CN108815215B - Lu Li grass root water extract and application thereof in reducing blood sugar - Google Patents
Lu Li grass root water extract and application thereof in reducing blood sugar Download PDFInfo
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Abstract
The invention discloses a water extract of a root of a ruelia chinensis bunge and application thereof in reducing blood sugar. The invention firstly discloses a water extract of the roots of the ruelia chinensis bunge, which is obtained by extracting the roots of the ruelia chinensis bunge with water as an extraction solvent. The established diabetes animal model proves that the water extract of the root tuber of the ruelia chinensis has obvious hypoglycemic effect. The invention further adopts a macroporous resin technology and a step-by-step alcohol precipitation method to collect the water extract of the roots of the Trigonella foenum-graecum in sections to obtain four different fractions, wherein compared with the other three fractions, the fraction 3(LLC-3) has an extremely obvious blood sugar reduction effect and is an effective part of the water extract of the roots of the Trigonella foenum-graecum for reducing blood sugar; therefore, the invention further provides the hypoglycemic effective part of the water extract of the roots of the ruelia chinensis. The invention further provides the application of the water extract of the roots of the ruelia chinensis and the effective part of the water extract of the roots of the ruelia chinensis in preparing the hypoglycemic medicament.
Description
Technical Field
The invention relates to a water extract of Trigonella foenum-graecum (Ruellia tuberosa Linn) and pharmacological application thereof, in particular to a water extract of the root of the Trigonella foenum-graecum and application thereof in reducing blood sugar, belonging to the field of the water extract of the Trigonella foenum-graecum and the pharmacological application thereof.
Background
Rheum palmatum (Ruellia tuberosa Linn) belongs to the order of Convolvulaceae (Acanthaceae) Dicotyledoneae, subclass Hemerophyceae, Siphonophyceae. The ruelia is a perennial plant mainly distributed in tropical and subtropical regions, and is widely distributed in southeast Asia, subtropical regions in China and other regions. In the traditional Chinese medicine, the leaf of the ruelia chinensis is mainly used as the medicine, and the traditional Chinese medicine has the effects of promoting urination, treating psychosis, relieving pain, reducing blood pressure, quenching thirst, detoxifying and the like.
Diabetes is a metabolic disease characterized by hyperglycemia, which is caused by defects in insulin secretion or impaired biological action, or both. Hyperglycemia often causes chronic damage and dysfunction of various tissues, particularly eyes, kidneys, heart, blood vessels and nerves, often with the following hazards:
firstly, the damage of the kidney, due to hyperglycemia, hypertension and hyperlipidemia, glomerular microcirculation filtration pressure is abnormally increased, and the occurrence and the development of diabetic nephropathy are promoted. The early stage of the disease is manifested by proteinuria and edema, and the late stage of the disease is renal failure, which is the most main cause of death of diabetes.
Secondly, the damage of the cardiovascular and cerebrovascular, cardiovascular and cerebrovascular complications is fatal complications of diabetes. Mainly manifested by aortic artery, coronary artery, cerebral atherosclerosis, and microvascular diabetic lesions with extensive small vessel endothelial hyperplasia and capillary basement membrane thickening. The blood vessel contraction and expansion are not coordinated, blood platelets are cohered, and lipid is deposited on the blood vessel wall to form hyperglycemia, hyperlipidemia, hyperviscosity and hypertension, so that the morbidity and mortality of the diabetic cardiovascular and cerebrovascular diseases are raised.
And thirdly, acute complications, the diabetes harm also comprises acute complications, and the urine disease is infected together: the incidence rate is high, and the two are causal, and must be treated at the same time. Common infections include respiratory infections and tuberculosis, urinary infections and skin infections. The harm also lies in diabetic hyperosmolar syndrome: most of the cases occur in the middle-aged and the elderly, most of the cases have no history of diabetes, the clinical manifestations include severe dehydration, sometimes can be misdiagnosed as cerebrovascular accidents due to hemiplegia, coma and other clinical manifestations, and the death rate is as high as fifty percent.
Fourthly, lactic acidosis is harmful, patients mostly have the history of heart, liver and kidney diseases or shock, infection, anoxia, drinking and taking glucose lowering medicine in large quantities, symptoms are not specific, and death rate is high.
The traditional Chinese medicine extract or the traditional Chinese medicine composition is an important medicine for treating diabetes, but the traditional Chinese medicine extract has the defects of insignificant blood sugar reducing effect and the like in different degrees and needs to be improved.
Disclosure of Invention
One of the purposes of the invention is to provide a water extract of the roots of the Trigonella foenum-graecum and a hypoglycemic effective part of the water extract of the roots of the Trigonella foenum-graecum;
the invention also aims to provide a method for preparing the water extract of the roots of the Trigonella foenum-graecum and the hypoglycemic effective parts of the water extract of the roots of the Trigonella foenum-graecum;
the invention also aims to apply the water extract of the roots of the Trigonella foenum-graecum and the hypoglycemic effective part of the water extract of the roots of the Trigonella foenum-graecum to reduce blood sugar.
The above object of the present invention is achieved by the following technical solutions:
the invention firstly provides a water extract of the roots of the ruelia chinensis bunge, and the extraction method comprises the following steps:
extracting the tuberous roots of the ruelia chinensis with water as an extraction solvent; the water extract of the root tuber of the ruelia chinensis is obtained.
As a preferred extraction method, comprising: (1) pulverizing root tuber of Trillium procumbens into coarse powder; (2) adding 3-12 times of water into the coarse powder, soaking, extracting under reflux, and filtering; (3) extracting the filter residue with water under reflux; (4) mixing the filtrates, and recovering under reduced pressure to obtain dry extract of radix Trigonellae water extract; wherein, preferably, the soaking time in the step (2) is more than 1 hour; the reflux extraction time can be 20-80min, and most preferably 40 min; the reflux extraction time in the step (3) can be 1-4 times, preferably 2 times, and the time of each reflux extraction is preferably 30-50 min.
The invention further adopts a macroporous resin technology and a step-by-step alcohol precipitation method to carry out a segmentation experiment on the water extract of the roots of the Trifolium repens and obtain 4 different fractions; mixing the alcohol eluates with different concentrations to obtain 95% ethanol eluate to obtain fraction 1(LLC-1), and detecting to obtain 95% ethanol extract of Trifolium tuberose root mainly containing phenolic components, saponin, cardiac glycoside and its lactone components and anthraquinone effective components. Mixing the effluent and water eluate, concentrating under reduced pressure to obtain water eluate, dissolving the water eluate in water (about 0.5g crude drug/mL), gradually adding 95% ethanol into the water eluate solution to make ethanol concentration reach 70%, stirring for 10min to contact fully, centrifuging at 5000rpm for 30min, and decanting the supernatant to obtain 70% ethanol precipitate as fraction 2 (LLC-2); adding 95% ethanol into the supernatant to 84% ethanol concentration, precipitating, centrifuging to obtain 84% ethanol precipitate as fraction 3(LLC-3), and concentrating the supernatant to obtain fraction 4 (LLC-4).
Through detection, the water extract of the roots of the Trigonella foenum-graecum mainly contains active ingredients such as polysaccharide ingredients, saponin, organic acid, iridoid and the like. The 95% ethanol extract of the roots of the ruelia reuteri root mainly comprises: phenolic component, saponin, cardiac glycoside, lactone component, anthraquinone, etc.
In order to observe whether the water extract of the roots of the ruelia reuteri has a definite blood sugar reducing effect, the diabetes model of a mouse is caused by Streptozotocin (STZ), the water extract of the roots of the ruelia reuteri is continuously administered by intragastric administration, the influence on the blood sugar of the STZ diabetic mouse is observed, and the fasting blood sugar measurement result shows that the water extract of the roots of the ruelia reuteri has an obvious blood sugar reducing effect.
The invention further uses Streptozotocin (STZ) to create a diabetic mouse model, ig. four different fractions (namely LLC-1, LLC-2, LLC-3 and LLC-4) of the tuberous root Trillia L aqueous extract are continuously given to the tuberous root for 3 weeks, the influence of each fraction on the blood sugar and related indexes of the STZ diabetic mouse is observed and detected, and the respective blood sugar reducing effect of each fraction is preliminarily evaluated. The fasting blood glucose measurement result of the continuous administration for 3 weeks shows that the blood glucose of the Trigonella foenum-graecum root aqueous extract fraction 3 is reduced by 38.3 percent (P is less than 0.01), and the blood glucose reducing effect of other 3 fractions is not obvious, so that the Trigonella foenum-graecum root aqueous extract fraction 3 provided by the invention has an obvious blood glucose reducing effect, and the fraction 3 is determined to be the main blood glucose reducing active part of the Trigonella foenum-graecum aqueous extract.
Therefore, the invention further provides the hypoglycemic effective part of the aqueous extract of the ruelia tuberosa, and the extraction method comprises the following steps:
(1) extracting the tuberous roots of the ruelia chinensis with water as an extraction solvent to obtain an aqueous extract of the tuberous roots of the ruelia chinensis;
(2) loading the water extract of the roots of the ruelia chinensis planch into a macroporous resin column, and collecting water eluate by using water as an eluent;
(3) dissolving the water eluate in water, adding 95% ethanol until ethanol concentration reaches 70%, stirring, centrifuging, collecting supernatant,
(4) adding 95% ethanol into the supernatant to 84% ethanol concentration for precipitation, centrifuging, and collecting 84% ethanol precipitate to obtain hypoglycemic effective component of the Trigonella foenum-graecum aqueous extract.
Preferably, the stirring time in the step (2) is preferably 5-20min, more preferably 10 min; the centrifugation is preferably carried out at a rotation speed of 2000-8000rpm for 20-40min, more preferably at a rotation speed of 5000rpm for 30 min.
The invention further discloses an application of the hypoglycemic effective part of the ruelia chinensis water extract or the ruelia chinensis water extract in preparing the hypoglycemic medicine. The medicine is prepared into a proper clinical preparation according to a conventional preparation method of traditional Chinese medicines, and preferably an oral preparation; the dosage form of the medicament comprises: tablet, granule, lyophilized powder, pill, capsule or oral liquid.
The invention also discloses a pharmaceutical composition for reducing blood sugar, which comprises: the water extract of the root of the ruelia chinensis berk with effective dose in treatment and pharmaceutically acceptable auxiliary materials.
The invention also discloses a pharmaceutical composition for reducing blood sugar, which comprises: effective amount of hypoglycemic effective part of the water extract of the roots of the ruelia chinensis and pharmaceutically acceptable auxiliary materials.
The pharmaceutical composition is prepared into a proper clinical preparation according to a conventional preparation method of traditional Chinese medicines, and preferably an oral preparation; the dosage form of the medicament comprises: tablet, lyophilized powder, granule, pill, capsule or oral liquid.
The adjuvant or carrier of the present invention refers to the conventional adjuvants or carriers in the pharmaceutical field, such as: diluents, disintegrants, lubricants, excipients, binders, glidants, fillers, surfactants, and the like; in addition, other adjuvants such as flavoring agents and sweeteners may be added to the composition. The diluent can be one or more components for increasing the weight and volume of the tablet, and common diluents comprise lactose, starch, pregelatinized starch, microcrystalline cellulose, sorbitol, mannitol, inorganic calcium salt and the like; the most common of them are lactose, starch, microcrystalline cellulose. The disintegrant can be one or more of crosslinked polyvinylpyrrolidone (with a total weight ratio of 2-6%), crosslinked sodium carboxymethylcellulose (with a total weight ratio of 2-6%), alginic acid (with a total weight ratio of 2-5%), and microcrystalline cellulose (with a total weight ratio of 5-15%). The lubricant comprises one or a mixture of stearic acid, sodium stearate, magnesium stearate, calcium stearate, polyethylene glycol, talcum powder and hydrogenated vegetable oil. The amount of lubricant used ranges (relative to the total weight) from 0.10 to 1%, typically from 0.25 to 0.75%. The binding agent can be one or more components which are beneficial to granulation; it may be starch slurry (10-30% by weight of the total binder), hydroxypropyl methylcellulose (2-5% by weight of the total binder), polyvinylpyrrolidone (2-20% by weight of the total binder), preferably an aqueous solution of polyvinylpyrrolidone in ethanol. The glidant can be one or a mixture of more of superfine silica gel powder, talcum powder and magnesium trisilicate. The surfactant may be one or more ingredients that improve wetting and increase drug dissolution, and is typically sodium lauryl sulfate (typically in the range of 0.2-6% by weight relative to the total weight).
Detailed Description
The invention is further described below in conjunction with specific embodiments, the advantages and features of which will become apparent from the description. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be within the scope of the invention.
Example 1 preparation of an aqueous extract of Trillia tuberosa roots
Cutting root tuber of Trillium Gratissima into small segments, pulverizing into coarse powder, weighing a certain amount of coarse powder, placing into a round-bottomed flask, adding 8 times of water, soaking for more than 2hr, reflux-extracting for 40min, filtering, and reflux-extracting with 5 times of water for 2 times and 40 min/time; mixing the filtrates, and recovering under reduced pressure to obtain dry extract, which has a yield of 68.43%.
Through detection, the water extract of the root tuber of the rueli grass mainly comprises the following components: polysaccharides, saponin, organic acid and iridoid.
Example 2 preparation of an aqueous extract of Trillia tuberosa roots
Cutting root tuber of Trillium Gratissima into small segments, pulverizing into coarse powder, weighing a certain amount of coarse powder, placing into a round-bottomed flask, adding 5 times of water, soaking for more than 1hr, reflux-extracting for 30min, filtering, and reflux-extracting with 5 times of water for 2 times and 50 min/time; mixing the filtrates, and recovering under reduced pressure to obtain dry extract, which has a yield of 67.43%.
Through detection, the water extract of the root tuber of the rueli grass mainly comprises the following components: polysaccharides, saponin, organic acid and iridoid.
Example 3 preparation of an aqueous extract of Trillia tuberosa root
Cutting root tuber of Trillium Gratissima into small segments, pulverizing into coarse powder, weighing a certain amount of coarse powder, placing into a round-bottomed flask, adding 10 times of water, soaking for more than 1hr, reflux-extracting for 50min, filtering, and reflux-extracting with 10 times of water for 2 times and 50 min/time; mixing the filtrates, and recovering under reduced pressure to obtain dry extract, which has a yield of 66.43%.
Through detection, the water extract of the root tuber of the rueli grass mainly comprises the following components: polysaccharides, saponin, organic acid and iridoid.
Example 4 preparation and characterization of various fractions of an aqueous extract of Trillia procumbens root tuber
Stage test of water extract D101 macroporous resin
Taking the water extract (1g crude drug/mL, density 1.20g/mL) of the root tuber of the ruelia tuberosa prepared in example 1, loading the water extract to a macroporous resin column, performing gradient elution by adopting water washing and ethanol with different concentrations, respectively collecting an effluent liquid, a water washing liquid and ethanol eluates with different concentrations (namely, sequentially performing elution by using water, 30% ethanol, 60% ethanol and 95% ethanol), determining to combine the effluent liquid and the water washing liquid because the proportion of the ethanol eluates with different concentrations is very low, and performing reduced pressure concentration to obtain an eluate; mixing the alcohol eluates with different concentrations to obtain 95% ethanol eluate, and detecting to obtain 95% ethanol extract of Trifolium tuberose root mainly containing: phenolic component, saponin, cardiac glycoside and its lactone component, and anthraquinone.
The yield of each part of the water extract of the roots of the tuberous roots of the Trigonella foenum-graecum by sections of the macroporous resin is as follows: the yield of water eluate was 93.6%; the yield of 95% ethanol eluate (LLC-1) was 6.4%.
(II) fractional alcohol precipitation experiment of water eluate
The water eluate was dissolved in water (about 0.5g crude drug/mL) and subjected to stepwise alcohol precipitation with 95% ethanol. Firstly, gradually adding 95% ethanol into water eluate solution to make ethanol concentration reach 70%, stirring for 10min for full contact, centrifuging at 5000rpm for 30min, and decanting the supernatant to obtain 70% ethanol precipitate (LLC-2); and continuously adding 95% ethanol into the supernatant to 84% ethanol concentration, then carrying out precipitation and centrifugation, taking 84% ethanol precipitate (LLC-3), concentrating the supernatant to obtain a sample LLC-4, respectively calculating the yield, and carrying out fractional ethanol precipitation on water eluate, wherein the results are shown in table 1 (each part is a fraction used for research on pharmacological activity).
TABLE 1 fractional alcohol precipitation test results of water eluate
The results of the preliminary analysis experiments show that: the Lilium Candidum root tuber contains polysaccharides, saponin, organic acid, iridoid, phenols, cardiac glycoside and its lactone, anthraquinone, etc.; wherein the medium molecular weight and small molecular weight polysaccharides are the main components (about 70-80% of the extractable components).
EXAMPLE 5 preparation of tablets
Taking 100g of the water extract thick paste of the roots of the rillic grass blocks prepared in the example 1, adding 25g of mannitol and 25g of cane sugar, carrying out wet granulation by using 5% povidone ethanol solution, carrying out granulation by using a 24-mesh sieve, drying for 4 hours at the temperature of 60 ℃, carrying out granulation by using a 20-mesh sieve, adding 0.5g of magnesium stearate, uniformly mixing, and tabletting to prepare 1000 tablets. Each tablet contains 100mg of water extract of the root of the Trigonella foenum-graecum.
Experimental example 1 Effect of water extract of Trigonella foenum-graecum root on blood sugar of streptozotocin-induced diabetic mice
1. Purpose of experiment
A mouse diabetes model is formed by Streptozotocin (STZ), and the influence on the blood sugar of an STZ diabetic mouse is observed by continuously intragastrically administering the oryza pulvialis aqueous extract.
2. Experimental Material
2.1 animals:
ICR mice, 22-24 g, male, 4 weeks old, 94, purchased from Beijing Huafukang Biotechnology GmbH, license number: SCXK Jing 2009-. Feeding the strain in an SPF animal house (facility license number: SYXK (Kyoto) 2003-.
2.2 reagent:
streptozotocin (streptazocin, STZ), Sigma S0130, available from boai port trade company ltd, beijing, 1 g/bottle, lot number: 110908
2.3 test paper and kit
Blood glucose test paper: luokang full activity type, lot number: 23445234, expiration date to 2013/03, Roche diagnostics GmbH, Germany.
Glucose assay kit (glucose oxidase method), purchased from Zhongsheng north controlled Biotechnology GmbH, product batch number: 122891, effective period to 2013/05.
2.4 test drugs
The aqueous extract of roots of ruelia tuberosa prepared in example 1: the brown liquid is frozen and preserved after being subpackaged, and a proper amount of the brown liquid is taken before use to prepare the brown liquid with corresponding concentration.
2.5 Positive control
Phenformin (phenformin hydrochloride tablets), Shandong Kenren and Tang pharmaceutical Co., Ltd., national Standard H37021531, product batch No.: 101104, effective period to 2013.10.
3. Experimental methods
The ICR mice are randomly divided into groups of normal control, model control, positive drug hypoglycemic agent (100mg/kg), tuberous root erethistle aqueous extract 5g/kg, 2.5g/kg, 1.25g/kg and 0.625g/kg, 10 normal controls per group and the rest 14 normal controls per group. Adaptive feeding is carried out for 3 days, after all mice are fasted and water is not forbidden for 12 hours, the mice of the model and each administration group are injected with 125mg/kg STZ in the abdominal cavity, and the mice of the normal group are injected with citric acid buffer solution with equal ip dosage. The blood from tail tip of each group of mice was continuously collected for 3 days after 7 days of molding to determine the fasting blood glucose level. And performing grouping adjustment according to the blood sugar measurement result. The administration group is administered by intragastric administration according to the above dosage, the administration volume is 0.2ml/10g, and the normal and model groups mice are administered by intragastric administration with distilled water of equal volume for 25 days continuously. Blood from the tail tip is taken at 7 days, 14 days and 25 days after administration to determine fasting blood glucose value, and the blood glucose reducing effect of the water extract of the root of the Trigonella foenum-graecum is evaluated.
4. Statistical method
Data processing was performed using Microsoft Excel statistical software and comparisons of differences between groups were performed using the t-test method.
5. Results of the experiment
After 25 days of administration, the four dose groups of the water extract of the roots of the Trifolium tuberosum L have obvious blood sugar reducing effects, and compared with the model group, the four dose groups can respectively reduce the blood sugar by 45 percent, 33 percent, 44 percent and 32 percent (P is less than 0.01), which is shown in table 2.
TABLE 2 Effect of water extract of root tuber of Trigonella foenum-graecum on fasting plasma glucose in STZ diabetic mice (X + -S, mmol/L)
Comparison with normal control group: p < 0.05; p <0.01
Comparison with model control group:▲P<0.05；▲▲P<0.01
in the experiment, an STZ is injected into the abdominal cavity to cause a mouse diabetes model, after the model is made, the fasting blood sugar of the mouse is increased, the weight of the mouse is obviously reduced, and the diabetes symptom of polydipsia and diuresis appears, which indicates that the model is successfully established. After the model is formed, the stomach is continuously infused for 25 days, and the result of fasting blood sugar measurement shows that the water extract of the roots of the Trigonella foenum-graecum has obvious blood sugar reducing effect.
Experimental example 2 Effect of different fractions of aqueous extract of Trillia aurea root tuber on streptozotocin-induced diabetic mice
1. Purpose of experiment
Using Streptozotocin (STZ) to create a diabetic mouse model, ig. different fractions of the water extract of the tuberous root of the Trifolium reuteri L are continuously given for 3 weeks, the influence of each fraction on the blood sugar and related indexes of the STZ diabetic mouse is observed and detected, and the fraction with the hypoglycemic activity is preliminarily confirmed.
2. Experimental Material
2.1 Experimental animals
ICR mice, 22-24 g, male, 4 weeks old, 150, from Beijing Huafukang biotech GmbH, license number: SCXK Jing 2009-. Feeding the strain in an SPF animal house (license number: SYXK (Kyoto) 2003-.
2.2 reagents
Streptozotocin (streptazocin, STZ), Sigma S0130, available from boai port trade company ltd, beijing, 1 g/bottle, lot number: 110908
2.3 test paper and kit
Blood glucose test paper: luokang full activity type, lot number: 23445234, expiration date to 2013/03, Roche diagnostics GmbH, Germany, 50 packs.
Glucose assay kit (glucose oxidase method), purchased from Zhongsheng north controlled Biotechnology GmbH, product batch number: 120551, effective period to 2013/08.
2.4 test drugs
Tuberous root ruelia water extract tuberous root fraction 1(LLC fraction 1): tan powder, aqueous extract fraction 2(LLC fraction 2): tan cake solid, aqueous extract fraction 3(LLC fraction 3): brown liquid, aqueous extract fraction 4(LLC fraction 4): light brown liquid. Refrigerating and sealing LLC fraction 1 and LLC fraction 2, and dissolving with distilled water to obtain corresponding concentration when it is used; LLC fraction 3 and LLC fraction 4 were stored frozen and diluted with water to the appropriate concentrations for use.
2.5 Positive control
Phenformin (phenformin hydrochloride tablets), Shandong Kenren and Tang pharmaceutical Co., Ltd., national Standard H37021531, product batch No.: 101104, effective period to 2013.10.
3. Experimental methods
The ICR mice are randomly divided into normal control, model control, positive medicine hypoglycemic agent (100mg/kg), LLC fraction 1(2.5g/kg), LLC fraction 2(2.5g/kg), LLC fraction 3(2.5g/kg), LLC fraction 4(2.5g/kg), 10 normal control groups, 14 model groups and 14 administration groups. Adaptive feeding is carried out for 2 days, after all mice are fasted and are not forbidden for 12 hours, the model and the mice of each administration group are injected with 125mg/kg STZ in the abdominal cavity for molding, and the mice of a normal control group have ip. equal-volume citric acid buffer solution. The blood glucose value of the fasting blood glucose value of each group of mice is measured by a glucose kit after 7 days of molding and 3 days of continuous tail tip blood taking. Grouping according to blood sugar measurement results. The administration group is administered by intragastric administration according to the above dosage, the administration volume is 0.2ml/10g, and the normal and model groups mice are administered by intragastric administration of distilled water with equal volume for 21 days continuously. Blood from the tail cusp was collected at 7, 14 and 21 days after administration to determine fasting blood glucose level, and the effect of different fractions of the water extract of the root tuber of ruelia reuteri on fasting blood glucose of STZ diabetic mice was observed.
4. Statistical method
Data processing was performed using Microsoft Excel statistical software and comparisons of differences between groups were performed using the t-test method.
5. Results of the experiment
After 3 weeks of continuous administration, the aqueous extract of Trillia tuberosa root LLC fraction 3 exhibited a significant hypoglycemic effect, reducing blood glucose by 38.3% (P < 0.01) compared to model mice, see Table 3.
TABLE 3 Effect of different fractions of the aqueous extract of Trifolium repens on fasting plasma glucose in STZ diabetic mice (X. + -. S, mmol/L)
Comparison with normal control group: p < 0.05; p <0.01
Comparison with model control group:▲P<0.05；▲▲P<0.01
6. conclusion of the experiment
The STZ is applied to the experiment to cause the diabetes model of the mouse, after the model is made, the fasting blood sugar of the mouse is increased, the weight of the mouse is reduced to some extent, and the diabetes symptom of polydipsia and diuresis is generated, which indicates that the model is successfully established. After the model is formed, 4 different fractions of the water extract of the roots of the Trigonella tuberosa are intragastrically administered, and the fasting blood glucose measurement result of continuous administration for 3 weeks shows that the fraction 3 of the water extract of the roots of the Trigonella tuberosa has obvious blood glucose reducing effect, and the blood glucose effect of the other three fractions is not obvious, so that the fraction 3 is primarily determined to be the main active part of the water extract of the Trigonella tuberosa for reducing the blood glucose.
Claims (6)
1. The hypoglycemic effective part of the water extract of the Trigonella foenum-graecum is characterized in that the extraction method comprises the following steps:
(1) extracting the tuberous roots of the ruelia chinensis with water as an extraction solvent to obtain an aqueous extract of the tuberous roots of the ruelia chinensis;
(2) loading the water extract of the roots of the ruelia chinensis planch into a macroporous resin column, adopting water as an eluent, collecting effluent liquid and washing liquid, and combining the effluent liquid and the washing liquid into water eluate;
(3) dissolving the water eluate in water, adding 95% ethanol until ethanol concentration reaches 70%, stirring, centrifuging, collecting supernatant,
(4) adding 95% ethanol into the supernatant until reaching 84% ethanol concentration, precipitating, centrifuging, and collecting 84% ethanol precipitate to obtain the hypoglycemic effective component of the water extract of Trigonella tuberosa L.
2. The hypoglycemic effective part of the tuberous root ruelia aqueous extract according to claim 1, characterized in that the stirring time in the step (3) is 5-20 min; the centrifugation is carried out for 20-40min at the rotating speed of 2000-8000 rpm.
3. The hypoglycemic effective part of the tuberous root ruelia aqueous extract according to claim 2, characterized in that the stirring time in the step (3) is 10 min; the centrifugation is carried out for 30min at the rotating speed of 5000 rpm.
4. The hypoglycemic active part of the tuberous root ruelia aqueous extract according to claim 1, characterized in that the macroporous resin column is a D101 macroporous resin column.
5. Use of the effective fraction of the aqueous extract of trefoil roots according to any of claims 1 to 4 for the preparation of a hypoglycemic medicament.
6. A pharmaceutical composition for treating diabetes, comprising: a therapeutically effective amount of the effective part of the aqueous extract of the roots of ruelia reuteri of any one of claims 1 to 4 and pharmaceutically acceptable excipients.
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