CN102302737B - Traditional Chinese medicine composition for treating gastric cancer - Google Patents

Traditional Chinese medicine composition for treating gastric cancer Download PDF

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CN102302737B
CN102302737B CN 201110262095 CN201110262095A CN102302737B CN 102302737 B CN102302737 B CN 102302737B CN 201110262095 CN201110262095 CN 201110262095 CN 201110262095 A CN201110262095 A CN 201110262095A CN 102302737 B CN102302737 B CN 102302737B
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chinese medicine
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gastric cancer
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曹鹏
霍介格
王小宁
蔡雪婷
秦凤霞
朱京红
丁蓉
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Jiangsu Provincial Insititute of Traditional Chinese Medicine
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Abstract

The invention discloses a traditional Chinese medicine composition for treating gastric cancer. The traditional Chinese medicine composition is prepared from the following components in parts by weight: 10-20 parts of pilose asiabell root, 15-35 parts of fresh rhizoma atractylodis macrocephalae, 10-20 parts of zedoary, 10-15 parts of rhizoma pinelliae preparata, 10-30 parts of Chinese actinidia root and 10-30 parts of rhodiola rosea. The traditional Chinese medicine composition exerts advantages of comprehensive therapy of traditional Chinese medicine, and dialectics is combined with differentiation of disease; and according to etiology, pathogenesis and pathology features of gastric cancer, experimental observation on an in-vitro experiment for inhibiting gastric cancer SGC7901 cell proliferation by medicines and an in-vivo experiment for inhibiting transplanted tumor of gastric cancer SGC7901 cells in nude mice by medicines shows that the traditional Chinese medicine composition in the invention can induce the SGC7901 cells to produce autophagy so as to inhibit the proliferation of the gastric cancer SGC7901 cells, thereby opening a road for hope for treating patients with gastric cancer.

Description

A kind of Chinese medicine composition for the treatment of gastric cancer
Technical field
The invention belongs to the Chinese medicine compound technical field, be specifically related to a kind of Chinese medicine composition for the treatment of gastric cancer.
Background technology
The sickness rate of gastric cancer a lot of countries in the whole world is all very high, is number four in modal malignant tumor.China is the High Risk For Gastric Cancer area, and gastric cancer often just obtains diagnosis to late period, and occupies critical role take chemotherapy as main Comprehensive Treatment in the gastric cancer medical treatment.But at present chemotherapy of gastric cancer CR leads lowly, and prolong not remarkable life cycle, and the untoward reaction such as digestive tract reaction, bone marrow depression are larger, adds that the late gastric cancer patient has degradation under weak, poor appetite, the body constitution mostly, and immunologic function is poor, and is poor to chemotherapy tolerance.Present stage, the Therapeutic Method of gastric cancer has: operation, chemotherapy, radiotherapy, Biotherapeutics etc., but curative effect is all unsatisfactory, there is no at present effective Western medicine and Chinese patent medicine.
Along with the development of medical skill, motherland's traditional medicine obtains the favor of more and more patients with gastric cancer.Traditional Chinese Medicine for Gastric Carcinoma is mainly reflected in these 3 aspects of toxic and side effects of improving precancerous lesion, postoperative righting auxiliary treatment and alleviating chemotherapy.The primary treatment method comprises that Chinese medicine for oral administration compound decoction, powder, capsule, ball are loose etc., external fumigation and washing method, application, rinsing the mouth method, clysmata, the methods such as foot bath method.At present, the effect mechanism that utilizes modern molecular biology technique to study Traditional Chinese Medicine for Gastric Carcinoma becomes the focus of research, is mainly reflected in: the several respects such as cytotoxicity, the impact on apoptosis of tumor cells, the transfer of inhibition tumor invasion, inhibition tumor-blood-vessel growth.
In recent years, many scholar's research are observed, and discovery plurality of Chinese and effective ingredient thereof and Chinese medicine compound decoction can be induced apoptosis in gastric cancer by number of ways, bring into play anticancer effect:
1, the Rhizoma Atractylodis Macrocephalae has the inhibition tumor cell growth, reduces the invasive ability of tumor cell, improves the body anti-tumor capacity.
2, Oleum Curcumae and extract thereof by direct cytotoxicity, inducing apoptosis of tumour cell, affect the cancerous cell nucleic acid metabolism and affect cancer cell membrane current potential etc. and suppress tumor growth;
3, Radix Actinidiae Chinensis Chinensis extrat Radix Actinidiae Chinensis n-butyl alcohol and total flavonoid glycoside all have antitumor action, and can reduce the release of the tumor factor, find that cell cycle arrest may be one of mechanism of Radix Actinidiae Chinensis inhibition tumor growth;
4, Radix Rhodiolae extract has growth inhibited, induces the effect of breaking up and promoting apoptosis kinds of tumor cells, can reach antitumor action by immunomodulating and inhibition angiogenesis etc. simultaneously, when processing human breast cancer in nude mice transplanted tumor with Radix Rhodiolae, MMP-2 albumen, CathD protein expression are starkly lower than contrast, point out it may have the effect of the infiltration metastasis that suppresses tumor.
Gastric cancer relates to the topical manifestations of systemic disease, weakness of the spleen and stomach through disease all the time, thereby cause that the stagnation of QI, expectorant are solidifying, stasis blocking, poison knot, the pathogenesis of gastric cancer is summarized as weakness of the spleen and stomach, give birth in the evil poison, and drafted Chinese medicine compound stomach recovering particles according to this pathogenic characteristic, in the side take the Radix Codonopsis invigorating the spleen and benefiting QI as monarch, it is minister that the Rhizoma Atractylodis Macrocephalae helps the Radix Codonopsis invigorating the spleen to clear away damp pathogen, Rhizoma Pinelliae dissipating phlegm and resolving masses, regulating qi-flowing for harmonizing stomach, Radix Rhodiolae benefiting QI for activating blood circulation, Radix Actinidiae Chinensis clearing heat, detoxicating, dispelling dampness, the Rhizoma Curcumae removing blood stasis is adjuvant drug altogether, and all medicines share and gather altogether the merit of replenishing QI to invigorate the spleen, blood circulation promoting and blood stasis dispelling, powder detoxifcation.Modern study shows, Rhizoma Atractylodis Macrocephalae, Rhizoma Curcumae, Radix Actinidiae Chinensis, Radix Rhodiolae all have in various degree inhibitory action to cancerous cell among the we.
Summary of the invention
Technical problem to be solved by this invention provides the Chinese medicine composition of the treatment gastric cancer that a kind of efficient is high, toxic and side effects is little.
The technical problem that the present invention also will solve provides the preparation method of above-mentioned Chinese medicine composition.
The technical problem that the present invention will solve at last provides the application of above-mentioned Chinese medicine composition in preparation treatment gastric cancer medicine.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
A kind of Chinese medicine composition for the treatment of gastric cancer, it is made by the component of following parts by weight: 10~20 parts of Radix Codonopsis, 15~35 parts of Rhizoma Atractylodis Macrocephalae, 10~20 parts of Rhizoma Curcumae, 10~15 parts of Rhizoma Pinelliae Preparatum, 10~30 parts of Radix Actinidiae Chinensis, 10~30 parts of Radix Rhodiolaes.
As a kind of preferred technical scheme of the present invention, the Chinese medicine composition of described treatment gastric cancer is made by the component of following parts by weight: 10~15 parts of Radix Codonopsis, 15~20 parts of Rhizoma Atractylodis Macrocephalae, 10~15 parts of Rhizoma Curcumae, 10~15 parts of Rhizoma Pinelliae Preparatum, 15~20 parts of Radix Actinidiae Chinensis, 15~20 parts of Radix Rhodiolaes.
As a kind of most preferred technical scheme of the present invention: the Chinese medicine composition of described treatment gastric cancer is made by the component of following parts by weight: 13 parts of Radix Codonopsis, 20 parts of Rhizoma Atractylodis Macrocephalae, 13 parts of Rhizoma Curcumae, 13 parts of Rhizoma Pinelliae Preparatum, 20 parts of Radix Actinidiae Chinensis, 20 parts of Radix Rhodiolaes.
The preparation method of above-mentioned Chinese medicine composition with Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Rhizoma Curcumae, Rhizoma Pinelliae Preparatum, Radix Actinidiae Chinensis and the Radix Rhodiolae mixing of formula ratio, is soaked, and big fire is fried open rear little fire and decocted 2~4 hours, and extracting juice is filtered in cooling, concentrates and get final product.
Wherein, adding water weight is 16~20 times of medical material weight.
Wherein, described soak time is 30 minutes~2 hours.
The application of above-mentioned Chinese medicine composition in preparation treatment gastric cancer medicine.
Above-mentioned Chinese medicine composition is induced application in the dead medicine of stomach cancer cell generation autophagy in preparation.
Wherein, described stomach cancer cell is the SGC7901 cell.
Beneficial effect: Chinese medicine composition of the present invention has been brought into play the advantage that Chinese medicine is comprehensively cured the disease, dialectically combine with differential diagnosis of diseases, etiology and pathogenesis, pathological characteristic according to gastric cancer, suppress the experiment in vitro of gastric cancer SGC7901 cell proliferation and medicine by medicine and suppress the laboratory observation of testing in the body of gastric cancer SGC7901 cell transplanted tumor in nude mice, find that Chinese medicine composition of the present invention can induce the SGC7901 cell to produce autophagy, reach the propagation that suppresses gastric cancer SGC7901 cell, thereby opened up the road of hope for the treatment patients with gastric cancer.
Description of drawings
Fig. 1 is Western blot testing result.
Fig. 2 is that embodiment 1 Chinese medicine composition is on the impact of gastric cancer SGC7901 cell autophagy correlative protein expression.
The specific embodiment
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, process conditions and result thereof only are used for explanation the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1:
A kind of Chinese medicine composition for the treatment of gastric cancer is made by the component of following parts by weight: 13 parts of Radix Codonopsis, 20 parts of Rhizoma Atractylodis Macrocephalae, 13 parts of Rhizoma Curcumae, 13 parts of Rhizoma Pinelliae Preparatum, 20 parts of Radix Actinidiae Chinensis, 20 parts of Radix Rhodiolaes.
Concrete preparation method is: Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Rhizoma Curcumae, Rhizoma Pinelliae Preparatum, Radix Actinidiae Chinensis and Radix Rhodiolae mixing with above-mentioned formula ratio, add 18 times of water soakings 30 minutes, and big fire is fried open rear little fire and decocted 2 hours, and extracting juice is filtered in cooling, concentrates and get final product.
Embodiment 2:
A kind of Chinese medicine composition for the treatment of gastric cancer is made by the component of following parts by weight: 10 parts of Radix Codonopsis, 15 parts of Rhizoma Atractylodis Macrocephalae, 10 parts of Rhizoma Curcumae, 10 parts of Rhizoma Pinelliae Preparatum, 10 parts of Radix Actinidiae Chinensis, 10 parts of Radix Rhodiolaes.
Concrete preparation method is: Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Rhizoma Curcumae, Rhizoma Pinelliae Preparatum, Radix Actinidiae Chinensis and Radix Rhodiolae mixing with above-mentioned formula ratio, add 16 times of water soakings 1 hour, and big fire is fried open rear little fire and decocted 3 hours, and extracting juice is filtered in cooling, concentrates and get final product.
Embodiment 3:
A kind of Chinese medicine composition for the treatment of gastric cancer is made by the component of following parts by weight: 20 parts of Radix Codonopsis, 35 parts of Rhizoma Atractylodis Macrocephalae, 20 parts of Rhizoma Curcumae, 15 parts of Rhizoma Pinelliae Preparatum, 30 parts of Radix Actinidiae Chinensis, 30 parts of Radix Rhodiolaes.
Concrete preparation method is: Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Rhizoma Curcumae, Rhizoma Pinelliae Preparatum, Radix Actinidiae Chinensis and Radix Rhodiolae mixing with above-mentioned formula ratio, add 20 times of water soakings 2 hours, and big fire is fried open rear little fire and decocted 4 hours, and extracting juice is filtered in cooling, concentrates and get final product.
Embodiment 4:
1 Chinese medicine composition of the present invention suppresses gastric cancer SGC7901 cell proliferation experiment
1.1MTT colorimetry:
Embodiment 1 Chinese medicine composition is concentration and time-dependent inhibition gastric cancer SGC7901 and gastric cancer MGC803 Growth of Cells.
Experiment takes embodiment 1 Chinese medicine composition of variable concentrations and the 5-FU of variable concentrations to cooperate with gastric cancer SGC7901 cell 24h, and its cell proliferation inhibition rate increases and increases along with concentration.Draw by statistical analysis, embodiment 1 Chinese medicine composition is time dependence and concentration dependent to the inhibited proliferation of gastric cancer SGC7901 cell and MGC803 cell, and strengthen with its suppression ratio after the 5-FU coupling, illustrate after embodiment 1 Chinese medicine composition and the 5-FU coupling to have synergism.Table 1 shows that embodiment 1 Chinese medicine composition is concentration and time-dependent inhibition gastric cancer SGC7901 Growth of Cells.Table 2 shows that variable concentrations embodiment 1 Chinese medicine composition (0.02g/ml-0.03g/ml) and variable concentrations 5-FU (0-1000 μ M) cooperate with gastric cancer SGC7901 cell 24h, and its cell proliferation inhibition rate increases and increases along with concentration.Table 3 shows that variable concentrations embodiment 1 Chinese medicine composition (0.005g/ml-0.05g/ml) acts on the cell proliferation inhibition rate of Gastric Cancer MGC 803 24h-72h, its IC50 is 0.035g/ml respectively, 0.026g/ml, 0.017g/ml prompting embodiment 1 Chinese medicine composition also is concentration and the growth of time-dependent inhibition Gastric Cancer MGC 803.
Experimental result shows: embodiment 1 Chinese medicine composition has the growth inhibited effect to gastric cancer SGC7901 cell, but cell death way has apoptosis, cell autophagy and necrocytosis.Difference maximum between the three is: detect the significant molecule L C3 protein expression that cell produces the autophagy phenomenon, so take the Western-blot method to detect whether embodiment 1 Chinese medicine composition exists autophagy specific markers LC3 protein expression on gastric cancer SGC7901 cell impact.
The suppression ratio of table 1 variable concentrations different time sections embodiment 1 Chinese medicine composition treatment S GC7901 cell
Figure BDA0000089303030000041
Figure BDA0000089303030000042
Figure BDA0000089303030000051
Compare Concentraton gradient with matched group: * P<0.05, △ P<0.01;
Compared time gradient with 24 hours: ★ P<0.05, ▲ P<0.01.
The suppression ratio of table 2 variable concentrations embodiment 1 Chinese medicine composition associating 5-FU effect SGC7901 cell 24h
Figure BDA0000089303030000052
Figure BDA0000089303030000053
Compare Concentraton gradient with matched group: * P<0.05, △ P<0.01
Table 3 variable concentrations different time sections embodiment 1 Chinese medicine composition is processed the suppression ratio of MGC803 cell
Figure BDA0000089303030000055
Figure BDA0000089303030000061
Compare Concentraton gradient with matched group: * P<0.05, △ P<0.01;
Compared time gradient with 24 hours: ★ P<0.05, ▲ P<0.01.
1.2Western-blot detect:
Embodiment 1 Chinese medicine composition can be induced SGC7901 cell generation autophagy, and the impact of LC3 protein expression is time and concentration dependent.But under the embodiment of variable concentrations 1 Chinese medicine composition is intervened, the Bcl-2 of gastric cancer SGC7901 cell, Bcl-xL, Bax, Mcl-1, the P53 protein expression does not significantly change, prompting embodiment 1 Chinese medicine composition to Bcl-2 family and P53 albumen in the gastric cancer SGC7901 cell without remarkable regulating and controlling effect.Fig. 1 shows Western blot testing result: gastric cancer SGC7901 cell is at the embodiment of variable concentrations 1 Chinese medicine composition (0g/ml, 0.02g/ml, 0.025g/ml, 0.03g/ml) intervene down Bcl-2, Bcl-xL, Bax, Mcl-1, the P53 protein expression does not significantly change.Prompting embodiment 1 Chinese medicine composition to Bcl-2 family and P53 albumen in the gastric cancer SGC7901 cell without remarkable regulating and controlling effect.
1.3GFP-LC3 fluorescent staining method detects:
After adding embodiment 1 Chinese medicine composition 2h, the positive is painted in gastric cancer SGC7901 nucleus peripheral region, and the autophagy vesicle increases, and shows that embodiment 1 Chinese medicine composition can induce gastric cancer SGC7901 cell generation autophagy.Has concordance with the Western-blot testing result.Prompting embodiment 1 Chinese medicine composition can be induced gastric cancer SGC7901 cell autophagy, reaches the propagation that suppresses gastric cancer SGC7901 cell.Fig. 2 shows that variable concentrations embodiment 1 Chinese medicine composition produces the impact of the significant molecule L C3 protein expression of autophagy on gastric cancer SGC7901 cell.Establish 0.02g/ml for upper one group, 0.025g/ml, three concentration groups of 0.03g/ml and matched group, along with concentration increases, the LC3-I protein expression reduces, the LC3-II increasing expression; Next picture group is established 0.02g/ml embodiment 1 Chinese medicine composition effect 0h, 2h, 4h, 6h and matched group HCQ (50 μ M) 24h, the phenomenon that LC3-I transforms to LC3-II occurs equally.
2. Chinese medicine composition of the present invention suppresses the interior experiment of body of gastric cancer SGC7901 cell transplanted tumor in nude mice
2.1 Nude Mouse Model is set up
Select 5 of healthy nude mices, after extremity are fixing, select nude mice right fore oxter as the plantation position, behind the 75% ethanol routine disinfection, with No. 7 entry needle percutaneous punctures, (contain cell about 2 * 10 in the subcutaneous slow SGC7901 cell suspension that injects 0.2ml 7Individual/ml, single-cell suspension is in 1640 culture medium).Nude mice is after continuing to raise 20 days under the SPF condition, 5 nude mices are put to death in dislocation, skin degerming, under the aseptic condition, take out tumor piece (normal weight is 1-1.8g) in the right fore oxter, place the culture dish of a small amount of normal saline, (normal tumor piece is white to remove slough, redness is slough, and general skin is normal tumor piece, and inner redness is slough), weigh, by tumor piece weight, add normal saline, in Potter-Elvehjem Tissue Grinders, grind, the ratio of tissue and normal saline is 1g:4ml, be made into suspension, filtered through gauze, concentration is about 2 * 10 7Individual/ml, No. 7 syringe needles are given experiment mice right fore oxter inoculation oncocyte, and 0.2ml/ only.
2.2 grouping and administration
Inoculation SGC7901 cell will become the tumor nude mice to be divided at random 5 groups by the tumor size after 16 days, 9 every group: model control group, and positive drug control group, the high, medium and low dosage group of embodiment 1 Chinese medicine composition begins administration next day.The high, medium and low dosage group of embodiment 1 Chinese medicine composition is respectively crude drug 9.6,4.8,2.4g/kg, positive drug group lumbar injection 5-fluorouracil 15mg/kg, dosage is 10ml/kg, model group nude mice gavage gives the distilled water with volume, successive administration 25 days, weighed in per 3 days, and measured one time the tumor volume, observe nude mice hair color, active state and feed situation.Took off vertebra behind the medicine in 25 days and put to death nude mice, peel off tumor and weigh, and be calculated as follows tumor control rate (%): tumor control rate (%)=(model control group-administration group) weight in wet base average * 100%/model control group weight in wet base average.
2.3 experimental result
Nude mice is after being administered to the embodiment 1 traditional Chinese medicine composition water decocting liquid with variable concentrations, and each administration group and positive drug group nude mice body weight, volume all increase to some extent, but between each group significant difference (P>0.05) is not arranged.Along with the increase of drug level, tumor weight descends gradually, and tumour inhibiting rate obviously improves, and gross tumor volume changes and to reduce, at middle dosage (4.8g/kg) and high dose group (9.6g/kg) tumour inhibiting rate apparently higher than the positive drug group.Table 4 shows the increase along with drug level, and tumor weight descends gradually, and tumour inhibiting rate obviously improves, and gross tumor volume changes and reduces, at middle dosage (4.8g/kg) and high dose group (9.6g/kg) tumour inhibiting rate apparently higher than the positive drug group.
Table 4 various dose " embodiment 1 Chinese medicine composition " suppresses the effect of people's gastric cancer in nude mice Implanted and compares
Figure BDA0000089303030000071
Figure BDA0000089303030000072
Figure BDA0000089303030000081
Compare * P<0.05, △ P<0.01 with model control group.

Claims (8)

1. a Chinese medicine composition for the treatment of gastric cancer is characterized in that, it is made by the component of following parts by weight: 10~20 parts of Radix Codonopsis, 15~35 parts of Rhizoma Atractylodis Macrocephalae, 10~20 parts of Rhizoma Curcumae, 10~15 parts of Rhizoma Pinelliae Preparatum, 10~30 parts of Radix Actinidiae Chinensis, 10~30 parts of Radix Rhodiolaes.
2. Chinese medicine composition according to claim 1 is characterized in that, it is made by the component of following parts by weight: 10~15 parts of Radix Codonopsis, 15~20 parts of Rhizoma Atractylodis Macrocephalae, 10~15 parts of Rhizoma Curcumae, 10~15 parts of Rhizoma Pinelliae Preparatum, 15~20 parts of Radix Actinidiae Chinensis, 15~20 parts of Radix Rhodiolaes.
3. the preparation method of Chinese medicine composition claimed in claim 1 is characterized in that, with Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Rhizoma Curcumae, Rhizoma Pinelliae Preparatum, Radix Actinidiae Chinensis and the Radix Rhodiolae mixing of formula ratio, soak, big fire is fried open rear little fire and decocted cooling 2~4 hours, filter extracting juice, concentrate and get final product.
4. the preparation method of Chinese medicine composition according to claim 3 is characterized in that, adds water weight and be 16~20 times of medical material weight.
5. the preparation method of Chinese medicine composition according to claim 3 is characterized in that, soak time is 30 minutes~2 hours.
6. the application of Chinese medicine composition claimed in claim 1 in preparation treatment gastric cancer medicine.
7. Chinese medicine composition claimed in claim 1 is induced application in the dead medicine of stomach cancer cell generation autophagy in preparation.
8. application according to claim 6 is characterized in that, described gastric cancer, and its stomach cancer cell is the SGC7901 cell.
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