherein the ratio of ganglioside, vitamin B6 and vitamin C is 5-6: 0.8-1.2: 0.8-1.2; particularly preferably 5: 1: 1.

6. use of a medicament according to any one of claims 1 to 5 for the manufacture of a medicament for the treatment of acute myelitis.

7. The use of claim 6, comprising restoring exercise capacity in patients with acute myelitis.