CN117045739A - Composition for treating type 2 diabetes and preparation method and application thereof - Google Patents
Composition for treating type 2 diabetes and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of diabetes medicine research, and particularly relates to a composition for treating type 2 diabetes, a preparation method and application thereof. The composition is prepared from the following raw materials in parts by weight: 10-18 parts of bupleurum, 8-12 parts of cortex moutan, 8-16 parts of red paeony root, 15-24 parts of radix scrophulariae, 15-30 parts of dwarf lilyturf tuber, 5-15 parts of peach seed, 6-13 parts of dried rehmannia root, 6-15 parts of Chinese yam and 6-12 parts of hemerocallis. The composition provided by the invention has the effects of reducing blood glucose and improving insulin metabolism in vivo.
Description
Technical Field
The invention belongs to the technical field of diabetes medicine research, and particularly relates to a composition for treating type 2 diabetes, a preparation method and application thereof.
Background
"diabetes" is a disease in which glucose in blood tends to accumulate too much. Epidemiological investigation report in 2019 shows that the number of diabetics in China is 1.164 hundred million, and with the increasing population aging, the probability of suffering from diabetes rises year by year, wherein 60% -80% of diabetics are obese people before the onset of the disease, have poor constitution and need to be controlled by taking medicine for a long time. It is predicted that during 1995-2025, the incidence of diabetes in developed countries abroad will break through 27%. Diabetes has become an epidemic worldwide in the 21 st century. The traditional Chinese medicine is a chronic non-infectious disease which is the third most serious threat to human health after tumor and vascular lesions, has the characteristics of high mortality rate, high disability rate and high medical cost, and becomes a public health problem commonly faced by countries in the world. Diabetes mellitus is a serious situation in China.
China is a large country of traditional Chinese medicine resources and has a long history of treating diabetes by traditional Chinese medicines, so that the development of traditional Chinese medicine preparations for effectively treating diabetes is a wish of many pharmaceutical workers. In recent years, it has also been proved that various compounds having pharmacological activity can be extracted and isolated from natural medicines. The action mechanism of the traditional Chinese medicine composition on diabetes and complications thereof has been discussed by students at home and abroad from various angles. Proved by researches, baicalin is an effective Aldose Reductase (AR) inhibitor, and can obviously reduce the AR activity of kidney rats with kidney diseases; quercetin also has certain inhibition effect on AR; the extract of radix Sophorae Flavescentis mainly contains isoflavone, and can prevent cataract of diabetic patient by inhibiting activity of key enzyme AR in glucose metabolism; guava She Mugan can enhance capillary permeability of diabetics. Because the flavonoid compounds in the natural plants or the medicaments have low toxicity, not only can reduce blood sugar, but also can inhibit the development of complications, the development of the medicament sources is very important at home and abroad, and new effective components are extracted. However, most of the pharmacological actions of flavonoids against diabetes and complications thereof are still in the experimental stage, and only a small part of the clinical trials are entered. Some of the other traditional Chinese medicine components act similarly to alpha-glucosidase inhibitors. For example, foreign studies have found that a variety of polyhydroxy alkaloids are present in Morus plants, and that some polyhydroxy alkaloids isolated from Morus alba, mori folium and Mori fructus have good glycosidase inhibitory activity; the researchers in China also successfully extract alkaloids (mainly 1-deoxynojirimycin) with similar effects from tender mulberry twigs and silkworm excrement. It is pleasant that the Chinese medicine insulin sensitizer which is independently developed in China is about to enter clinical research. Experts generally consider that developing a new way for treating diabetes by using traditional Chinese medicines and ethnic medicines with clear effective parts, clear mechanism and definite curative effect is an important task to be solved urgently by medical workers. Western medicines such as dapagliflozin, metformin, sitagliptin and the like are combined with insulin to be used in combination mainly in foreign countries. In recent years, with the deep application research of traditional Chinese medicines for treating diabetes, a plurality of traditional Chinese medicines and compound preparations thereof show good curative effects. The Chinese medicinal preparations of Xiaokeling tablet, kelening tablet, jinqi Jiangtang tablet, yuye Xiaokejie granule, yangyangjiang Jiapian, jiangtangsu capsule, xiaokejiangjiang tablet, jiangtangjiao capsule, xiaokejiangjiang capsule, xiaokejiangjiangjiang capsule, xiaokewan and the like are accepted by the pharmacopoeia of people's republic of China and the ministry of force standard. The traditional Chinese medicine has the characteristics of single effect, can reduce blood sugar, improve insulin secretion in vivo and prevent and treat diabetic complications. Therefore, developing an innovative traditional Chinese medicine preparation with good curative effect, convenient administration and safe use is an urgent need of drug researchers.
Disclosure of Invention
The invention aims to overcome the defect of single action of the traditional Chinese medicine or ethnic medicine for treating type 2 diabetes, and develops a composition with the functions of reducing blood sugar and improving insulin metabolism in vivo.
The aim of the invention can be achieved by the following measures:
the invention provides a composition for treating type 2 diabetes, which is prepared from the following raw materials in parts by weight: 10-18 parts of bupleurum, 8-12 parts of cortex moutan, 8-16 parts of red paeony root, 15-24 parts of radix scrophulariae, 15-30 parts of dwarf lilyturf tuber, 5-15 parts of peach seed, 6-13 parts of dried rehmannia root, 6-15 parts of Chinese yam and 6-12 parts of hemerocallis.
Preferably, the composition is prepared from the following raw materials in parts by weight: 10 parts of bupleurum, 10 parts of cortex moutan, 10 parts of red paeony root, 15 parts of radix scrophulariae, 15 parts of radix ophiopogonis, 7.5 parts of peach kernel, 10 parts of radix rehmanniae, 9 parts of Chinese yam and 10 parts of hemerocallis.
In a second aspect of the present invention, there is provided a process for preparing the composition comprising the steps of:
weighing bupleurum, cortex moutan, red paeony root, radix scrophulariae, dwarf lilyturf tuber, peach kernel, dried rehmannia root, chinese yam and hemerocallis in parts by weight, mixing, extracting with 40-45% ethanol solution by mass fraction, and collecting an extracting solution to obtain the composition.
Preferably, the extraction is carried out by adding ethanol solution with mass fraction of 40-45% which is 8-10 times of total weight of mixed medicinal materials, reflux extracting for 3 times, each time for 2.5-3 hours.
Preferably, the composition is obtained by concentrating the extract and drying.
In a third aspect, the invention provides the use of the composition in the manufacture of a medicament for the treatment of type 2 diabetes.
Preferably, the composition is used for preparing a hypoglycemic drug.
Preferably, the composition is used for the preparation of a medicament for improving insulin metabolism in vivo.
In a fourth aspect, the present invention provides a pharmaceutical formulation for the treatment of type 2 diabetes comprising the composition, and pharmaceutically acceptable excipients.
The traditional Chinese medicine composition provided by the invention has the following effects and characteristics of the components of each medicinal material:
bupleurum root: bitter in property and slightly cold in nature, enter liver and gallbladder meridians. Has the effects of relieving exterior and interior syndrome, soothing liver and raising yang. Researches show that bupleurum root has the effects of relieving fever, resisting inflammation, resisting virus and protecting liver, wherein flavonoid compounds have strong antioxidant capacity, polysaccharide has an immunosuppressive effect on mice with delayed hypersensitivity, and the action mechanism is possibly related to the reduction of lymphocyte numbers in intestinal peyer's patch of the mice and the inhibition of lymphocyte interferon-gamma level and the increase of Gao Baijie hormone-4 level.
Cortex moutan: bitter and pungent, and slightly cold. Has effects of cooling blood, clearing heat, promoting blood circulation, removing blood stasis, and has various pharmacological effects such as protecting liver and kidney, relieving inflammation, tranquilizing, cooling, relieving fever, relieving pain, and relieving spasm. The main chemical component is paeonol, which has obvious anti-inflammatory and analgesic activities and plays an important role in the treatment of arthritis, skin inflammation, organ injury, periodontitis, stomatitis and colonitis. Paeonol has certain neuroprotective effect, and is a potential therapeutic agent for improving nervous system related diseases (such as neurotoxicity, parkinson disease, depression, cerebral ischemic injury, epilepsy, senilism, etc.).
Radix paeoniae rubra: bitter and slightly cold. Enter liver meridian. Has effects of clearing heat, cooling blood, promoting blood circulation, and removing blood stasis. Can be used for treating heat entering nutrient blood, toxic heat, speckle, hematemesis, epistaxis, conjunctival congestion, swelling and pain, liver Yu Xie pain, amenorrhea, dysmenorrhea, abdominal mass, abdominal pain, traumatic injury, carbuncle, swelling and sore. The radix paeoniae rubra has been identified as a plurality of effective chemical components such as radix paeoniae rubra total glycosides, gallic acid, radix paeoniae rubra total flavonoids, neolignan glycoside compounds and the like, and experiments prove that the effective components have the effects of protecting nerves, resisting depression, regulating blood sugar and blood fat, resisting inflammation and bacteria, resisting tumors, resisting oxidation, regulating immunity and the like.
Radix scrophulariae: sweet, bitter and salty in taste, slightly cold in nature, and has the effects of clearing heat and cooling blood, nourishing yin and reducing fire, and detoxifying and resolving masses. The active ingredients of the composition mainly comprise iridoid glycoside, phenylpropanoid, flavone, polysaccharide, triterpene, sesquiterpene, benzyl alcohol glycoside, phenethyl alcohol glycoside and the like, and have the effects of protecting cardiovascular system, resisting tumor, resisting oxidation, resisting inflammation, reducing blood sugar, protecting liver and the like. The high-dose radix scrophulariae polysaccharide can obviously improve the glycolipid metabolism function of the type 2 diabetes rat, improve the oxidation capacity of the organism and increase the insulin secretion. The glucose-lowering effect of the radix scrophulariae extract is detected by adopting HepG2 glucose consumption and MTT experiments, and the harpagoside has the strongest glucose consumption capacity and the best glucose-lowering activity.
Radix Ophiopogonis: sweet, slightly bitter and slightly cold. It enters heart, lung and stomach meridians. Nourishing yin, promoting salivation, moistening lung, and relieving cough. Can be used for treating thirst, dry cough and hemoptysis due to deficiency of body fluid caused by deficiency of lung-yin and stomach-yin; palpitations due to deficiency of heart yin, and impairment of body fluids due to heat in the late stage of fever. The radix Ophiopogonis has abundant chemical components, and contains steroid saponins, high isoflavone, polysaccharides, etc., and has the pharmacological effects of protecting cardiovascular system, protecting digestive system, resisting tumor, protecting nervous system, improving liver and lung injury, resisting inflammation and oxidation, lowering blood sugar, resisting aging, regulating immunity, etc. The radix Ophiopogonis single medicine has 22 chemical components which can directly act on 20 diabetes related targets, wherein the main targets comprise protein kinase B1 (Akt 1), cannabinoid receptor 1 (CNR 1) and dopamine receptor D2 (DRD 2), and the main components comprise radix Ophiopogonis methyl homoisoflavone A and radix Ophiopogonis methyl homoisoflavone B.
Peach kernel: bitter, sweet and flat. It enters heart, liver and large intestine meridians. Has effects of promoting blood circulation, removing blood stasis, loosening bowel to relieve constipation, relieving cough and asthma. Can be used for treating amenorrhea dysmenorrhea, abdominal mass, pulmonary abscess, acute appendicitis, traumatic injury, constipation due to intestinal dryness, cough and asthma. The semen Persicae contains multiple chemical components including volatile oil, cyanogenic glycoside, amino acid and protein, flavone and its glycoside, sterol and its glycoside, aromatic glycoside, fatty acid, phenylpropanoid, nucleoside, trace element and other compounds. Studies show that peach kernel can reduce the expression of serine/threonine kinase-like protein (TRIB 3), messenger RNA (mRNA) and Toll-like receptor, protein kinase B (AKT) and mitogen-activated protein kinase (MAPK) signal paths of diabetic macrovascular fibrosis rats, inhibit the expression of c-Jun amino terminal kinase JNK1 gene in femoral artery blood vessels, reduce the generation and expression of JNK1 protein, and reduce the expression of ERK1/2, p-ERK1/2 and MAPK m RNA, thereby effectively inhibiting diabetic vascular lesions and facilitating the recovery of peripheral neuropathy of diabetes.
Radix rehmanniae: sweet and cold, enter heart meridian, liver meridian and kidney meridian, and has the effects of clearing heat, cooling blood, nourishing yin and promoting fluid production. Can be used for treating fever and polydipsia caused by pyrexia. The chemical components of the medicine mainly comprise iridoid glycosides, amino acids, phenethyl alcohol glycosides, trace elements, polysaccharides, ionones and the like, and the medicine has pharmacological effects of influencing blood systems, resisting aging and tumors, enhancing immunity, preventing and treating osteoporosis, reducing blood sugar, improving cardiovascular system, improving memory, calming, treating parkinsonism, resisting inflammation, resisting fatigue and the like. The rehmannia root extract can inhibit glycogen synthase kinase-3 beta (AKT/GSK-3 beta) signal pathway activation and relieve podocyte Epithelial Mesenchymal Transition (EMT) process, so as to realize the protection of DKD rats, wherein rehmannia root monomer, namely, rehmannia root glycoside D, has the function of reducing blood sugar. The rehmannia root oligosaccharide can effectively relieve weight loss of model mice, and can play a role in diabetes and complications thereof by reducing blood sugar and blood lipid levels.
Chinese yam: sweet, warm, flat and nontoxic. In treating injury, tonifying deficiency, eliminating pathogenic cold and heat, invigorating middle warmer, invigorating qi, growing muscle, and strengthening yin. The Chinese yam has the effects of reducing blood pressure and blood fat, resisting tumor, resisting inflammation, regulating immunity and the like. The Chinese yam polysaccharide, chinese yam protein, saponin and the like have good preventive effects on chronic kidney disease, diabetes, coronary heart disease, atherosclerosis and the like. The microelements in the yam have positive significance for preventing and treating diabetes and complications thereof. The aqueous extract of the mountain tea polysaccharide can obviously reduce the blood sugar content of a dexamethasone-induced diabetes model mouse.
Daylily: sweet and cool. Clearing heat, promoting urination, cooling blood and stopping bleeding. Can be used for treating parotitis, jaundice, cystitis, hematuria, dysuria, galactorrhea, menoxenia, epistaxis, and hematochezia. It can be used for treating mastitis by external application. The hemerocallis contain various active ingredients including flavonoids, alkaloids, anthraquinones, polyphenols, terpenes, triterpenes and their glycosides, naphthalosides, steroids, phenethyl alcohols, lignans, carotenoids and trace elements; has sedative, hypnotic, antidepressant, antioxidant, liver protecting, antiinflammatory, antitumor, antibacterial, and insecticidal effects. The hemerocallis extract has a great difference in the liver toxicity results induced by different components, has a protective effect on the liver toxicity induced by Palmitic Acid (PA) and acetaminophen, and has no effect on the liver toxicity induced by Oleic Acid (OA).
Modern diabetes is a chronic metabolic disease, usually occurring after the age of 35-40 years, accounting for more than 90% of diabetics. Patients are characterized by hyperglycemia, relative lack of insulin, insulin resistance, etc. Common symptoms include polydipsia, frequent urination, weight loss due to unknown causes, and may also include overeating, tiredness, or soreness. Diabetes mellitus is generally called diabetes in traditional Chinese medicine, and diabetes may be caused by improper diet, emotional disorder, deficiency of kidney yin and other factors. Therefore, the basic treatment method for clearing heat and moistening lung, clearing stomach and purging fire, nourishing yin and strengthening kidney and promoting the production of body fluid and quenching thirst is formed. Also has the functions of resolving phlegm, promoting blood circulation, removing blood stasis, dispersing stagnated liver qi, clearing heat, invigorating spleen, purging pathogenic fire, and removing toxic substances from different aspects such as turbid phlegm, blood stasis, liver fire, spleen deficiency, and interior toxin. In the traditional Chinese medicine composition provided by the invention, bupleurum and cortex moutan are monarch drugs, and are used for soothing liver, raising yang, clearing heat, cooling blood, activating blood and dissolving stasis; radix paeoniae rubra, radix scrophulariae, radix ophiopogonis and peach kernel are ministerial drugs, and the effect of assisting the monarch drug in clearing heat and cooling blood, and activating blood circulation to dissipate blood stasis; the dried rehmannia root, the Chinese yam and the hemerocallis are taken as adjuvant drugs, the effect of clearing heat and cooling blood of the whole prescription is enhanced, and meanwhile, the dried rehmannia root and the ministerial drug ophiopogon root are matched in a synergistic way, so that the dried orange peel-root-containing Chinese yam-hemerocallis have the effects of nourishing yin and promoting the production of body fluid.
Drawings
FIG. 1 is a graph showing the change in body weight of rats in each group;
FIG. 2 is a graph of model group (A) and normal group (B) pancreatic HE staining;
FIG. 3 is a graph of pancreatic HE staining of the low dose group (A) of the composition and the dose group (B) of the composition;
FIG. 4 is a graph of pancreatic HE staining of the high dose group (A) and metformin group (B) of the compositions;
FIG. 5 is a graph of insulin immunohistochemistry for normal group (A) and model group (B);
FIG. 6 is a graph of composition high dose group (A), composition low dose group (B) insulin immunohistochemistry;
fig. 7 is a graph of insulin immunohistochemistry of dose group (a) and metformin group (B) in a composition.
Detailed Description
The following specific examples are given to illustrate the technical aspects of the present invention, but the scope of the present invention is not limited thereto.
The following test is a conclusion test of summarized research and development personnel based on the technical scheme to be protected by the invention on the basis of multiple creative tests.
Example 1
A composition for treating type 2 diabetes, which is prepared from the following raw materials: 10g of bupleurum, 8g of cortex moutan, 8g of red paeony root, 15g of radix scrophulariae, 15g of radix ophiopogonis, 5g of peach seed, 6g of radix rehmanniae, 6g of Chinese yam and 6g of hemerocallis.
Example 2
A composition for treating type 2 diabetes, which is prepared from the following raw materials: 18g of bupleurum, 12g of cortex moutan, 16g of red paeony root, 24g of radix scrophulariae, 30g of radix ophiopogonis, 15g of peach kernel, 13g of radix rehmanniae, 15g of Chinese yam and 12g of hemerocallis.
Example 3
A composition for treating type 2 diabetes, which is prepared from the following raw materials: 12g of bupleurum, 12g of cortex moutan, 12g of red paeony root, 22g of radix scrophulariae, 28g of radix ophiopogonis, 10g of peach kernel, 12g of radix rehmanniae, 12g of yam and 10g of hemerocallis.
Example 4
A composition for treating type 2 diabetes, which is prepared from the following raw materials: 15g of bupleurum, 15g of cortex moutan, 10g of red paeony root, 20g of radix scrophulariae, 25g of radix ophiopogonis, 12g of peach seed, 10g of radix rehmanniae, 10g of Chinese yam and 8g of hemerocallis.
Example 5
A composition for treating type 2 diabetes, which is prepared from the following raw materials: 18g of bupleurum, 12g of cortex moutan, 12g of red paeony root, 20g of figwort root, 28g of dwarf lilyturf tuber, 12g of peach seed, 8g of dried rehmannia root, 6g of Chinese yam and 8g of hemerocallis.
Example 6
A composition for treating type 2 diabetes, which is prepared from the following raw materials: 10g of bupleurum, 10g of cortex moutan, 10g of red paeony root, 15g of radix scrophulariae, 15g of radix ophiopogonis, 7.5g of peach kernel, 10g of radix rehmanniae, 9g of Chinese yam and 10g of hemerocallis.
The compositions provided in examples 1-6 above were prepared according to the following procedure:
according to the compatibility ratio of each medicinal material in the formula, nine medicinal materials of bupleurum, cortex moutan, red paeony root, figwort root, dwarf lilyturf tuber, peach seed, dried rhizome of rehmannia, chinese yam and hemerocallis are sequentially weighed, mixed, extracted for 3 hours by reflux with 10 times of 40% ethanol, extracted for two times, and the filtrates are combined. Concentrating the filtrate under reduced pressure to obtain paste, lyophilizing (-54 deg.C) to remove solvent.
Example 7
A composition for treating type 2 diabetes, which differs from example 6 only in the preparation method, specifically:
according to the compatibility ratio of each medicinal material in the formula, nine medicinal materials of bupleurum, cortex moutan, red paeony root, figwort root, dwarf lilyturf tuber, peach seed, dried rhizome of rehmannia, chinese yam and hemerocallis are sequentially weighed, mixed, extracted for 2.5h by reflux with ethanol with the mass fraction of 45 percent of 8 times, extracted for 3 times, and the filtrates are combined. Concentrating the filtrate under reduced pressure to obtain paste, lyophilizing (-54 deg.C) to remove solvent.
Since the therapeutic effects of the compositions provided in examples 1 to 7 are substantially the same, the effects will be described below by taking the composition provided in example 6 as an example.
The composition with the effects of reducing blood sugar and improving insulin metabolism in vivo obtained by the invention is subjected to the following efficacy test, and the curative effect of the composition on treating type 2 diabetes is verified, wherein the experimental process and the result are as follows:
1. model building
150 Wistar rats were randomly divided into a normal group and a model group, the normal group rats were given 10ml/kg of distilled water by lavage (ig), the model group rats were given 10ml/kg of fat emulsion on the basis of a normal diet, and after 4 weeks, the normal group and the model group randomly took 12 rats with blood collected from the tail vein to detect the content of serum TC, TG, FFA. Then, the animals of each group were fasted without water inhibition for 12 hours, the normal group was sublingually intravenous injected (iv) with 0.1mol/L, PH 4.2.2 citric acid-sodium citrate buffer, the model group was sublingually iv STZ (streptozotocin) with 30mg/kg, the model group rats ig 25% glucose solution for 4 hours after administration to avoid hypoglycemia reaction, and rats with fasting blood glucose values >11.1mmol/L were selected from model rats after 72 hours as model rats for type 2 diabetes.
2. Grouping and administration methods
Model type 2 diabetes rats were randomly divided into 8 groups of 15 rats each, namely: model control group, composition small, medium, large dose group and metformin group. The specific dosing scheme is as follows:
normal control group: administering 0.5% CMC-Na in an amount of 10ml/kg;
model control group: CMC-Na was administered at 0.5% and at 10ml/kg.
Metformin group: administering metformin in an amount of 100mg/kg;
low dose group of compositions: a small dose of 500mg/kg was administered to the composition prepared in example 6;
dose group in composition: a medium dose of 1000mg/kg was administered to the composition prepared in example 6;
high dose group of compositions: a large dose of 2000mg/kg was administered to the composition prepared in example 6;
animals of each group were dosed 1 time daily for 4 weeks following protocol ig described above. Type 2 diabetic rats ig were given 10ml/kg of fat milk 1 time every 3 days. Blood was taken from the tail vein weekly, fasting blood glucose was measured once with a glucometer, and group comparisons were made as measured values or percent change. The animals are fasted for 12h before the last 1 dose of each group of animals on the fourth week, the fasting blood glucose value is measured, the ig dose is given, 10% chloral hydrate is injected for 30mg/kg for anesthesia in the abdominal cavity 1h after the administration, the abdominal aorta is used for blood collection, serum is separated, and various biochemical indexes and release indexes are measured. Livers of the same part of each mouse are collected, and the content of liver glycogen is measured. The pancreas of each mouse was collected and fixed with 4% formaldehyde.
3. Observation index and measurement method
3.1 general State viewing
The general state, weight, food intake, water intake, etc. of each group of rats during the experiment were monitored, and daily food intake and water intake of each group of rats were recorded.
3.2 blood glucose and liver glycogen determination
Before blood glucose measurement, rats in each group were fasted without water control for 12 hours, blood was taken from the tail vein the next day, and fasting blood glucose values were measured with a glucometer and a York glucose test paper 1 time a week. The liver of the same part of each mouse is collected on the 28 th day of administration, and the glycogen content of the liver is measured according to the instruction of a liver glycogen kit.
3.3 determination of diabetes-related factors
Taking about 6ml of abdominal aorta blood, injecting into a clean test tube, centrifuging at 2000rpm for 10min at 4 ℃, separating serum, and strictly operating according to the specification of a corresponding kit, wherein a part of serum is used for measuring the contents of TC, TG, HDL-c and LDL-c, SOD, MDA in the serum; another part of serum was analyzed by radioimmunoassay to determine IL-1β, IL-6, TNF- α, INS, and C peptide contents, and Insulin Sensitivity Index (ISI) and insulin resistance Index (IR) were calculated by Homa Model method.
Isi=1/(fasting insulin level x fasting blood glucose level)
Ir=ln (fasting insulin level x fasting blood glucose level/22.5)
3.4 morphological observations of islet cells
Pancreas of each mouse was collected, fixed with 4% formaldehyde, and subjected to immunohistochemistry to observe morphology of islet cells.
4. Experimental results
4.1 model construction
Compared with the normal group, the rats in the model group showed lipid metabolism disorder after 4 weeks of continuous ig of the lipid emulsion: the TC, TG, FFA content is obviously increased (P is less than 0.01 or P is less than 0.05), and the characteristics similar to those of clinical type 2 diabetes patients are shown in table 1.
Table 1 influence of the composition on the lipid profile of normal ratsn＝12)
Note that: in comparison with the control group, # P＜0.05， ## P＜0.01。
4.2 Effect of the composition on the body weight of the rat model of type 2 diabetes
Compared with the normal control group, the weight of rats in the model control group is obviously reduced (P is less than 0.01) in weeks 0, 1, 2, 3 and 4; the composition-dosed groups had elevated body weight at weeks 0, 1, 2, 3, and 4 compared to the model control group, but no significant differences (P > 0.05), see fig. 1.
4.3 Effect of the composition on the model FBG and hepatic glycogen for diabetes type 2 in rats
After the sublingual iv 30mg/kg STZ appears on the basis of the blood lipid metabolic disorder of the rats in the model group, the blood glucose values of the rats in the model control group at weeks 0, 1, 2, 3 and 4 are obviously increased (P < 0.01) compared with the normal control group, and the liver glycogen content at week 4 is obviously reduced (P < 0.01) compared with the normal control group, which indicates that the establishment of the T2DM model of the rats is successful. Compared with the model control group, the compositions 500, 1000 and 2000mg/kg dose groups have obviously reduced FBG (P < 0.05 or P < 0.01) at 1, 2, 3 and 4 weeks after the medicine, but have no obvious change in liver glycogen content (P > 0.05) at 4 weeks, which suggests that the compositions can obviously improve hyperglycemia symptoms of T2DM rats, but have no obvious influence on liver glycogen content, as shown in tables 2 and 3.
Table 2 effects of compositions on blood glucose levels in a rat model of type 2 diabetes
Note that: # P＜0.05， ## p < 0.01vs normal group; * P＜0.05， ** model group with P < 0.01vs
Table 3 Effect of compositions on liver glycogen in rat type 2 diabetes model
## P < 0.01, vs normal group
4.4 Effect of the composition on lipid metabolism in the model of type 2 diabetes in rats
Compared with the normal group, the model control group TG, TC, LDL-c has obviously increased FFA content value (P < 0.05 or P < 0.01) and obviously decreased HDL-c content (P < 0.05); compared with the model control group, the Mongolian medicine composition 1000, the 2000mg/kg dose group TC, TG, LDL-c and the FFA content value are obviously reduced (P is less than 0.01 or P is less than 0.05); the HDL-c content value of the composition 1000 and 2000mg/kg dose group is obviously increased (P is less than 0.01), which shows that the composition can improve the blood lipid metabolism disorder of T2DM rats, and the blood lipid metabolism disorder is shown in Table 4.
Table 4 Effect of compositions on lipid metabolism in a model of type 2 diabetes in rats
# P＜0.05， ## P is less than 0.01, vs normal group; * P＜0.05， ** p < 0.01, vs model group
4.5 Effect of the composition on insulin secretion function and insulin resistance in the rat type 2 diabetes model
Compared with the normal group, the serum insulin and C-peptide contents and insulin secretion index ISI of the model control group are obviously reduced (P is less than 0.01), and Homa-IR is obviously increased (P is less than 0.01); the insulin and C-peptide content of the composition 1000, 2000mg/kg dose group was significantly increased (P < 0.05 or P < 0.01) compared to the model control group; the content of the metformin group C-peptide and ISI are obviously increased (P is less than 0.01), homa-IR is obviously reduced (P is less than 0.01), which proves that the composition can improve the functions of T2DM islet beta cells, promote insulin secretion and enhance the sensitivity of tissues to insulin. See table 5.
Table 5 Effect of compositions on insulin secretion function and insulin resistance in rat type 2 diabetes model
## P is less than 0.01, vs normal group; * P＜0.05， ** p < 0.01, vs model group
4.6 Effect of the composition on the serum SOD Activity and MDA content of T2DM rats
Compared with the normal group, the SOD activity of the model control group is obviously reduced (P is less than 0.01), and the MDA content is obviously increased (P is less than 0.01); the SOD activity of the compositions 1000, 2000mg/kg dose group and the metformin group were significantly improved (P < 0.05 or P < 0.01) and the MDA content of the compositions 500, 1000, 2000mg/kg dose group and the metformin group was significantly reduced (P < 0.05 or P < 0.01) compared with the model control group, as shown in Table 6.
TABLE 6 influence of the compositions on the SOD Activity and MDA content of serum of T2DM rats
## P is less than 0.01, vs normal group; * P＜0.05， ** p < 0.01, vs model group
4.7 Effect of the compositions on T2DM rat serum IL-1 beta, IL-6, TNF-alpha
Compared with the normal group, the IL-1 beta, IL-6 and TNF-alpha contents of the model control group are obviously increased (P is less than 0.05 or P is less than 0.01); the IL-6 content of the composition 1000, the dose group of 2000mg/kg and the metformin group is obviously reduced (P < 0.05 or P < 0.01) compared with a model control group; composition 1000, 2000mg/kg dose group and metformin group TNF-alpha content significantly reduced (P < 0.05 or P < 0.01); the IL-1β content tended to decrease, but was not significantly different, as shown in Table 7.
TABLE 7 Effect of compositions on T2DM rat serum IL-1 beta, IL-6, TNF-alpha
# P＜0.05， ## P is less than 0.01, vs normal group; * P＜0.05， ** p < 0.01, vs model group
4.8 Effect of the compositions on serum GHb content of T2DM rats
Compared with the normal group, the serum GHb content of the model control group is obviously increased (P is less than 0.01); compared with the model control group, the GHb content of the composition 1000, the 2000mg/kg dose group and the metformin group is obviously reduced (P < 0.05 or P < 0.01), and the GHb content is shown in Table 8.
Effect of table 8 composition on serum GHb content in T2DM rats
# P＜0.05， ## P is less than 0.01, vs normal group; * P is less than 0.05, ** p < 0.01, vs model group
4.9 Effect of the composition on CAT Activity and GSH-PX Activity of liver tissue of T2DM rat
Compared with the normal group, the liver tissue CAT and GSH-PX activities of the model control group are obviously reduced (P is less than 0.01); the CAT and GSH-PX activities of the 1000, 2000mg/kg dose groups were significantly increased (P < 0.05 or P < 0.01) and the GSH-PX activity of the metformin group was significantly increased (P < 0.05) compared to the model control group, as shown in Table 9.
Table 9 Effect of compositions on T2DM rat liver tissue CAT Activity and GSH-PX Activity
# P＜0.05， ## P < 0.01vs normal group; * P＜0.05， ** model group with P < 0.01vs
5.0 effects of the composition on pancreatic pathomorphology
Pancreatic HE staining results show that compared with a normal control group, islet atrophy of a model group is reduced, the number of the model group is reduced, endocrine cells are subjected to vacuolation degeneration, and punctate necrosis is visible; compared with the model group, the composition 1000, the 2000mg/kg dose group and the metformin group have the advantages that the number of islets is increased, the islet structure tends to be complete, and the damage of islets is better improved (see figures 2-4).
5.1 Effect of the compositions on insulin expression
The immunohistochemical result shows that compared with a normal control group, the positive expression quantity of the insulin in the model group is reduced, and the islet cells are subjected to cavitation denaturation; compared with the model group, the positive expression level of insulin in the composition 1000, the 2000mg/kg dose group and the metformin group is increased, and the islet structure tends to be complete (see figures 5-7).
The above results indicate that:
(1) the composition can obviously reduce the blood sugar of the T2DM rat, and the blood sugar value reduction is most obvious at the beginning of the 2 nd week after the administration;
(2) the composition can improve lipid metabolism disorder of T2DM rat;
(3) the composition can obviously raise the serum C-peptide level and insulin content of T2DM rats;
(4) the composition can obviously reduce the MDA content of serum of a T2DM rat and increase the SOD activity;
(5) the composition can obviously reduce the content of IL-6 and TNF-alpha in serum of a T2DM rat, and can lead the content of IL-1 beta to have a decreasing trend;
(6) the composition can obviously reduce the serum GHb content of the T2DM rat, which suggests that the composition not only can stably control the blood sugar level, but also can prevent the complications of type 2 diabetes;
(7) the composition can obviously raise the activity of CAT and GSH-PX of liver tissues of T2DM rats;
(8) the composition can improve pancreatic injury of T2DM rat.
In conclusion, the composition has the effect of reducing blood sugar. The mechanism of action may be related to the antioxidant, anti-inflammatory and lipid metabolism disorder regulating effects of the composition.
The embodiments described in the present specification are merely examples of implementation forms of the inventive concept, and the scope of protection of the present invention should not be construed as being limited to the specific forms set forth in the embodiments, and the scope of protection of the present invention and equivalent technical means that can be conceived by those skilled in the art based on the inventive concept. Although the following embodiments of the present invention are described, the present invention is not limited to the specific embodiments and fields of application described above, which are illustrative, instructive, and not limiting. Those skilled in the art, having the benefit of this disclosure, may effect numerous forms of the invention without departing from the scope of the invention as claimed.
Claims (9)
1. The composition for treating type 2 diabetes is characterized by being prepared from the following raw materials in parts by weight: 10-18 parts of bupleurum, 8-12 parts of cortex moutan, 8-16 parts of red paeony root, 15-24 parts of radix scrophulariae, 15-30 parts of dwarf lilyturf tuber, 5-15 parts of peach seed, 6-13 parts of dried rehmannia root, 6-15 parts of Chinese yam and 6-12 parts of hemerocallis.
2. The composition according to claim 1, wherein the composition is prepared from the following raw materials in parts by weight: 10 parts of bupleurum, 10 parts of cortex moutan, 10 parts of red paeony root, 15 parts of radix scrophulariae, 15 parts of radix ophiopogonis, 7.5 parts of peach kernel, 10 parts of radix rehmanniae, 9 parts of Chinese yam and 10 parts of hemerocallis.
3. A method of preparing the composition of claim 1, comprising the steps of:
weighing bupleurum, cortex moutan, red paeony root, radix scrophulariae, dwarf lilyturf tuber, peach kernel, dried rehmannia root, chinese yam and hemerocallis in parts by weight, mixing, extracting with 40-45% ethanol solution by mass fraction, and collecting an extracting solution to obtain the composition.
4. The method according to claim 3, wherein the extraction is performed by adding an ethanol solution with a mass fraction of 40-45% which is 8-10 times the total weight of the mixed medicinal materials, and reflux-extracting for 2-3 times, each time for 2.5-3 hours.
5. A method according to claim 3, wherein the composition is obtained by concentrating the extract and drying.
6. Use of the composition of claim 1 for the preparation of a medicament for the treatment of type 2 diabetes.
7. The use according to claim 6, wherein the composition is for the preparation of a hypoglycemic agent.
8. The use according to claim 6, wherein the composition is for the preparation of a medicament for improving insulin metabolism in vivo.
9. A pharmaceutical formulation for the treatment of type 2 diabetes comprising the composition of claim 1 and a pharmaceutically acceptable adjuvant.
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