CN109528744A - Gentiamarin and its application - Google Patents
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- CN109528744A CN109528744A CN201811653689.2A CN201811653689A CN109528744A CN 109528744 A CN109528744 A CN 109528744A CN 201811653689 A CN201811653689 A CN 201811653689A CN 109528744 A CN109528744 A CN 109528744A
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- 230000000638 stimulation Effects 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 210000003684 theca cell Anatomy 0.000 description 1
- 229930192474 thiophene Natural products 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 208000009852 uremia Diseases 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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Abstract
The invention discloses gentiamarins to prepare the application in drug that is hypoglycemic and/or improving diabetic vascular complications；And application of the gentiamarin in the drug of preparation reducing blood lipid；The invention also discloses a kind of methods for extracting gentiamarin.Gentiamarin is demonstrated in the present invention can be substantially reduced the glycometabolisms index such as blood glucose, glycosylated hemoglobin, haemocyanin of diabetic mice, the exception for adjusting model mice blood lipid and oxidative stress index significantly improves the exception of model mice renal function and renal pathology, inflammatory fibrous chemical conversion point；And gentiamarin improve LDL-C, for 24 hours in terms of work well, it follows that gentiamarin is to reduce blood glucose, comprehensive improve the potential traditional Chinese medicine monomer drug of diabetic nephropathy.
Description
Technical field
The present invention relates to a kind of Traditional Chinese medicine for decreasing blood sugar extract, especially gentiamarin and its in preparing hypoglycemic drug
Using.
Background technique
Diabetes are one of fastest-rising diseases of disease incidence in global range, are after cardiovascular disease, tumour
Three big high mortality diseases.World Health Organization's estimation, to the year two thousand thirty, global diabetic's number can exceed that 3.6 hundred million people,
In 90%~95% be diabetes B.China is second-biggest-in-the-world diabetes country, internal authority periodical " New England's medicine
Magazine " " the Chinese population diabetes epidemic status " of publications in 2010 years show that standardized diabetes morbidity of Chinese age is
9.7% (male 10.6%, female 8.8%)；The disease incidence of diabetes increases with the age, and 20~39 years old, 40~59 years old and more than 60 years old
Population, diabetes morbidity is respectively 3.2%, 11.5% and 20.4%.The disease incidence of urban population is higher than people in the countryside
(11.4%vs.8.2%).Troubling, diabetes are no longer middle-aged and the old's disease, the oriented widened trend of teenager.
With the use of the drugs such as insulin, under diabetes merge the disease incidence of the Diabetic Acutes complication such as ketoacidosis substantially
Drop, and diabetes merge the major complications as diabetes such as chronic vascular complications, are diabetes disability rate, death rate liter
The main reason for high.
Diabetic nephropathy (DN) is also known as diabetic glomerulosclerosis, is common and refractory chronic micro- of diabetes (DM)
Vascular complication, diffused glomerulosclerosis are the specific injury of kidney of diabetes.Clinical manifestation is albuminuria, oedema etc.,
Azotemia can be formed by further developing, uremia, and disability rate is higher with the death rate.With the raising of diabetes morbidity, sugar
It urinates sick nephrosis and has become the main reason for leading to end stage renal failure.It is at present to take control blood glucose, ACEI, calcium antagonist, thiophene more
The complex treatments such as oxazolidinedione class antidiabetic drug, lipid-lowering medicine, still lacking to reduce blood glucose and improve the curative effect of kidney injury expires
The targeted drug of meaning.
Therefore, research and development have the drug for reducing blood glucose, the special effect for improving diabetic nephropathy, will greatly improve product
The market competitiveness, fill a hole in the market, generate good social benefit, obtain considerable economic benefit.
Gentiamarin (Gentiopicrosid) is widely present in dragon as one of main pharmacodynamics ingredient gentianaceous
The drying root and rhizome of the plants such as gallbladder grass, gentianae macrophyllae.Gentiamarin belongs to secoiridoid glycosides compound, molecular formula in structure
For C16H20O9, it is easily soluble in water, methanol and ethyl alcohol equal solvent, gentiamarin is distributed rapidly in vivo, and kidney concentration is high, is arranged through kidney
It lets out, is not easy to accumulate.Gentiamarin molecular structural formula is as follows:
Previously research shows that: gentiamarin has anti-inflammatory and antalgic, Hepatoprotective cholagogue, stomach invigorating, adjusts the pharmacological actions such as blood pressure, but
There is not been reported for application of the gentiamarin in diabetes and its diabetes chronic kidney trouble.
Summary of the invention
Based on the above issues, a kind of reduction is provided it is an object of the invention to overcome above-mentioned the deficiencies in the prior art place
Blood glucose, the drug that kidney injury can be improved again.
To achieve the above object, the technical solution that the present invention takes includes following aspects:
In the first aspect, the present invention provides gentiamarin prepare it is hypoglycemic and/or improve diabetic vascular it is concurrent
Application in the drug of disease.
Preferably, the vascular complication is nephrosis.
Preferably, the symptom of the nephrosis is one of oxidativestress damage, renal hypertrophy, renal fibrosis or more
Kind.
In the second aspect, the application the present invention provides gentiamarin in the drug of preparation reducing blood lipid.
In the third aspect, the present invention provides drug that is a kind of hypoglycemic and/or improving diabetic vascular complications, institutes
It states and contains gentiamarin in drug.
Preferably, the symptom of the vascular complication is one of oxidativestress damage, renal hypertrophy, renal fibrosis
Or it is a variety of.
In the fourth aspect, the present invention provides a kind of drug of reducing blood lipid, contain gentiamarin in the drug.
Preferably, the pharmaceutical dosage form is oral agents or injection, and the oral agents are tablet or capsule, the tablet
For coating tablet, dispersible tablet or sustained release tablets, the capsule is hard capsule, soft capsule or spansule.
At the 5th aspect, the present invention provides a kind of methods for extracting gentiamarin, include the following steps:
(1) radix gentianae is ground into coarse powder；
(2) ingredient in the coarse powder as obtained by ethanol refluxing process or pure water method extraction step (1), the concentration of gained filtrate decompression,
Obtain concentrate；
(3) concentrate obtained by step (2) separated using macroreticular resin absorbing method, purified, then freeze-dried
The gentiamarin of purity > 95%.
In conclusion the invention has the benefit that
Blood glucose, glycosylated hemoglobin, blood that gentiamarin can be substantially reduced diabetic mice are demonstrated in the present invention
The glycometabolisms index such as albumin adjusts the exception of model mice blood lipid and oxidative stress index, significantly improves model mice kidney function
The exception of energy and renal pathology, inflammatory fibrous chemical conversion point；And gentiamarin improve LDL-C, for 24 hours in terms of imitate
Fruit is good, it follows that gentiamarin is to reduce blood glucose, the comprehensive improvement potential traditional Chinese medicine monomer drug of diabetic nephropathy.
Detailed description of the invention
Fig. 1 is detection gentiamarin to model mice hepatic glycogen, liver organization glucokinase (GCK) and low-density lipoprotein
The result figure that polymeric immunoglobulin receptor (LDLR) protein expression influences, wherein * is compared with normal group, P < 0.05；# and model group ratio P < 0.05；
Fig. 2 is detection gentiamarin to model mice blood and kidney superoxide dismutase (SOD), malonaldehyde (MDA) shadow
Loud result figure, wherein * is compared with normal group, P < 0.05；# and model group ratio P < 0.05；
Fig. 3 is the result figure for detecting gentiamarin and influencing on diabetic mice glomerulus pathology HE, PAS, Masson dyeing,
Wherein, * * is compared with normal group, P < 0.01；# and model group ratio P < 0.05；
Fig. 4 is to detect the result figure that influences on diabetic mice glomerulus FN immunohistochemistry of gentiamarin, wherein * * and just
Normal group is compared, P < 0.01；# and model group ratio P < 0.05；
Fig. 5 be detection gentiamarin to diabetic mice fibronectin (FN), Intercellular Adhesion Molecule (ICAM-1),
The result figure of the inflammatory fibrous composition influences such as transforming growth factor (TGF-β 1), wherein * is compared with normal group, P < 0.05；#
With model group ratio P < 0.05.
Specific embodiment
Diabetic nephropathy lesion mechanism is not yet fully apparent from, and be presently considered to be multifactor functioning as a result, such as glycolipid
The stimulation of the factors such as metabolic disorder, the change of renal blood flow dynamics, protein non-enzyme glycosylation, oxidative stress results in cause polyalcohol
The activation of the signal paths such as access, mitogen-activated protein kinase (MAPK), results in Renal Structure and function under diabetic disease states
Pathological change.It is more and more in recent years the study found that glucose -lipid metabolism disorder, oxidative stress and its inflammatory reaction of mediation also
The occurrence and development process of DN (diabetic nephropathy) is taken part in, someone is even additionally considered that DN is a kind of chronic inflammation.Diabetic disease states
Under, the overexpression of inflammatory factor such as cell adhesion molecule (ICAM-1), transforming growth factor (TGF-β) causes to continue or put
Big inflammatory reaction, the secretion for promoting the extracellular matrixs such as kidney fibrous connection albumen (FN) increases, in diabetic nephropathy
It plays an important role in occurrence and development.Therefore, glucose -lipid metabolism disorder is adjusted, anti-oxidant, inhibition kidney inflammatory reaction will have
Conducive to generation, the development for suppressing diabetic nephropathy.
The present invention provides a kind of reduction blood glucose, improve the traditional Chinese medicine monomer drug of diabetic nephropathy, mainly by from radix gentianae
The effective component gentiamarin extracted in equal plant medicines.Gentiamarin is the reduction blood glucose with good market potential, changes
The traditional Chinese medicine monomer drug of kind diabetic nephropathy.
In some embodiments, the present invention also provides the extracting method of gentiamarin, include the following steps:
1) by pulverizing medicinal materials such as radix gentianaes at coarse powder；
2) by ingredient in a certain concentration ethanol refluxing process or pure water-swollen squid coarse powder, merging filtrate is concentrated under reduced pressure；Using
Macroreticular resin absorbing method carries out the separation of gentiamarin, purifying to filtrate, and freeze-drying obtains purity > 95% gentiamarin raw material
Medicine.
In some embodiments, appropriate amount of auxiliary materials is added in gentiamarin, related peroral dosage form or injection type is made.In order to
Influence of the gentiamarin to diabetic mouse model blood glucose, blood lipid, oxidative stress and kidney injury is studied, in some embodiments
In, the diabetic mouse model of low dose of streptozotocin (STZ) induction is fed, added using high glucose and high fat；Observe gentiamarin
Model mice blood lipid, oxidation are answered in influence to the glycometabolisms index such as model mice blood glucose, glycosylated hemoglobin, haemocyanin
The influence for swashing index, the influence to model mice renal function and renal pathology, inflammatory fibrous chemical conversion point, with clear gentiamarin
The pharmacodynamics effect that blood glucose is reduced to diabetic mice, improves Diabetic Nephropathy.
To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with the drawings and specific embodiments pair
The present invention is described further.Unless otherwise instructed, the experimental method in the present invention is all made of conventional method.
Necessary explanation is carried out to experimental material involved in the present invention, instrument and method first:
(1) trial drug: gentiamarin is right by pharmaceutical college, Zhongshan University medicament laboratory separation and Extraction (purity > 95%)
It applies valuable pharmaceutical Co. Ltd by Sino-U.S. Shanghai according to drug metformin hydrochloride tablet to provide, lot number 101002, specification 0.5g/ piece.
(2) experimental animal: healthy male C57 mouse, SPF grades, weight is 7-8 weeks, 60, by Zhongshan University experimental animal
Center provides.Zoopery carries out in Zhongshan University's Experimental Animal Center Animal Experimental barrier environment laboratory.
(3) reagent: streptozotocin (streptozotocin, STZ), sodium citrate: Sigma Co., USA；Serum is total
Cholesterol (TC), serum levels of triglyceride (TG), glycated serum protein (GSP), glycosylated hemoglobin (HbA1c), serum creatinine (Cr),
The assay kits such as urea nitrogen (BUN), hepatic glycogen: Bioengineering Research Institute is built up in Nanjing；Superoxide dismutase (SOD), the third two
The assay kits such as aldehyde (MDA): the green skies；Dehydrated alcohol, methanol, the concentrated sulfuric acid (AR): Guangzhou Chemical Reagent Factory；Anti- mouse secondary antibody,
Anti-rabbit secondary antibody: promega company, the U.S.；LDLR rabbit monoclonal antibody: Proteintech company；GCK rabbit is mostly anti-: U.S. Santa Cruz
Biotechnology company；FN, ICAM-1, TGF-β rabbit are mostly anti-: abcam company；α-Tubulin mouse monoclonal antibody: Sigma company.
(4) instrument: Beck-manCX-5 full-automatic biochemical analysis；Gel protein analyzes software (UVP company, the U.S.)；PVDF
Film (millipore company, the U.S.)；General microplate reader (Bio-Tek company, the U.S.)；Desk-top high-speed refrigerated centrifuge
(Centrifuge 5417R) (German Eppendorf company)；Milli.PROTEAN II electrophoresis apparatus, Mini Trans.Blot
Electrophoresis transfer device, Gel DoeXR gel frame (Bio.rad company, the U.S.).
(5) preparation of experimental high glucose and high fat diabetic mice animal model: SPF grades of C57 mouse, using SPF grades conventional
It full nutrition feed adaptive feeding two days, takes 10 to be only used as Normal group at random, gives conventional SPF always in experimentation
Grade full nutrition feed is fed.Remaining C57 mouse, after high glucose and high fat forage feed 2 weeks, empty stomach hour, by the agent of 40mg/kg
Amount intraperitoneal injection streptozotocin (STZ is diluted with sodium citrate buffer solution, PH=4.0, Fresh in ice chest).Continuous note
It penetrates 5 days.After C57 mouse peritoneal injects STZ, continue to give high glucose and high fat forage feed, after 10 days after mouse blood sugar is stablized, use
Blood glucose meter measures blood glucose value, is diabetic mouse model with fasting 6-8h Xue Tang Zhi≤11.1mmol/L person.Selection Model mouse
50, it is divided into model group, gentiamarin low dose group, middle dose group, high dose group, melbine group.
(6) experimental administration and grouping: Normal group, diabetic model group, gentiamarin low dose group group (25mg
kg-1·d-1), gentiamarin middle dose group (50mgkg-1·d-1), gentiamarin high dose group (100mgkg-1·d-1)、
Melbine group (195mgkg-1·d-1), every group 10.Daily gastric infusion, is administered according to mouse weight, and administered volume is
0.1mL/10g mouse.Administration time: 9:00-10:30am, weekly administration 6 days, totally 8 weeks.During experiment in addition to Normal group,
Remaining mouse continues to give high glucose and high fat feed, mouse ad lib, drinking-water.
It makes a collection of specimens after observing 8 weeks, urine for 24 hours is accurately collected in experiment with metabolic cage on the day before terminating, carry out Urine proteins for 24 hours
Detection；Mouse after 8 hours, plucks eyeball and takes blood 2ml on an empty stomach, and serum is left and taken after centrifugation, makees biochemical kidney function test；After mouse is put to death
Kidney is taken out, rapidly with pre-cooling normal saline flushing, removes envelope, dissociate kidney, and double kidney weights are claimed after blotting bloodstain with filter paper simultaneously
Record is observed Kidney Size, color, quality, longitudinally splits left kidney along median sagittal plane using blade, left kidney half is put in
Marked in embedded box be put into togerther in 4% formalin again it is fixed after carry out PAS dyeing processing and immunohistochemistry detection；
Left kidney the other half and right kidney separate cortex renis, by resulting whole cortex renis be quickly charged with cryopreservation tube be put into it is quick-frozen in liquid nitrogen, after
- 80 DEG C of preservations are transferred to, are detected for western blot.
1 Indexs measure of embodiment
(1) detection of the biochemical indicators such as blood glucose, blood lipid and renal function
Fasting blood-glucose (FBG) takes blood using tail vein, measures blood glucose value using blood glucose meter (Johnson Co.)；Serum is total
COD-PAP method (single reagent type), GPO-PAP method (single reagent is respectively adopted in cholesterol (T-CHO), serum levels of triglyceride (TG)
Type)；Low density cholesterol lipoprotein (LDL-C), high density cholesterol lipoprotein (HDL-C) are detected using enzymic colorimetric；Blood flesh
Acid anhydride, urea nitrogen, the renal function index such as Urine proteins are respectively adopted enzymic creatinine assay method, urease method, CBB method and are detected for 24 hours, phase
It closes the kit that detection uses and Bioengineering Research Institute's offer is built up by Nanjing.
(2) detection of glycated serum protein and hemoglobin
Glycated serum protein (GSP), glycosylated hemoglobin (HbA1c) are detected with fructosamine method, enzymic colorimetric respectively,
The kit that coherent detection uses builds up biological study by Nanjing and is provided.
(3) detection of hepatic glycogen, liver glucose kinases (GCK) and LDL receptor (LDLR)
Hepatic glycogen content is detected with enzyme linked immunosorbent assay in hepatic tissue, and the kit which uses builds up bioengineering by Nanjing and grinds
Study carefully and is provided；The protein expression situation of glucokinase (GCK) and LDL receptor (LDLR) are used in liver organization
The detection of Wester blotting method.
(4) detection of blood and kidney SOD, MDA
Serum and renal tissue superoxide dismutase (SOD) activity, (WST-8, sulphur is respectively adopted in MDA content to malonaldehyde
For barbital acid system (TBA), the kit that coherent detection uses is provided by the green skies.
(5) renal hypertrophy index (weight, kidney compare again), pathological examination
It weekly routine weighing and records each group mouse weight and observes its variation, reflected at the end of experiment with kidney weight/weight
Renal hypertrophy index.Renal tissue is dehydrated, is embedded, routine paraffin wax flaking after 10% formalin solution is fixed, slice thick
About 3-4 μm of degree, HE dyeing, PAS dyeing and Masson then are carried out to paraffin section and dyed, with cell imaging system (EVOS
FL Auto) carry out histological observation.Light microscopic observation Pathological histological change can be carried out after mounting a couple of days, emphasis is seen
Examine the pathological change of glomerulus.
(6) kidney fibrous connection albumen (FN), transforming growth factor (TGF-β 1), intercellular adhesion molecule (ICAM-1) etc.
Inflammatory fibrous composition detection
The protein expression situation of FN, ICAM-1 and TGF-β 1 is detected with Western blot method in renal tissue.
The experimental result of the present embodiment is indicated using mean ± standard error (Mean ± SEM), is united with Graphpad Prism5
Meter software carries out analysis comparison, compares in two groups of comparison among groups, group and is examined with t；Multiple-group analysis one-way analysis of variance (one-
Way ANOVA, Bonferroni method).Think if P < 0.05 statistically significant.SpecificallyExperimental result is as follows:
1)Influence of the gentiamarin to model mice fasting blood-glucose
As seen from Table 1, the blood glucose of each composition mould mouse is apparently higher than Normal group (P < 0.05) before administration, and locates
In phase same level, gentiamarin and positive control drug melbine are given after two weeks, compared with model group, the low middle height of gentiamarin
Three dosage groups and melbine group can reduce mouse FBG (P < 0.05) to a certain extent；With the extension of the administration time,
The hypoglycemic effect of gentiamarin administration group is remarkably reinforced, after administration 8 weeks, compared with model group, and gentiamarin middle dosage and height
Dosage group can significantly reduce mouse FBG (P < 0.05), and gentiamarin high dose group (100mg/kg) and melbine group
The FBG of (195mg/kg) is closer to, and gentiamarin is prompted to have significant hypoglycemic effect.
1 gentiamarin of table to the STZ diabetic mice fasting blood-glucose induced influence (N=10)
*P < 0.05vs. Normal group,#P < 0.05vs. model group
2)Influence of the gentiamarin to diabetic mice glycosylated hemoglobin (HbA1c) and glycated serum protein (GSP)
As seen from Table 2, HbA1c the and GSP content of model group mouse is apparently higher than Normal group (P < 0.05).Administration
After 8 weeks, compared with model group, the middle and high dosage of gentiamarin can be substantially reduced GSP content (P < 0.05), gentiamarin high dose
Group can be substantially reduced HbA1c content (P < 0.05).The exception of HbA1c is to lead to chronic vascular including diabetic nephropathy simultaneously
An important factor for sending out disease prompts gentiamarin to play the role of improvement diabetic vascular complications (such as diabetic nephropathy).
2 gentiamarin of table to the STZ diabetic mice glycosylated hemoglobin and glycated serum protein induced influence (N=10)
| HbA1C (%) | GSP(mmol/L) | |
| Normal group | 8.66±0.79 | 2.79±0.08 |
| Model group | 14.52±2.68* | 3.66±0.14* |
| Low dose group | 13.31±2.12 | 3.41±0.20 |
| Middle dose group | 11.59±1.67 | 3.26±0.08# |
| High dose group | 9.94±1.36# | 3.11±0.09# |
| Melbine group | 10.45±1.19# | 3.25±0.22# |
*P < 0.05vs. Normal group,#P < 0.05vs. model group
3)Influence of the gentiamarin to model mice fasting plasma lipid four
As seen from Table 3, model group mice serum TC, TG, LDL-C level with normal group compared to apparent increase (P <
0.05).Compared with model group, each administration group of gentiamarin and melbine group can be substantially reduced TG, LDL-C of each group mouse
Horizontal (P < 0.05), meanwhile, gentiamarin high dose group and melbine group energy apparent increase HDL-C are horizontal (P < 0.05), and
Gentiamarin high dose group is slightly better than melbine to the opsonic action trend of four items of blood lipid tests, and the reducing blood lipid of gentiamarin is prompted to make
With clearly.
3 gentiamarin of table to the STZ diabetic mice four items of blood lipid tests induced influence (N=10)
*P < 0.05vs. Normal group,#P < 0.05vs. model group
4)Gentiamarin is to model mice hepatic glycogen, liver organization glucokinase (GCK) and LDL receptor (LDLR) influence of protein expression
As shown in Fig. 1 .A, compared with normal group, model group liver glycogen content is substantially reduced (P < 0.05).Gentiamarin
Compared with model group, hepatic glycogen content is obviously increased (P < 0.05) for middle and high dosage group and melbine group, prompts gentiamarin tool
It is improved the effect of hepatic glycogen.As shown in Fig. 1 .B, compared with normal group, the protein expression of GCK is obvious in model group liver organization
It reduces (P < 0.05), the middle and high dosage of gentiamarin and melbine group can significantly improve GCK albumen compared with model group
It expresses (P < 0.05), gentiamarin is prompted to can be improved the protein expression of GCK in rigid dirty tissue.The result of Fig. 1 .A and Fig. 1 .B into
One step specifies that gentiamarin may raise liver GCK protein expression, play and adjust glycometabolism by improving hepatic glycogen content
Effect.
As shown in Fig. 1 .C, compared with normal group, the albumen of LDL receptor (LDLR) in model group liver organization
Expression is substantially reduced (P < 0.05), and each dosage group of gentiamarin and melbine group can significantly improve compared with model group
The protein expression (P < 0.05) of LDLR prompts gentiamarin that can participate in the adjusting of lipid metaboli by raising the protein expression of LDLR.
5)Influence of the gentiamarin to model mice blood and kidney superoxide dismutase (SOD), malonaldehyde (MDA)
As shown in Fig. 2 .A, B, compared with normal group, the activity of superoxide dismutase (SOD) is obvious in model group serum
It reduces (P < 0.05), the content of metabolism of lipid peroxide product malonaldehyde (MDA) dramatically increases (P < 0.05)；Gentiamarin
Middle and high dosage group and melbine group are compared with model group, and the activity of SOD obviously increases in serum, and the content of malonaldehyde is significant
It reduces.Likewise, the middle and high dosage group of gentiamarin and melbine group can also obviously increase renal tissue compared with model group
Middle SOD is active (P < 0.05), substantially reduces MDA content (P < 0.05).Oxidativestress damage is the center for leading to diabetic nephropathy
Link prompts gentiamarin that may participate in the Renoprotective Effect to diabetic mice by anti-oxidation stress.
6)Influence of the gentiamarin to diabetic mice renal function
As seen from Table 4, compared with normal group, the renal hypertrophy index of model group mouse obviously increases (P < 0.05), blood
Creatinine (Cr), urea nitrogen (BUN), the raw functional parameter such as Urine proteins (UP/24h) obviously rises (P < 0.05) for 24 hours；Gentiamarin is each
Group is compared with melbine with model group, can be substantially reduced renal hypertrophy index (P < 0.05), and Cr, BUN, UP/24h (P are reduced
<0.05).Prompt gentiamarin can be obviously improved the renal function of diabetic mice.
4 gentiamarin of table to the STZ Diabetic Nephropathy mouse renal function induced influence (N=10)
*P < 0.05vs. Normal group,#P < 0.05vs. model group
7)Influence of the gentiamarin to diabetic mice Pathological
Experimental animal renal glomerulus lesion situation is further looked at, as shown in HE and PAS coloration result in Fig. 3, and just
Normal mouse is compared, and the glomerular volume of diabetic mice increases, mesangial region expansion obvious with matrix secreted, basilar memebrane increasing
Thickness, and epithelial cell and capsula glomeruli adhesion are obvious in glomerulus, extracellular matrix index is significantly raised (P < 0.01).Gentiamarin and
After Or Metformin In Treating 8 weeks, the middle and high dosage of gentiamarin and melbine can be substantially reduced extracellular matrix index (P <
0.01), hence it is evident that improve kidney morphology damage, including glomerulus inner cell number significantly reduces, glomerular basement membrane thickening and is
Theca cell hyperplasia is substantially reduced, and colleague also mitigates the adhesion between glomerulus and capsula glomeruli.Masson dyeing prompt, diabetes are small
Collagenous fibres obviously increase in the glomerulus of mouse, and can be changed after the basic, normal, high dosage of gentiamarin and Or Metformin In Treating
It is kind.
Fibronectin (FN) is the important component outside messangial cell, and excessive FN generation is to promote glomerulus fiber
Change the one of the major reasons of lesion.As shown in FN ImmunohistochemistryResults Results in Fig. 4, FN protein level is bright in diabetic mice glomerulus
It is aobvious to increase (P < 0.01), and the middle and high dosage of gentiamarin and melbine can be reduced protein level in glomerulus (P < 0.01).
Fig. 3 and Fig. 4 prompts gentiamarin that diabetic mice Pathological can be delayed to damage, and reduces renal fibrosis.
8)Gentiamarin gives birth to diabetic mice fibronectin (FN), Intercellular Adhesion Molecule (ICAM-1), conversion The influence of the inflammatory fibrous such as the long factor (TGF-β 1) chemical conversion point
FN and ICAM-1 is the important component of messangial cell epimatrix, and excessive FN and ICAM-1 are glomerulus fibrosis
The one of the major reasons of lesion, TGF-β 1 are the main rush fibrosis factors of mesangial cell secretion, promote mesangial cell
Proliferation and loose, the final accumulation for aggravating messangial cell epimatrix.We further extract renal tissue and pass through Western
FN, ICAM-1 and 1 protein level of TGF-β of the method detection kidney of Blotting, as shown in Fig. 5 A, B, C., the sugar of STZ induction
The protein level of FN, ICAM-1 and TGF-β 1 for urinating sick mouse kidney tissue obviously increase (P < 0.05), and gentiamarin is middle and high
Dosage and melbine can significantly reduce FN, ICAM-1 and 1 protein level of TGF-β (P < 0.05) of diabetic mice kidney.
Result above further proves that gentiamarin reduces the generation of diabetic mice glomerulus inflammation fibrotic component, and it is fine to improve kidney
Dimensionization.
Experiment conclusion
Gentiamarin 50mg/kg, 100mg/kg dosage group can be substantially reduced the blood glucose of diabetic mice, HbAle
The glycometabolisms index such as albumen, haemocyanin adjusts the exception of model mice blood lipid and oxidative stress index, it is small to significantly improve model
The exception of mouse renal function and renal pathology, inflammatory fibrous chemical conversion point.Class is acted on compared with melbine 195mg/kg dosage group
Seemingly, and improve LDL-C, for 24 hours in terms of show certain Action advantage.In conclusion gentiamarin is to reduce
Blood glucose, the comprehensive improvement potential traditional Chinese medicine monomer drug of diabetic nephropathy.
Finally it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention
The limitation of range, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should be managed
Solution, can with modification or equivalent replacement of the technical solution of the present invention are made, without departing from technical solution of the present invention essence and
Range.
Claims (9)
1. gentiamarin is preparing the application in drug that is hypoglycemic and/or improving diabetic vascular complications.
2. application according to claim 1, which is characterized in that the vascular complication is nephrosis.
3. application according to claim 2, which is characterized in that the symptom of the nephrosis is oxidativestress damage, kidney fertilizer
Greatly, one of renal fibrosis or a variety of.
4. application of the gentiamarin in the drug of preparation reducing blood lipid.
5. drug that is a kind of hypoglycemic and/or improving diabetic vascular complications, which is characterized in that contain rough gentian in the drug
Bitter glycosides.
6. drug according to claim 5, which is characterized in that the symptom of the vascular complication be oxidativestress damage,
One of renal hypertrophy, renal fibrosis are a variety of.
7. a kind of drug of reducing blood lipid, which is characterized in that contain gentiamarin in the drug.
8. the drug according to claim 5 or 7, which is characterized in that the pharmaceutical dosage form be oral agents or injection, it is described
Oral agents are tablet or capsule, and the tablet is coating tablet, dispersible tablet or sustained release tablets, and the capsule is hard capsule, flexible glue
Capsule or spansule.
9. a kind of method for extracting gentiamarin, which comprises the steps of:
(1) radix gentianae is ground into coarse powder；
(2) ingredient in the coarse powder as obtained by ethanol refluxing process or pure water method extraction step (1), gained filtrate decompression concentration, obtains
Concentrate；
(3) concentrate obtained by step (2) separated using macroreticular resin absorbing method, purified, then freeze-dried purity >
95% gentiamarin.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111067914A (en) * | 2020-01-03 | 2020-04-28 | 吉林大学 | Application of gentiopicroside in preparation of medicine for treating hyperlipidemia |
| CN113041276A (en) * | 2019-12-26 | 2021-06-29 | 西安远大德天药业股份有限公司 | Tibetan capillary artemisia extract and preparation method and application thereof |
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| CN1437953A (en) * | 2003-03-21 | 2003-08-27 | 孙文基 | Gentiamarin powder infection for muscle injection and intravenous injection |
| CN103848875A (en) * | 2012-11-30 | 2014-06-11 | 哈尔滨誉衡药业股份有限公司 | Preparation method of gentiopicroside raw material |
| CN105012329A (en) * | 2015-07-27 | 2015-11-04 | 中南民族大学 | Drug for treatment of type II diabetes |
| CN107929304A (en) * | 2017-12-19 | 2018-04-20 | 西北大学 | Application of the gentiamarin in non-alcohol fatty liver treatment |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1437953A (en) * | 2003-03-21 | 2003-08-27 | 孙文基 | Gentiamarin powder infection for muscle injection and intravenous injection |
| CN103848875A (en) * | 2012-11-30 | 2014-06-11 | 哈尔滨誉衡药业股份有限公司 | Preparation method of gentiopicroside raw material |
| CN105012329A (en) * | 2015-07-27 | 2015-11-04 | 中南民族大学 | Drug for treatment of type II diabetes |
| CN107929304A (en) * | 2017-12-19 | 2018-04-20 | 西北大学 | Application of the gentiamarin in non-alcohol fatty liver treatment |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113041276A (en) * | 2019-12-26 | 2021-06-29 | 西安远大德天药业股份有限公司 | Tibetan capillary artemisia extract and preparation method and application thereof |
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