CN101172120B - Anti-liver cancer gecko Chinese actinidia root compound preparations - Google Patents
Anti-liver cancer gecko Chinese actinidia root compound preparations Download PDFInfo
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- CN101172120B CN101172120B CN2006101143065A CN200610114306A CN101172120B CN 101172120 B CN101172120 B CN 101172120B CN 2006101143065 A CN2006101143065 A CN 2006101143065A CN 200610114306 A CN200610114306 A CN 200610114306A CN 101172120 B CN101172120 B CN 101172120B
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- actinidiae chinensis
- radix actinidiae
- gekko swinhonis
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- hepatocarcinoma
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Abstract
The invention relates an anti-liver cancer gecko rattan pear root compound preparation, which is formed by performing aqueous extract to fried rice horse lizard dry powder body and a rattan pear root and performing the separation of macro pore adsorptive resin to aqueous extract. The experiment of inhibiting tomour cells is performed to the anti-liver cancer gecko rattan pear root compound preparation, the result show: the part has inhibitory effect to liver cancer cell lines Bel-7402, the inhibition rate under the high concentration can reach 93.4 percent, and the invention can be used for preparing the medicine for remedying the liver cancer.
Description
Technical field
The invention belongs to the field of Chinese medicines, specifically relate to a kind of Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma.
Background technology
The mankind find in the long process of exploring the treatment cancer drug, and the anticancer effective active composition of extraction separation from animal, plant and the microorganism of nature tool antitumaous effect is the most effective and the most feasible research method of screening cancer therapy drug.Hepatocarcinoma is one of China's common malignancy, and because of its grade of malignancy height, PD is fast, and the treatment difficulty is big, weak curative effect, person " king in the cancer ".
Radix Actinidiae Chinensis is the root of Actinidiaceae Actinidia arguta Sieb.et Zucc, energy heat-clearing and toxic substances removing, appetizing pain relieving.House liyard powder does not have all, salty-cold slightly poisonous of web Gekko Swinhonis for the Gekkonidae animal, energy eliminating stagnation removing blood stasis, and the detoxifcation antitumor, both all belong to eliminating evil medicine.The Radix Actinidiae Chinensis Herb is rich in actinidine, root contains ursolic acid, oleanolic acid, succinic acid, daucosterol and arginine, lysine, leucine, alanine, aspartic acid etc. and is rich in elemental composition (Wei Jinyu etc. such as Fe, Cu, Mn, Se, in the experimentation of Radix Actinidiae Chinensis extract antitumor action more, the Guangxi Chinese medicine, 2003,26 (6): 49).Contain protein, fat in the house liyard powder, organize amine, aminoacid, also contain and the similar noxious substance of hornet poison, in recent years, it is widely used in the multiple disease of clinical treatment, particularly, definite curative effect is arranged all in multiple difficult miscellaneous diseases such as the various malignant tumor of treatment, tuberculosis, myelopathy and surgery infection.
Chinese medicine compound is the elite place of the traditional Chinese medical science, separates obtaining its effective site and active ingredient from Chinese medicine compound, can effectively remove invalid or virose composition in the compound extract.The extraction separation method of Chinese medicine is a lot, and extracting method commonly used has: water is carried, organic solution or utilize acid-base solution extraction, supercritical extraction etc.Lixiviating solution need separate again, and separation method has: extraction, fractional distillation, precipitation, chromatographic column (silica gel, aluminium oxide, polyamide, resin, gel) etc.At China be scattered a lot of single, proved recipe among the people, these prescriptions are usually having good effect aspect the treatment cancer.
Summary of the invention
The objective of the invention is to utilize macroporous adsorbent resin, with folk remedy---extraction separation work is carried out in the active ingredient in the Radix Actinidiae Chinensis house liyard powder, thereby obtains a kind of have obvious anti-hepatoma carcinoma cell and active higher, Gekko Swinhonis Radix Actinidiae Chinensis compound preparation that toxicity is lower.
The objective of the invention is to realize by the following technical solutions:
The Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma provided by the invention is that rice stir-fry back Gekko Swinhonis dry powder body and Radix Actinidiae Chinensis are carried out water extraction together, aqueous extract is carried out the macroporous adsorbent resin separation obtain, and its concrete preparation method comprises the steps:
1) rice is fried the preparation of house liyard powder:
With commercially available house liyard powder and rice according to weight ratio 3～6: 1 mix homogeneously, put into marmite then and stir-fry, until the rice buff, remove rice, take out the house liyard powder cold of drying in the air;
2) preparation of house liyard powder and Radix Actinidiae Chinensis water extract:
After the house liyard powder of step 1) and Radix Actinidiae Chinensis mixed by weight 1: 3～9, add distilled water, reflux is carried out lixiviate, obtains the aqueous extract of Gekko Swinhonis Radix Actinidiae Chinensis;
Adding the volume of entry and the part by weight of Gekko Swinhonis Radix Actinidiae Chinensis mixture during lixiviate is 10～15ml: 1g;
Preferred 3～6 hours of reflux extracting time;
Preferred 1～3 time of reflux, extract, number of times;
3) water extract carries out the macroporous adsorbent resin separation:
At first with step 2) the Gekko Swinhonis Radix Actinidiae Chinensis water extraction product of gained, slowly pour in macroporous adsorbent resin (model the is AB-8) pillar of handling well adsorption equilibrium into behind 5～10 times of the dilute with waters.With the continuous eluting pillar of pure water, behind 5～6 times of column volumes of eluting, collect eluent; Eluent removes at 30～45 ℃ of rotary evaporations and desolvates, and until obtaining the light brown paste, vacuum drying 24h obtains the water elution part that water is carried the Gekko Swinhonis Radix Actinidiae Chinensis, is the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma;
The Gekko Swinhonis Radix Actinidiae Chinensis compound preparation (water is put forward Gekko Swinhonis Radix Actinidiae Chinensis water elution part) of anti-hepatocarcinoma provided by the invention is suppressed the tumor cell experiment, the result shows, this part has the inhibition effect to hepatoma cell strain Bel-7402, suppression ratio can reach 93.4% under high concentration, shows that it can be used to prepare the medicine for the treatment of hepatocarcinoma.
The specific embodiment
The water extract I of embodiment 1, preparation Gekko Swinhonis Radix Actinidiae Chinensis
Commercially available house liyard powder and rice according to 4: 1 mix homogeneously of mass ratio, are put into frying pan then and stir-fried,, remove rice, take out the house liyard powder cold of drying in the air, obtain rice and fry a house liyard powder until the rice buff.
Weighing 315 gram Radix Actinidiae Chinensis and 45 restrain house liyard powders, mix homogeneously respectively.Add 4500ml water, refluxed 4 hours, extract; Backflow lixiviate 3 times merges lixiviating solution, obtains the water extract of Gekko Swinhonis Radix Actinidiae Chinensis.
The Gekko Swinhonis Radix Actinidiae Chinensis lixiviating solution that water extraction is obtained dilutes 5 times, slowly pours into (AB-8) in the macroporous adsorptive resins of handling well, constantly uses the pure water eluting, collects 5 column volumes.The eluent collected at 38 ℃ of rotary evaporations, is removed and desolvates, and until obtaining brown paste, vacuum drying 24h obtains the water elution part I that water is carried the Gekko Swinhonis Radix Actinidiae Chinensis.
The water extract II of embodiment 2, preparation Gekko Swinhonis Radix Actinidiae Chinensis
Commercially available house liyard powder and rice according to 3: 1 mix homogeneously of mass ratio, are put into frying pan then and stir-fried,, remove rice, take out the house liyard powder cold of drying in the air, obtain rice and fry a house liyard powder until the rice buff.
Weighing 135 gram Radix Actinidiae Chinensis and 45 restrain house liyard powders, mix homogeneously respectively.Add 1800ml water, refluxed 3 hours, extract; Backflow lixiviate 2 times merges lixiviating solution, obtains the water extract of Gekko Swinhonis Radix Actinidiae Chinensis.
The Gekko Swinhonis Radix Actinidiae Chinensis lixiviating solution that water extraction is obtained dilutes 10 times, slowly pours into (AB-8) in the macroporous adsorptive resins of handling well, constantly uses the pure water eluting, collects 6 column volumes.The eluent collected at 30 ℃ of rotary evaporations, is removed and desolvates, and until obtaining brown paste, vacuum drying 24h obtains the water elution part II that water is carried the Gekko Swinhonis Radix Actinidiae Chinensis.
The water extract III of embodiment 3, preparation Gekko Swinhonis Radix Actinidiae Chinensis
Commercially available house liyard powder and rice according to 6: 1 mix homogeneously of mass ratio, are put into frying pan then and stir-fried,, remove rice, take out the house liyard powder cold of drying in the air, obtain rice and fry a house liyard powder until the rice buff.
Weighing 405 gram Radix Actinidiae Chinensis and 45 restrain house liyard powders, mix homogeneously respectively.Add 6750ml water, refluxed 6 hours, extract, obtain the water extract of Gekko Swinhonis Radix Actinidiae Chinensis.
The Gekko Swinhonis Radix Actinidiae Chinensis lixiviating solution that water extraction is obtained dilutes 7 times, slowly pours into (AB-8) in the macroporous adsorptive resins of handling well, constantly uses the pure water eluting, collects 6 column volumes.The eluent collected at 45 ℃ of rotary evaporations, is removed and desolvates, and until obtaining brown paste, vacuum drying 24h obtains the water elution part III that water is carried the Gekko Swinhonis Radix Actinidiae Chinensis.
Embodiment 4, water are carried Gekko Swinhonis Radix Actinidiae Chinensis water elution part cell experiment
Cell strain: hepatoma cell strain Bel-7402 is provided by the Shanghai cell
Screening technique: sulphonyl rhodamine B (Sulforhodamine B, SRB) protein staining method;
Action time: 72 hours
Experimental unit: The National Center for Drug Screening
The water that embodiment 1 obtains is carried the cell experiment that Gekko Swinhonis Radix Actinidiae Chinensis pure water eluting part I suppresses tumor.The method of experiment is: the hepatoma cell strain Bel-7402 of the trophophase of taking the logarithm is inoculated in 96 well culture plates, and cell is placed 37 ℃, 5%CO
2Pre-after adherent 24 hours in the incubator, abandoning supernatant, the Gekko Swinhonis Radix Actinidiae Chinensis water elution part of adding variable concentrations.Each concentration is provided with three multiple holes.And the normal saline solvent that is provided with respective concentration contrasts and acellular withered hole.Tumor cell is at 37 ℃, 5%CO
2Cultivated 72 hours under the condition, the culture fluid that inclines adds SRB solution and trihydroxy methyl ammonia and methane (Tris) solution successively, measures each hole absorbance under microplate reader under the 515nm wavelength.And calculate by following formula, the results are shown in table 1.
Table 1, water are proposed the inhibition experiment of Gekko Swinhonis Radix Actinidiae Chinensis pure water eluting part to hepatoma cell strain Bel-7402
The above results shows that water provided by the invention is put forward Gekko Swinhonis Radix Actinidiae Chinensis pure water eluting part hepatoma cell strain Bel-7402 is had the good restraining effect, and cell inhibitory rate reaches 93.36% under high concentration.
At the preliminary stage of this test, attempt water is carried Gekko Swinhonis Radix Actinidiae Chinensis total extract, and interior animal experiment (the zoopery unit: Xiyuan Hospital, Chinese Medicine Academy of China) of body carry out cell in vitro (test cell line unit: The National Center for Drug Screening).The cell in vitro test shows that total extract has very strong specific aim and greater activity to hepatoma carcinoma cell.Animal test results shows in the body, and this water extract is same biologically active in the mice body, and the tumor of administration group is heavy significantly to reduce (P＜0.05) with the tumor matched group.
Total extraction product is biologically active in vivo, but has dosage drawback bigger than normal.So being carried product, water carried out further macroporous adsorbent resin separating experiment, confirm through test cell line, separate the pure water eluting that obtains and partly have higher biological activity, and in to this part, in the further separation and evaluation work of contained effective ingredient, find that it mainly contains saccharide and glycoside material.This class material not only water solublity is fabulous but also have very high physiologically active, and getting a good chance of developing becomes novel anti-cancer agent.
Claims (6)
1. the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of an anti-hepatocarcinoma is that rice stir-fry back Gekko Swinhonis dry powder body and Radix Actinidiae Chinensis are carried out water extraction together, aqueous extract is carried out the macroporous adsorbent resin separation obtain, and its concrete preparation method comprises the steps:
1) rice is fried the preparation of house liyard powder:
With commercially available house liyard powder and rice according to weight ratio 3～6: 1 mix homogeneously, put into marmite then and stir-fry, until the rice buff, remove rice, take out the house liyard powder cold of drying in the air;
2) preparation of house liyard powder and Radix Actinidiae Chinensis water extract:
After the house liyard powder of step 1) and Radix Actinidiae Chinensis mixed by weight 1: 3～9, add distilled water, reflux is carried out lixiviate, obtains the aqueous extract of Gekko Swinhonis Radix Actinidiae Chinensis;
3) water extract carries out the macroporous adsorbent resin separation:
At first with step 2) the Gekko Swinhonis Radix Actinidiae Chinensis water extraction product of gained, slowly pour in macroporous adsorptive resins of handling well adsorption equilibrium into behind 5～10 times of the dilute with waters; With the continuous eluting pillar of pure water, behind 5～6 times of column volumes of eluting, collect eluent; Eluent removes at 30～45 ℃ of rotary evaporations and desolvates, and until obtaining the light brown paste, vacuum drying 24h obtains the water elution part that water is carried the Gekko Swinhonis Radix Actinidiae Chinensis, is the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma.
2. the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma as claimed in claim 1 is characterized in that: add the volume of entry during lixiviate described step 2) and the part by weight of Gekko Swinhonis Radix Actinidiae Chinensis mixture is 10～15ml: 1g.
3. the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma as claimed in claim 1 is characterized in that: reflux extracting time is 3～6 hours described step 2).
4. the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma as claimed in claim 1 is characterized in that: the reflux, extract, number of times is 1～3 time described step 2).
5. the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of anti-hepatocarcinoma as claimed in claim 1, it is characterized in that: the model of macroporous adsorptive resins in the described step 3) is AB-8.
6. the Gekko Swinhonis Radix Actinidiae Chinensis compound preparation of the described anti-hepatocarcinoma of one of claim 1 to 5 is in the purposes of preparation in the medicines resistant to liver cancer.
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| CN101172120B true CN101172120B (en) | 2010-06-09 |
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| 陈德轩,潘立群.抗肿瘤植物药藤梨根.中药材27 2.2004,27(2),86-87. |
| 陈德轩,潘立群.抗肿瘤植物药藤梨根.中药材27 2.2004,27(2),86-87. * |
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Granted publication date: 20100609 Termination date: 20131103 |