CN110496148B - Eucommia ulmoides extract composition for treating gestational hypertension and application thereof - Google Patents
Eucommia ulmoides extract composition for treating gestational hypertension and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of traditional Chinese medicine extracts, and particularly relates to an eucommia ulmoides extract composition for treating gestational hypertension and application thereof. The composition comprises: 20-40 parts of ursolic acid, 10-25 parts of chlorogenic acid, 20-60 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 30-50 parts of total flavone. The eucommia ulmoides extract composition is prepared by the steps of enzymolysis, fractional extraction, dissolution and the like, the preparation method is simple and efficient, and the prepared eucommia ulmoides extract composition has a good effect on treating gestational hypertension and simultaneously reduces side effects.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicine extracts, and particularly relates to an eucommia ulmoides extract composition for treating gestational hypertension and application thereof.
Background
Pregnancy hypertension refers to the condition of hypertension, edema, proteinuria, etc. after 20 weeks of pregnancy, which is transient and disappears after delivery. Belongs to specific diseases of pregnancy, and is one of the main causes of diseases and deaths of pregnant and lying-in women and perinatal infants at present. In the later stage of pregnancy, the systolic pressure is above 140 mmHg (18.7 kPa) and the diastolic pressure is above 90 mmHg (12.0 kPa), and hypertension must be considered. The cause of pregnancy-induced hypertension is thought to be related to genetic factors, and many theories generally suggest that certain substances from placenta enter maternal blood to cause immune factor changes in the body of a pregnant woman, so that systemic arteriospasm of the pregnant woman can cause hypertension. The main clinical manifestations are:
hypertension: the diagnosis can be made when the blood pressure reaches 140/90 mmHg. Proteinuria: the midstream urine should be taken for examination, and the abnormality is found when the quantity of urine protein is more than 0.5g in 24 hours. Simultaneously has hypertension and urine protein as preeclampsia. Edema. Subjective symptoms: there are symptoms such as headache, dim eyesight, chest distress, nausea and vomiting. The occurrence of these subjective symptoms indicates that the disease has progressed to the preeclampsia stage, and should be examined and treated in time. Twitching and coma: convulsions and coma are the manifestations of this disease in the severe stage of development, and should be treated immediately with medical attention to terminate pregnancy at the right moment.
Women with gestational hypertension or preeclampsia have an increased risk of hypertension in the later years of age, and even up to 50 years later, have a high probability of hospitalization for hypertension-related disorders, with women with a history of preeclampsia having a 4-fold higher risk of stroke than women with normotensive pregnancy.
And the antihypertensive drug can enter the fetus through the placenta or appear in the milk, so that toxic and side effects are generated on the fetus or the baby. Therefore, the traditional Chinese medicines are mostly adopted clinically, but some Chinese patent medicines actually contain a certain amount of western medicine components which are unfavorable for the fetus, and the pregnant women who adopt the Chinese patent medicines and the Chinese herbal medicines also pay attention to the blood pressure reduction.
Chinese patent application CN 102670791A discloses eucommia ulmoides traditional Chinese medicine oral liquid for relieving hypertension, which is prepared from the following raw material medicines in parts by weight: the oral liquid comprises 300 parts of eucommia bark, 300 parts of angelica sinensis, 300 parts of morinda officinalis, 300 parts of polygonum multiflorum and 300 parts of lotus leaf, can quickly relieve symptoms such as tinnitus, dizziness, headache, palpitation, short breath and insomnia caused by hypertension, but because the components are numerous, the separation and purification process of the effective components is not carried out, and impurities and harmful components in the traditional Chinese medicine components can not be effectively removed.
The Chinese patent application CN 103750156A discloses a processing method of dried noodles which utilize eucommia ulmoides leaves, are low in cost, rich in active substances such as geniposide acid and aucubin, also rich in nutrient substances such as amino acid and glucose, and can reduce gutta percha, can be applied to prevention and diet, but needs to be further improved in the effect of treating pregnancy hypertension.
Disclosure of Invention
In order to overcome the technical problems, the invention provides the eucommia ulmoides extract composition for treating the gestational hypertension, the composition can effectively treat the gestational hypertension, the preparation method is simple and efficient, and the purity of effective components can be effectively improved, the impurity content is reduced, and the side effect is reduced.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows:
an eucommia ulmoides extract composition for treating gestational hypertension comprises the following components in parts by weight: 20-40 parts of ursolic acid, 10-25 parts of chlorogenic acid, 20-60 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 30-50 parts of total flavone.
Preferably, the composition comprises, in parts by weight: 25-30 parts of ursolic acid, 15-20 parts of chlorogenic acid, 30-50 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 35-45 parts of total flavone.
The invention also aims to provide a preparation method of the eucommia ulmoides extract composition for treating gestational hypertension, which comprises the following steps:
(1) pulverizing Eucommiae cortex, adding into water, performing enzymolysis, and filtering to obtain enzymolysis solution and residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in cold ethanol, performing ultrasonic treatment, filtering to obtain an alcohol solution 1, adding water into the alcohol solution 1, and separating out a solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid;
eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a polyethylene glycol aqueous solution and a dextran saline solution into the mixed solution, adjusting the pH value, stirring, standing, layering, taking a lower layer solution, concentrating, adding ethanol for extraction to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain the total flavone;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside, and total flavone in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition.
Preferably, in the step (1), the crushed mesh number is 20-40 meshes;
preferably, in the step (1), the amount of the added water is 5-8 times of the mass of the eucommia ulmoides;
preferably, in step (1), the enzyme for enzymolysis is: a mixture of cellulase and polymethylgalacturonase;
preferably, the mass ratio of the cellulase to the polymethylgalacturonase is 1: 1-3;
preferably, in the step (2), the temperature of the cold ethanol is 1-10 ℃;
preferably, in the step (2), the temperature of the ultrasonic treatment and the filtering is not more than 10 ℃;
preferably, in the step (2), the dosage of the ethanol is 5-8 times of the mass of the filter residue 1.
Preferably, in the step (4), the molecular weight of the polyethylene glycol is M1500-; preferably 2000;
preferably, in the step (4), the concentration of the polyethylene glycol aqueous solution is 30-40%;
preferably, in the step (4), the addition amount of the polyethylene glycol aqueous solution is 5-6% of the total mass of the alcohol-water mixed solution of polyethylene glycol;
preferably, in the step (4), the molecular weight of the glucan is M40000-50000; preferably M ═ 45000;
preferably, in the step (4), the aqueous solution of dextran is prepared by dissolving dextran in physiological saline; the dextran concentration of the dextran saline solution is 20-30%;
preferably, in the step (4), the addition amount of the aqueous solution of dextran salt is 3-4% of the total mass of the mixed solution of alcohol and water;
preferably, in the step (4), the pH value is 6-7; preferably the pH value is 6;
preferably, in step (4), the stirring is performed at 40 to 50 ℃.
The invention also aims to provide the eucommia ulmoides extract composition for treating gestational hypertension or the application of the eucommia ulmoides extract composition in preparing a medicine for treating gestational hypertension.
Preferably, the drug for treating gestational hypertension is any one of tablets, capsules, pills, oral liquid or syrup of eucommia ulmoides extract composition.
Compared with the prior art, the invention has the technical advantages that:
(1) according to the invention, the eucommia ulmoides extract composition with a good curative effect on treating gestational hypertension is effectively prepared by selecting and applying a plurality of effective components in the eucommia ulmoides extract, so that the side effect is reduced;
(2) through the processes of step-by-step extraction, purification and the like, the mixed extract of the effective components of chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside is respectively prepared by macroporous adsorption resin, so that the purity of the extract is improved; sequentially adding the rest components into aqueous solution of polyethylene glycol and aqueous solution of dextran saline, extracting total flavonoids by aqueous two-phase extraction technology, and further purifying the total flavonoids after extraction; so that impurities in the whole composition are reduced; the purity of the eucommia ulmoides extract is improved;
(3) in the process of extracting the total flavone, the dosage, the pH value and the temperature value of the polyethylene glycol and the glucan have great influence on the extraction rate, and the total flavone can be effectively extracted better by reasonably controlling various factor conditions and synergistic action; the change of single factor can cause the change of the extraction effect of the general flavone, so that the extraction rate is greatly reduced or the impurity content in the flavone is increased; the dosage, pH value and temperature value range of the polyethylene glycol and the glucan given by the invention can better exert the extraction effect of the total flavone, reduce impurities and improve the curative effect of the total flavone in the gestational hypertension.
(4) The residual components of the eucommia bark after separation and purification can also be used for preparing other medicines, thereby improving the medicinal value of the eucommia bark, saving the resources of the treasure medicinal materials and reducing the waste.
Detailed Description
Example 1
In this embodiment, the eucommia ulmoides extract composition for treating gestational hypertension includes, by weight: 25 parts of ursolic acid, 15 parts of chlorogenic acid, 50 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 35 parts of total flavone;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) pulverizing eucommia ulmoides to 30 meshes, adding water 7 times of the mass of the eucommia ulmoides into the eucommia ulmoides, and mixing the eucommia ulmoides with water according to a mass ratio of 1: 2, performing enzymolysis on the mixture of the cellulase and the polymethylgalacturonase, and filtering to obtain an enzymolysis liquid and filter residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in 6 times of 2 deg.C cold ethanol, performing ultrasonic treatment at the same temperature, filtering to obtain alcohol solution 1, adding water into the alcohol solution 1, and separating out solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid;
eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 35% polyethylene glycol (M ═ 2000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 6% of the total mass of the alcohol-water mixed solution), a glucan (M ═ 45000) aqueous salt solution (the glucan aqueous salt solution is added according to the amount of the glucan accounting for 3% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 6, stirring at 45 ℃, standing, layering, taking a lower layer solution, concentrating, adding 35% ethanol by mass to extract to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the aqueous dextran salt solution has a dextran concentration of 25%;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside, and total flavone in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition.
Example 2
In this embodiment, the eucommia ulmoides extract composition for treating gestational hypertension includes, by weight: 40 parts of ursolic acid, 10 parts of chlorogenic acid, 60 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 30 parts of total flavone;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) pulverizing Eucommiae cortex to 20 mesh, adding into water 5 times of Eucommiae cortex, and mixing with water in a weight ratio of 1: 1, performing enzymolysis on a mixture of cellulase and polymethylgalacturonase, and filtering to obtain an enzymolysis liquid and filter residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in 5 times of 1 deg.C cold ethanol, performing ultrasonic treatment at the same temperature, filtering to obtain alcohol solution 1, adding water into the alcohol solution 1, and separating out solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid;
eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 30% polyethylene glycol (M ═ 1500) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 5% of the total mass of the alcohol-water mixed solution), a glucan (M ═ 40000) saline solution (the glucan saline solution is added according to the amount of the glucan accounting for 4% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 7, stirring at 40 ℃, standing, layering, taking a lower layer solution, concentrating, adding 30% ethanol for extraction to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the dextran concentration of the dextran saline solution is 20%;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside, and total flavone in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition.
Example 3
In this embodiment, the eucommia ulmoides extract composition for treating gestational hypertension includes, by weight: 20 parts of ursolic acid, 25 parts of chlorogenic acid, 20 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 50 parts of total flavone;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) pulverizing Eucommiae cortex to 40 mesh, adding into water 8 times of Eucommiae cortex, and mixing with water in a weight ratio of 1: 3, performing enzymolysis on the mixture of the cellulase and the polymethylgalacturonase, and filtering to obtain an enzymolysis liquid and filter residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in 8 times of 10 deg.C cold ethanol, performing ultrasonic treatment at the same temperature, filtering to obtain alcohol solution 1, adding water into the alcohol solution 1, and separating out solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid;
eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 40% polyethylene glycol (M ═ 3000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 6% of the total mass of the alcohol-water mixed solution) and a glucan (M ═ 50000) aqueous salt solution (the glucan aqueous salt solution is added according to the amount of the glucan accounting for 3% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 6, stirring at 50 ℃, standing, layering, taking a lower layer solution, concentrating, adding 40% ethanol for extraction to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the dextran concentration of the dextran saline solution is 30%;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside, and total flavone in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition.
Example 4
In this embodiment, the eucommia ulmoides extract composition for treating gestational hypertension includes, by weight:
30 parts of ursolic acid, 20 parts of chlorogenic acid, 30 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 45 parts of total flavone.
The preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) pulverizing Eucommiae cortex to 20 mesh, adding into water 6 times of Eucommiae cortex, and mixing with water in a weight ratio of 1: 2, performing enzymolysis on the mixture of cellulase and polymethylgalacturonase, and filtering to obtain an enzymolysis liquid and filter residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in 6 times of 5 deg.C cold ethanol, performing ultrasonic treatment at the same temperature, filtering to obtain alcohol solution 1, adding water into the alcohol solution 1, and separating out solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid;
eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 30% polyethylene glycol (M ═ 2000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 5% of the total mass of the alcohol-water mixed solution) and a glucan (M ═ 40000) saline solution (the glucan saline solution is added according to the amount of the glucan accounting for 3% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 6, stirring at 50 ℃, standing, layering, taking a lower layer solution, concentrating, adding 40% ethanol for extraction to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the dextran concentration of the dextran saline solution is 20%;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside, and total flavone in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition.
Comparative example 1
Compared with example 1, the difference is only in the weight parts of the components of the composition;
in this embodiment, the eucommia ulmoides extract composition for treating gestational hypertension includes, by weight: 15 parts of ursolic acid, 15 parts of chlorogenic acid, 70 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 25 parts of total flavonoids;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension is the same as that of example 1.
Comparative example 2
Compared with example 1, the difference is only in the weight parts of the components of the composition;
in this embodiment, the eucommia ulmoides extract composition for treating gestational hypertension includes, by weight: 5 parts of ursolic acid, 15 parts of chlorogenic acid, 50 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 55 parts of total flavonoids;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension is the same as that of example 1.
Comparative example 3
The difference compared to example 1 is only the pH difference during the extraction of the active principle.
The eucommia ulmoides extract composition for treating gestational hypertension in the embodiment has the same weight parts as those in embodiment 1;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) - (3) Steps (1) to (3) of example 1;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 35% polyethylene glycol (M ═ 2000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 6% of the total mass of the alcohol-water mixed solution), a glucan (M ═ 45000) aqueous salt solution (the glucan aqueous salt solution is added according to the amount of the glucan accounting for 3% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 3, stirring at 45 ℃, standing, layering, taking a lower layer solution, concentrating, adding 35% ethanol by mass to extract to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the aqueous dextran salt solution has a dextran concentration of 25%;
(5) same as in step (5) of example 1.
Comparative example 4
The difference compared to example 1 is only the pH difference during the extraction of the active principle.
The eucommia ulmoides extract composition for treating gestational hypertension in the embodiment has the same weight parts as those in embodiment 1;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) - (3) Steps (1) to (3) of example 1;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 35% polyethylene glycol (M ═ 2000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 6% of the total mass of the alcohol-water mixed solution), a glucan (M ═ 45000) aqueous salt solution (the glucan aqueous salt solution is added according to the amount of the glucan accounting for 3% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 9, stirring at 45 ℃, standing, layering, taking a lower layer solution, concentrating, adding 35% ethanol by mass to extract to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the aqueous dextran salt solution has a dextran concentration of 25%;
(5) same as in step (5) of example 1.
Comparative example 5
Compared with example 1, the difference is only the amount of polyethylene glycol and dextran used in the extraction process of the effective ingredients.
The eucommia ulmoides extract composition for treating gestational hypertension in the embodiment has the same weight parts as those in embodiment 1;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) - (3) Steps (1) to (3) of example 1;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 35% polyethylene glycol (M ═ 2000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 3% of the total mass of the alcohol-water mixed solution), a glucan (M ═ 45000) aqueous salt solution (the glucan aqueous salt solution is added according to the amount of the glucan accounting for 6% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 6, stirring at 45 ℃, standing, layering, taking a lower layer solution, concentrating, adding 35% ethanol by mass to extract to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the aqueous dextran salt solution has a dextran concentration of 25%;
(5) same as in step (5) of example 1.
Comparative example 6
Compared with example 1, the difference is only the amount of polyethylene glycol and dextran used in the extraction process of the effective ingredients.
The eucommia ulmoides extract composition for treating gestational hypertension in the embodiment has the same weight parts as those in embodiment 1;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) - (3) Steps (1) to (3) of example 1;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a 35% polyethylene glycol (M ═ 2000) aqueous solution (the polyethylene glycol aqueous solution is added according to the amount of the polyethylene glycol accounting for 3% of the total mass of the alcohol-water mixed solution), a glucan (M ═ 45000) aqueous salt solution (the glucan aqueous salt solution is added according to the amount of the glucan accounting for 3% of the total mass of the alcohol-water mixed solution), adjusting the pH value to 6, stirring at 45 ℃, standing, layering, taking a lower layer solution, concentrating, adding 35% ethanol by mass to extract to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain total flavone;
the preparation method of the aqueous solution of the glucan is to dissolve the glucan in physiological saline to prepare the aqueous solution of the glucan; the aqueous dextran salt solution has a dextran concentration of 25%;
(5) same as in step (5) of example 1.
Examples of effects
SD rat, 240 plus 280g, SPF grade, provided by Qinglongshan animal breeding farm in Jiangning district of Nanjing;
dissolving N-nitro-L-arginine methyl ester (L-NAME) with normal saline to obtain final concentration of 0.05 g/ml;
the samples of examples 1-4 and comparative examples 1-6 were suspended and dissolved in 0.5% CMC-Na to a final concentration of 0.1 g/ml;
after the rats were acclimatized for one week, the female rats and the male rats were housed for three days at a ratio of 2:1, marking the first day of pregnancy as the obvious open and red swelling of vaginal suppository or vaginal orifice, removing non-pregnant female rats in the ninth day of pregnancy, dividing 120 successfully pregnant female rats into 12 groups which are respectively a blank control group, a model group, groups of examples 1-4 and groups of comparative examples 1-6, starting administration on the 10 th day, and measuring blood pressure of pregnant mouse before administration (blood pressure is measured for several times, and average value is taken), feeding physiological saline 0.5g/kg to blank control group, the other groups are injected with a modeling agent L-NAME according to 0.25g/kg per day, the modeling agent is filled with one group of physiological saline with the gavage amount of 0.5g/kg, the suspensions prepared in the other groups according to the 0.5g/kg intragastric administration examples 1-4 and the comparative examples 1-6; measuring blood pressure after administration on day 19, collecting blood after measurement, anesthetizing, killing, dissecting, and collecting placenta.
(1) Blood index changes of rats before and after administration (Table 1)
TABLE 1 rat blood index changes before and after dosing: (
n＝10)
| Test group | Blood viscosity (cp) | Blood flow (ml/min) |
| Blank control group | 3.81±0.62a | 7.24±1.13d |
| Hypertension model group | 5.53±0.81c | 3.56±0.54f |
| Example 1 | 4.12±0.72a | 5.52±0.47e |
| Example 2 | 4.09±1.03a | 5.74±0.69e |
| Example 3 | 4.21±0.55a | 5.38±0.85e |
| Example 4 | 4.15±0.61a | 5.59±0.76e |
| Comparative example 1 | 4.73±1.23b | 4.35±1.25f |
| Comparative example 2 | 4.91±0.93b | 4.12±1.04f |
| Comparative example 3 | 4.67±0.77b | 4.26±0.69f |
| Comparative example 4 | 4.86±1.31b | 4.54±0.77f |
| Comparative example 5 | 5.13±1.28c | 3.92±0.96f |
| Comparative example 6 | 5.34±0.88c | 4.06±1.11f |
Note: in the same column, P between different letters is less than 0.05;
the experimental data show that compared with a blank control group, the blood viscosity of a hypertension model group is obviously increased, the blood flow is obviously reduced, and the success of molding is shown, and the experiments of examples 1-4 show that the eucommia ulmoides extract has a good inhibition effect on pregnancy high viscosity and can improve the blood flow;
the experimental data of the comparative examples 1 to 6 show that the proportion of each effective component and the specific parameters in the extraction process of the effective components have significant influence on the drug effect of the eucommia ulmoides extract.
(2) Effect of eucommia ulmoides Oliver extract on average arterial pressure of pregnant hypertension rats (Table 2)
TABLE 2 mean arterial pressure changes in rats before and after administration: (
n＝10)
| Test group | Before administration (mmHg) | After administration (mmHg) |
| Blank control group | 105.6±8.9 | 109.3±9.4a |
| Hypertension model group | 103.7±10.4 | 142.7±3.3b |
| Example 1 | 106.2±7.2 | 114.2±9.1a |
| Example 2 | 108.1±10.1 | 115.3±5.2a |
| Example 3 | 106.5±9.6 | 112.8±11.3a |
| Example 4 | 107.2±5.7 | 113.5±6.7a |
| Comparative example 1 | 105.4±6.4 | 126.5±13.2c |
| Comparative example 2 | 106.9±9.2 | 122.3±9.4c |
| Comparative example 3 | 108.4±13.1 | 127.5±11.4c |
| Comparative example 4 | 107.5±8.6 | 123.6±7.6c |
| Comparative example 5 | 107.9±5.3 | 130.1±10.8c |
| Comparative example 6 | 108.6±11.5 | 135.5±16.4c |
Note: in the same column, P between different letters is less than 0.05;
according to experimental data, the mean arterial pressure of rats has no significant difference before administration; after administration, compared with a blank control group, the hypertension model group has obviously raised blood pressure, which indicates successful molding, and experiments of examples 1-4 show that the eucommia ulmoides extract has better inhibitory effect on gestational hypertension;
the experimental data of the comparative examples 1 to 6 show that the proportion of each effective component and the specific parameters in the extraction process of the effective components have significant influence on the drug effect of the eucommia ulmoides extract.
The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.
Claims (8)
1. An eucommia ulmoides extract composition for treating gestational hypertension comprises the following components in parts by weight: 20-40 parts of ursolic acid, 10-25 parts of chlorogenic acid, 20-60 parts of a mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 30-50 parts of total flavonoids;
the preparation method of the eucommia ulmoides extract composition for treating gestational hypertension comprises the following steps of:
(1) pulverizing Eucommiae cortex, adding into water, performing enzymolysis, and filtering to obtain enzymolysis solution and residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in cold ethanol, performing ultrasonic treatment, filtering to obtain an alcohol solution 1, adding water into the alcohol solution 1, and separating out a solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid; eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a polyethylene glycol aqueous solution and a dextran saline solution into the mixed solution, adjusting the pH value, stirring, standing, layering, taking a lower layer solution, concentrating, adding ethanol for extraction to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain the total flavone;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside and total flavonoids in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition;
wherein, in the step (4), the molecular weight of the polyethylene glycol is M = 1500-; the addition amount of the polyethylene glycol aqueous solution is 5-6% of the total mass of the alcohol-water mixed solution of polyethylene glycol;
the molecular weight of the glucan is M =40000 and 50000; the addition amount of the glucan salt water solution is 3-4% of the total mass of the alcohol-water mixed solution of glucan;
the pH value is 6-7; the stirring is carried out at 40-50 ℃.
2. The eucommia ulmoides extract composition for treating gestational hypertension according to claim 1, wherein the composition comprises, in parts by weight: 25-30 parts of ursolic acid, 15-20 parts of chlorogenic acid, 30-50 parts of mixed extract of pinoresinol diglucoside and syringaresinol diglucoside and 35-45 parts of total flavone.
3. The preparation method of the eucommia ulmoides extract composition for treating gestational hypertension according to claim 1 or 2, comprising the steps of:
(1) pulverizing Eucommiae cortex, adding into water, performing enzymolysis, and filtering to obtain enzymolysis solution and residue 1;
(2) washing the filter residue 1 with petroleum ether, placing in cold ethanol, performing ultrasonic treatment, filtering to obtain an alcohol solution 1, adding water into the alcohol solution 1, and separating out a solid which is ursolic acid;
(3) sequentially adsorbing the enzymolysis solution by XDA-1 type macroporous resin and S-8 type macroporous adsorption resin; eluting the adsorbed XDA-1 type macroporous resin with ethanol, concentrating, and drying to obtain chlorogenic acid; eluting the adsorbed S-8 type macroporous adsorption resin with ethanol, concentrating, and drying to obtain mixed extract of pinoresinol diglucoside and syringaresinol diglucoside;
(4) mixing the alcoholic solution remaining after the solid is separated out in the step (2) with the enzymolysis solution remaining after the macroporous resin adsorption in the step (3) to obtain an alcohol-water mixed solution, sequentially adding a polyethylene glycol aqueous solution and a dextran saline solution into the mixed solution, adjusting the pH value, stirring, standing, layering, taking a lower layer solution, concentrating, adding ethanol for extraction to obtain an alcoholic solution 2, concentrating and drying the alcoholic solution 2 to obtain the total flavone;
(5) dissolving the obtained mixed extract of ursolic acid, chlorogenic acid, pinoresinol diglucoside and syringaresinol diglucoside and total flavonoids in ethanol, and adding lecithin to obtain Eucommiae cortex extract composition;
wherein, in the step (4), the molecular weight of the polyethylene glycol is M = 1500-; the addition amount of the polyethylene glycol aqueous solution is 5-6% of the total mass of the alcohol-water mixed solution of polyethylene glycol;
the molecular weight of the glucan is M =40000 and 50000; the addition amount of the glucan salt water solution is 3-4% of the total mass of the alcohol-water mixed solution of glucan;
the pH value is 6-7; the stirring is carried out at 40-50 ℃.
4. The method for preparing the eucommia ulmoides extract composition for treating gestational hypertension according to claim 3, wherein in the step (1), the crushed mesh number is 20-40 mesh; the amount of the added water is 5-8 times of the mass of the eucommia ulmoides.
5. The method for preparing the eucommia ulmoides extract composition for treating gestational hypertension according to claim 3, wherein in the step (1), the enzyme for enzymolysis is: a mixture of cellulase and polymethylgalacturonase; the mass ratio of the cellulase to the polymethylgalacturonase is 1: 1-3.
6. The method for preparing the eucommia ulmoides extract composition for treating gestational hypertension according to claim 3, wherein the temperature of the cold ethanol in the step (2) is 1-10 ℃; the temperature of the ultrasonic treatment and the filtration do not exceed 10 ℃.
7. The method for preparing the eucommia ulmoides extract composition for treating gestational hypertension according to claim 3, wherein in the step (2), the amount of the ethanol is 5-8 times of the mass of 1 of the residue.
8. Use of the eucommia ulmoides extract composition for treating gestational hypertension according to any one of claims 1 to 2 in the preparation of a medicament for treating gestational hypertension.
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| 杜仲的化学成分与功能;杜仲综合利用技术国家地方联合工程实验室;《https://duzhong.jsu.edu.cn/info/1017/1169.htm》;20170516;第1-6页 * |
| 杜仲颗粒结合常规疗法治疗妊娠期高血压疾病的临床疗效;彭红梅等;《重庆医学》;20121130;第41卷(第31期);第3262-3264页 * |
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