Pharmaceutical composition for preventing and treating diabetic peripheral neuropathy and application thereof

Abstract

The invention relates to an innovative medicine composition of a medicine for treating diabetic peripheral neuropathy (DPN), and belongs to the technical field of medicines. The composition takes 6-gingerol and polygonatum polysaccharide as key active ingredients, and shows the advantage of synergistic interaction clinically through unique ingredient compatibility. Researches find that when the two components are combined for use, the two components show a synergistic treatment effect superior to that of a single component in the aspects of reducing blood sugar, improving nerve conduction speed and relieving core symptoms such as sensory disorders of both lower limbs. Although statistical significant difference is not achieved among administration groups, the composition is superior to single-drug treatment in the aspects of overall symptom improvement rate, onset speed and safety. According to the invention, the limitation of traditional single-component treatment is broken through, and a DPN treatment scheme with more comprehensive curative effect and safety is provided through optimal combination of 6-gingerol and polygonatum sibiricum polysaccharide. The composition not only can intervene the pathological process of DPN in a multi-target manner, but also provides an important thought for developing a novel synergistic treatment drug.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and particularly relates to a pharmaceutical composition for preventing and treating diabetic peripheral neuropathy and its application. Background Art

[0002] Diabetic peripheral neuropathy (DPN), as one of the most harmful microvascular complications in the course of diabetes, has significant complexity and progression characteristics in its pathological evolution process. Globally, this disease is not only the primary risk factor leading to foot ulcers and amputations in diabetic patients, but also the main inducement causing a serious decline in the quality of life of patients. Against the background of metabolic disorders caused by persistent hyperglycemia, nerve tissues are continuously exposed to a glycotoxic environment, gradually forming oxidative stress damage and neuro-microcirculation (nutrition) disorders due to mitochondrial dysfunction, and ultimately leading to irreversible damage to peripheral nerve axonal degeneration and myelin sheath structure destruction.

[0003] Based on the development process of diabetic peripheral neuropathy, its treatment methods include three important development stages: in the early stage, the intervention target is simply blood glucose control, but clinical observations have found that strict blood glucose management can only delay but not prevent the progression of neuropathy; in the middle stage, it turns to metabolic pathway intervention strategies, and representative drugs include aldose reductase inhibitors, antioxidants, and neurotrophic factors, etc. Aldose reductase inhibitors play a role by inhibiting the abnormal activation of the polyol pathway, antioxidants such as α-lipoic acid are dedicated to scavenging oxygen free radicals, and neurotrophic factor drugs focus on repairing damaged nerve structures. Although these single-target drugs show intervention value in specific pathological links, due to the complexity of the multi-dimensional pathological mechanism of DPN, inherent defects such as a single action dimension and incomplete lesion coverage are exposed in clinical practice. Especially when irreversible nerve fiber damage occurs in the late stage of the disease course, the existing treatment system is more faced with the dilemma of a significant decline in the response rate.

[0004] The limitations of the current clinical treatment plan are mainly reflected in three aspects: First, it is difficult for chemically synthesized drugs to achieve multi-target synergistic regulation and cannot synchronously intervene in key pathological links such as inflammatory cascade reactions, oxidative stress damage, and neurotrophic support; second, there are phenomena of increased drug tolerance and reduced efficacy during long-term drug use, seriously affecting the continuity of treatment; third, although existing traditional Chinese medicine compound preparations have the potential advantage of multi-component synergistic effects, there are generally bottleneck problems such as unclear active ingredients and low standardization of quality control. Against this background, the innovative combination research based on active ingredients of traditional Chinese medicine has become an important direction to break through the existing treatment dilemma.

[0005] In addition, on the one hand, there are differences in the understanding among those skilled in the art; on the other hand, although the inventor studied a large number of documents and patents when making this invention, all details and content could not be listed in detail due to space limitations. However, this does not mean that this invention does not possess the features of these prior arts. On the contrary, this invention already has all the features of the prior arts, and the applicant reserves the right to add relevant prior arts in the background art. Summary of the Invention

[0006] Based on the above technical problems, the present invention relates to a pharmaceutical composition with a specific ratio, its preparation method and application. Specifically, the present invention provides a pharmaceutical composition composed of 6-gingerol and polygonatum polysaccharide in a weight ratio of 1:1. This composition has significant effects in regulating immune function, anti-inflammatory and other aspects.

[0007] The present invention innovatively integrates the pleiotropic characteristics of natural products through the compatibility of 6-gingerol and polygonatum polysaccharide, inheriting the traditional advantages of multi-target regulation of traditional Chinese medicine and meeting the precision requirements of modern drug research and development, providing a breakthrough solution for constructing a new DPN treatment system.

[0008] One of the purposes of the present invention is to provide a pharmaceutical composition for preventing and treating diabetic peripheral neuropathy, which contains 1-2 parts by weight of 6-gingerol and 1-2 parts by weight of polygonatum polysaccharide.

[0009] According to a preferred embodiment, the pharmaceutical composition contains 1 part by weight of 6-gingerol and 1 part by weight of polygonatum polysaccharide.

[0010] According to a preferred embodiment, 6-gingerol is a ginger extract.

[0011] According to a preferred embodiment, the pharmaceutical composition further contains a pharmaceutically or foodologically acceptable carrier. Preferably,

[0012] According to a preferred embodiment, the pharmaceutical composition is an oral preparation. Preferably, the oral preparation is a liquid dosage form, any one of pills, granules, ointments, powders, decoctions, capsules, and dripping pills.

[0013] According to a preferred embodiment, the purity of 6-gingerol is not less than 95%; the purity of the polygonatum polysaccharide is not less than 80%.

[0014] Another purpose of the present invention is to provide the application of the above pharmaceutical composition in the preparation of a drug for preventing, alleviating or treating diabetic peripheral neuropathy.

[0015] According to a preferred embodiment, the drug for preventing, alleviating or treating diabetic peripheral neuropathy can protect pancreatic tissue, reduce pain threshold and repair nerve fiber function damage.

[0016] According to a preferred embodiment, "repairing the functional injury of nerve fibers" is manifested as improving the nerve conduction velocity and repairing the demyelination pathological features of the sciatic nerve.

[0017] According to a preferred embodiment, the oral dosage of the pharmaceutical composition is 1.85 - 2.0 g / 60 Kg / day.

[0018] One of the objects of the present invention is also to provide a preparation method of a pharmaceutical composition for preventing and treating diabetic peripheral neuropathy, which comprises the following steps:

[0019] S1: Mix 6-gingerol with a purity of not less than 95% and polygonatum polysaccharide with a purity of not less than 80% obtained from Polygonatum sibiricum in a weight ratio of 1:1;

[0020] S2: After fully dissolving the solvent, obtain the pharmaceutical composition powder through spray drying or freeze drying technology.

[0021] The beneficial effects of the present technical solution are as follows:

[0022] The specific proportion combination of gingerol and polygonatum polysaccharide exerts technical advantages through a synergistic effect in the treatment of diabetic peripheral neuropathy. The combined application of the two can improve microcirculation disorders and slow down the decline of nerve conduction velocity through multiple pathways. After optimizing the proportion, it can not only enhance the targeted regulation of the key pathological links of neuropathy, but also reduce the potential risks of single-component large-dose use through the compatibility of natural ingredients, providing a drug combination plan that takes into account both safety and comprehensiveness of the mechanism of action for the treatment of diabetic peripheral neuropathy. Description of the Drawings

[0023] Figure 1 This is the HE staining (50 μm) of the pancreatic tissue of the present invention, where Control: normal group; Model: model group; 6-Gin+PRP: 6-gingerol combined with polygonatum polysaccharide group; 6-Gin: 6-gingerol group; PRP: polygonatum polysaccharide group;

[0024] Figure 2 This is the observation of the effect of 6-gingerol + polygonatum polysaccharide on the ultrastructure of the sciatic nerve of DPN model rats by transmission electron microscopy of the present invention, where Control: normal group; Model: model group; 6-Gin+PRP: 6-gingerol combined with polygonatum polysaccharide group; 6-Gin: 6-gingerol group; PRP: polygonatum polysaccharide group;

[0025] Figure 3 This is the effect of 6-gingerol + polygonatum polysaccharide on the blood lipid levels of DPN model rats of the present invention, where Control: normal group; Model: model group; 6-Gin+PRP: 6-gingerol combined with polygonatum polysaccharide group; 6-Gin: 6-gingerol group; PRP: polygonatum polysaccharide group;

[0026] Figure 4 The effect of 6-gingerol + polygonatum polysaccharide of the present invention on inflammatory factors in DPN model rats, where Control: normal group; Model: model group; 6-Gin+PRP: 6-gingerol combined with polygonatum polysaccharide group; 6-Gin: 6-gingerol group; PRP: polygonatum polysaccharide group. Detailed implementation mode

[0027] In the description of the present invention, the terms are only for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the quantity of the indicated technical features. In the description of the present invention, "a plurality" means two or more, unless otherwise specifically defined.

[0028] Unless otherwise specified, the test methods used in the following examples are all conventional methods; the materials, reagents or instruments used without indicating the manufacturer are all reagents and materials that can be obtained commercially; for those not indicating specific conditions in the examples, they are all carried out according to conventional conditions or the conditions recommended by the manufacturer. At the same time, the present invention does not limit the source of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are all common commercially available products in the technical field.

[0029] The experimental group (6-Gin+PRP) in the following efficacy verification obtains the complex according to the ratio shown in Example 1.

[0030] The raw materials of the composition involved in the present invention are commercially available products. 6-gingerol is a product with a purity of 80%, purchased from Xi'an Ruilin Biotechnology Co., Ltd. This product is prepared through specific extraction and purification processes, with stable chemical properties and meeting relevant quality standards, providing a reliable active ingredient basis for the subsequent preparation of the composition. Polygonatum polysaccharide is also a product with a purity of 80%, purchased from Xi'an Ruilin Biotechnology Company. This product is obtained through advanced extraction techniques, retaining the effective biological activity of polygonatum polysaccharide, and having good quality and availability in the market. During the preparation of the composition, they are mixed in a ratio of 1:1 to construct a composition system with specific efficacy.

[0031] Example 1

[0032] This example provides a pharmaceutical composition and its preparation method, specifically including the following parts by mass of pharmaceutical raw materials:

[0033] 6-gingerol and polygonatum polysaccharide are mixed in a weight ratio of 1:1.

[0034] The preparation method of the pharmaceutical composition includes the following steps:

[0035] Precisely weigh 6-gingerol with a purity of not less than 80% and polygonatum polysaccharide with a purity of not less than 80% obtained from polygonatum sibiricum red. by modern extraction technology in a ratio of 1:1 by weight, that is, take 1 part by weight each, and mix them in an appropriate amount of distilled water or ethanol (just make the solvent dissolve fully), and ensure that the two are fully dissolved and mixed evenly by stirring or ultrasonic treatment. Subsequently, obtain a dry and uniform powder of the 6-gingerol and polygonatum polysaccharide composition by spray drying or freeze drying technology.

[0036] Example 2

[0037] This example provides a pharmaceutical composition and its preparation method, specifically including the pharmaceutical raw materials in parts by mass:

[0038] 6-Gingerol and polygonatum polysaccharide are mixed in a ratio of 1:2 by weight.

[0039] The preparation method of the pharmaceutical composition includes the following steps:

[0040] Precisely weigh 6-gingerol with a purity of not less than 80% and polygonatum polysaccharide with a purity of not less than 80% obtained from polygonatum sibiricum red. by modern extraction technology in a ratio of 1:1, that is, take 1 part by weight each, and mix them in an appropriate amount of distilled water or ethanol (just make the solvent dissolve fully), and ensure that the two are fully dissolved and mixed evenly by stirring or ultrasonic treatment. Subsequently, remove the solvent by spray drying or freeze drying technology to obtain a dry and uniform powder of the 6-gingerol and polygonatum polysaccharide composition.

[0041] Example 3

[0042] This example provides a pharmaceutical composition and its preparation method, specifically including the pharmaceutical raw materials in parts by mass:

[0043] 6-Gingerol and polygonatum polysaccharide are mixed in a ratio of 2:1 by weight.

[0044] The preparation method of the pharmaceutical composition includes the following steps:

[0045] Precisely weigh 6-gingerol with a purity of not less than 80% and polygonatum polysaccharide with a purity of not less than 80% obtained from polygonatum sibiricum red. by modern extraction technology in a ratio of 2:1 by weight, mix them in an appropriate amount of distilled water or ethanol (just make the solvent dissolve fully), and ensure that the two are fully dissolved and mixed evenly by stirring or ultrasonic treatment. Subsequently, remove the solvent by spray drying or freeze drying technology to obtain a dry and uniform powder of the 6-gingerol and polygonatum polysaccharide composition.

[0046] I. Pharmacodynamic verification

[0047] 1. Experimental animals and modeling grouping

[0048] 80 male Sprague-Dawley rats, weighing 180 - 200 g, were randomly divided into a normal control group of 10 rats and a model group of 70 rats. Among the 70 rats in the model group, 60 rats successfully developed diabetes. Among these 60 diabetic rats, 15 were randomly assigned to the 6-gingerol + polygonatum polysaccharide (6-Gin + PRP) treatment group, 15 to the 6-gingerol (6-Gin) treatment group, 15 to the polygonatum polysaccharide (PRP) treatment group, and 15 to the diabetic peripheral neuropathy (DPN) group.

[0049] 2. Modeling method

[0050] Rats in the model group were fed a high-fat diet and allowed free access to food and water. Each rat was intraperitoneally injected with streptozotocin (STZ) solution. Before use, STZ was dissolved in 0.1 mol / L sodium citrate buffer at pH 4.2 to prepare a 1% concentration solution, and the rats were intraperitoneally injected with the STZ solution at a dose of 35 mg / kg. After 8 weeks, tail vein blood was taken from the rats in the model group to measure blood glucose. If the fasting blood glucose remained above 16.7 mmol / L, the sciatic nerve conduction velocity was significantly slower than that of the blank group, and the heat pain threshold decreased by 10%, the diabetic peripheral neuropathy model was considered successful.

[0051] 60 successfully modeled rats were randomly selected and then randomly divided into a 6-gingerol + polygonatum polysaccharide (6-Gin + PRP) treatment group of 15 rats, a 6-gingerol (6-Gin) treatment group of 15 rats, a polygonatum polysaccharide (PRP) treatment group of 15 rats, and a diabetic peripheral neuropathy (DPN) group of 15 rats.

[0052] 10 rats in the normal control group were only injected with normal saline and fed a normal diet. The treatment groups were gavaged with 6-gingerol + polygonatum polysaccharide (1:1) once a day, 6-gingerol (6-Gin) once a day, and polygonatum polysaccharide (PRP) once a day for 8 consecutive weeks. The DPN group and the normal control group were gavaged with normal saline once a day for 8 consecutive weeks.

[0053] 3. Observe the effect of 6-gingerol + polygonatum polysaccharide on blood glucose in DPN rats

[0054] Since the occurrence and development of DPN are closely related to abnormal body metabolism, especially the continuous increase in blood glucose levels and the exacerbation of insulin resistance, which will further promote the development of the disease. Therefore, this example aims to detect the effect of the complex composed of 6-gingerol and polygonatum polysaccharide on the blood glucose of DPN model rats, providing new ideas for the prevention and treatment of DPN.

[0055] Appropriately establish DPN model rats. Drug intervention: The DPN model rats were randomly divided into an experimental group and a control group. The experimental group was given gavage or injection intervention with the complex of 6-gingerol and polygonatum polysaccharide, while the control group was given an equal amount of normal saline or other placebo.

[0056] Blood glucose detection: At specific time points (such as before intervention, every week after intervention, etc.), the fasting blood glucose level of rats was detected by tail vein blood sampling or other appropriate methods.

[0057] Statistical analysis was used to analyze the differences between the two groups to evaluate the effect of 6-gingerol + polygonatum polysaccharide complex on improving blood glucose in DPN model rats. As shown in Table 1, after 6-gingerol + polygonatum polysaccharide intervention in DPN rats induced by high-fat diet combined with intraperitoneal injection of STZ, the fasting blood glucose of rats was significantly improved, suggesting that 6-gingerol + polygonatum polysaccharide could overall improve the blood glucose of DPN rats.

[0058] Table 1

[0059] Group 6 weeks 8 weeks 10 weeks 12 weeks 14 weeks Control group 4.75±0.65 5.28±0.4 5.78±0.94 4.55±0.90 4.93±0.32 Model group 20.10±2.64 20.77±2.51 22.28±3.17 23.32±3.17 25.03±2.85 6-Gin + PRP group 18.78±1.23 16.87±0.78 15.62±0.68 13.67±0.92 12.22±0.66 6-Gin group 19.55±0.93 17.98±1.05 16.42±0.72 15.03±1.21 13.75±0.98 PRP group 18.53±1.23 16.82±1.69 15.63±1.48 14.57±1.18 13.23±1.29

[0060] 4. Observe the effect of 6-gingerol + polygonatum polysaccharide on the pancreatic tissue of DPN rats

[0061] HE staining was used to observe the pathological changes of pancreatic tissues in rats of each group. Figure 1 The green arrows indicate islets of Langerhans, which are located in the endocrine part of the pancreas and secrete hormones such as insulin and glucagon. They are round cell clusters with clear boundaries and evenly distributed cell nuclei. The yellow dotted line box indicates acinar cells, the main cells of the exocrine part of the pancreas, which are responsible for secreting digestive enzymes. The cells are arranged in a bunch of grapes, and there is a tiny acinar lumen in the center of each acinus, occupying most of the pancreatic tissue. The pancreatic tissue structure of rats in the normal group was intact, the acini were closely arranged, the islet morphology was normal, the intercalated ducts and ducts were clearly structured, and no obvious pathological changes were seen. The pancreatic tissue of rats in the model group showed significant pathological damage, mainly manifested as: 1) pancreatic tissue edema; 2) obvious atrophy of islets of Langerhans and significant reduction in the number of islet cells; 3) destruction of acinar structure, visible acinar atrophy and cell swelling; 4) inflammatory cell infiltration was seen around islets of Langerhans and acini; 5) obvious thickening of blood vessel walls.

[0062] The observation results of the drug administration group showed that the pathological changes of pancreatic tissues in the 6-gingerol and polygonatum polysaccharide combined administration group were significantly improved, specifically manifested as: 1) the number of islets of Langerhans was basically the same as that in the normal group, and only slight atrophy was seen; 2) the acinar structure was basically intact and no obvious atrophy was seen; 3) the degree of inflammatory cell infiltration was significantly reduced; 4) the thickness of blood vessel walls returned to normal. In contrast, although the single drug administration group had a certain improvement effect, there were still pathological changes such as: 1) slight atrophy of islets of Langerhans; 2) mild atrophy of acini; 3) a small amount of inflammatory cell infiltration; 4) varying degrees of thickening of blood vessel walls.

[0063] As Figure 1 shown, the combined use of 6-gingerol and polygonatum polysaccharide has a significant protective effect on pancreatic tissue damage in diabetic rats, and its effect is significantly better than that of the single-component administration group.

[0064] Improvement Effect of 5.6-Gingerol and Polygonatum sibiricum Polysaccharide Complex on Thermal Pain Threshold in DPN Model Rats

[0065] Since the occurrence and development of DPN are closely related to nerve dysfunction and are often accompanied by symptoms such as hyperalgesia or hypoalgesia. Among them, the change of thermal pain threshold is one of the important indicators for evaluating the degree of neuropathy. This example aims to detect the effect of the complex composed of 6-gingerol and Polygonatum sibiricum polysaccharide on the thermal pain threshold of DPN model rats, providing a new choice of drug combination for the treatment of DPN.

[0066] Detection of thermal pain threshold: Before and after experimental intervention, a professional hot plate instrument was used to detect the thermal pain threshold of rats. The specific operation was to place the rats on the test platform, stimulate the soles of the rats' feet with a heat source of a specific intensity, and record the time from the rats feeling the stimulus to showing obvious avoidance reactions (such as leg retraction, foot licking, etc.), which was the thermal pain threshold.

[0067] Data recording and analysis: Record and compare the changes in the thermal pain threshold of rats in the experimental group and the control group before and after experimental intervention, and use appropriate statistical methods for data analysis to evaluate.

[0068] The experimental results are shown in Table 2. Compared with the control group, the thermal pain threshold of rats in the experimental group was significantly shortened after experimental intervention, indicating that the 6-gingerol and Polygonatum sibiricum polysaccharide complex has a significant improvement effect on the thermal pain threshold of DPN model rats.

[0069] This example detected the effect of the 6-gingerol and Polygonatum sibiricum polysaccharide complex on the thermal pain threshold of DPN model rats, providing a new choice of drug combination for the treatment of DPN. This complex may improve the symptoms of hyperalgesia or hypoalgesia in DPN model rats by reducing nerve inflammatory responses and other mechanisms, and has high clinical application value.

[0070] Table 2

[0071]

[0072]

[0073] 6. Observe the Effect of 6-Gingerol + Polygonatum sibiricum Polysaccharide on Nerve Conduction Velocity (NCV) in DPN Rat Model

[0074] Since DPN, as a common diabetic complication, its occurrence and development are closely related to the damage and dysfunction of nerve fibers, and nerve conduction velocity is an important indicator for measuring nerve function status. This example aims to detect the effect of the complex composed of 6-gingerol and Polygonatum sibiricum polysaccharide on the nerve conduction velocity of DPN model rats, providing a new choice of drug combination for the treatment of DPN.

[0075] Experimental method: First, a model was established by using a high-fat diet combined with intraperitoneal injection of STZ. After the model was successfully established, the rats were intervened with the 6-gingerol + polygonatum polysaccharide composition provided by the present invention, and the effect of drug intervention on nerve conduction velocity (NCV) was observed.

[0076] Nerve conduction velocity detection: The rats were fixed in the prone position after being anesthetized with isoflurane. The rats were placed at room temperature of 24 °C. After disinfecting the proximal and distal parts of the lower limbs with alcohol, electrodes were placed. To determine the motor nerve conduction velocity (NCV), the sciatic nerve was stimulated with a single super-large square wave pulse (5 - 10 mA and 40 μs duration) through a fine needle electrode inserted percutaneously at the sciatic nerve incision and the ankle. The distance between the two stimulation sites was 2 mm. The NCV was calculated by subtracting the distal latency from the proximal latency, and the result was divided by the distance between the stimulating electrode and the recording electrode. All measurements were performed in triplicate for each group. The nerve conduction velocity (m / s) was substituted into the formula = distance between the stimulating electrode and the recording electrode / latency, and then the conduction velocity of the sciatic nerve was calculated.

[0077] Data recording and analysis: The changes in nerve conduction velocity of rats in the experimental group and the control group before and after experimental intervention were recorded and compared, and appropriate statistical methods were used for data analysis to evaluate the improvement effect of the 6-gingerol and polygonatum polysaccharide complex on the nerve conduction velocity of DPN model rats.

[0078] The experimental results are shown in Table 3. Compared with the control group, the nerve conduction velocity of rats in the experimental group was significantly increased after experimental intervention, indicating that the 6-gingerol and polygonatum polysaccharide complex has a significant improvement effect on the nerve conduction velocity of DPN model rats.

[0079] In this example, by detecting the effect of the 6-gingerol and polygonatum polysaccharide complex on the nerve conduction velocity of DPN model rats, the effectiveness of this complex in improving the nerve function of DPN rats was confirmed. This complex may improve the nerve conduction velocity of DPN model rats by promoting the repair and regeneration of nerve fibers, improving the transmission of neurotransmitters and other mechanisms, providing a new drug combination option for the treatment of DPN.

[0080] Table 3

[0081] Group Nerve conduction velocity (NCV) Control 71.24±11.26 Model <![CDATA[39.82±7.04 a > 6-Gin + PRP <![CDATA[59.66±5.35 b > 6-Gin <![CDATA[56.38±9.51 bb > PRP 57.48±5.98

[0082] 7. Observe the effect of 6-gingerol + polygonatum polysaccharide on the ultrastructure of the sciatic nerve in DPN rat models

[0083] Since DPN is a serious complication of diabetes, it often causes changes in the ultrastructure of peripheral nerves, including myelin sheath damage and axonal degeneration. The complex formed by 6-gingerol and polygonatum polysaccharide has a positive effect on improving the ultrastructure of the sciatic nerve in DPN model rats. This example aims to explore the effects of the 6-gingerol and polygonatum polysaccharide complex on the myelin sheath and axonal ultrastructure of the sciatic nerve in DPN model rats through transmission electron microscopy (TEM) observation.

[0084] Experimental method: First, a model was established by using a high-fat diet combined with intraperitoneal injection of STZ. After the model was successfully established, the rats were intervened with the 6-gingerol + polygonatum polysaccharide composition provided by the present invention, and the effect of drug intervention on the pathology of the sciatic nerve was observed.

[0085] Sampling and preparation of sciatic nerve: After the treatment was completed, the rats were sacrificed and the sciatic nerves were quickly removed. The sciatic nerve samples were fixed, dehydrated, embedded, etc., and prepared into ultra-thin sections suitable for transmission electron microscopy observation.

[0086] Observation by transmission electron microscopy: The ultra-thin sections of the sciatic nerve were observed using a transmission electron microscope. Focus was placed on the integrity, hierarchical structure of the myelin sheath, as well as the morphology of the axon and the changes of Schwann cells. Representative images were taken and recorded.

[0087] Data analysis and statistics: Qualitative and quantitative analyses were performed on the results of transmission electron microscopy observation. The improvement effect of the 6-gingerol and polygonatum polysaccharide complex on the myelin sheath and axonal ultrastructure of the sciatic nerve in DPN model rats was evaluated. Statistical methods were used to compare and analyze the experimental data.

[0088] As Figure 2 shown, the green dotted-line box in the figure indicates the myelin sheath, which wraps the axon; the yellow dotted-line box indicates the axon, which is a circular or oval area in the center of the myelin sheath, containing microtubules and neurofilaments, and is the nerve signal conduction channel; the red box indicates the neurofilaments in the axon. In the sciatic nerve of the model group rats, the myelin sheath lamellar structure of a large number of myelinated nerve fibers was blurred, the lamellae were separated, there were a large number of vacuolar defects, the microtubules and microfilaments inside the axon were significantly broken and dissolved, and some areas were fragmented, which was in line with the demyelination pathological characteristics caused by DPN. In the 6-gingerol + polygonatum polysaccharide group rats, the neurofilaments in the axon were arranged tightly, the continuity was significantly improved, the fracture phenomenon was significantly reduced, the myelin sheath thickness was uniform, and the lamellar structure was clear. This indicates that the 6-gingerol and polygonatum polysaccharide complex has a significant improvement effect on the ultrastructure changes of the sciatic nerve in DPN model rats.

[0089] In this example, the effects of the 6-gingerol and polygonatum polysaccharide complex on the myelin sheath, axons, and Schwann cells of the sciatic nerve in DPN model rats were detected. The experimental results showed that the complex had a significant neuroprotective effect and could reduce the ultrastructural damage of the sciatic nerve in DPN model rats, providing new ideas and methods for the treatment of DPN. At the same time, this example also provided strong experimental evidence for the application of the 6-gingerol and polygonatum polysaccharide complex in the field of neuroprotection.

[0090] 8. Observe the effects of 6-gingerol + polygonatum polysaccharide on blood lipids in DPN rat models

[0091] As Figure 3 shown, the composition of the present invention has a significant regulatory effect on DPN-related blood lipid metabolism disorders. Compared with the normal group, the model group of rats showed obvious dyslipidemia, characterized by significantly increased serum total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL-C), and significantly decreased high-density lipoprotein (HDL-C).

[0092] After treatment with the composition of the present invention, the above abnormal blood lipid indexes were significantly improved, manifested as significantly decreased levels of TC, TG, and LDL-C, and significantly increased levels of HDL-C. In particular, the improvement effects of using 6-gingerol and polygonatum polysaccharide alone were significantly lower than those of the composition of the present invention. These results fully demonstrated that the composition of the present invention could effectively improve DPN-related blood lipid metabolism disorders, and its therapeutic effect was significantly better than that of single-component treatment.

[0093] 9. Effects of 6-gingerol + polygonatum polysaccharide on inflammatory factors in DPN rat models

[0094] As Figure 4 shown, the experimental data indicated that the composition of the present invention had a significant regulatory effect on DPN-related inflammatory responses. Compared with the normal group, the levels of pro-inflammatory factors IL-6, TNF-α, and IL-1β in the serum of the model group of rats were significantly increased, while the level of the anti-inflammatory factor IL-10 was significantly decreased, indicating an obvious imbalance of inflammatory factors in the model group. After intervention with the 6-gingerol + polygonatum polysaccharide of the present invention, the levels of pro-inflammatory factors IL-6, TNF-α, and IL-1β in the serum of rats were significantly decreased, and the level of the anti-inflammatory factor IL-10 was significantly increased, and the inflammatory response was significantly improved.

[0095] These results confirmed that the composition of the present invention could more effectively regulate the balance of pro-inflammatory / anti-inflammatory cytokines, reduce the nerve inflammatory response, and thus play a significant therapeutic role in DPN

[0096] It should be noted that the above specific embodiments are exemplary. Those skilled in the art can come up with various solutions inspired by the disclosed content of the present invention, and these solutions also fall within the scope of the disclosure of the present invention and within the protection scope of the present invention. Those skilled in the art should understand that the specification and drawings of the present invention are illustrative and do not constitute a limitation on the claims. The protection scope of the present invention is defined by the claims and their equivalents.

Claims

1. A pharmaceutical composition for preventing and treating diabetic peripheral neuropathy, characterized in that, It contains 1-2 parts of 6-gingerol and 1-2 parts of polygonatum polysaccharide by weight.

2. The pharmaceutical composition according to claim 1, wherein It contains 1 part of 6-gingerol and 1 part of polygonatum polysaccharide by weight.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that, The 6-gingerol is a ginger extract.

4. The pharmaceutical composition according to any one of claims 1 to 3, characterized in that, The pharmaceutical composition further comprises a pharmaceutically or food acceptable carrier.

5. The pharmaceutical composition according to claim 1, wherein The purity of the 6-gingerol is not less than 95%; the purity of the polygonatum polysaccharide is not less than 80%.

6. Use of a pharmaceutical composition in the preparation of a medicament for preventing, alleviating or treating diabetic peripheral neuropathy, characterized in that, The pharmaceutical composition is the pharmaceutical composition according to any one of claims 1-5.

7. The application according to claim 6, characterized in that, The drug capable of preventing, alleviating or treating diabetic peripheral neuropathy can protect pancreatic tissue, reduce pain threshold and repair nerve fiber function damage.

8. The application according to claim 6, characterized in that, "Repairing nerve fiber function damage" is manifested as increasing nerve conduction velocity and repairing the demyelination pathological features of the sciatic nerve.

9. The application according to claim 6, wherein The oral dosage of the pharmaceutical composition is 1.85-2.0 g / 60 Kg / day.

10. A preparation method of a pharmaceutical composition for preventing and treating diabetic peripheral neuropathy, characterized in that, It comprises the following steps: S1: Mix 6-gingerol with a purity of not less than 95% and polygonatum polysaccharide with a purity of not less than 80% obtained from polygonatum sibiricum in a weight ratio of 1:1; S2: After fully dissolving the solvent, obtain the pharmaceutical composition powder by spray drying or freeze drying technology.

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