CN101649004B - Marine oligosaccharide compound with type II diabetes resisting activity and preparation method thereof - Google Patents
Marine oligosaccharide compound with type II diabetes resisting activity and preparation method thereof Download PDFInfo
- Publication number
- CN101649004B CN101649004B CN2009101777100A CN200910177710A CN101649004B CN 101649004 B CN101649004 B CN 101649004B CN 2009101777100 A CN2009101777100 A CN 2009101777100A CN 200910177710 A CN200910177710 A CN 200910177710A CN 101649004 B CN101649004 B CN 101649004B
- Authority
- CN
- China
- Prior art keywords
- diabetes
- chromium
- type
- control group
- pmc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The invention relates to a marine oligosaccharide compound with type II diabetes resisting activity, which is a compound which is formed by following steps: taking D-polymannose aldehydic oligosaccharide which comes from sea and contains carboxyl in each oligosaccharide ring, reacting with dilute alkali and chromium salt solution and introducing trivalent chromium ions which is closely relative to the generation and the development of diabetes into oligosaccharide molecules. A pharmacological experiment proves that the product has remarkable effect on accelerating insulin secretion, is not influenced by amylin and has effects on lightening glucose load, improving blood-lipoid metabolism, insulin sensitivity and certain kidney protection and lightening pancreatic injury for type II diabetes rats and mouse. The product of the invention comes from marine natural species, has the advantages of good security, unique structure, low molecular weight, high chromium binding ratio, good oral absorption effect, and the like, can play a hypoglycemic role in a plurality of links and has favorable market application prospect on the aspect of preventing and treating type II diabetes.
Description
Technical field
The present invention relates to a kind of compound with anti-diabetic activity.Specifically, relate to a kind of POLYMANNURONATE chromic compound that derives from the ocean brown alga and preparation method thereof and the application in anti-type ii diabetes.
Background technology
Mellitus are common endocrine metabolism property diseases of a kind of multi-pathogenesis.According to IDF's statistics in 2007, global diabetic subject's total number of persons reached 2.46 hundred million, and wherein type ii diabetes accounts for more than 90% of total glycosuria patient.According to estimates, global PM has 6 people to die from mellitus approximately, and the number dead because of mellitus and complication thereof is only second to cancer and cardiovascular and cerebrovascular diseases, has become the third-largest killer who influence human life's health.At present, the antidiabetic medicine of using clinically mainly contains medicines such as biguanides, sulfourea, alpha-glucosidase inhibitor and thiazolidinediones.The mechanism of action of these medicines has nothing in common with each other: biguanides mainly is through decomposition that suppresses liver glycogen and increase peripheral tissues blood sugar reducing function to be brought into play in the utilization of glucose; Sulfonylurea drugs has the effect of insulin secretion accelerating, puts on weight and is prone to spinoff such as generation hypoglycemia but exist; Alpha-glucosidase inhibitor is the absorption that comes delay glucose through the activity of epichorial starch of competitive inhibition small intestine epithelium and disaccharide-hydrolysing enzymes, is prone to cause GI untoward reaction; The thiazolidinediones medicine has the effect that increases insulin sensitivity, but is difficult to bring into play the ideal curative effect for the patient of insulin function defective.All in all, though above-mentioned antidiabetic medicine has significantly hypoglycemic effect, they have in various degree toxic side effect and untoward reaction mostly, or have the comparatively single shortcoming of the mode of action.Therefore, the newtype drug of research and development with good hypoglycemic activity is still pressing for clinically.
Big quantity research shows that chromium has the glycometabolic effect of obvious improvement, and trivalent chromium is the important component that constitutes glucose tolerance factor (GIF), also is that Regular Insulin is normally brought into play the necessary cofactor of physiological function.Chromium plays a role through combine formation GTF to work in coordination with Regular Insulin with nicotinic acid or amino acid, and the transphosphorylase and the succinodehydrogenase that can act in the glucose metabolism increase sugared utilization.The activity of the content of chromium and Regular Insulin is proportionate in the serum, when chromium deficiency, can cause sugar tolerance impaired, and the active and tissue of Regular Insulin descends to the susceptibility of Regular Insulin, can cause carbohydrate metabolism disturbance when serious, causes or increase the weight of mellitus.In addition, chromium has also been participated in the regulating effect of metabolism of fat, Proteometabolism and nucleic acid metabolism, also can cause hyperlipidaemia, atherosclerotic lesion and growth retardation etc. during chromium deficiency.Therefore, trivalent chromium is for mellitus, and the prevention and the treatment of the type ii diabetes that is especially caused by insulin resistant etc. have great importance.Chrome uptake and its chemically bound form are closely related, and inorganic chromium not only toxicity is big, and specific absorption extremely low (＜0.5%), but the specific absorption of organic chromium such as CrP, nicotinic acid chromium etc. can reach 10-25%.Toxicologic study result shows that trivalent chromium can not see through red cell membrane, and its toxicity is far below sexavalent chrome, and chromic toxic dose is bigger at least 1000 times than safety intake every day of recommending, thereby is one of element the safest in the micro elements needed by human.
The research and development of carbohydrate medicine are one of the focuses of biochemical drug research field in this century.At present existing more saccharide compound has demonstrated good prospects for application aspect the preventing and treating of mellitus; As discover the squash polyoses and the bitter melon polysaccharide of plant origin; The tremella polysaccharide of originated from fungus, ganoderan and Cordyceps polysaccharide; Chitosan, algal polysaccharide and the spirulina polysaccharide etc. in marine animal and plant source all have hypoglycemic activity in various degree; But, thereby and exist shortcomings such as oral absorption is poor, bioavailability is low to limit its application clinically at present because the action target spot of these polysaccharide is indeterminate.As to polysaccharide compound; Particularly derive from the polysaccharide compound of ocean; Carry out suitable degraded and obtain oligose or the oligosaccharide compound that structure is clear and definite, oral absorption is good; And carry out rational molecular modification according to the morbidity mechanism of action of mellitus, and will improve the hypoglycemic effect of saccharide compound greatly, be expected to become one type of novel antidiabetic medicine.
Summary of the invention
The objective of the invention is with the POLYMANNURONATE oligosaccharides that derives from the ocean brown alga is molecular skeleton; Itself and trivalent chromium are carried out ligand complex; So that a kind of marine oligosaccharide chromic compound with good hypoglycemic activity to be provided, satisfy the needs that prevent and treat mellitus clinically.
A kind of ocean POLYMANNURONATE oligosaccharide chromic compound; The saccharide residue that it is characterized in that it is made up of the D-mannuronic acid, and with β-1, the 4-glycosidic link connects; Carry out the ligand complex preparation with trivalent chromic ion; The quality percentage composition of chromium is 0.1-15%, and weight-average molecular weight is≤10kD that its molecular formula can be expressed as [C
6H
8-mO
6Cr
m]
n, m=0.1-1 wherein, n=1-50.
The preparation method of a kind of ocean POLYMANNURONATE oligosaccharide chromic compound is characterized in that POLYMANNURONATE solution under pH=5-12 and temperature 45-75 ℃ condition, dropwise adding an amount of diluted alkaline and chromium salt solution; Insulation reaction 1h filters, with the organic solvent deposit of 2-4 times of volume; Leave standstill; Collecting precipitation, with organic solvent washing, dehydration, drying under reduced pressure promptly gets.
A kind of ocean POLYMANNURONATE oligosaccharide chromic compound is characterized in that being used for the prevention and the treatment of type ii diabetes.
Chromic salts of the present invention is meant chromium chloride, chromium acetate and chromium nitrate; Described diluted alkaline is meant dilute sodium hydroxide, Pottasium Hydroxide, yellow soda ash, salt of wormwood, sodium hydrogencarbonate, saleratus.
The present invention is a raw material with this sea life oligosaccharides that in each sugar encircles, all contains carboxyl of POLYMANNURONATE; With itself and mellitus take place and evolution in closely-related trivalent chromic ion carry out ligand complex, obtained a kind of active marine oligosaccharide compound of good resistance type ii diabetes that has.Compared with similar products, rule of origin Yu Haiyang natural biological of the present invention, it is good to have a security; Characteristics such as structure is unique, molecular weight is low, and the chromium combination rate is high, and oral absorption is good; And can be at a plurality of links performance blood sugar reducing functions, aspect the preventing and treating of type ii diabetes, have better market prospect.
Embodiment
Come the present invention is done further explanation through some concrete embodiment below.
Embodiment 1
Taking by weighing 10g POLYMANNURONATE (weight-average molecular weight is 7350 dalton) is dissolved in the 500ml pure water; Regulate pH=10 with sodium hydroxide solution; In 45 ℃ of water-baths, dropwise add 2mol/L sodium hydroxide and 2mol/L chromium chloride solution; When just green flocks having occurred in the solution, stop to add sodium hydroxide and chromium chloride, continue insulation 1h.Take out reaction solution, filter, filtrating is concentrated into 95% ethanol that adds 3 times of volumes behind the 200mL, deposition leaves standstill, and collecting precipitation with the absolute ethyl alcohol dehydration, at 45 ℃ of following drying under reduced pressure, obtains chrome content and is about 15% POLYMANNURONATE chromic compound.
Embodiment 2
Take by weighing 10g POLYMANNURONATE (weight-average molecular weight is 5200 dalton) and be dissolved in the 500ml pure water, regulate pH=8, in 70 ℃ of water-baths, dropwise add 2mol/L yellow soda ash and 2mol/L chromium acetate solution 1mL, insulation reaction 1h with sodium carbonate solution.Take out reaction solution, filter, filtrating is concentrated into the acetone that adds 3 times of volumes behind the 200mL, deposition leaves standstill, and collecting precipitation with the acetone dehydration, at 45 ℃ of following drying under reduced pressure, obtains chrome content and is about 1% POLYMANNURONATE chromic compound.
Embodiment 3
Take by weighing 10g POLYMANNURONATE (weight-average molecular weight is 1500 dalton) and be dissolved in the 500ml pure water, regulate pH=5, in 60 ℃ of water-baths, dropwise add 2mol/L sodium hydroxide and 2mol/L chromium nitrate solution 5mL, insulation reaction 1h.Take out reaction solution, filter, filtrating is concentrated into the acetone that adds 4 times of volumes behind the 200mL, deposition leaves standstill, and collecting precipitation with the acetone dehydration, at 45 ℃ of following drying under reduced pressure, obtains chrome content and is about 5% POLYMANNURONATE chromic compound.
Embodiment 4
Take by weighing 10g POLYMANNURONATE (weight-average molecular weight is 3000 dalton) and be dissolved in the 500ml pure water, regulate pH=9, in 55 ℃ of water-baths, dropwise add 2mol/L salt of wormwood and 2mol/L chromium chloride solution 10mL, insulation reaction 1h with solution of potassium carbonate.Take out reaction solution, filter, filtrating is concentrated into 95% ethanol that adds 4 times of volumes behind the 200mL, deposition leaves standstill, and collecting precipitation with the absolute ethyl alcohol dehydration, at 45 ℃ of following drying under reduced pressure, obtains chrome content and is about 10% POLYMANNURONATE chromic compound.
Embodiment 5
Take by weighing 10g POLYMANNURONATE (weight-average molecular weight is 9500 dalton) and be dissolved in the 500ml pure water, regulate pH=11, in 50 ℃ of water-baths, dropwise add 2mol/L Pottasium Hydroxide and 2mol/L chromium acetate solution 3mL, insulation reaction 1h with potassium hydroxide solution.Take out reaction solution, filter, filtrating is concentrated into the acetone that adds 2 times of volumes behind the 200mL, deposition leaves standstill, and collecting precipitation with the acetone dehydration, at 45 ℃ of following drying under reduced pressure, obtains chrome content and is about 3% POLYMANNURONATE chromic compound.
Embodiment 6
Take by weighing 10g POLYMANNURONATE (weight-average molecular weight is 2400 dalton) and be dissolved in the 500ml pure water, regulate pH=7, in 65 ℃ of water-baths, dropwise add 2mol/L sodium hydrogencarbonate and 2mol/L chromium nitrate solution 0.5mL, insulation reaction 1h with sodium hydrogen carbonate solution.Take out reaction solution, filter, filtrating is concentrated into the acetone that adds 4 times of volumes behind the 200mL, deposition leaves standstill, and collecting precipitation with the acetone dehydration, at 45 ℃ of following drying under reduced pressure, obtains chrome content and is about 0.5% POLYMANNURONATE chromic compound.
The ocean POLYMANNURONATE oligosaccharide chromic compound of the present invention's preparation is amorphous powder of light green to deep green or crystal; Water-soluble; Be insoluble to organic solvents such as ethanol, acetone, EC, the quality percentage composition of chromium is 0.1-15%, weight-average molecular weight≤10kD.
The pharmacodynamic experiment of the anti-type ii diabetes of ocean POLYMANNURONATE oligosaccharide chromic compound (PMC)
1PMC is to the promoting insulin secretion research of islet cells
Be below weight-average molecular weight be the PMC of 3000Da under different concns, there are down the result of study that influences to the PMC promoting insulin secretion in the influence of RIN-5F islet cells insulin secretion effect and pancreas opsonin.
Experimental technique: islet cells RIN-5F is inserted in 96 orifice plates every hole about 2 * 10
5Individual cell is at the CO of 37 ℃ of temperature and 5%
2Cultivate 72h in the incubator.Change the RPMI RPMI-1640 and cultivate 24h again, remove nutrient solution, after cell is washed with fresh medium, add high concentration glucose solution, add the PMC sample solution of different concns again through the sterilization of 0.22 μ m membrane filtration.With the pure water is blank control group, with the positive control group of the U26452 of 1mmol/L, at the CO of 37 ℃ of temperature and 5%
2Cultivate 3h in the incubator.Get each hole nutrient solution 300 μ L, the centrifugal cell of removing with 100 times of nutrient solution dilutions, adopts ELISA kit measurement concentration of insulin with supernatant, investigates the promoting insulin secretion of PMC to islet cells.
Other gets the RIN-5F islet cells with after the fresh medium washing, and each pancreas opsonin that adds 100 μ mol/L is hatched 30min.Except that the pure water blank control group, each group adds the PMC solution of high concentration glucose solution and 100 μ mol/L, with the positive control group of the U26452 of 100 μ mol/L, at the CO of 37 ℃ of temperature and 5%
2Cultivate 3h in the incubator.Get each hole nutrient solution 300 μ L, the centrifugal cell of removing with 100 times of nutrient solution dilutions, adopts ELISA kit measurement concentration of insulin with supernatant, investigates high density pancreas opsonin and has down the influence to the PMC promoting insulin secretion.
Experimental result: (1) is compared with the blank group, and high sugar group secretion of insulin amount significantly increases, and positive controls secretion of insulin amount has the increase of highly significant, shows that experimental model is credible.PMC promptly has the effect of significant stimulation islet cells excreting insulin under 30 μ mol/L concentration, and along with the increase of PMC concentration, it stimulates the effect of RIN-5F islet cells excreting insulin to strengthen, and is dose-dependence.
(2) in the positive control group of U26452, through the RIN-5F islet cells that the pancreas opsonin was hatched, its secretion of insulin amount obviously reduces.But in the PMC experimental group, before and after the pancreas opsonin is hatched the RIN-5F islet cells, secretion of insulin amount no significant difference.
Experiment conclusion: PMC and U26452 all have the effect of tangible stimulation islet cells excreting insulin.The existence of high density pancreas opsonin can obviously reduce the promoting insulin secretion of U26452, but the promoting insulin secretion of PMC not had obvious influence.
The pancreas opsonin is by a kind of normal hormone of islet cells excretory, and its secretion and metabolism occur unusually under pathological conditions, can assemble forming the formation that fibril is participated in islet amyloid appearance proteinosis.The deposition of pancreas opsonin in pancreas islet, making beta cell generation apoptosis and then causing secretion of insulin to reduce is the typical pathogenesis of type ii diabetes, the pancreas opsonin plays an important role in the generation of type ii diabetes and evolution.The pancreas opsonin obviously reduces the promoting insulin secretion of U26452 through the deposition at cell surface, and this is one of reason of sulfonylurea drugs secondary failure.This experiment is found; The existence of high density pancreas opsonin does not have obvious influence to the promoting insulin secretion of PMC; This shows that PMC can be through disturbing its deposition at cell surface with the interaction of pancreas opsonin; The toxicity that opposing pancreas opsonin produces islet cells, thus in the prevention of type ii diabetes and treatment, demonstrated good prospects for application.
2PMC is to the therapeutic action of Wistar type ii diabetes rat model
Experimental technique: get 112 of Wistar rats, male and female half and half, body weight 200～250g.Animal is raised in advance and is used for test after 7 days.12 of picked at random are as the normal control group, and male and female half and half are fed with basal feed, ad lib, and all the other 100 rats feed with high glucose and high fat feed (in the basal feed with sucrose 10%, lard 10%, SUV 5%) and raised for 4 weeks, freely drink water simultaneously.After 4 weeks, hello high glucose and high fat feed rat is all injected low dose of streptozotocin (STZ) and induces mellitus, presses 30mgkg
-1Once abdominal cavity injection 1%STZ solution faces with preceding with citric acid-Trisodium Citrate BP damping fluid preparation.The mouse tail is got hematometry and is respectively organized rat fasting blood-glucose behind the 72h, and blood sugar >=11.1mmol/L person can think that modeling is successful.The rat successful to modeling is divided into 5 groups at random by body weight, is respectively model control group, positive controls, the basic, normal, high dose groups of PMC.More than the administration respectively of each treated animal, normal control group and model control group rat give zero(ppm) water 10mlkg
-1D
-1, positive controls gives Dimethyldiguanide hydrochloride enteric solubility tablet 150mgkg
-1D
-1, the basic, normal, high dose groups of PMC gives PMC solution respectively, and dosage is respectively 17mgkg
-1D
-1, 35mgkg
-1D
-1, 70mgkg
-1D
-1, administration volume 10mlkg
-1, 1 time/d, successive administration 30 days.2h measures blood sugar behind the 1st day medicine of administration; Observe whether have acute blood sugar reducing function; The set time was measured body weight and food ration weekly after administration began; Administration was measured fasting plasma glucose and FPI in the 15th day and the 30th day, carried out oral glucose tolerance test, calculated area (AUC) and insulin sensitivity index (ISI) under the glucose tolerance curve.1h behind the last medicine; With 3% vetanarcol anesthetized rat; Abdominal aortic blood; Centrifuging and taking serum, the content of free lipid acid (FFA), triglyceride level (TG), BSA (ALB), total protein (TP), total cholesterol (CHOL), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) in the mensuration rat blood serum.Put to death animal, separate liver, kidney, pancreas, spleen, heart, weigh, and calculate organ coefficient, with animal kidney, pancreas, fix with 4% formaldehyde solution, cut into slices behind the paraffin embedding, histopathologic examination is carried out in H-E dyeing.
Symptoms such as experimental result: (1) duration of test animal general state and changes of weight situation: the hair color tarnish appears in the model control group animal, and hydrouria is few moving, and it is for sleeping in to curl up.Positive controls, PMC are basic, normal, high, and the dose groups animal also has similar performance, but its symptom has clear improvement than model control group; Each treated animal changes of weight there was no significant difference (P＞0.05).
(2) food ration changes: duration of test, each treated animal in different times, food ration there was no significant difference (P＞0.05).Administration 3 and 4wk, the average food ration of the basic, normal, high dose groups of PMC is low than positive controls and model control group.
(3) acute blood sugar reducing function test: the preceding positive controls of medicine, basic, normal, high each group of PMC and model control group be unknown significance difference (P＞0.05) relatively, and respectively organizing blood sugar behind the medicine has certain reduction trend with the dosage increase.
(4) oral glucose tolerance test: administration the 15th day, model control group animal fasting plasma glucose is higher, give glucose after, blood sugar increasing is obvious, AUC obviously increases than the blank group; Positive control treated animal fasting plasma glucose is low than model control group; Blood sugar also has rising after giving glucose; But each time point blood glucose value and AUC are starkly lower than model control group, and 30 relatively have significant difference (P＜0.05) with 60min blood sugar and AUC and model control group; PMC low dose group animal fasting plasma glucose is consistent with positive controls with blood sugar increasing trend, and is low than model control group, but its numerical value compares no difference of science of statistics a little more than positive controls with model control group; Increase with dosage, the middle and high dose groups blood sugar of PMC reduces effect and weakens each time point blood glucose value of high dose group and AUC and model control group basically identical.The model control group animal raises than blank group FPI content, and insulin sensitivity index reduces; The positive control treated animal reduces than model control group FPI content, and insulin sensitivity index raises, and the insulin sensitivity index statistical analysis has significant difference (P＜0.05); PMC low dose group animal is similar with positive controls, with model control group tangible reduction FPI content, the effect of rising insulin sensitivity index is arranged relatively, and significant difference (P＜0.05) is arranged; Increase with dosage, the middle and high dosage of PMC reduces FPI content, the effect of rising insulin sensitivity index weakens.Administration the 30th day, the 15th day result is consistent with administration; Positive controls and PMC low dose group animal fasting blood sugar with give the blood sugar increasing that glucose causes and be starkly lower than model control group; PMC increases the effect of inhibition blood sugar increasing with dosage and obviously weakens.Positive controls and PMC are low, the high dose group insulin sensitivity index is high than model control group, and statistical analysis has significant difference (P＜0.05).The high dose group insulin sensitivity index is relatively higher than low dose group and middle dose groups, does not increase trend but dose-dependently appears in basic, normal, high dose groups.
(5) PMC is to the influence of rat fat: administration 30d; Model control group, positive controls, the basic, normal, high dose groups FFA of PMC and TG all increase than blank control group, explain that basic, normal, high dose groups all exists insulin resistant in model control group, positive controls, PMC.TP there was no significant difference between each group of model control group, positive controls, the basic, normal, high dose groups of PMC, blank group, and model control group, the middle and high dose groups ALB of PMC reduce than the blank group, the prompting renal function has to a certain degree damage.Model control group, positive controls, the basic, normal, high dose groups CHOL of PMC, LDL-C all increase than the blank group, and along with the increase of PMC dosage, CHOL, LDL-C have increase trend.There was no significant difference between each group of HDL-C.
(6) PMC is to the influence of Rats Organs and Tissues coefficient: administration finishes back model control group, positive controls, the basic, normal, high dose groups liver of PMC coefficient to be increased than blank control group; PMC low dose group liver coefficient is relatively low, with model control group significant difference (P＜0.05) is arranged.Heart coefficient, spleen coefficient, pancreas coefficient are respectively organized there was no significant difference.Kidney coefficient model control group, positive controls, PMC are basic, normal, high, and dose groups increases than blank control group, and blank control group left and right sides kidney coefficient and model control group relatively have significant difference (P＜0.05).With the increase of PMC dosage, liver coefficient, pancreas coefficient, left and right sides kidney coefficient have increase trend.
(7) pathological examination result: 1. the boundary of blank group rat kidney skin, medullary substance is clear, and each segment structure of uriniferous tubules is clear, interstitial fibers tissue less, renal glomerulus is evenly distributed, volume is normal, it is narrow that glomerular capsule does not have adhesion; Mostly pancreas islet is circular or oval-shaped cell mass, is scattered between the pancreatic acini, and quantity is more, and the boundary of pancreas islet is clear, no coating, and cell mass not of uniform size, the kytoplasm of islet cells is abundant, and light the dying of endochylema is baby pink, examines to be mostly circular engrain; 2. the model control group rat is compared with the blank control group rat, kidney renal cells vacuolar degeneration, and individually with protein cast, renal cortex internal glomerulus capillary loops reduces, and matrix increases, and the renal capsule blister cavities increases; The pancreas islet volume reduces in the pancreas, and quantity reduces, and boundary is unclear, and islet cells quantity also reduces, cellular swelling, kytoplasm understain, karyopyknosis; 3. also there is kidney renal tubular epithelial vacuolar degeneration in the positive controls rat, and the glomerular capillary loop reduces, and matrix increases, and the renal capsule blister cavities increases, and alleviates but compare degree with the model control group rat; Pancreas islet quantity reduces in the pancreas, and islet cells quantity reduces, cellular swelling, and the kytoplasm understain, karyopyknosis is not seen notable difference with the model control group rat; 4. low dose group rat, kidney renal tubular epithelial vacuolar degeneration, cortex internal glomerulus capillary loops reduces, matrix increases, the renal capsule blister cavities increases, but with the model control group rat relatively degree alleviate, do not see protein cast in the uriniferous tubules; Pancreas islet quantity reduces in the pancreas, and islet cells quantity reduces, cellular swelling, and the kytoplasm understain, karyopyknosis is not seen notable difference with the model control group rat; 5. middle dose groups rat, kidney do not see obviously unusual, do not see notable difference with the blank control group rat; Pancreas islet quantity increases than model control group to some extent in the pancreas, but islet cells quantity is still less, cellular swelling, kytoplasm understain, karyopyknosis; 6. high dose group rat, kidney do not see obviously unusual, do not see notable difference with the blank control group rat; Pancreas islet quantity increases in the pancreas, and islet cells quantity increases, and the cellular swelling degree alleviates, and compares with the model control group rat, and injury of pancreas alleviates.
Experiment conclusion: under this test conditions; PMC does not have obvious influence to type ii diabetes rat body weight and food ration, does not have acute blood sugar reducing function, the danger that does not exist the property a crossed blood sugar to reduce; PMC can alleviate glucose load; Improve blood lipid metabolism, improve type ii diabetes rat insulin Sensitivity Index through reducing FFA, and have certain kidney protection and alleviate the injury of pancreas effect.
3PMC is to the therapeutic action of KM type ii diabetes model mice
Experimental technique: get 112 of KM mouse, male, body weight 18～22g.12 mouse of picked at random feed with basal feed as the normal control group, freely drink water, and all the other 100 mouse feed with high glucose and high fat feed (containing sucrose 10%, lard 10%, SUV 5%) and raised for 6 weeks, freely drink water simultaneously.After 6 weeks, feed high glucose and high fat feed mouse peritoneal injection 150mgkg
-1The STZ of body weight divides to give for 5 times, and continues to give the high glucose and high fat feed after 1 week, measures and respectively organizes the mouse fasting plasma glucose, and concentration>=11.1mmol/L person is as the type ii diabetes mouse model.The mouse successful to modeling is divided into 5 groups at random by body weight, is respectively model control group, positive controls, the basic, normal, high dose groups of PMC, and not doing 12 mouse of model is the normal control group.Normal control group and model control group mouse give zero(ppm) water 20mlkg
-1D
-1, positive controls gives Dimethyldiguanide hydrochloride enteric solubility tablet 225mgkg
-1D
-1, the basic, normal, high dose groups of PMC gives PMC solution respectively, and dosage is respectively 25mgkg
-1D
-1, 50mgkg
-1D
-1, 100mgkg
-1D
-1, administration volume 20mlkg
-1, 1 time/d, successive administration 30 days.2h measures blood sugar behind the 1st day medicine of administration, observes whether to have acute blood sugar reducing function.1h behind the last medicine; The tail intravenous needle is adopted the hematometry fasting plasma glucose; Extract eyeball of mouse again and get blood; Centrifuging and taking serum, the content of Regular Insulin, free fatty acids (FFA), triglyceride level (TG), BSA (ALB), total protein (TP), total cholesterol (CHOL), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) in the mensuration mice serum.With animal kidney, pancreas, fix with 4% formaldehyde solution, to cut into slices behind the paraffin embedding, histopathologic examination is carried out in H-E dyeing.
Experimental result: (1) acute blood sugar reducing function test-results: before the blank group medicine with medicine after 2h blood sugar and model control group relatively have significant difference (P＜0.05); 2h blood sugar and model control group relatively have certain reduction trend after positive controls, the basic, normal, high dose groups animal drugs of PMC; But compare there was no significant difference with model control group; Each organizes before and after the administration own control relatively, and the N1,N1-Dimethylbiguanide group reduces 0.72mmol/L before than medicine to be had significant difference (P＜0.05), low dose group and reduce 0.46mmol/L before than medicine and have significant difference (P＜0.05), middle dose groups and reduce 0.59mmol/L, high dose group before than medicine and reduce 0.37mmol/L before than medicine.
(2) each dose groups animal fasting blood sugar dose-dependently of fasting plasma glucose and FPI: PMC raises behind the last medicine; Positive controls, low, the middle dose groups fasting blood sugar of PMC are compared with model control group all and are reduced, and have significant difference (P＜0.05), but still are higher than the blank group, and high dose group and model control group be unknown significance difference (P＞0.05) relatively.
Positive controls, the basic, normal, high dose groups FPI of PMC level all are lower than the blank group; More all have significant difference (P＜0.05) with the blank group, positive controls, the basic, normal, high dose groups FPI of PMC and model control group be unknown significance difference (P＞0.05) relatively.
Positive controls, low, the middle dose groups animal insulin of PMC Sensitivity Index raise than model control group, have significant difference (P＜0.05).Positive controls insulin sensitivity index and blank group be there was no significant difference (P＞0.05) relatively, and model control group, the basic, normal, high dose groups of PMC are all low than the blank group, have significant difference (P＜0.05).
(3) PMC significantly raises to the influence of KM mouse blood fat: model control group FFA, TG, LDL-C, CHOL, and HDL-C significantly reduces, and relatively has significant difference (P＜0.05) with the blank group, and the hints model control group exists hyperlipidaemia and insulin resistant.Model control group TP and blank group be there was no significant difference relatively, but ALB points out renal function possibly have certain damage than the reduction of blank group significance.Compare with model control group, positive controls, low dose group FFA reduce, and have significant difference (P＜0.05), and the effect of prompting insulin resistant reduces.Basic, normal, high each the dose groups TG of PMC all reduces (P＜0.05) than the model control group significance; Low, the middle dose groups CHOL of PMC reduces (P＜0.05) than the model control group significance; In addition; PMC low dose group HDL-C raises than the model control group significance, the effect of pointing out the PMC tool to have some improvement blood lipid metabolism.
(4) pathological examination result: 1. blank group, kidney skin, medullary substance boundary are clear, each segment structure of uriniferous tubules is clear, interstitial fibers tissue less, renal glomerulus is evenly distributed, volume is normal, it is narrow that glomerular capsule does not have adhesion.Mostly pancreas islet is circular or oval-shaped cell mass, is scattered between the pancreatic acini, and quantity is more, and the boundary of pancreas islet is clear, no coating, and cell mass not of uniform size, the kytoplasm of islet cells is abundant, and light the dying of endochylema is baby pink, examines to be mostly circular engrain; 2. model control group compares kidney renal cells vacuolar degeneration with blank control group; The pancreas islet volume reduces, and quantity reduces, and it is sparse to distribute, islet cells swelling, and kytoplasm is painted shallow, and cavity increases, karyopyknosis; 3. positive controls, kidney are not seen obviously unusual; Pancreas islet and model control group relatively volume increase, and quantity increases, and islet cells swelling alleviates; 4. low dose group, kidney are not seen obviously unusual; Pancreas islet and model control group relatively volume increase, and quantity increases, and islet cells swelling alleviates; 5. middle dose groups and high dose group, kidney are not seen obviously unusual; The pancreas islet volume increases, and quantity increases, and compares with the model control group rat, and injury of pancreas recovers to some extent.
Experiment conclusion: under this test conditions; PMC does not have acute blood sugar reducing function to the type ii diabetes mouse; The danger that does not exist the property a crossed blood sugar to reduce, PMC can improve type ii diabetes mouse blood lipid metabolism, reduces the plain opposing of type ii diabetes mouse islets; Have the effect of certain enhancing insulin sensitivity, and have certain kidney protection and alleviate the injury of pancreas effect.
Claims (4)
1. one kind has the active marine oligosaccharide compound of anti-type ii diabetes, it is characterized in that its saccharide residue is made up of the D-mannuronic acid; And with β-1; The 4-glycosidic link connects, and carries out the ligand complex preparation with trivalent chromic ion, and the quality percentage composition of chromium is 0.1-15%; Weight-average molecular weight≤10kD, its molecular formula is expressed as [C
6H
8-mO
6Cr
m]
n, m=0.1-1 wherein, n=1-50.
2. the described preparation method with the active marine oligosaccharide compound of anti-type ii diabetes of claim 1 is characterized in that, with POLYMANNURONATE solution under pH=5-12 and temperature 45-75 ℃ condition; Dropwise add an amount of diluted alkaline and chromium salt solution, insulation reaction 1h filters; Organic solvent deposit with 2-4 times of volume leaves standstill, collecting precipitation; With organic solvent washing, dehydration, drying under reduced pressure promptly gets.
3. the described preparation method with the active marine oligosaccharide compound of anti-type ii diabetes of claim 2 is characterized in that chromic salts is meant chromium chloride, chromium acetate or chromium nitrate.
4. the described preparation method with the active marine oligosaccharide compound of anti-type ii diabetes of claim 2 is characterized in that, diluted alkaline is dilute sodium hydroxide, Pottasium Hydroxide, yellow soda ash, salt of wormwood, sodium hydrogencarbonate or saleratus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101777100A CN101649004B (en) | 2009-09-18 | 2009-09-18 | Marine oligosaccharide compound with type II diabetes resisting activity and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101777100A CN101649004B (en) | 2009-09-18 | 2009-09-18 | Marine oligosaccharide compound with type II diabetes resisting activity and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101649004A CN101649004A (en) | 2010-02-17 |
| CN101649004B true CN101649004B (en) | 2012-08-22 |
Family
ID=41671346
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009101777100A Active CN101649004B (en) | 2009-09-18 | 2009-09-18 | Marine oligosaccharide compound with type II diabetes resisting activity and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101649004B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103059154B (en) * | 2012-11-29 | 2015-03-11 | 滨州学院 | Fungal glucan oligomer chrome complex and preparation method thereof |
| CN106349298B (en) * | 2015-07-17 | 2021-02-09 | 上海绿谷制药有限公司 | Application of algin oligosaccharide and derivatives thereof in improving sleep disorder |
| CN108164613A (en) * | 2018-01-22 | 2018-06-15 | 中国海洋大学 | A kind of preparation and its application with the green algae polysaccharide chromium for preventing diabetes |
| CN110812364A (en) * | 2019-10-22 | 2020-02-21 | 中国海洋大学 | Application of galactooligosaccharide and derivatives thereof as SGLTs inhibitor |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1289936B1 (en) * | 2000-06-02 | 2004-10-20 | Telik, Inc. | Substituted stilbenes as glucose uptake enhancers |
| CN1557301A (en) * | 2004-01-15 | 2004-12-29 | 高春平 | Compositions for senile diabetes patient blood sugar and blood fat reduction |
| WO2007060924A1 (en) * | 2005-11-22 | 2007-05-31 | Ajinomoto Co., Inc. | PROTECTING AGENT FOR PANCREATIC β-CELL |
-
2009
- 2009-09-18 CN CN2009101777100A patent/CN101649004B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1289936B1 (en) * | 2000-06-02 | 2004-10-20 | Telik, Inc. | Substituted stilbenes as glucose uptake enhancers |
| CN1557301A (en) * | 2004-01-15 | 2004-12-29 | 高春平 | Compositions for senile diabetes patient blood sugar and blood fat reduction |
| WO2007060924A1 (en) * | 2005-11-22 | 2007-05-31 | Ajinomoto Co., Inc. | PROTECTING AGENT FOR PANCREATIC β-CELL |
Non-Patent Citations (1)
| Title |
|---|
| EP1289936B1B1 2004.10.20 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101649004A (en) | 2010-02-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR100674672B1 (en) | Chitosan-containing polysaccharide, process for producing the same and use thereof | |
| CN101597389B (en) | Mixture of momordica saponins and polysaccharides and extraction process and application thereof | |
| CN102860451B (en) | Composite with function of assisting in decreasing blood glucoses and products thereof | |
| CN101649004B (en) | Marine oligosaccharide compound with type II diabetes resisting activity and preparation method thereof | |
| CN102994305B (en) | Method for preparing health-care food (therapy) product (nutrient juice wine) by use of extracts from cordyceps militaris and cocoon | |
| CN101229316A (en) | Rhizoma anemarrhenae extrac and applications as type 2 diabetes-curing medicine thereof | |
| Eleazu et al. | Biochemical basis of the use of cocoyam (Colocassia esculenta L.) in the dietary management of diabetes and its complications in streptozotocin induced diabetes in rats | |
| CN107475338A (en) | The extracting method of albumen and dietary fiber in a kind of tealeaves | |
| CN101691410B (en) | Marine oligosaccharide chromic compound having function of preventing and treating insulin resistance | |
| CN107343889A (en) | A kind of peacilomyce hepiahi bacterium fermentation culture medium for being used to treat the 1st type or diabetes B | |
| JP6386550B2 (en) | Use of cinnamon pentacyclic triterpenoid saponins compounds for the preparation of hypoglycemic drugs | |
| KR100894911B1 (en) | Sugar food composition containing sbp extract | |
| CN103494997A (en) | Traditional Chinese sophora flower medicine for treating hyperglycemia and diabetes | |
| CN102266388A (en) | Pharmaceutical composition for preventing and treating type 2 diabetes and complication thereof | |
| CN110090209A (en) | Application of the formoononetin in treatment nonalcoholic fatty liver | |
| CN114052062A (en) | Health food for assisting in reducing blood sugar | |
| CN104906145B (en) | Medical application of the Paecilomyces hepiali chen mutagenic strain PH40 in treatment diabetes | |
| CN101417130A (en) | Medicine combination for treating II type diabetes and complicating diseases thereof | |
| CN101073596B (en) | Alpha-glycosidase inhibitor, its extraction and use | |
| CN109223735B (en) | Use of active compounds isolated from secondary metabolites of aspergillus versicolor | |
| CN113663043A (en) | Composition with alpha-glucosidase inhibition effect and preparation method and application thereof | |
| CN115068559B (en) | Plant-based compound oral liquid with auxiliary blood sugar reducing effect and preparation method thereof | |
| TW202034790A (en) | Bitter gourd peptide composition | |
| CN105030806B (en) | A kind of medical composition and its use for treating diabetes | |
| CN114304225A (en) | Health food for reducing blood sugar and blood fat |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |