CN108452073B - Composite plant lectin and preparation method and application thereof - Google Patents
Composite plant lectin and preparation method and application thereof Download PDFInfo
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- CN108452073B CN108452073B CN201810457585.8A CN201810457585A CN108452073B CN 108452073 B CN108452073 B CN 108452073B CN 201810457585 A CN201810457585 A CN 201810457585A CN 108452073 B CN108452073 B CN 108452073B
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- phytohemagglutinin
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- lectin
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- 108010089814 Plant Lectins Proteins 0.000 title claims abstract description 17
- 239000003726 plant lectin Substances 0.000 title claims abstract description 17
- 239000002131 composite material Substances 0.000 title claims description 15
- 238000002360 preparation method Methods 0.000 title abstract description 14
- 108010047620 Phytohemagglutinins Proteins 0.000 claims abstract description 33
- 230000001885 phytohemagglutinin Effects 0.000 claims abstract description 33
- 150000001875 compounds Chemical class 0.000 claims abstract description 29
- 239000003814 drug Substances 0.000 claims abstract description 26
- 206010020772 Hypertension Diseases 0.000 claims abstract description 13
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- 241000512259 Ascophyllum nodosum Species 0.000 claims abstract description 11
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- 241000304195 Salvia miltiorrhiza Species 0.000 claims abstract description 11
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- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 claims abstract description 5
- 229930003944 flavone Natural products 0.000 claims abstract description 5
- 150000002212 flavone derivatives Chemical class 0.000 claims abstract description 5
- 235000011949 flavones Nutrition 0.000 claims abstract description 5
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229930189092 Notoginsenoside Natural products 0.000 claims abstract description 3
- 239000002253 acid Substances 0.000 claims abstract description 3
- KBPHJBAIARWVSC-RGZFRNHPSA-N lutein Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\[C@H]1C(C)=C[C@H](O)CC1(C)C KBPHJBAIARWVSC-RGZFRNHPSA-N 0.000 claims abstract description 3
- 235000012680 lutein Nutrition 0.000 claims abstract description 3
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- ORAKUVXRZWMARG-WZLJTJAWSA-N lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C ORAKUVXRZWMARG-WZLJTJAWSA-N 0.000 claims abstract description 3
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims abstract description 3
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- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 claims abstract description 3
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 claims abstract description 3
- 108090001090 Lectins Proteins 0.000 claims description 31
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- DBTMGCOVALSLOR-DEVYUCJPSA-N (2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](CO)O[C@H](O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-DEVYUCJPSA-N 0.000 abstract 1
- 229920001543 Laminarin Polymers 0.000 abstract 1
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- 235000003143 Panax notoginseng Nutrition 0.000 abstract 1
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- HTIQEAQVCYTUBX-KRWDZBQOSA-N (S)-amlodipine Chemical compound CCOC(=O)C1=C(COCCN)NC(C)=C(C(=O)OC)[C@@H]1C1=CC=CC=C1Cl HTIQEAQVCYTUBX-KRWDZBQOSA-N 0.000 description 9
- 230000001603 reducing effect Effects 0.000 description 9
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 8
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- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 4
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/168—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/46—Eucommiaceae (Eucommia family), e.g. hardy rubber tree
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/488—Pueraria (kudzu)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
- A61K36/537—Salvia (sage)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/80—Scrophulariaceae (Figwort family)
- A61K36/804—Rehmannia
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
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- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Botany (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
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- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Alternative & Traditional Medicine (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the field of natural medicines, in particular to a compound plant lectin and a preparation method and application thereof. The compound phytohemagglutinin is prepared from notoginseng, eucommia bark, kudzu root, rehmannia root, salvia miltiorrhiza, kelp and potentilla anserine leaf, wherein the compound phytohemagglutinin comprises danshenic acid, danshenflavone, notoginsenoside, rehmannia root saponin, lutein, laminarin, pueraria flavone and eucommia terpene. The pharmacodynamic experiment proves that the compound phytohemagglutinin has ideal treatment effect on hypertension, when the compound phytohemagglutinin is combined with the antihypertensive drug, the antihypertensive effect is further enhanced, the compound phytohemagglutinin is obviously superior to that of the antihypertensive drug which is singly used, and the toxic and side effects of edema and the like of the antihypertensive drug after the combination are greatly reduced. In addition, the compound phytohemagglutinin is a pure natural preparation, is safe and reliable to use, is suitable for various hypertension patients, and is very suitable for the current clinical requirements.
Description
Technical Field
The invention relates to the field of natural medicines, in particular to a compound plant lectin and a preparation method and application thereof.
Background
Hypertension (hypertension) is a clinical syndrome characterized by an increase in systemic arterial blood pressure (systolic pressure and/or diastolic pressure) (systolic pressure not less than 140 mm hg, diastolic pressure not less than 90 mm hg), which may be accompanied by functional or organic damage to organs such as heart, brain, kidney, etc. Hypertension is the most common chronic disease and also the most major risk factor for cardiovascular and cerebrovascular diseases.
The medicine for treating hypertension mainly comprises western medicines and a plurality of traditional Chinese medicine formulas such as a renin-angiotensin converting enzyme inhibitor, a calcium ion antagonist, a diuretic, a vasodilator, a sympathetic nerve inhibitor and the like. However, the pharmacotherapy has some disadvantages, for example, western medicines are mostly used for symptomatic treatment, which treats the symptoms but not the root causes, and the disease condition is easy to relapse, has many adverse reactions, is easy to generate drug resistance, and can generate dependence and withdrawal reaction after long-term use; although the oral traditional Chinese medicine can fundamentally comprehensively regulate diseases, the effect is slow and the curative effect is not exact, in addition, the traditional Chinese medicine needs to be treated according to syndrome differentiation, the effect is not good and even the disease condition is aggravated if the traditional Chinese medicine is taken randomly, and the tolerance and the compliance of patients are reduced to a great extent due to the defects of the medicine treatment.
Lectin (Lectin) is a glycoprotein or carbohydrate-binding protein extracted from plant and animal cells and is known as Lectin because it agglutinates red blood cells. Lectins play an important role in cell recognition and adhesion reactions, and have multiple binding sites for sugars, and can participate in cell recognition and adhesion, linking different cells.
Phytohemagglutinin is a non-enzymatic protein derived from plants and capable of agglutinating cells and precipitating monosaccharide or polysaccharide complexes, and has important roles in various physiological processes such as signal transduction, immune response, plant defense and the like due to the specific binding capacity of the phytohemagglutinin to the monosaccharide or polysaccharide complexes. Meanwhile, the plant agglutinin has various capacities of cell agglutination, virus resistance, fungus resistance, cell apoptosis induction or autophagy induction and the like, so the plant agglutinin has high research value and good application prospect in the aspects of life science, medicine and agriculture.
The inventor discovers a G6 lectin extracted from traditional Chinese medicines and having the effect of reducing blood pressure in previous researches, and provides an invention patent application ('G6 lectin extracted from traditional Chinese medicines and application thereof in detoxifying, stabilizing pressure, softening blood vessels and preventing complications', application number: 201110321989.2). pharmacological experiments prove that the G6 lectin has a good effect of reducing blood pressure, is safe to take and has small toxic and side effects. On the basis of the results, the inventor carries out further research and screening, properly adjusts the formula of the raw materials, and purifies to obtain a novel compound lectin, and compared with the prior G6 lectin, the blood pressure reducing effect of the compound lectin is obviously enhanced.
Disclosure of Invention
The invention provides a compound plant lectin and a preparation method and application thereof, and the compound plant lectin has the advantages of simple formula and definite curative effect, and is suitable for various hypertension patients.
All raw materials of the composite phytohemagglutinin are natural medicines, and the use safety is very high.
The composite phytohemagglutinin of the invention is prepared by the following raw materials by weight:
1 part of pseudo-ginseng, 1 part of eucommia bark, 1 part of kudzu root, 1 part of rehmannia root, 1 part of salvia miltiorrhiza, 1 part of kelp and 1.5 parts of potentilla anserina leaf;
the compound phytohemagglutinin comprises the following components:
danshenic acid, Saviae Miltiorrhizae radix flavone, notoginsenoside, rehmanniae radix saponin, lutein, laminarine, radix Puerariae flavone, and Eucommiae cortex terpene.
Further, the composite plant lectin is prepared by the following method:
(1) weighing dry medicinal materials of pseudo-ginseng, eucommia, kudzu root, rehmannia root, salvia miltiorrhiza, kelp and potentilla anserine leaf, mixing and grinding into medicinal powder of 50-100 meshes;
(2) adding 5 times volume of Tris buffer solution into the medicine powder, homogenizing, soaking at 4 ℃ for 12 hours, centrifuging, and taking supernatant;
(3) filtering the obtained supernatant by a hypertonic agglutination method, and removing impurities to obtain a filtrate containing phytohemagglutinin;
(4) and dialyzing, concentrating and drying the obtained filtrate to obtain the compound phytohemagglutinin.
The purification process of the composite phytohemagglutinin adopts a high-permeability agglutination technology, has stronger selectivity, can quickly and accurately identify, purify and agglutinate active ingredients in the raw medicinal materials, and the composite phytohemagglutinin purified by the technology has extremely high permeability, is convenient for absorption and utilization by a human body, and can further obviously improve the auxiliary treatment effect of the composite phytohemagglutinin on diseases.
Further preferably, the Tris buffer solution in the preparation step (2) of the composite phytohemagglutinin of the invention is 50mmol/L Tris-HCl, 1mmol/L MgCl2, 1mmol/L CaCl2 and pH 8.0.
Further preferably, the rotation speed of the centrifugation in the preparation step (2) of the composite phytohemagglutinin of the invention is 3000rpm, the time is 15min, and the temperature is 4 ℃.
Further preferably, the dialysis in the complex lectin preparation step (4) of the present invention is a dialysis with 50 volumes of 0.5mol/L PBS buffer solution of pH7.4 at 4 ℃ for 24 hours.
In addition, the compound plant agglutinin can be further prepared into tablets, capsules or granules according to the conventional method for preparing tablets, capsules or granules in the field.
Meanwhile, the invention also relates to application of the compound phytohemagglutinin in preparation of a medicine for treating hypertension.
In the examples of the present invention, the inventors have studied the blood pressure lowering effect of the above-mentioned complex phytohemagglutinin and the inhibitory effect on the toxic and side effects of the conventional hypotensive drugs. The result shows that the compound phytohemagglutinin of the invention shows ideal blood pressure reducing effect when used alone; when the antihypertensive drug is used in combination with the antihypertensive drug, the antihypertensive effect is further enhanced, the antihypertensive drug is obviously superior to the antihypertensive drug used alone, and the toxic and side effects of edema and the like of the antihypertensive drug after combination are greatly reduced. In addition, the compound phytohemagglutinin of the invention is a pure natural preparation, and achieves the purposes of stabilizing blood pressure and preventing cardiovascular and cerebrovascular diseases by comprehensively conditioning physiological functions of patients, so that the treatment effect on hypertension and cardiovascular and cerebrovascular complications is gradually and stably exerted, thereby reducing the occurrence of drug withdrawal reaction to a certain extent. In conclusion, the composite phytohemagglutinin of the invention is very suitable for the current clinical requirements.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
Example 1
A composite plant lectin is prepared from the following raw materials in parts by weight:
1 part of pseudo-ginseng, 1 part of eucommia bark, 1 part of kudzu root, 1 part of rehmannia root, 1 part of salvia miltiorrhiza, 1 part of kelp and 1.5 parts of potentilla anserina leaf.
The preparation method comprises the following steps:
(1) weighing dry medicinal materials of pseudo-ginseng, eucommia, kudzu root, rehmannia root, salvia miltiorrhiza, kelp and potentilla anserine leaf, mixing and grinding into 50-mesh medicinal powder;
(2) adding 5 times volume of Tris buffer solution (50mmol/L Tris-HCl, 1mmol/L MgCl2, 1mmol/L CaCl2, pH8.0) into the above medicinal powder, homogenizing, soaking at 4 deg.C for 12 hr, centrifuging at 4 deg.C for 15min at 3000rpm, and collecting supernatant;
(3) filtering the supernatant by hyperosmotic agglutination method to remove macromolecular substances such as protein, starch, pectin, tannin, etc. and microorganisms to obtain filtrate containing plant lectin;
(4) the obtained filtrate was dialyzed against 50 volumes of 0.5mol/L PBS buffer solution of pH7.4 at 4 ℃ for 24 hours, concentrated and dried to obtain a complex lectin. The quantitative analysis of the protein obtained revealed that 272mg of the complex lectin protein was obtained from 375g of the drug powder.
Example 2
A composite plant lectin is prepared from the following raw materials in parts by weight:
1 part of pseudo-ginseng, 1 part of eucommia bark, 1 part of kudzu root, 1 part of rehmannia root, 1 part of salvia miltiorrhiza, 1 part of kelp and 1.5 parts of potentilla anserina leaf.
The preparation method comprises the following steps:
(1) weighing dry medicinal materials of pseudo-ginseng, eucommia, kudzu root, rehmannia root, salvia miltiorrhiza, kelp and potentilla anserine leaf, mixing and grinding into medicinal powder of 100 meshes;
(2) adding 5 times volume of Tris buffer solution (50mmol/L Tris-HCl, 1mmol/L MgCl2, 1mmol/L CaCl2, pH8.0) into the above medicinal powder, homogenizing, soaking at 4 deg.C for 12 hr, centrifuging at 4 deg.C for 15min at 3000rpm, and collecting supernatant;
(3) filtering the supernatant by hyperosmotic agglutination method to remove macromolecular substances such as protein, starch, pectin, tannin, etc. and microorganisms to obtain filtrate containing plant lectin;
(4) the obtained filtrate was dialyzed against 50 volumes of 0.5mol/L PBS buffer solution of pH7.4 at 4 ℃ for 24 hours, concentrated and dried to obtain a complex lectin. The quantitative analysis of the obtained protein showed that 375g of the drug powder contained 303mg of the complex lectin protein.
Comparative example
The G6 lectin is prepared from the following raw materials in parts by weight:
1 part of pseudo-ginseng, 1 part of eucommia bark, 1 part of kudzu root, 1 part of rehmannia root, 1 part of salvia miltiorrhiza and 1 part of kelp.
The preparation method comprises the following steps:
(1) weighing dried medicinal materials of pseudo-ginseng, eucommia, kudzu root, rehmannia root, salvia miltiorrhiza and kelp, mixing and grinding into medicinal powder of 50 meshes;
(2) adding 5 times volume of Tris buffer solution (50mmol/L Tris-HCl, 1mmol/L MgCl2, 1mmol/L CaCl2, pH8.0) into the above medicinal powder, homogenizing, soaking at 4 deg.C for 12 hr, centrifuging at 4 deg.C for 15min at 3000rpm, and collecting supernatant;
(3) filtering the supernatant by hyperosmotic agglutination method to remove macromolecular substances such as protein, starch, pectin, tannin, etc. and microorganisms to obtain filtrate containing G6 agglutinin;
(4) the filtrate was dialyzed against 50 volumes of 0.5mol/L PBS buffer (pH7.4) at 4 ℃ for 24 hours, and the G6 lectin was obtained after concentration and drying. The protein thus obtained was quantitatively analyzed, and as a result, 252mg of the lectin G6 protein was obtained from 300G of the drug powder.
Pharmacodynamic experiment
The compound phytohemagglutinin can be used for blood pressure lowering experiment of hypertension rat model and inhibition experiment of adverse side effects caused by hypotensor.
[ purpose of experiment ]
Observing the influence of the composite phytohemagglutinin on the blood pressure level of the hypertensive rat;
the combined application of the compound lectin and the levamlodipine is observed to inhibit the side effect of edema caused by water-sodium retention caused by the compound lectin and the levamlodipine.
[ Experimental materials ]
Experimental animals: spontaneous hypertensive SHR rats, each half male and female, weighing 250-300 g.
Experimental drugs: the complex phytohemagglutinin prepared in example 1.
Control drugs: g6 lectin, prepared by comparative example.
Positive drug: levamlodipine besylate.
Blank control: 0.9% physiological saline.
[ Experimental methods ]
Grouping experiments: 60 spontaneous hypertensive SHR rats with similar body weights, half of males and females, and a body weight of 250-300g, were selected and randomly divided into 6 groups, and the grouping manner is shown in Table 1.
The administration method comprises the following steps: the oral gavage is adopted for administration, and the administration is continuously carried out for 4 weeks.
The determination method comprises the following steps: the indirect blood pressure (systolic pressure) of the rat in the waking state was measured by a rat electronic blood pressure meter and a tail volume method, and the systolic pressure of the tail artery was measured before, 2 weeks, and 4 weeks after administration, respectively, and the edema condition was observed.
TABLE 1SHR rat grouping
[ Experimental results ]
The results of blood pressure measurement before and after administration of SHR rats in each group are shown in Table 2.
TABLE 2 measurement of blood pressure before and after administration of SHR rats in each group
Note: compared to the blank control group:**P≤0.01；***P≤0.001.
as can be seen from Table 2, the blood pressure values of the rats in each group were decreased to different degrees at the end of the experiment compared with the blank control group, indicating that the hypertension of the rats in each group was decreased. In contrast, the blood pressure reducing effect of the compound phytohemagglutinin is better than that of the positive medicament of the levamlodipine besylate and the control medicament of G6 lectin; the blood pressure reducing effect of the compound lectin-positive drug levamlodipine besylate is further enhanced after the compound lectin-positive drug is combined, and the blood pressure reducing effect is superior to an experimental group which singly uses the compound lectin or the levamlodipine besylate, and is also superior to a combined group of the levamlodipine besylate and G6 lectin. In the side effect aspect of edema, the positive drug levamlodipine besylate group has a 30% edema incidence rate, and the blank control group, the experimental drug group, the control drug group and the combined application group have no edema phenomenon, which shows that the combined application of the compound lectin or the G6 lectin is beneficial to reducing the side effects of edema and the like caused by the sodium-water retention of the levamlodipine besylate.
While the preferred embodiments and examples of the present invention have been described in detail, the present invention is not limited to the embodiments and examples, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art.
Claims (6)
1. The composite plant lectin is characterized by being prepared from the following raw materials in parts by weight:
1 part of pseudo-ginseng, 1 part of eucommia bark, 1 part of kudzu root, 1 part of rehmannia root, 1 part of salvia miltiorrhiza, 1 part of kelp and 1.5 parts of potentilla anserina leaf;
the compound phytohemagglutinin comprises the following components: danshenic acid, Saviae Miltiorrhizae radix flavone, notoginsenoside, rehmanniae radix saponin, lutein, laminarine, radix Puerariae flavone, and Eucommiae cortex terpene;
the compound plant lectin is prepared by the following method:
(1) weighing dry medicinal materials of pseudo-ginseng, eucommia, kudzu root, rehmannia root, salvia miltiorrhiza, kelp and potentilla anserine leaf, mixing and grinding into medicinal powder of 50-100 meshes;
(2) adding 5 times volume of Tris buffer solution into the medicine powder, homogenizing, soaking at 4 ℃ for 12 hours, centrifuging, and taking supernatant;
(3) filtering the obtained supernatant by a hypertonic agglutination method, and removing impurities to obtain a filtrate containing phytohemagglutinin;
(4) and dialyzing, concentrating and drying the obtained filtrate to obtain the compound phytohemagglutinin.
2. The complex lectin as claimed in claim 1, wherein the Tris buffer in step (2) is 50mmol/LTris-HCl, 1mmol/LMgCl2 ，1mmol/LCaCl2 ，pH8.0。
3. The complex lectin as claimed in claim 1, wherein the centrifugation in step (2) is carried out at 3000rpm for 15min at 4 ℃.
4. The complex lectin as claimed in claim 1, wherein the dialysis in step (4) is performed by dialysis with 50 volumes of 0.5mol/L PBS buffer (pH7.4) at 4 ℃ for 24 hours.
5. The complex phytohemagglutinin of any one of claims 1 to 4, wherein the complex phytohemagglutinin is further processed into the complex phytohemagglutinin tablet, capsule or granule according to methods conventional in the art for preparing tablets, capsules or granules.
6. Use of the complex plant lectin as claimed in any one of claims 1 to 4 in the manufacture of a medicament for the treatment of hypertension.
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