CN117959394A - Application of ginseng and astragalus pulse-invigorating preparation in preparation of medicine for preventing and treating diabetic neuropathy - Google Patents

Abstract

The invention belongs to the field of traditional Chinese medicines, and in particular relates to an application of a ginseng and astragalus pulse-invigorating preparation in preparation of a medicine for treating diabetic neuropathy. The ginseng astragalus pulse-invigorating preparation contains plant medicines and animal medicines, the traditional Chinese medicine composition in pharmacological and clinical cases can reduce blood sugar level, regulate nerve conduction rate and improve peripheral neuropathy caused by painful diabetic neuropathy, especially diabetic foot, has quick treatment effect and high total effective rate, and has higher market value.

Description

Application of ginseng and astragalus pulse-invigorating preparation in preparation of medicine for preventing and treating diabetic neuropathy

Technical Field

The invention relates to an application of a ginseng and astragalus pulse-invigorating preparation in preparing a medicament for preventing and treating diabetic neuropathy, belonging to the field of traditional Chinese medicines.

Background

Diabetes-induced peripheral neuropathy (DPN) is one of the common complications of diabetes. According to statistics, the DPN pathogenicity rate caused by diabetes mellitus can reach 24.4% -78.8% in global patients; of the chinese patients, about 80% of diabetes-induced DPN, of which about 25% of patients clinically manifest as painful diabetic neuropathy. Typical diabetic neuropathy patients are characterized by spontaneous pain, hyperalgesia and allodynia due to disease invasion of sensory nerves, and can be characterized by ant crawling, burning, needling, stabbing, jumping, cutting, tearing, etc., accompanied by muscle weakness, directly affecting peripheral nerves, central nerves, evident pain at night, severely affecting the quality of daily life, limiting daily activities, and possibly causing emotional depression of the patient, affecting social contact.

Clinical manifestations of diabetic foot are neurological and vascular lesions, including sensory changes (stocking-like manifestations, light touch, proprioception, joint attenuation of temperature sensation and pain sensation), intrinsic muscular atrophy of the foot; skin sweat normally and temperature and blood transport regulating function are lost: local ulcers, infections, osteomyelitis, charcot's joint disease, and the like.

In recent years, researches show that the application of traditional Chinese medicine to diabetics can reduce the blood sugar accidents of the diabetics, and can improve the clinical symptoms of the diabetics and prevent and treat chronic complications of the diabetics; the traditional Chinese medicine is used for reducing blood sugar, and the effect is mostly inferior to western medicines, but has remarkable effect on treating vascular complications of diabetics. At the same time, traditional Chinese medicine can have various approaches to the treatment of AGEs-RAGE systems, such as inhibiting the formation of AGEs and inhibiting the signal transduction associated with the expression of RAGE downstream thereof.

In view of the above, there is a great need in clinic to provide a medicament for treating diabetic neuropathy with little side effects, remarkable curative effect and low recurrence rate.

Disclosure of Invention

Aiming at the defects of the prior art, the ginseng-astragalus pulse-invigorating decoction is comprehensively researched and totally innovated, and the original 5 medicines are changed into 12 medicines.

The invention aims at providing a new type of ginseng and astragalus pulse-invigorating preparation. Can be an oral preparation, preferably, the oral preparation is decoction, pill, tablet, mixture, capsule, granule, powder or paste, more preferably, tablet or pill, and most preferably, tablet.

On the basis of clinical experiments, the same or similar therapeutic effects can be achieved by the preparation obtained by adopting the following technical scheme.

Specifically, the technical purpose of the invention is realized by the following technical scheme:

a preparation comprising radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, saviae Miltiorrhizae radix, caulis Spatholobi, flos Lonicerae, radix Codonopsis, lumbricus, scorpio and Glycyrrhrizae radix for preventing and treating diabetic neuropathy is provided.

The traditional Chinese medicine composition comprises the following components:

preferably, the ginseng astragalus pulse-invigorating preparation comprises the following components:

a second object of the present invention is to provide a pharmaceutical use of the ginseng and astragalus pulse-invigorating preparation. The neuropathy is painful diabetic peripheral neuropathy and lower limb vascular nerve.

Furthermore, the diabetic neuropathy is mainly lower limb vascular neuropathy, the ginseng and astragalus pulse-invigorating preparation contains various traditional Chinese medicinal materials of plant medicines and animal medicines for synergistic effect, and the traditional Chinese medicinal composition can reduce blood sugar level, regulate nerve conduction rate, improve peripheral neuropathy caused by painful diabetic neuropathy, particularly diabetic foot, has quick treatment effect and high total effective rate and has higher market value in pharmacological and clinical cases.

Further, the diabetes mellitus of the invention is type 1 diabetes or type 2 diabetes.

Further, the vascular nerve of the lower limb is a diabetic foot.

Furthermore, the diabetic foot refers to a disease state that the foot of a diabetic patient suffers from lower limb protection function decline due to neuropathy, and ulcers and gangrene occur due to microcirculation disturbance caused by insufficient arterial perfusion due to macrovascular and microvascular lesions, and is a complication of diabetes.

The third object of the present invention is to provide a preparation of the above traditional Chinese medicine composition, wherein the traditional Chinese medicine composition is prepared into a traditional Chinese medicine oral preparation or an external preparation by using the traditional Chinese medicine components, preferably, the ginseng and astragalus pulse-invigorating preparation is an oral preparation, preferably, the oral preparation is a decoction, a pill, a tablet, a mixture, a capsule, a granule, a powder or a paste, and more preferably, a tablet.

Further, the invention also provides a preparation method of the ginseng astragalus pulse-invigorating preparation, which comprises the following steps:

Pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 65-85% ethanol for 1-2 times, each time for 1.5-3.5 hr, filtering, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.05-1.10; extracting the residue with 2-5 times of water for 1-2 times, filtering, and collecting water extractive solution;

Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, adding 1/2 of prescription amount of Lumbricus and Scorpio into the rest medicinal liquid and residue, decocting with water for 1-2 times, each time for 1.5-3.5 hr, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 40-50%, standing at normal temperature for 24-48 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.15-1.20;

Mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into clinically acceptable dosage form by directly or adding pharmaceutically acceptable excipient.

Further, the preparation method of the ginseng astragalus pulse-invigorating preparation provided by the invention comprises the following steps:

Pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 75% ethanol for 2 times and 2.5 hr each time, filtering, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.08; extracting the residue with 3 times of water for 2 times, filtering, and collecting water extractive solution;

Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, adding 1/2 of Lumbricus and Scorpio with the rest medicinal liquid and residue, decocting with water for 2 times and 2.5 hr each time, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 45%, standing at normal temperature for 48 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.18;

Mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into clinically acceptable dosage form by directly or adding pharmaceutically acceptable excipient.

The invention has perfect monarch, minister, assistant and guide, proper compatibility and a middle-jiao-regulating mechanism, and the monarch drug in the formula is as follows: radix astragali, atractylodis rhizoma. These herbs are the main components of the pulse-invigorating prescription that play a central role. The radix astragali has the effects of benefiting qi, nourishing blood, invigorating qi and consolidating superficial resistance; atractylodis rhizoma has effects of invigorating spleen, relieving diarrhea, invigorating spleen, and stopping bleeding; ministerial drugs: radix Cyathulae, caulis Spatholobi, lumbricus, and Scorpio. These medicinal materials play roles in assisting treatment and enhancing effects in the pulse-invigorating prescription. Adjuvant drug: coix seed, chinese angelica, red sage root, honeysuckle flower and pilose asiabell root have the efficacy of inducing diuresis to alleviate edema and clearing heat and detoxicating; the angelica can promote blood circulation, regulate menstruation, enrich blood and nourish blood; the red sage root has the functions of promoting blood circulation to remove blood stasis, relaxing tendons and activating collaterals; caulis Spatholobi has effects in promoting blood circulation, dispelling blood stasis, and relieving swelling and pain; the honeysuckle has the effects of clearing heat, detoxicating, removing toxicity and relieving swelling; the radix codonopsis has the effects of replenishing qi to invigorate the spleen and improving immunity; the earthworm can promote blood circulation to remove meridian obstruction, dispel wind and remove dampness; scorpio has effects of promoting blood circulation, removing blood stasis, and relieving pain. And (3) enabling the mixture to be subjected to the following steps: glycyrrhrizae radix has effects of regulating drug property, increasing drug stability and promoting absorption in the preparation of the composition.

Compared with the prior art, the invention has the following advantages:

The study is to adopt oral administration of the Shenqi pulse-invigorating tablets for patients with diabetic ulcers with qi deficiency and blood stasis, detect changes of downstream factors of AGEs-RAGE of different time nodes of the patients, healing conditions of wound surfaces and the like, further verify the effectiveness and safety of the Shenqi pulse-invigorating tablets for treating the patients with diabetic ulcers, and carry out clinical popularization. The traditional Chinese medicine composition disclosed by the invention is reasonable in formula and has a remarkable curative effect on neuropathic pain.

Drawings

FIG. 1 shows the wound healing analysis of example 1 and comparative examples 1-3.

FIG. 2 shows the change in wound surface area of example 1 and comparative examples 1 to 3.

Figure 3 shows the wound area comparison at 3 weeks of treatment for example 1 and comparative examples 1-3.

Figure 4 shows the wound area comparison at 4 weeks of treatment for example 1 and comparative examples 1-3.

FIG. 5 shows the modification of TNF-a of example 1 and comparative examples 1-3.

FIG. 6 shows the IL-6 modification of example 1 and comparative examples 1-3.

FIG. 7 shows the change of the hypersensitive C-reactive protein of example 1 and comparative examples 1 to 3.

Detailed Description

In order to verify the efficacy of the ginseng and astragalus pulse-invigorating preparation, the inventor develops pharmacodynamic test research. The medicine selected by the pharmacodynamic test research of the invention is a representative formula and a medicine obtained by the preparation method thereof. The tests and results related to other formulations and to the pharmaceutical products obtained by the preparation method, which are encompassed by the present invention, are not exhaustive.

Example 1: preparation of ginseng astragalus pulse-invigorating tablet

The formula comprises the following components:

The preparation method comprises the following steps:

A) Pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

B) Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 65 ethanol for 1 time for 1.5 hr, filtering, recovering ethanol from filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.05; extracting the filtered residues with 2 times of water for 1 time, filtering, and collecting water extractive solution;

c) Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, decocting the rest medicinal liquid and residue with 1/2 of Lumbricus and Scorpio with water for 1 time and 1.5 hr each time, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 40%, standing at normal temperature for 24 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.15;

D) Mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into tablet directly or by adding pharmaceutically acceptable excipient.

Example 2: preparation of ginseng astragalus pulse-invigorating tablet

The formula comprises the following components:

The preparation method comprises the following steps:

A) Pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

b) Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 85% ethanol for 2 times and 3.5 hr each time, filtering, recovering ethanol from filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.10; extracting the filtered residues with 5 times of water for 2 times, filtering, and collecting water extractive solution;

c) Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, adding 1/2 of Lumbricus and Scorpio with the rest medicinal liquid and residue, decocting with water for 3.5 hr each time, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 50%, standing at normal temperature for 48 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.20;

D) Mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into tablet directly or by adding pharmaceutically acceptable excipient.

Example 3: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method comprises the following steps:

A) Pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

B) Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 75% ethanol for 2 times and 2.5 hr each time, filtering, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.08; extracting the residue with 3 times of water for 2 times, filtering, and collecting water extractive solution;

c) Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, adding 1/2 of Lumbricus and Scorpio with the rest medicinal liquid and residue, decocting with water for 2 times and 2.5 hr each time, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 45%, standing at normal temperature for 48 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.18;

D) Mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into tablet directly or by adding pharmaceutically acceptable excipient.

Example 4: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method is the same as in example 3.

Example 5: preparation of ginseng astragalus pulse-invigorating pill

The preparation method is the same as in example 3, and auxiliary materials are added according to the conventional process to prepare pills.

Example 6: preparation of ginseng astragalus external ointment for invigorating pulse

The preparation method is the same as that of example 3, and auxiliary materials are added according to the conventional process to prepare the external preparation.

Comparative example 1: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method is the same as in example 3.

Comparative example 2: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method is the same as in example 3.

Comparative example 3: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method is the same as in example 3.

Comparative example 4: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method is the same as in example 3.

Comparative example 5: preparation of ginseng astragalus pulse-invigorating tablet

The preparation method is the same as in example 3.

Comparative example 6: preparation of ginseng astragalus external emulsifiable paste for invigorating pulse

The preparation method comprises the following steps:

A) Pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

B) Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 75% ethanol for 2 times and 2.5 hr each time, filtering, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.08; standby;

C) Decocting flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi in water for 2 times and 2.5 hr each time, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 45%, standing at normal temperature for 48 hr, recovering ethanol from the filtrate, and concentrating to obtain extract with relative density of 1.18;

d) Mixing Lumbricus and Scorpio with the ethanol extract of step B and the extract of step C, and making into topical cream directly or by adding pharmaceutically acceptable excipient.

Comparative example 7: preparation of ginseng astragalus external emulsifiable paste for invigorating pulse

The preparation method comprises the following steps:

A) Extracting Lumbricus, scorpio, radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 75% ethanol for 2 times each for 2.5 hr, filtering, recovering ethanol from filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.08; extracting the residue with 3 times of water for 2 times, filtering, and collecting water extractive solution;

b) Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, decocting the rest medicinal liquid and residues with water for 2 times and 2.5 hr each time, adding ethanol into the filtrate and the water extract obtained in step B to make ethanol content reach 45%, standing at normal temperature for 48 hr, recovering ethanol from the filtrate, and concentrating to obtain extract with relative density of 1.18;

C) Mixing the water extract with the ethanol extract of step B and the water extract of step C, and directly or adding pharmaceutically acceptable excipient to obtain topical cream.

1. Pharmacological experiments-experiments and results of the formulations of the invention on the wounded surface of mice

1. Material

1.1 Animals:

SPF-class Kunming mice, 18-22 g, experimental animal license number: SYXK (robust) 20180004, and the animals were subjected to adaptive feeding for 7 days before the experiment.

1.2 Test article

1.2.1 Medicaments

The formulations and preparation methods of the creams prepared according to example 6 and comparative examples 6-7 of the present invention.

Moisten burn cream group (national medicine standard Z20000004).

1.2.2 Methods of use

The paste of the present invention: is externally applied. Is coated on the wound surface or the surface to be healed (foam thickness is 1-2cm, cream thickness is less than 1 mm), and new medicine is replaced every 2-4 hours. Foam or liquid remained on the wound surface is wiped off before dressing change. The wound surface is exposed, and then the administration is continued.

Wet burn cream group: is externally applied. The product is applied to wound surfaces (thickness is less than 1 mm) such as burn, scald, and burn, and new medicine is replaced every 2-4 hours. Before changing the dressing, the medicine and the liquefied substance remained on the wound surface should be wiped off. The wound surface is exposed, and then the administration is continued.

2. Modeling, grouping, and administration

2.1 Method of Forming mold

On day 1, the back of the mouse is dehaired, the dehairing area is about 3cm multiplied by 4cm, and the unhairing area is observed for 24 hours, so that the unhairing position is determined to have no abnormal conditions such as redness, inflammation, breakage and the like; on day 2, after the abdominal cavity of the mouse is anesthetized at room temperature, the limbs are stretched and fixed on an operation table to enable the back skin to be flat, the back hair of the mouse is pushed out, the range is about 6cm, the area is larger than the expected burn area, a weight of 20g is placed in a container and heated to be boiling, the temperature is kept for 10min, 75% ethanol is used for disinfecting the skin of a mouse experiment area, the heated weight is rapidly placed on the skin of the mouse, and a slight pressure is applied to form a shallow II-degree scald model with the scald area of about 2cm multiplied by 2 cm. The local skin after the scald is porcelain white, the epidermis is wrinkled, the common local skin is broken, the normal area and the scald part are clear and distinguishable, and the subcutaneous tissue edema is obvious in 4 hours of the scald. 1 mouse is randomly extracted, wound tissues of the mouse are taken for pathological histology examination, the epidermis of a scalding area of the mouse can be seen to disappear under a lens, the whole epidermis layer and the dermis superficial layer are infiltrated, and the scald is in line with shallow II-degree scalding.

2.2 Grouping and administration

Mice 50 successfully molded were randomly divided into 10 groups of model, wet burn cream, example 6, and comparative examples 6-7.

Experimental groups: the wet burn cream set, example 6 set, and comparative examples 6 to 7, the application areas were respectively applied with the corresponding medicines according to the above application methods, and the model set was applied with 0.9% physiological saline. The administration was continued for 14 days.

3. Observation index

3.1 Rate of healing of skin lesions local areas were measured by tracing and weighing at days 7, 10 and 14 after treatment. Firstly, marking the skin damage range by using a transparent film, and then cutting the transparent film corresponding to the size of the skin damage area to obtain the quality of the transparent film, wherein the quality replaces the area.

3.2 Statistical method

SPSS17.0 software was used for data processing. The data obtained are expressed as mean ± standard deviationThe normal distribution test and the variance homogeneity test of each group of data are in accordance with the requirement of the Pearson test, and the single-factor variance analysis is used for the comparison between the groups. The difference of P < 0.05 is statistically significant.

4. Results and conclusions

The skin lesions of the inventive example 6 and the comparative examples 5-6 and the wet burn cream groups were repaired to different extents on days 7, 10 and 14 after the treatment, and the healing was better as the time was longer, wherein the healing effect was best in the inventive example 6 group.

Compared with the model group, the healing rate of mice in each administration group on days 7, 10 and 14 is higher, and the difference has statistical significance (P is less than 0.01), so that the success of the model is further demonstrated; compared with the moist burn ointment group, the mice in the group 6 of the invention have higher skin injury healing rate, and the difference has statistical significance (P is less than 0.05).

TABLE 1 comparison of skin loss healing Rate for mice of each groupn＝10)

Group of	Day 7 of treatment	Day 10 of treatment	Day 14 of treatment
				Model group	32.34±3.11	47.47±5.83	58.27±7.11
Moist burn ointment set	42.43±5.82&	57.74±6.14&	74.92±8.19&
				Example 6	44.56±6.11&	63.09±6.72&#	84.33±7.75&#
Comparative example 6	39.67±3.40	52.36±7.89	64.77±8.28
				Comparative example 7	38.33±5.71	53.46±5.11	65.98±6.42

Note that: ^*P＜0.05,^& P <0.01 in comparison with the model group;

^# P < 0.05 in contrast to moist burn cream.

2. Antibacterial test of the pharmaceutical composition of the present invention

2.1 Materials and methods

1) Strain and culture medium: culture medium for salmonella, staphylococcus aureus, candida albicans, escherichia coli and pseudomonas aeruginosa

Bacteria (the above strains are commercially available).

2) Medicament: according to the invention of examples 3-4, the raw materials and the method of comparative examples 4-5 produced pharmaceutical compositions (raw materials before adding the auxiliary materials).

3) The method comprises the following steps: the sand culture medium was prepared and filled into Erlenmeyer flasks and sterilized by heat and humidity. Each of examples 3 to 4 according to the present invention, comparative examples 4 to 5 and a 1:20 dilution of sterilized physiological saline were aseptically filled into sterilized dishes with 10 dishes (10 cm in diameter) per dose group, and then inoculated with the above four bacteria (approximately 0.5mm diameter pellet) upon pouring into a sand culture. Incubation (37 ℃) for 6 days. Colony growth status and presence or absence of growth of the bacteria were observed, and colony diameter (mm) was measured and compared with a blank group.

TABLE 2 antibacterial effect of the groups of examplesn＝10)

Note that: in contrast to comparative example 41, ^*P＜0.05,^& P < 0.01;

in contrast to comparative example 44, ^#P＜0.05,^@ P < 0.01;

As is clear from Table 2, the examples 3-4 of the present invention have remarkable antibacterial effect, and have remarkable inhibitory or killing effect on pathogenic bacteria such as Staphylococcus aureus, candida albicans, enterobacter, pseudomonas aeruginosa, etc.

2. Pharmacodynamic experiments:

1. Clinical study: the study randomly divided 120 cases of diabetic ulcer patients meeting the standard into 5 groups by selecting 120 cases of diabetic ulcer patients meeting the standard from the clinic or hospitalization patients of traditional Chinese medicine hospitals in the medical science of Jinan of 7 months 2021 to 2023, wherein the gender, age and wound area of the groups are not significantly different (P is more than 0.05).

2. Grouping and administration

The tablets of example 1 and comparative examples 1-3 were taken in 24 cases each, 6 tablets each day, and twice a day with a hollow warm dose. The preparation method comprises the steps of taking a treatment course for 1 week, and continuously taking 4 treatment courses, wherein a positive control group (Chinese medicinal quasi-word Z32020535 Tongsaimai tablet) is taken according to the specification, and selecting a proper hypoglycemic medicament according to the blood sugar level of a patient and the actual condition of the patient, and performing oral hypoglycemic medicament or subcutaneous insulin injection treatment to maintain the fasting blood sugar below 8.0 mmol/L. Each group was given local normal saline care if needed.

Diagnostic criteria

3.1 According to the diagnostic standard of Chinese diabetes foot diagnosis and treatment guidelines (2020 edition) of the Chinese society of Chinese medicine:

(1) Patients with type 2 diabetes mellitus present with limb vascular and/or neuropathy and/or with infections;

(2) The limb of the type 2 diabetic has clinical manifestations and signs of wet gangrene or dry gangrene, and accords with the level 1-5 gangrene standard;

(3) ABI index at rest is less than 0.9;

(4) Ultrasonic color Doppler examination prompts patients with limb vascular stenosis, limb blood supply reduction and ischemia or gangrene;

(5) Angiography confirms that CTA and MRA prompt vascular lumen stenosis or blockage and has clinical manifestations;

(6) Electrophysiological examination, it can be seen that peripheral nerve conduction velocity is slow or electromyography and somatosensory evoked potential are abnormally changed;

(7) X-ray examination can show osteoporosis decalcification, bone destruction, osteomyelitis or arthropathy, hand and foot deformity, and summer joint change;

meanwhile, the two (1) and (2) are met, and 1 piece of any one of the (3) to (7) is added for clear diagnosis.

3.2 Diagnostic criteria for TCM

Reference is made to the gangrene diagnostic standard in the "traditional Chinese medicine disease diagnosis efficacy Standard" issued by the national administration of Chinese medicine in 2017:

(1) Is often applied to one or both sides of the lower limb. The patients have a history of diabetes, cold, trauma, and long-term massive smoking. The first toe and cold finger pain, the lower limb soreness, numbness and pain, the aggravation when walking, the relief when resting, intermittent claudication and the weakening of the fuyang pulse. The pain is continuous, the skin at the limb end is cool, the skin is dark red, blue and purple, the skin is dry, the vellus falls off, the toenail is deformed and thickened, the muscle is atrophic, and the Fuyang pulse disappears. Further necrosis, severe pain, and sleep, tuck and sitting. When the ulcer is infected, wet necrosis occurs, the limb ends are red, swollen, hot and painful, and the whole body is heated;

(2) Blood vessel stasis type: the toes feel warm and feel cold, the tingling and swelling pain is caused by the severe pain caused by cold, the walking is unfavorable, the feet are heavy and weak, the pain is aggravated when the feet walk more, the skin color is dark red and bluish purple, and the pain of the patient with the ulcer is continuously aggravated and can not be fall asleep for a long time. A dark red tongue or ecchymosis with white coating, wiry or astringent pulse, and vanishing of the pulse of the Fuyang.

3.3 Grading Standard

The most classical diabetic foot classification, wagner classification, was used.

There is a risk factor for foot ulcers at grade 0, but currently there are no ulcers

Grade 1 superficial foot ulcers, with no signs of infection, appear prominently as neuropathic ulcers

Deep ulcers of grade 2, frequent soft tissue infections, no osteomyelitis or deep abscess

Deep ulcers of grade 3, including abscess or osteomyelitis

Grade 4 localized gangrene (toe, heel or forefoot dorsum), characterized by ischemic gangrene, usually combined with neuropathy

Grade 5 full foot gangrene

3.4 Inclusion criteria

(1) Meets the diagnosis standard of type 2 diabetes; (2) meets the Western diagnosis standard of diabetic foot ulcer;

(3) Meets the diagnostic standard of blood stasis in traditional Chinese medicine; (4) Wagner classification belongs to class 1-3;

(5) Ankle humerus index (ABI) is more than or equal to 0.4; (6) age 18-80 years, unlimited gender;

3.5 exclusion criteria

(1) Critical limb ischemia, patients requiring emergency vascular bypass or intervention;

(2) Amputees need to perform localized necrosis or severe infection of the limb;

(3) Ankle humerus index (ABI) < 0.4;

(4) Cancerous, tubercular, and other specific ulcers;

(5) Serious illness and unstable vital signs;

(6) Severe malnutrition;

(7) Combining psychotic disorders, alcoholism or drug addiction;

(8) Ozone therapy contraindications such as hyperthyroidism, hypotension, hypocalcemia, hypoglycemia, ozone allergy, citrus allergy (with sodium citrate), coagulation disorder (such as hemophilia), and fava bean disease;

(9) Simultaneously taking part in other test patients;

3.6 reject and drop criteria

(1) Mismatching, and failing to meet the inclusion standard;

(2) Poor compliance, and failure to receive a prescribed treatment regimen;

(3) Data is lost, and the validity and security judgment cannot be performed;

(4) Serious adverse reactions to the drug occur, and the test must be stopped;

(5) Exacerbation or death cases, resulting in termination of the study;

4. Clinical observation index

Observing the wound surface area at 0 week, 1 week, 2 weeks, 3 weeks and 4 weeks of treatment; serum (TNF-a, IL-6, hypersensitive C reactive protein (hs-CRP)) was tested at 4 weeks of treatment.

5. Efficacy assessment criteria

Referring to the clinical research guidelines formulated in the clinical research guidelines (2022) of new traditional Chinese medicines, and combining with the clinical characteristics of diabetic foot ulcer, the clinical efficacy evaluation criteria are formulated as follows:

and (3) healing: the wound surface is basically healed, and clinical symptoms and signs are basically disappeared. The integral index of the wound surface syndrome is more than or equal to 90 percent.

The effect is shown: the area (depth) of the wound surface is obviously reduced, the local pain of the wound surface is disappeared or obviously reduced, the secretion is obviously reduced, the granulation is fresh, and the periphery is free from reddening and swelling. The integral index of the wound surface symptoms is less than or equal to 60 percent and less than 90 percent.

The method is effective: the area (depth) of the wound surface is reduced, the local pain of the wound surface is reduced, secretion is reduced, granulation is fresh, and peripheral redness and swelling are reduced. The integral index of the wound surface symptoms is less than or equal to 30 percent and less than 60 percent.

Invalidation: the area (depth) of the wound surface is not reduced or increased, the local pain of the wound surface is not reduced or aggravated, secretion is not reduced, and the condition of the wound surface is not improved. The integral index of wound surface symptoms is less than 30 percent.

Analysis of results

FIG. 1 shows the analysis of the healing of the wound surface of example 1 and comparative examples 1-3, and it can be seen that the treatment effect of example 1 is optimal, and the healing and significant effect of the individual is far greater than that of other examples.

Fig. 2 shows the change in wound area for example 1 and comparative examples 1-3, and it can be seen that there was a significant change between groups and across weeks 0-2, and significant stratification began to occur at the beginning of treatment for 3 weeks, with the treatment effect of example 1 being significantly better than that of comparative examples 1-3.

Figure 3 shows a comparison of the wound area at 3 weeks of treatment for example 1 and comparative examples 1-3, and it can be seen that there is a significant difference between examples and other comparative examples.

Fig. 4 shows a comparison of the wound surface areas of example 1 and comparative examples 1 to 3 at 4 weeks of treatment, and it can be seen that there is a significant difference between the examples and the other comparative examples, and the difference in the therapeutic effects is significantly increased.

FIG. 5 shows the modification of TNF-a of example 1 and comparative examples 1-3.

FIG. 6 shows the IL-6 modification of example 1 and comparative examples 1-3.

FIG. 7 shows the change of the hypersensitive C-reactive protein of example 1 and comparative examples 1 to 3.

Claims (10)

1. The application of the ginseng and astragalus pulse-invigorating preparation in preparing medicaments for preventing and treating diabetic neuropathy is characterized in that the ginseng and astragalus pulse-invigorating preparation is prepared from raw astragalus, bighead atractylodes rhizome, medicinal cyathula root, coix seed, chinese angelica, red sage root, suberect spatholobus stem, honeysuckle, dangshen, earthworm, scorpion and raw licorice.

2. The use according to claim 1, wherein the ginseng-astragalus pulse-invigorating preparation comprises the following components:

10-70 parts of raw astragalus root, 5-30 parts of bighead atractylodes rhizome and 8-40 parts of medicinal cyathula root

10-60 Parts of coix seed, 10-50 parts of angelica, and 5-30 parts of red sage root

5-30 Parts of spatholobus stem, 5-30 parts of honeysuckle, and 10-50 parts of dangshen

5-30 Parts of earthworm, 1-6 parts of scorpion and 5-20 parts of raw licorice.

3. The use according to claim 2, wherein the ginseng-astragalus pulse-invigorating preparation comprises the following components:

30 parts of raw astragalus, 15 parts of bighead atractylodes rhizome and 18 parts of medicinal cyathula root

30 Parts of coix seed, 20 parts of angelica, 15 parts of red sage root

15 Parts by weight of suberect spatholobus stem, 15 parts by weight of honeysuckle, and 30 parts by weight of dangshen

15 Parts of earthworm, 3 parts of scorpion and 10 parts of raw licorice.

4. A use according to any one of claims 1 to 3, wherein the diabetes is type 1 diabetes or type 2 diabetes.

5. The use according to claim 4, wherein the neuropathy is painful diabetic peripheral neuropathy, lower extremity vascular nerve.

6. The use according to claim 5, wherein the vascular nerve of the lower limb is a diabetic foot.

7. The use according to claim 6, wherein the diabetic foot is a disease state in which the foot of a diabetic patient suffers from a decrease in lower limb protection due to neuropathy, and ulcers and gangrene due to a microcirculation disturbance caused by insufficient arterial perfusion due to macrovascular and microvascular lesions, which is a complication of diabetes.

8. The use according to claim 6, wherein the ginseng and astragalus pulse-invigorating preparation is an oral preparation, preferably the oral preparation is a decoction, a pill, a tablet, a mixture, a capsule, a granule, a powder or a paste, more preferably a tablet.

9. The use of claim 8, wherein the preparation method of the ginseng astragalus pulse-invigorating preparation is as follows:

A. pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

B. Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 65-85% ethanol for 1-2 times, each time for 1.5-3.5 hr, filtering, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.05-1.10; extracting the residue with 2-5 times of water for 1-2 times, filtering, and collecting water extractive solution;

C. Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, adding 1/2 of prescription amount of Lumbricus and Scorpio into the rest medicinal liquid and residue, decocting with water for 1-2 times, each time for 1.5-3.5 hr, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 40-50%, standing at normal temperature for 24-48 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.15-1.20;

D. mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into clinically acceptable dosage form by directly or adding pharmaceutically acceptable excipient.

10. The use of claim 8, wherein the preparation method of the ginseng astragalus pulse-invigorating preparation is as follows:

A. pulverizing Lumbricus and Scorpio to particle size smaller than 100 μm;

B. Extracting radix astragali, atractylodis rhizoma, radix Cyathulae, coicis semen, radix Angelicae sinensis, and radix Codonopsis with 75% ethanol for 2 times and 2.5 hr each time, filtering, recovering ethanol from the filtrate, and concentrating under reduced pressure to obtain extract with relative density of 1.08; extracting the residue with 3 times of water for 2 times, filtering, and collecting water extractive solution;

C. Extracting volatile oil from flos Lonicerae, saviae Miltiorrhizae radix, glycyrrhrizae radix and caulis Spatholobi with water, adding 1/2 of Lumbricus and Scorpio with the rest medicinal liquid and residue, decocting with water for 2 times and 2.5 hr each time, adding ethanol into the filtrate and the water extractive solution obtained in step B to make ethanol content reach 45%, standing at normal temperature for 48 hr, recovering ethanol from the filtrate, concentrating to obtain extract with relative density of 1.18;

D. mixing the rest 1/2 of the prescription amount of Lumbricus and Scorpio with the ethanol extract of step B and the water extract of step C, and making into clinically acceptable dosage form by directly or adding pharmaceutically acceptable excipient.