CN104208210B - A kind of Chinese medicine composition treating insulin resistant and relevant disease and preparation method thereof and Chinese medicine preparation - Google Patents
A kind of Chinese medicine composition treating insulin resistant and relevant disease and preparation method thereof and Chinese medicine preparation Download PDFInfo
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Abstract
The invention discloses a kind of Chinese medicine composition treating insulin resistant and relevant disease thereof, described Chinese medicine composition is mainly made up of the raw material of Chinese medicine of following weight proportion: Fructus Crataegi 100 300, Radix Polygoni Multiflori Preparata 90 300, the Radix Paeoniae Alba 120 360, Radix Salviae Miltiorrhizae 100 500.Correspondingly, the invention also discloses the preparation method of above-mentioned Chinese medicine composition, and the Chinese medicine preparation containing above-mentioned Chinese medicine composition.Using the present invention, described Chinese medicine composition can treat insulin resistant and relevant disease thereof simultaneously, plays the many sick effects of medicine treatment, and toxic and side effects is little, price is low.
Description
Technical field
The present invention relates to field of medicaments, particularly to a kind of Chinese medicine composition treating insulin resistant and relevant disease thereof
And preparation method thereof, and the Chinese medicine preparation containing above-mentioned Chinese medicine composition.
Background technology
Insulin resistant (insulin resistance；IR) refer to that a variety of causes makes the target tissue (master of insulin action
Liver to be, fat, skeletal muscle, vascular endothelial cell etc.) insulin is promoted that its picked-up and the efficiency utilized and effect reduce,
In early days, beta Cell of islet is the insulin (i.e. hyperinsulinemia) making blood glucose still maintain normal and compensatory supersecretion.
Insulin resistant is prevalent in multiple physiology and pathological state, in adolescence, geratic period, trimester of pregnancy,
There is insulin sensitivity index reduction i.e. IR in various degree.Normal population only has 15%-20% and there is IR, but body weight increases,
Blood glucose increases, blood pressure increases, blood fat disorder, hyperuricemia time, incidence rate and the degree of IR all dramatically increase.
Insulin resistant is the basic feature of type 2 diabetes mellitus, is the key link of metabolism syndrome, is also fat, blood fat
Disorder, vascular endothelial dysfunction, cardiovascular disease and blood height stick the pathologic basis of state.
The insulin of normal level has promotion glycogen, fat and protein synthesis in vivo, promotes glucose utilization and energy
Amount generates, suppresses steatolysis, reinforcement cell proliferation and differentiation, maintenance inner skin cell function, suppresses lipase thus lipotropism solution
Etc. many effects.
Insulin resistant occurs, and the biological action of insulin weakens so that tissue is to a certain amount of insulin
Biologically utilizes minimizing less than anticipated normal level, the sugar ultimately resulting in muscle and fat, and liver glyconeogenesis increases, fat
Cell release free fatty increases, thus blood glucose, dyslipidemia occurs；The insulin long duration of action of high concentration is in Ink vessel transfusing
The synthesis secretions such as chrotoplast, may result in vasodilatory material and reduce, pressor material such as Endothelin (ET) increase, thus
Appearance dysarteriotony (Chen Linxi, Qin Xuping, yellow Qiu Lin etc., vascular endothelial cell pharmacology and clinic, People's Medical Officer Press,
2012, P164-166.)；High-caliber insulin makes the inhibitor synthesis of plasma tissue type activator of plasminogen and release increase
Add, suppress fibrin degradation, thus (Yang Sheng, Xu Xiaoguang, Yue Guiying etc., propylene glycol alginate sodium sulfate is to 2 type glycosurias to promote thrombosis
Patient's blood fat lectin from hemolymph and the impact of insulin resistant, the practical internal medicine journal of China, 2004,24 (3): 183.)；Long-term blood
Liquid high insulin levels may result in blood viscosity rising, and (dragon is for army building, Zhao Aiting, Luo Bingquan etc., Hypertensive Disease Insulin Resistance
With the comprehensive study of microcirculation disturbance relation, China hemorheology magazine, 2001,11 (2): 111-114.).
Occur that insulin resistant is closely related with the abnormal expression of related gene.After insulin is combined with its receptor, signal
To intracellular transduction and amplify step by step, make insulin play Metabolism regulation effect.IRS-1 (IRS-1) and islets of langerhans
Element receptor substrate-2 (IRS-2) plays an important role, when IRS-1 and IRS-2 producer during insulin signal transduction
When sudden change or abnormal expression, may result in insulin signaling and be obstructed at endocellular transduction and cause insulin resistant；Glucose transport
Albumen 4 (GLUT4) is present in muscle and adipose cell, IRS-1 phosphorylation under insulin action, makes GLUT4 indexing to thin
Endochylema film, accelerates the facilitory transport process of glucose, makes muscle increase glucose uptake.GLUT4 abnormal expression or indexing are subject to
Resistance, may result in insulin resistant after receptor；The quantity of glucose transporter 2 (GLUT2) reduces or activity reduces, it is possible to decrease liver
Glucose uptake, increase hepatic glycogen output (insulin resistant) (Wang Jiyao, internal medicine, People's Health Publisher, 2008,
P974-975).Peroxisome proliferator-activated receptors γ (PPAR-γ) is mainly expressed in fatty tissue, regulates adipose cell
Differentiation and propagation.Activation energy suppression fatty tissue secreting tumor necrosis factor α (TNF-α) of PPAR-γ, blocks TNF-α and exists
To Insulin receptor INSR and the inhibitory action of IRS phosphorylation in signal transduction pathway, be conducive to improving insulin and support
Anti-.The steatolysis of beta 3 adrenoreceptor (β 3AR) mainly regulation body and exothermal process, the β 3AR of low expression may result in
Fatty tissue heat production reduces, and causes lipopexia.Lipid, at Intracellular retention, causes interior fat to increase, and causes visceral fertilizer
Fat, cause insulin resistant (An Yali, Guangwei LI etc., beta 3 adrenoreceptor gene pleiomorphism and insulin resistant, fertilizer
Fat, China-Japan Friendship Hospital's journal, 2004,18 (6): 366-368.).
Clinical treatment insulin resistant mainly applies euglycemic agent at present.Metformin passes through liver plasma membrane G egg
The white recovery insulin suppression to adenyl cyclase, reduces hepatic gluconeogenesis and output, promotes anaerobic glycolysis, increase muscle
Deng peripheral tissues, the picked-up of glucose and utilization, suppression or delay glucose are improved carbohydrate metabolism in gastrointestinal absorption etc..But
Taking metformin it is possible that many side effect such as gastrointestinal reaction, anaphylaxis etc., severe patient may induction lactate
Acidosis；Rising person notable to serum insulin levels, have ketoacidosis tendency person, severe depletion of oxygen, heart failure, severe liver disease,
Kidney patient is not suitable for using metformin.Thiazolidinediones can increase the insulin sensitivity in peripheral tissues, alleviates pancreas
Insulin resistance, is a kind of euglycemic agent.Thiazolidinediones is not suitable for insulin and definitely lacks, in ketosis acid
Poison, serious and acute heart failure patient；Common adverse effect has headache, dizziness, weak, nauseating, diarrhoea, light anemia, edema, body
Heavily increase, hypercholesterolemias etc., some patients is it is possible that hepatic injury.
And, the most clinical in treating the lysis centered by insulin resistant, due to sending out of insulin resistant
The most adjoint raw multiple relevant disease is (such as hyperinsulinemia, diabetes, hypertension, hyperlipidemia, metabolism syndrome, metabolic arthritis
Mass formed by blood stasis, blood viscosity increase), need to take simultaneously depressor, antidiabetic drug, lipid lowerers, antithrombotic, even with insulin
Deng, to take medicine of a great variety, dosage is big, and majority is Western medicine, and toxic and side effects is more, especially gastrointestinal reaction and hepatic and renal function
Infringement.It addition, existing medicine needs long-term taking, medical expense is expensive.
Summary of the invention
The technical problem to be solved is, it is provided that the Chinese medicine composition that a kind of toxic and side effects is little, price is low, can
To treat insulin resistant and relevant disease thereof simultaneously, play the many sick effects of medicine treatment.
The technical problem to be solved also resides in, it is provided that the method preparing above-mentioned Chinese medicine composition.
The technical problem to be solved also resides in, it is provided that containing the Chinese medicine preparation of above-mentioned Chinese medicine composition.
For reaching above-mentioned technique effect, the invention provides a kind of Chinese drug-treated group treating insulin resistant and relevant disease thereof
Compound, described Chinese medicine composition is mainly made up of the raw material of Chinese medicine of following weight proportion:
Fructus Crataegi 100-300,
Radix Polygoni Multiflori Preparata 90-300,
Radix Paeoniae Alba 120-360,
Radix Salviae Miltiorrhizae 100-500.
As the improvement of such scheme, described Chinese medicine composition is mainly made up of the raw material of Chinese medicine of following weight proportion:
Fructus Crataegi 180-240,
Radix Polygoni Multiflori Preparata 180-240,
Radix Paeoniae Alba 120-300,
Radix Salviae Miltiorrhizae 120-360.
Correspondingly, present invention also offers the preparation of a kind of Chinese medicine composition treating insulin resistant and relevant disease thereof
Method, including:
A, take Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae by formula, pulverized；
B, extract with methanol, ethanol or acetone and concentrate, obtain the first extracting solution；
C, will extract after medicinal residues water extraction and concentrate, obtain the second extracting solution；
D, by the first extracting solution, second extracting solution merge.
As the improvement of such scheme, described step B includes:
Add 6-10 times amount, concentration is the methanol of 60-80%, ethanol or acetone soak 20-40min；
Reflux, extract, 1-3 time, each 1-3h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
As the improvement of such scheme, described step B includes:
Add 8 times amount, concentration be 70% methanol, ethanol or acetone soak 30min；
Reflux, extract, 2 times, each 2h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
As the improvement of such scheme, described step C includes:
Medicinal residues 6-10 times amount water soaking 20-40min after extracting；
Water carries 1-3 time, for the first time 45-75min, for the second time 30-60min；
United extraction liquid concentrating under reduced pressure, obtain the second extracting solution.
As the improvement of such scheme, described step C includes:
Medicinal residues 8 times amount water soaking 30min after extracting；
Water carries 2 times, 60min for the first time, for the second time 45min；
United extraction liquid concentrating under reduced pressure, obtain the second extracting solution.
Correspondingly, present invention also offers the system of the Chinese medicine composition of another kind for the treatment of insulin resistant and relevant disease thereof
Preparation Method, including:
A, weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae by formula；
B, added water no medical material, soaks 20-40min；
C, being heated to water and open, little fire continues to decoct 10-30min, filters, obtains the first medicinal liquid；
D, continued to add water no medical material, water after opening little fire continue to decoct 10-30min, filter, obtain the second medicinal liquid；
E, merge the first medicinal liquid, the second medicinal liquid.
Correspondingly, the present invention also provides for a kind of Chinese medicine preparation treating insulin resistant and relevant disease thereof, described Chinese medicine
Preparation contains any of the above-described Chinese medicine composition.
As the improvement of such scheme, described Chinese medicine preparation is decoction, powder, capsule, tablet, granule or pill.
Enforcement there is advantages that
The composition of Chinese medicine composition of the present invention is the active component of the medicines such as Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, passes through
Drug effect and study on mechanism to said composition find, it can be treated insulin resistant and relevant disease thereof simultaneously, play
The one many sick effects of medicine treatment, and toxic and side effects is little, price is low, specific as follows:
One, the compositions drug action of the present invention is notable, has insulin resistant and relevant disease thereof and preferably treats work
With.Mechanism of action of the present invention is clear and definite, and research discovery said composition targeting is in the related pathways of glucose-lipid metabolism, by regulation and control
The expression of IRS IRS-1, IRS-2 gene, improves the body sensitivity to insulin；Turned by upregulation of glucose
Fortune Protein G LUT2, the expression of GLUT4mRNA, promote the body picked-up to glucose, increases the body consumption to glucose；Logical
Cross rise PPAR-γ, the expression of β 3AR mRNA, promote the catabolism of adipose cell；Preventing and treating insulin is played from gene level
The effect of opposing.Meanwhile, said composition also has improves hemorheology, reduces endothelin-1 (ET-1) level, plays preventing and treating
Thrombosis and the effect of protection vascular endothelial function.Said composition plays treatment insulin resistant and phase thereof by many aspects
The effect of related disorders.
Two, the present invention is centered by improving insulin resistant, by regulating the signal path relevant to insulin resistant, with
Treatment insulin resistant is starting point, and then plays the treatment hyperinsulinemia relevant to insulin resistant, diabetes, high blood
The multiple diseases such as pressure, hyperlipidemia, metabolism syndrome, hyperuricemia, blood viscosity increase, reach the effect of " the many diseases of medicine ", carry
High patient compliance, alleviates economy and the physiological load of patient.
Three, the present invention is Chinese medicine preparation, and medicine composition is simple, and dose is little, and supports insulin through effect experiment checking
Anti-and relevant disease has obvious therapeutical effect, and toxic and side effects is little.
Four, the present invention is the extract of natural plant crude drugs, and crude drug is cheap, wide material sources, and extraction process is simple,
Large-scale industrialization application can be realized.
Accompanying drawing explanation
Fig. 1 is the flow chart of preparation method one embodiment of Chinese medicine composition of the present invention；
Fig. 2 is the flow chart of another embodiment of preparation method of Chinese medicine composition of the present invention.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearer, below in conjunction with accompanying drawing, the present invention is made into one
Step ground describes in detail.
The invention provides a kind of Chinese medicine composition treating insulin resistant and relevant disease thereof, described Chinese medicine composition
Mainly it is made up of the raw material of Chinese medicine of following weight proportion:
Fructus Crataegi 100-300,
Radix Polygoni Multiflori Preparata 90-300,
Radix Paeoniae Alba 120-360,
Radix Salviae Miltiorrhizae 100-500.
Wherein, insulin resistant relevant disease of the present invention, refer to the relevant disease caused by insulin resistant, as
Hyperinsulinemia, diabetes, hypertension, hyperlipidemia, metabolism syndrome, hyperuricemia, blood viscosity increase.
Preferably, described Chinese medicine composition is mainly made up of the raw material of Chinese medicine of following weight proportion:
Fructus Crataegi 180-240,
Radix Polygoni Multiflori Preparata 180-240,
Radix Paeoniae Alba 120-300,
Radix Salviae Miltiorrhizae 120-360.
Fructus Crataegi, warm in nature, sour in the mouth, sweet, return spleen, stomach, Liver Channel, function invigorating the stomach and promoting digestionization is stagnant, promoting flow of QI and blood dissipating blood stasis.Modern pharmacology
Research finds, Fructus Crataegi mainly contains flavones ingredient, including hyperin, Quercetin, flavone polymer；Organic acid, including
Citric acid, Crataegolic acid etc., possibly together with lactone, saccharide and glycoside etc..Modern pharmacological research find Fructus Crataegi have promotion fat digestion,
Cardioprotection, blood pressure lowering, blood fat reducing, antioxidation improve the effect such as immunity, antibacterial, antitumor.
Radix Polygoni Multiflori Preparata, bitter in the mouth, sweet, puckery, warm in nature, return liver, the heart, kidney channel.Function invigorating the liver and kidney, benefiting essence-blood, crow beard and hair, strong muscle
Bone.Modern pharmacological research finds that the main component of Radix Polygoni Multiflori has phospholipid, Anthraquinones, glucose glycoside, gallic acid and multiple micro-
Secondary element.Have endocrine regulation function, improve immunity of organisms, blood fat reducing, atherosclerosis, antioxidation protect the liver etc. makees
With.
Radix Paeoniae Alba bitter in the mouth, acid, cold nature.Return liver, spleen channel.Function suppressing the hyperactive liver pain relieving, nourishing blood for regulating menstruation, astringing YIN to stop sweating.Main component
There are peoniflorin, paeonol, flavone, monoterpene, triterpene and glycosides compound thereof.Modern pharmacological research find to have protecting the liver, ease pain, anti-
The effects such as scorching, regulation immunity, calmness, anti-bacteria and anti-virus.
Radix Salviae Miltiorrhizae, bitter in the mouth, cold nature, GUIXIN, Liver Channel.Function stasis-dispelling and pain-killing, promoting blood circulation to restore menstrual flow, clear away heart-fire relieving restlessness.Main pharmacological becomes
Divide and have Radix Salviae Miltiorrhizae acid, salvianolic acid, Tanshinone I, tanshinone ⅡA, cryptotanshinone I, iso tanshinone I etc. more than 40 to plant modern pharmacology research
Find Radix Salviae Miltiorrhizae have antithrombotic, change hemorheological property, resist myocardial ischemia, the effect such as antibacterial, antiinflammatory antitumor.
In clinical practice, the dosage of Chinese medicine composition of the present invention is 0.4-3g kg-1·d-1.Preferably, Chinese medicine
The dosage of compositions is 0.6-2g kg-1·d-1。
It should be noted that raw material of Chinese medicine of the present invention can be substituted by the Chinese medicine with same or similar function,
And these medical materials all can be concocted according to method in " Chinese Pharmacopoeia " or " Chinese medicine voluminous dictionary ".
Correspondingly, see Fig. 1, present invention also offers a kind of Chinese medicine composition treating insulin resistant and relevant disease thereof
The preparation method of thing, including:
S101, take Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae by formula, pulverized；
The Chinese medicine composition that Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae form formula in parts by weight is as follows:
Fructus Crataegi 100-300 part,
Radix Polygoni Multiflori Preparata 90-300 part,
Radix Paeoniae Alba 120-360 part,
Radix Salviae Miltiorrhizae 100-500 part.
Preferably, the Chinese medicine composition that Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae form formula in parts by weight is as follows:
Fructus Crataegi 180-240 part,
Radix Polygoni Multiflori Preparata 180-240 part,
Radix Paeoniae Alba 120-300 part,
Radix Salviae Miltiorrhizae 120-360 part.
S102, extract with methanol, ethanol or acetone and concentrate, obtain the first extracting solution；
As step S102 mono-preferred embodiment, including:
Add 6-10 times amount, concentration is the methanol of 60-80%, ethanol or acetone soak 20-40min；
Reflux, extract, 1-3 time, each 1-3h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
As step S102 more preferably embodiment, including:
Add 8 times amount, concentration be 70% methanol, ethanol or acetone soak 30min；
Reflux, extract, 2 times, each 2h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
As the embodiment that step S102 is optimal, including:
Add 8 times amount, concentration is soak with ethanol 30min of 70%；
Reflux, extract, 2 times, each 2h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
S103, will extract after medicinal residues water extraction and concentrate, obtain the second extracting solution；
As step S103 mono-preferred embodiment, including:
Medicinal residues 6-10 times amount water soaking 20-40min after extracting；
Water carries 1-3 time, for the first time 45-75min, for the second time 30-60min；
United extraction liquid concentrating under reduced pressure, obtain the second extracting solution.
As step S103 more preferably embodiment, including:
Medicinal residues 8 times amount water soaking 30min after extracting；
Water carries 2 times, 60min for the first time, for the second time 45min；
United extraction liquid concentrating under reduced pressure, obtain the second extracting solution.
S104, by the first extracting solution, second extracting solution merge.
Preferably, the first extracting solution, the second extracting solution merging are concentrated into extractum.
The present invention is the extract of natural plant crude drugs, and crude drug is cheap, wide material sources, and extraction process is simple, can
Realize large-scale industrialization application.
It should be noted that the methanol described in the preparation method shown in Fig. 1, ethanol or acetone, refer to methanol, ethanol,
Acetone or their aqueous solution.
Correspondingly, see Fig. 2, present invention also offers another kind for the treatment of insulin resistant and the Chinese drug-treated group of relevant disease thereof
The preparation method of compound, including:
S201, weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae by formula.
The Chinese medicine composition that Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae form formula in parts by weight is as follows:
Fructus Crataegi 10-30 part,
Radix Polygoni Multiflori Preparata 9-30 part,
Radix Paeoniae Alba 12-36 part,
Radix Salviae Miltiorrhizae 10-50 part.
Preferably, the Chinese medicine composition that Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae form formula in parts by weight is as follows:
Fructus Crataegi 12-24 part,
Radix Polygoni Multiflori Preparata 12-24 part,
Radix Paeoniae Alba 18-30 part,
Radix Salviae Miltiorrhizae 15-36 part.
S202, added water no medical material, soaks 20-40min；
Preferably, added water no medical material, soaks 30min.
S203, being heated to water and open, little fire continues to decoct 10-30min, filters, obtains the first medicinal liquid；
Preferably, being heated to water and open, little fire continues to decoct 15min, filters, obtains the first medicinal liquid.
S204, continued to add water no medical material, water after opening little fire continue to decoct 10-30min, filter, obtain the second medicinal liquid；
Preferably, continued to add water no medical material, and water is little fire continuation decoction 15min after opening, and filters, obtains the second medicinal liquid.
S205, merge the first medicinal liquid, the second medicinal liquid.
Correspondingly, the present invention also provides for a kind of Chinese medicine preparation treating insulin resistant and relevant disease thereof, described Chinese medicine
Preparation contains the Chinese medicine composition of any of the above-described embodiment.
Described Chinese medicine preparation is decoction, powder, capsule, tablet, granule or pill.
For making above-mentioned dosage form be achieved, the adjuvant that can use in pharmaceutical preparation need to be added in preparation process: as filled out
Fill agent, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, preservative etc..Filler includes: starch, lactose,
Mannitol, microcrystalline Cellulose, sucrose, xylitol etc.；Disintegrating agent includes: starch, microcrystalline Cellulose, carboxymethyl starch sodium, low take
For HPMC etc.；Lubricant includes: Pulvis Talci, silicon dioxide, sodium lauryl sulphate, magnesium stearate etc.；Suspending
Agent includes: polyvinylpyrrolidone, microcrystalline Cellulose, agar, hydroxymethyl cellulose etc.；Binding agent includes: starch slurry, polyethylene
Ketopyrrolidine, hydroxymethyl cellulose etc.；Sweeting agent includes: saccharin sodium, aspartame, sucrose, cyclamate, enoxolone etc.；Rectify
Taste agent includes: sweeting agent and various essence；Preservative includes: parabens, benzoic acid, sodium benzoate, benzalkonium bromide, acetic acid chlorine
Calmly, eucalyptus oil, sorbic acid and its esters etc..
It should be noted that the Chinese medicine composition contained by described Chinese medicine preparation, and the preparation side of this Chinese medicine composition
Method is consistent with the above, does not repeats them here.
The present invention is expanded on further below with specific embodiment
Embodiment 1
The preparation of medicine capsule of the present invention
Formula: Fructus Crataegi 200g, Radix Polygoni Multiflori Preparata 180g, Radix Paeoniae Alba 240g, Radix Salviae Miltiorrhizae 300g
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 8 times amount 70% soak with ethanol 30min, backflow
Extracting 2 times, 2h/ time, united extraction liquid also concentrates；Medicinal residues 8 times amount water soaking 30min, water are carried twice, 1h for the first time, the
Secondary 45min, united extraction liquid, extracting solution is merged, is spray-dried, add 1% magnesium stearate, load capsule and make 1000
Capsule.
Embodiment 2
The preparation of medicinal tablet of the present invention
Formula: Fructus Crataegi 180g, Radix Polygoni Multiflori Preparata 240g, Radix Paeoniae Alba 300g, Radix Salviae Miltiorrhizae 240
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 8 times amount 70% soak with ethanol 30min, backflow
Extracting 2 times, 2h/ time, united extraction liquid also concentrates；Medicinal residues 8 times amount water soaking 30min, water are carried twice, 1h for the first time, the
Secondary 45min, united extraction liquid, extracting solution is merged, is spray-dried, adds 1% magnesium stearate, 1000 tablets of medicines made by tabletting
Sheet.
Embodiment 3
The preparation of medicinal granule of the present invention
Formula: Fructus Crataegi 180g, Radix Polygoni Multiflori Preparata 300g, Radix Paeoniae Alba 360g, Radix Salviae Miltiorrhizae 120
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 8 times amount 70% soak with ethanol 30min, backflow
Extracting 2 times, 2h/ time, united extraction liquid also concentrates；Medicinal residues 8 times amount water soaking 30min, water are carried twice, 1h for the first time, the
Secondary 45min, united extraction liquid, extracting solution is merged, is spray-dried, add soluble starch, make granule.
Embodiment 4
The preparation of medicine concentrated pill of the present invention
Formula: Fructus Crataegi 240g, Radix Polygoni Multiflori Preparata 240g, Radix Paeoniae Alba 180g, Radix Salviae Miltiorrhizae 360g
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 8 times amount 70% soak with ethanol 30min, backflow
Extracting 2 times, 2h/ time, united extraction liquid also concentrates；Medicinal residues 8 times amount water soaking 30min, water are carried twice, 1h for the first time, the
Secondary 45min, united extraction liquid, extracting solution is merged, is concentrated into extractum, add dextrin, make concentrated pill.
Embodiment 5
The preparation of decoction of the present invention
Formula: Fructus Crataegi 12g, Radix Polygoni Multiflori Preparata 12g, Radix Paeoniae Alba 18g, Radix Salviae Miltiorrhizae 15g
Preparation method: weigh above-mentioned medical material, added water no medical material, soaks 30min, is heated to water and opens, and little fire continues to decoct
15min, filters, and continued to add water no medical material, and water is little fire continuation decoction 15min after opening, and merges medicinal liquid and get final product.
Embodiment 6
The preparation of medicine capsule of the present invention
Formula: Fructus Crataegi 100g, Radix Polygoni Multiflori Preparata 90g, Radix Paeoniae Alba 120g, Radix Salviae Miltiorrhizae 100g
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 7 times amount 60% methanol and soaks 35min, backflow
Extracting 2 times, 2h/ time, united extraction liquid also concentrates；Medicinal residues 7 times amount water soaking 25min, water are carried twice, 1h for the first time, the
Secondary 45min, united extraction liquid, extracting solution is merged, is spray-dried, add 1% magnesium stearate, load capsule and make 1000
Capsule.
Embodiment 7
The preparation of medicinal tablet of the present invention
Formula: Fructus Crataegi 260g, Radix Polygoni Multiflori Preparata 200g, Radix Paeoniae Alba 200g, Radix Salviae Miltiorrhizae 280
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 6 times amount 80% soak with ethanol 20min, backflow
Extracting 2 times, 3h/ time, united extraction liquid also concentrates；Medicinal residues 6 times amount water soaking 20min, water are carried twice, 45min for the first time,
30min for the second time, united extraction liquid, extracting solution is merged, is spray-dried, adds 1% magnesium stearate, 1000 tablets of medicines made by tabletting
Sheet.
Embodiment 8
The preparation of medicinal granule of the present invention
Formula: Fructus Crataegi 300g, Radix Polygoni Multiflori Preparata 300g, Radix Paeoniae Alba 360g, Radix Salviae Miltiorrhizae 500
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 10 times amount 60% acetone soak 10min, backflow
Extracting 3 times, 3h/ time, united extraction liquid also concentrates；Medicinal residues 10 times amount water soaking 40min, water are carried 3 times, 75min for the first time,
60min for the second time, united extraction liquid, extracting solution is merged, is spray-dried, add soluble starch, make granule.
Embodiment 9
The preparation of medicine concentrated pill of the present invention
Formula: Fructus Crataegi 250g, Radix Polygoni Multiflori Preparata 220g, Radix Paeoniae Alba 200g, Radix Salviae Miltiorrhizae 320g
Preparation method: weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, adds 8 times amount 70% soak with ethanol 30min, backflow
Extracting 2 times, 2h/ time, united extraction liquid also concentrates；Medicinal residues 8 times amount water soaking 30min, water are carried twice, 1h for the first time, the
Secondary 45min, united extraction liquid, extracting solution is merged, is concentrated into extractum, add dextrin, make concentrated pill.
Embodiment 10
The preparation of decoction of the present invention
Formula: Fructus Crataegi 13g, Radix Polygoni Multiflori Preparata 13g, Radix Paeoniae Alba 20g, Radix Salviae Miltiorrhizae 16g
Preparation method: weigh above-mentioned medical material, added water no medical material, soaks 20min, is heated to water and opens, and little fire continues to decoct
10min, filters, and continued to add water no medical material, and water is little fire continuation decoction 10min after opening, and merges medicinal liquid and get final product.
Embodiment 11
The preparation of decoction of the present invention
Formula: Fructus Crataegi 15g, Radix Polygoni Multiflori Preparata 15g, Radix Paeoniae Alba 25g, Radix Salviae Miltiorrhizae 20g
Preparation method: weigh above-mentioned medical material, added water no medical material, soaks 40min, is heated to water and opens, and little fire continues to decoct
20min, filters, and continued to add water no medical material, and water is little fire continuation decoction 20min after opening, and merges medicinal liquid and get final product.
Embodiment 12
The preparation of decoction of the present invention
Formula: Fructus Crataegi 20g, Radix Polygoni Multiflori Preparata 20g, Radix Paeoniae Alba 30g, Radix Salviae Miltiorrhizae 25g
Preparation method: weigh above-mentioned medical material, added water no medical material, soaks 40min, is heated to water and opens, and little fire continues to decoct
20min, filters, and continued to add water no medical material, and water is little fire continuation decoction 20min after opening, and merges medicinal liquid and get final product.
Being further elaborated the present invention below in conjunction with experimental data, the present invention is with Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae
For crude drug, and being prepared by above-mentioned preparation method, hereinafter referred to as SHBD, its drug action and case study are as follows:
I, the SHBD Contained Serum impact on HepG2 cell insulin resistant model
One, experiment material
Cell: HepG2 cell strain, Tumor Hospital Attached to Zhongshan Univ. grants.
Animal: SD male rat, SPF level, body weight 180～220g, purchased from Guangdong Medical Lab Animal Center, produce and permitted
Can the number of card: SCXK (Guangdong) 0105817.
Medicine: Chinese medicine composition SHBD: Fructus Crataegi, Kangmei Pharmaceutical Co., Ltd's (product batch number: 120304861)；System
Radix Polygoni Multiflori, Kangmei Pharmaceutical Co., Ltd's (product batch number: 111203551)；The Radix Paeoniae Alba, Kangmei Pharmaceutical Co., Ltd is (raw
Product lot number: 120205421), Radix Salviae Miltiorrhizae, Guangzhou Lingnan prepared slices of Chinese crude drugs company limited (product batch number: 091008)
Metformin hydrochloride tablet, Shanghai Shi Guibao pharmaceutical Co. Ltd of Sino-U.S. (product batch number: 1204101)
Simvastatin blade, Hangzhou Mo Shadong pharmaceutical Co. Ltd (product batch number: 110593).
Reagent: RPMI 1640 culture medium, Hyclone；Hyclone, GIBCO；The pancreatin of 0.25%, Hyclone；Islets of langerhans
Element, Sigma；Glucose determination reagent box, Meikang Biotech Co., Ltd., Ningbo；G6Pase ELISA kit.
Instrument: carbon dioxide cell incubator, Thermo；IX71+DP72 type is inverted micro imaging system, Olympus；
ST16R frozen type common bench centrifuge, Thermo；Electric-heated thermostatic water bath, Shanghai hundred allusion quotation instrument and equipment company limited；Enzyme mark
Instrument, Bio-Rad；The multi-functional automatic biochemistry analyzer of ultramicron, Trilogy；Intelligent constant-temperature digital display electric jacket, Henan Ai Boteke
Skill Development Co., Ltd；Rotary Evaporators, IKA；
Two, experimental technique
(1) medicinal liquid extracts and concentrates
Medicinal liquid: prepare by the above-mentioned detailed description of the invention of the present invention, be settled to 0.864g ml-1。
Diformin tablet medicinal liquid: distilled water dissolves diformin tablet, and concentration is 20mg ml-1。
Simvastatin Tablets medicinal liquid: distilled water dissolves Simvastatin Tablets, and concentration is 1mg ml-1。
(2) preparation of Contained Serum
Male SD rat, 180-220g, by body weight be randomly divided into Normal group, simvastatin group, metformin group,
SHBD group, gives normal saline, simvastatin, metformin and SHBD medicinal liquid respectively, by every 100g body weight 1ml gavage, every day 2
Secondary administration, successive administration 3 days, after last 1 time is administered 1.5h, anesthetized rat, abdominal aortic blood, stand several minutes, centrifugal, take
Serum, each group serum mixes respectively, and 56 DEG C of inactivation 30min through 0.22 μm syringe needle micropore filter filtration sterilization, thus prepare each
Group Contained Serum ,-20 DEG C of Refrigerator stores, standby.
(3) HepG2 insulin resistant model is set up and pharmaceutical intervention processes
After being in the HepG2 cell dissociation of exponential phase, RPMI 1640 culture medium suspendible cell.Cultivate cell extremely
After cell attachment, with containing 1 × 10-8mol·L-1The culture medium of insulin acts on HepG2 cell 36h and sets up HepG2 cell tryptase
Insulin resistance model.
Experiment is grouped into SHBD large, medium and small dosage group (Contained Serum is diluted to 15%, 10%, 5%), metformin group
(Contained Serum is diluted to 15%), simvastatin group (Contained Serum is diluted to 15%), normal group and model group.HepG2 islets of langerhans
After element opposing model modeling success, give different Contained Serums respectively and intervene 24h.(4)3H-D-glucose participates in test inspection
Survey the impact on HepG2 cell tryptase insulin resistance Modeling glucose uptake ratio of the SHBD Contained Serum
After HepG2 insulin resistant model modeling success, give different Contained Serums respectively and intervene 24h.Add3H-D-
Glucose (1.3 μ Ci ml-1In culture fluid) hatch 2h, add 4 DEG C of phosphate buffer quick wash cells to terminate reaction.
After cell cracks with 20%KOH 0.5ml, it is transferred in test tube.Take the cell pyrolysis liquid of 100 μ l, use Coomassie Brilliant Blue to survey
Determine protein concentration.4 DEG C of ethanol solution wash 2 times.Room temperature 3000r min-1Abandoning supernatant after centrifugal 5min, by precipitate
(glycogen) is inverted and is deposited on the filter paper of diameter 2cm (repeatedly rinsing), and filter paper cold preservation is in the ethanol agitator treating 4 of 4 DEG C
Secondary, 3h altogether.Filter paper is dried and is placed in 10ml scintillation solution mensuration radiocounting.According to3The ratio radiation of H-D-glucose
Property, the protein concentration of cell pyrolysis liquid and the radioactivity of glycogen, calculate3The rate that participates in of H-D-glucose, unit is mmol
gPro·h-1.Formula is: the radiocounting (cpm) of the uptake ratio=glycogen of glucose/[3The specific activity of H-D-glucose
(Bq/mmol) protein concentration (the g l of × cell pyrolysis liquid-1) × cell pyrolysis liquid volume (ml) × incubative time (h)]
(5) the SHBD Contained Serum impact on HepG2 cell tryptase insulin resistance Modeling glucose consumption
With containing 1 × 10-8mol·L-1The culture medium of insulin acts on HepG2 cell 36h and sets up HepG2 cell insulin
Opposing model, after model is set up, after Contained Serum processes 24h, collects each group of cell culture medium supernatant, and ultramicron is multi-functional certainly
The glucose content of Automatic Biochemical Analyzer detection culture medium supernatant, and calculate the glucose utilization of each group of cell.
(6) SHBD Contained Serum to HepG2 cell insulin resistant model carbohydrate metabolism correlation factor IRS-1, IRS-2,
The expression regulation of GLUT2, GLUT4
After insulin resistant cell model modeling success, each Contained Serum effect 24h, discard culture medium.
(1) cell total RNA extracts
PBS 2 times, every 10cm2The cultivation cell of growth adds the TRIZOL reagent of 1ml in order to cell lysis, general
Interior celliferous lysate is transferred in centrifuge tube, liquid-transfering gun pressure-vaccum repeatedly, and room temperature stands 5min.Add chloroform, acutely vibrate
15sec, after solution is fully emulsified, room temperature stands 5min, and rear 12,000g, 4 DEG C of centrifugal 15min, Aspirate supernatant is transferred to separately
In one new centrifuge tube.Adding isopyknic isopropanol in supernatant, after mixing of turning upside down, 15～30 DEG C stand 10min.After
12,000g, 4 DEG C of centrifugal 10min.Careful supernatant discarded, adds the ethanol 1ml of 75%, the most lentamente along centrifugal tube wall
Reverse, carefully discard ethanol after 12,000g 4 DEG C of centrifugal 5min.Drying at room temperature precipitation 2～5min, adds appropriate RNase-
Free water dissolution precipitates.The absorbance of UV spectrophotometer measuring RNA, calculates content and the purity of total serum IgE.
(2) reverse transcription reaction:
1. the removing reaction of genomic DNA
Room temperature 5min, 4 DEG C.
2. reverse transcription reaction (reactant liquor is formulated in and carries out on ice)
37 DEG C of 15min, 85 DEG C of 5sec, 4 DEG C.
(3) Real Time PCR reaction
1. primer sequence
2. PCR reaction system
Reaction condition: Stage 1: denaturation, 95 DEG C, 30sec, 1Reps；Stage 2:PCR reacts, 95 DEG C, 5sec, and 60
DEG C, 34sec, 40Reps.
After PCR reaction terminates, verified amplification efficiency and the primer specific of primer by primer amplification curve and solubility curve
Property, according to 2-△△CtCalculating the relative expression quantity of each gene, wherein, △ Ct deducts reference gene equal to the Ct of genes of interest group
The Ct of group, wherein, β-actin is reference gene, and-△ △ Ct deducts the △ Ct of matched group equal to the △ Ct of Contained Serum group, with
Model group is comparison, then the 2 of model group-△△CtValue is 1.
(7) Contained Serum is to HepG2 cell insulin resistant model lipid metabolism correlation factor PPAR-γ and β3The table of AR
Reach regulation and control
Experimental procedure is with (six), and primer sequence is:
(9) statistical analysis
Experiment different batches cell is repeated 3 times above, carries out one factor analysis of variance, experimental data mean ± standard
DifferenceRepresent, with P ＜ 0.05 for statistically significant.
Three, experimental result
The impact on HepG2 cell tryptase insulin resistance Modeling glucose uptake ratio of the 1.SHBD Contained Serum
Insulin resistant cell model glucose uptake rate after medicine effect
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compared with normal group, model group glucose uptake rate significantly reduces (P < 0.05)；After pharmaceutical intervention, metformin
In group, SHBD heavy dose group and SHBD, dosage group glucose uptake rate significantly raises, have compared with model group significant difference (P <
0.05)。
The impact on HepG2 cell tryptase insulin resistance Modeling glucose consumption of the 2.SHBD Contained Serum
Insulin resistant cell model glucose utilization after medicine effect
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Comparing with normal group, model group glucose utilization significantly reduces (P < 0.05)；After pharmaceutical intervention, metformin
Group, SHBD big or middle dosage group glucose utilization relatively model group dramatically increases (P < 0.05).
3.SHBD Contained Serum is to HepG2 cell tryptase insulin resistance fasting insulin receptor substrate IRS-1, IRS-2mRNA
The impact expressed
Insulin resistant cell model IRS-1, IRS-2mRNA relative expression quantity after medicine effect
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compared with normal group, model group IRS-1, IRS-2mRNA relative expression quantity significantly reduce (P < 0.05)；Do through medicine
Prognosis, compared with model group, metformin group, SHBD heavy dose group with dosage group IRS-1, IRS-2mRNA in SHBD relative to table
The amount of reaching significantly raises (P < 0.05)；Simvastatin group IRS-2mRNA relative expression quantity also has notable rising compared with model group
(P < 0.05), but relatively SHBD heavy dose group significantly reduces (P < 0.05).
4.SHBD Contained Serum is to HepG2 cell tryptase insulin resistance Modeling glucose transport protein GLUT2, GLUT4mRNA
The impact expressed
Insulin resistant cell model GLUT2, GLUT4mRNA relative expression quantity after medicine effect
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compared with normal group, model group GLUT2, GLUT4mRNA relative expression quantity significantly reduce (P < 0.05)；Do through medicine
Prognosis, compared with model group, metformin group, SHBD big or middle dosage group GLUT2, GLUT4mRNA relative expression quantity significantly raise
(P<0.05)。
Four, experiment conclusion
Experimental result shows, SHBD Contained Serum can improve HepG2 cell tryptase insulin resistance Modeling glucose uptake ratio, increases
Add the insulin resistant model cell consumption to glucose；Meanwhile, SHBD Contained Serum can raise IRS
IRS-1, IRS-2 and glucose transporter GLUT2、GLUT4The expression of mRNA.Result shows, SHBD side's Contained Serum is by adjusting
Joint IRS is expressed, and increases the cell sensitivity to insulin；Simultaneously by the table of upregulation of glucose transport albumen
Reach, increase the cell picked-up to glucose, thus play the increase insulin resistant cell sensitivity to insulin, improve islets of langerhans
The element opposing cell effect to glucose consumption.
II, SHBD side's impact on insulin resistant model rat
One, experiment material
Laboratory animal: SD male rat, SPF level, body weight 180～220g, 100, purchased from Guangdong Province medical experiment animal
Center, production licence number: SCXK (Guangdong) 0105817.
High fat height sugar high salt emulsion formulations raw material: yolk powder, cholesterol, bovine bile, propylthiouracil, white sugar, food
Salt, Adeps Sus domestica.
Medicine: Chinese medicine composition SHBD: Fructus Crataegi, Kangmei Pharmaceutical Co., Ltd's (product batch number: 120304861)；System
Radix Polygoni Multiflori, Kangmei Pharmaceutical Co., Ltd's (product batch number: 111203551)；The Radix Paeoniae Alba, Kangmei Pharmaceutical Co., Ltd is (raw
Product lot number: 120205421)；Radix Salviae Miltiorrhizae, Guangzhou Lingnan prepared slices of Chinese crude drugs company limited (product batch number: 091008)
Metformin hydrochloride tablet, Shanghai Shi Guibao pharmaceutical Co. Ltd of Sino-U.S. (product batch number: 1204101)
Simvastatin Tablets, Hangzhou Mo Shadong pharmaceutical Co. Ltd (product batch number: 110593).
Reagent: pyrophosphoric acid diethylester (DEPC), Sigma；Chloroform (analytical pure), Guangzhou Chemical Reagent Factory；Isopropanol (is analyzed
Pure), Guangzhou Chemical Reagent Factory；Ethanol (analytical pure), Guangzhou Chemical Reagent Factory；PCR primer, TAKARA；Trizol Plus,
TAKARA, Lot:A8601-1；Prime Script RT reagent Kit with gDNA, TAKARA, Lot:BK1301；
SYBR Premix Ex Taq II, TAKARA, Lot:BK1303.
Instrument: intelligent constant-temperature digital display electric jacket, Henan Ai Bote development in science and technology company limited；Rotary Evaporators, SENCO is public
Department；Electronic balance, end Kohler Co；Biological functional system, Chengdu TME Technology Co., Ltd.；Ultramicron animal specific is many
Function automatic biochemistry analyzer, Trilogy；Automatically blood rheological instrument, Pulisheng Instruments Co., Ltd., Beijing；Microscope,
OLYMPUS。
Two, experimental technique
(1) foundation of Rat model of insulin-resistant and veterinary method
Healthy male SD rat 100, SPF level, body weight 180～220g, adaptability is raised 1 week, is randomly selected by body weight
10 are set to normal group, and remaining 90 use only for modeling.Before modeling group rat, 4 week every day gave the high fat without propylthiouracil
High sugar high salt Emulsion gavage 1ml/100g/d.From the beginning of the 5th week, give the high fat height sugar high salt Emulsion containing propylthiouracil
Gavage 1ml/100g/d, continuous 2 weeks.Normal group Isodose distilled water gavage, normal water.All feeding rats are commonly raised
Material, feedstuff and water are changed once every day.
High fat height sugar high salt Emulsion is prepared: Adeps Sus domestica 20%, cholesterol 5%, yolk powder 6%, bovine bile 1%, white sugar
20%, salt 10%.Take 20g Adeps Sus domestica, 5g cholesterol, 6g yolk powder, 1g bovine bile, 20g white sugar, 10g salt are put in beaker, add
Enter 100ml distilled water, put in constant temperature electric heating set and heat and be sufficiently agitated until and be completely dissolved, high fat height sugar high salt breast
Agent, continuous gavage, after 4 weeks, adds 0.1% propylthiouracil, then gavage 2 weeks continuously in Emulsion.4 DEG C of stored refrigerated of Emulsion, make
Melt with front 37 DEG C of heating in water bath.
In modeling the 6th weekend evenings fasting 12 hours, chloral hydrate intraperitoneal anesthesia next day 10%, eye socket only takes blood 1ml/,
3000rpm, centrifugal 15min take serum, and multi-functional automatic biochemistry analyzer detection rat blood sugar is, and use to put and exempt from method detection islets of langerhans
Cellulose content.Calculate insulin sensitivity index (insulin sensitivity index, ISI).ISI=Ln (FBG × FINS)-1
(FBG: fasting glucose, FINS: Diagnostic Value of Fasting Serum insulin).
(2) animal packet and administration
Choose the successful rat of modeling and be randomly divided into 6 groups, be model group, simvastatin group, metformin group, SHBD respectively
Large, medium and small dosage group, together with normal group totally 7 groups.
Model group and each medication group gavage in morning height fat height sugar high salt Emulsion 1ml/100g/d, each medication in afternoon group fills respectively
Stomach simvastatin, metformin and the large, medium and small dosage of SHBD medicinal liquid, model group and normal group gavage normal saline, normally drink
Water.All feeding rats normal diets, feedstuff and water are changed once every day, continuous 6 weeks.
Prepared by medicinal liquid: prepare by the above-mentioned detailed description of the invention of the present invention, be settled to 0.864g ml-1, heavy dose of for SHBD
Group；Middle dosage group dilute is 0.432g ml-1；Small dose group dilute is 0.216g ml-1；By 100g body weight 1ml
Gastric infusion.
Simvastatin medicinal liquid: distilled water dissolves Simvastatin Tablets, and liquor strength is 0.5mg ml-1, by 5mg kg-1·d-1Dosed administration.
Metformin medicinal liquid: distilled water dissolves diformin tablet, and liquor strength is 10mg ml-1, by 100mg kg-1·
d-1Dosed administration.
Animal dosage is according to " pharmacological experimental methodology " (third edition, Xu Shuyun, Bian Rulian, Chen Xiu chief editor, Ren Minwei
Raw publishing house, 2002) " between subordinate list 11-8 humans and animals, pressing the dose,equivalent ratio of body surface area conversion " (P1861), according to
The dosage of people is converted into rat dose,equivalent:
Rat dose,equivalent=(every consumption per day × 0.018 of people)/0.2
(3) collection of specimens
After medication 6 weeks, process the upper fasting evening before yesterday, m seq 10% chloral hydrate intraperitoneal anesthesia, abdominal aortic blood, divide
From serum, detection serum blood glucose and insulin level；Take whole blood and blood plasma detection hemorheology of rat and endothelin level water
Flat.
Taking rat liver, hind leg skeletal muscle and omental adipose tissue, put in centrifuge tube ,-80 DEG C of Refrigerator stores are standby.
For detecting the expression of IRS-1, IRS-2, GLUT2, GLUT4mRNA in tissue, and and PPAR-γ, β3The table of AR mRNA
Reach.
(4) detection method
1. IRS-1, IRS-2, GLUT in liver organization2、β3GLUT in AR mrna expression, skeletal muscle4Mrna expression is with big
The expression of PPAR-γ mRNA in omental adipose tissue
(1) extraction of liver total RNA:
-80 DEG C of refrigerators take out each group of rat liver, skeletal muscle and omental adipose tissue, PBS once, often
100mg liver adds the TRIZOL reagent of 1ml, and the centrifuge tube including rat tissue and lysate is put into Multifunctional biological sample
Homogenate in homogenizer, room temperature stands 5min.4 DEG C, 12,000g, centrifugal 5min, take supernatant, add the chloroform of supernatant 1/5 volume,
Acutely vibrate 15sec, and after solution is fully emulsified, room temperature stands 5min, rear 12,000g, 4 DEG C of centrifugal 15min, Aspirate supernatant
It is transferred in another new centrifuge tube.Isopyknic isopropanol is added in supernatant, after mixing of turning upside down, 15～30 DEG C of standings
10min.Rear 12,000g, 4 DEG C of centrifugal 10min.Careful supernatant discarded, adds the ethanol 1ml of 75% lentamente along centrifugal tube wall,
Turn upside down gently, after 12,000g4 DEG C of centrifugal 5min, carefully discard ethanol.Drying at room temperature precipitation 2～5min, adds appropriate
RNase-free water dissolution precipitates.
(2) reverse transcription reaction:
1. the removing reaction of genomic DNA
Room temperature 5min, 4 DEG C.
2. reverse transcription reaction (reactant liquor is formulated in and carries out on ice)
37 DEG C of 15min, 85 DEG C of 5sec, 4 DEG C.
(3) Real Time PCR reaction
1. primer sequence
2. PCR reaction system
Reaction condition: Stage 1: denaturation, 95 DEG C, 30sec, 1Reps；Stage 2:PCR reacts, 95 DEG C, 5sec, and 60
DEG C, 34sec, 40Reps.
After PCR reaction terminates, verified amplification efficiency and the primer specific of primer by primer amplification curve and solubility curve
Property, according to 2-△△CtCalculating the relative expression quantity of each gene, wherein, △ Ct deducts reference gene equal to the Ct of genes of interest group
The Ct of group, wherein, GAPDH is reference gene, and-△ △ Ct deducts the △ Ct of matched group equal to the △ Ct of medication group, with model group
For comparison, then the 2 of model group-△△CtValue is l.
Three, experimental result
1.SHBD side is on insulin resistant model rat blood sugar and the impact of serum insulin
Each experimental group rat fasting blood-glucose, serum insulin and insulin sensitivity index level
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compared with normal group, the significantly rising of model group rats fasting glucose (FBS) and serum insulin levels (Ins) (P <
0.05), insulin sensitivity index (ISI) reduces ((P < 0.05)), and Rat model of insulin-resistant modeling success is described；Through medicine
After intervention, compared with model group, metformin group and SHBD big or middle dosage group rat fasting blood-glucose (FBG) and serum insulin
Level (Ins) significantly reduces (P < 0.05), and insulin sensitivity index (ISI) raises；Compared with model group, simvastatin group,
SHBD small dose group FBS, Ins, ISI there are no significant difference (P > 0.05).
The impact on insulin resistant model hemorheology of rat of the 5.SHBD side
Each experimental group hemorheology of rat level
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compared with normal group, model group rats whole blood viscosity and plasma viscosity significantly raise (P < 0.05), illustrate through height
After fat height sugar high salt Emulsion is fed, hemorheology of rat changes, and blood viscosity increases；After pharmaceutical intervention, with mould
Type group is compared, and simvastatin group and SHBD big or middle dosage group whole blood viscosity and plasma viscosity all significantly reduce (P < 0.05).
The impact on insulin resistant model rat plasma Endothelin (ET-1) of the 6.SHBD side
Each experimental group rat plasma level of ET
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compared with normal group, model group plasma ET significantly raises (P < 0.05), illustrates that insulin resistant causes
Vascular endothelial function is impaired, and endothelin level synthesis secretion increases；After pharmaceutical intervention, compared with model group, metformin group,
In simvastatin group, SHBD heavy dose group and SHBD, dosage group plasma ET significantly reduces (P < 0.05).
4.SHBD side is to insulin resistant model rat liver IRS-1, IRS-2, GLUT2, β 3AR mrna expression level
Impact
Each experimental group rat liver IRS-1, IRS-2, GLUT2, β 3AR mRNA relative expression quantity
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compare with normal group, model group rats liver IRS-1, IRS-2, GLUT2, β3AR mRNA relative expression quantity is the most aobvious
Write and reduce (P < 0.05)；After pharmaceutical intervention, compared with model group, metformin group and SHBD big or middle dosage group IRS-1,
IRS-2, GLUT2mRNA relative expression quantity significantly raises (P < 0.05), and SHBD heavy dose group β 3AR mRNA relative expression quantity is notable
Raise (P < 0.05).
5.SHBD side is to insulin resistant model rat skeletal muscle GLUT4The impact of mrna expression level
Each experimental group rat skeletal muscle GLUT4mRNA relative expression quantity
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Comparing with normal group, model group rats skeletal muscle GLUT4mRNA relative expression quantity significantly reduces (P < 0.05)；Through medicine
After thing is intervened, compared with model group, metformin group and SHBD big or middle dosage group GLUT4mRNA relative expression quantity significantly raise
(P<0.05)。
The impact on insulin resistant model omental adipose tissue PPAR-γ mrna expression level of the 6.SHBD side
Each experimental group rat omental adipose tissue PPAR-γ mRNA relative expression quantity
Compared with normal group, △ P < 0.05；Compared with model group, * P < 0.05.
Compare with normal group, model group rats omental adipose tissue PPAR-γ mRNA relative expression quantity significantly reduce (P <
0.05)；After pharmaceutical intervention, compared with model group, simvastatin group dosage big or middle with SHBD group PPAR-γ mRNA is relative to table
The amount of reaching significantly raises (P < 0.05).
Four, experiment conclusion
Test result indicate that, SHBD can significantly reduce insulin resistant model rat fasting blood-glucose and serum insulin water
Flat, improve the insulin sensitivity index of rat model；The blood high viscosity state caused by insulin resistant can be reduced, reduce mould
Type rat whole blood viscosity and plasma viscosity；The vascular endothelial injury that energy reduction is caused by insulin resistant, reduction endothelin level-
1 level.After being intervened by SHBD side, rat model regulates glycometabolic gene IRS-1, IRS-2, GLUT2、GLUT4Mrna expression
Raise, regulate lipometabolic β 3AR and PPAR-γ mrna expression raises, demonstrate SHBD side's treatment insulin resistant further
And the effectiveness of the relevant disease caused.
In sum, the present invention is made up of Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae, has the following characteristics that
One, the compositions drug action of the present invention is notable, has insulin resistant and relevant disease thereof and preferably treats work
With.Mechanism of action of the present invention is clear and definite, and research discovery said composition targeting is in the related pathways of glucose-lipid metabolism, by regulation and control
The expression of IRS IRS-1, IRS-2 gene, improves the body sensitivity to insulin；Turned by upregulation of glucose
Fortune Protein G LUT2, the expression of GLUT4mRNA, promote the body picked-up to glucose, increases the body consumption to glucose；Logical
Cross rise PPAR-γ, the expression of β 3AR mRNA, promote the catabolism of adipose cell；Preventing and treating insulin is played from gene level
The effect of opposing.Meanwhile, said composition also has improves hemorheology, reduces endothelin-1 (ET-1) level, plays preventing and treating
Thrombosis and the effect of protection vascular endothelial function.Said composition plays treatment insulin resistant and phase thereof by many aspects
The effect of related disorders.
Two, the present invention is centered by improving insulin resistant, by regulating the signal path relevant to insulin resistant, with
Treatment insulin resistant is starting point, and then plays the treatment hyperinsulinemia relevant to insulin resistant, diabetes, high blood
The multiple diseases such as pressure, hyperlipidemia, metabolism syndrome, hyperuricemia, blood viscosity increase, reach the effect of " the many diseases of medicine ", carry
High patient compliance, alleviates economy and the physiological load of patient.
Three, the present invention is Chinese medicine preparation, and medicine composition is simple, and dose is little, and supports insulin through effect experiment checking
Anti-and relevant disease has obvious therapeutical effect, and toxic and side effects is little.
Four, the present invention is the extract of natural plant crude drugs, and crude drug is cheap, wide material sources, and extraction process is simple.
The above is the preferred embodiment of the present invention, it is noted that for those skilled in the art
For, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications are also considered as
Protection scope of the present invention.
Claims (10)
1. the Chinese medicine composition treating insulin resistant and relevant disease thereof, it is characterised in that described Chinese medicine composition by
The raw material of Chinese medicine of following weight proportion is made:
Fructus Crataegi 100-300,
Radix Polygoni Multiflori Preparata 90-300,
Radix Paeoniae Alba 120-360,
Radix Salviae Miltiorrhizae 100-500.
2. Chinese medicine composition as claimed in claim 1, it is characterised in that described Chinese medicine composition is by following weight proportion
Medicine raw material is made:
Fructus Crataegi 180-240,
Radix Polygoni Multiflori Preparata 180-240,
Radix Paeoniae Alba 120-300,
Radix Salviae Miltiorrhizae 120-360.
3. the method for the Chinese medicine composition prepared as described in any one of claim 1-2, it is characterised in that including:
A, take Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae by formula, pulverized；
B, extract with methanol, ethanol or acetone and concentrate, obtain the first extracting solution；
C, will extract after medicinal residues water extraction and concentrate, obtain the second extracting solution；
D, by the first extracting solution, second extracting solution merge.
4. preparation method as claimed in claim 3, it is characterised in that described step B includes:
Add 6-10 times amount, concentration is the methanol of 60-80%, ethanol or acetone soak 20-40min；
Reflux, extract, 1-3 time, each 1-3h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
5. preparation method as claimed in claim 4, it is characterised in that described step B includes:
Add 8 times amount, concentration be 70% methanol, ethanol or acetone soak 30min；
Reflux, extract, 2 times, each 2h；
United extraction liquid concentrating under reduced pressure recycling design, obtain the first extracting solution.
6. preparation method as claimed in claim 3, it is characterised in that described step C includes:
Medicinal residues 6-10 times amount water soaking 20-40min after extracting；
Water carries 1-3 time, for the first time 45-75min, for the second time 30-60min；
United extraction liquid concentrating under reduced pressure, obtain the second extracting solution.
7. preparation method as claimed in claim 6, it is characterised in that described step C includes:
Medicinal residues 8 times amount water soaking 30min after extracting；
Water carries 2 times, 60min for the first time, for the second time 45min；
United extraction liquid concentrating under reduced pressure, obtain the second extracting solution.
8. the method for the Chinese medicine composition prepared as described in any one of claim 1-2, it is characterised in that including:
A, weigh Fructus Crataegi, Radix Polygoni Multiflori Preparata, the Radix Paeoniae Alba, Radix Salviae Miltiorrhizae by formula；
B, added water no medical material, soaks 20-40min；
C, being heated to water and open, little fire continues to decoct 10-30min, filters, obtains the first medicinal liquid；
D, continued to add water no medical material, water after opening little fire continue to decoct 10-30min, filter, obtain the second medicinal liquid；
E, merge the first medicinal liquid, the second medicinal liquid.
9. the Chinese medicine preparation treating insulin resistant and relevant disease thereof, it is characterised in that described Chinese medicine preparation contain as
Chinese medicine composition described in any one of claim 1-2.
10. Chinese medicine preparation as claimed in claim 9, it is characterised in that described Chinese medicine preparation is decoction, powder, capsule, sheet
Agent, granule or pill.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101574423A (en) * | 2008-05-05 | 2009-11-11 | 天津天士力制药股份有限公司 | Application of medicine combination in preparing medicament for reducing insulin resistance |
| CN101618176A (en) * | 2008-07-04 | 2010-01-06 | 陈民 | Chinese medicament for treating dyslipidemia and atherosclerosis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101574423A (en) * | 2008-05-05 | 2009-11-11 | 天津天士力制药股份有限公司 | Application of medicine combination in preparing medicament for reducing insulin resistance |
| CN101618176A (en) * | 2008-07-04 | 2010-01-06 | 陈民 | Chinese medicament for treating dyslipidemia and atherosclerosis |
Non-Patent Citations (1)
| Title |
|---|
| 山楂叶总黄酮防治大鼠胰岛素抵抗及脂肪肝的实验研究;刘江等;《华东师范大学学报(自然科学版)》;20081130(第06期);127-132 * |
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