CN104523902B - A kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill - Google Patents

A kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill Download PDF

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CN104523902B
CN104523902B CN201510036927.5A CN201510036927A CN104523902B CN 104523902 B CN104523902 B CN 104523902B CN 201510036927 A CN201510036927 A CN 201510036927A CN 104523902 B CN104523902 B CN 104523902B
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ramulus cinnamomi
chinese medicine
medicine composition
drug resistance
cell
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CN104523902A (en
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韩立
郭晓娟
付卫云
王奎鹏
张韡
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Nanyang Institute of Technology
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Abstract

The invention discloses a kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, it is to be formed by Ramulus Cinnamomi, Poria, Cortex Moutan, Radix Paeoniae Rubra, Semen Persicae 5 taste traditional Chinese medicine.The present invention utilizes the multiple pharmacologically active that Ramulus Cinnamomi Poria pill is had, by changing the proportion of composing of the former side of Ramulus Cinnamomi Poria pill, become a kind of Chinese medicine composition based on Ramulus Cinnamomi Poria pill, prepare the concentrated solution of this Chinese medicine composition, and prepare Contained Serum according to this concentrated solution, it is experimentally confirmed this concentrated solution and Contained Serum can reverse the drug resistance of ovarian cancer cell of height drug resistance effectively, have the advantages that safety is high, toxicity is low, curative effect is lasting, treatment cost is low, can effectively reverse the drug resistance of the ovarian cancer cell of height drug resistance.

Description

A kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill
Technical field
The invention belongs to field of traditional Chinese medicine technology, be specifically related to a kind of based on Ramulus Cinnamomi Poria pill reverse oophoroma multidrug resistance
Chinese medicine composition.
Background technology
Ovarian cancer is the malignant tumor that in gynecological tumor, mortality rate is the highest, and wherein 90% is ovarian epithelial carcinoma.Owing to ovarian tumors is hidden, for late period (FIGO by stages: III phase and IV phase) when about 70% patient makes a definite diagnosis.The standard regimens that the chemotherapy based on paclitaxel/platinum class is current advanced ovarian cancer is carried out after surgical operation.At the chemotherapy initial stage, paclitaxel/platinum class Combination chemotherapy is highly effective to ovarian cancer, but the patient of 80% can be recurred, and to paclitaxel/platinum class drug resistance, causes 5 years survival rates of Patients with Advanced Ovarian Carcinoma to be only 18%～30%.Tumor cell once produces drug resistance to certain chemotherapeutics, also the medicine that other mechanism of action are different with chemical constitution can be produced drug resistance, here it is multidrug resistance (multidrug resistance, MDR).Tumor cell is while drug resistance, and its internal gene also can change, and such as tumor cell is expressed without P glycoprotein (P-glycoprotein, P-gp) before drug resistance, but after it occurs drug resistance, P-gp often expresses and increases.P-gp is a kind of " pump medicine " albumen, and after in tumor cell, P-gp expresses and increases, the ability pumping out antitumor drug strengthens.As a example by ovarian cancer, paclitaxel/platinum series antineoplastic medicament can kill normal ovarian tumor cell, and its reason is that normal tumor cell is expressed without P-gp, and antitumor drug can enter in tumor cell, reaches to kill the concentration of tumor cell.But ovarian cancer tumor cell P-gp expresses after increasing, and the antitumor drug being pumped out tumor cell by P-gp increases, the antitumor drug lowering of concentration in tumor cell, does not reaches the concentration that can kill tumor cell, which results in chemotherapy failure.Separately studies have found that, tumor cell P-gp expresses after increasing, and its wetting capacity and transfer ability strengthen, and the histological type of tumor tissues also can change.
The medicine of the research of reversion MDR mainly chemosynthesis the most both at home and abroad and Chinese medicine monomer.The MDR reversal agents of some chemosynthesis has been carried out I/II clinical trial phase, but these MDR reversal agents are all excessive due to toxicity or reversing drug resistance effect in clinical trial inconspicuous and terminate research, such as cyclosporin A ex hoc genus anne compound valspodar (Valspodar) etc., its reason is probably these MDR reversal agents simply single target spot and acts on tumor drug resistance cell, causes its clinical efficacy inconspicuous.Some research worker transfer to extract from the natural product including Chinese medicine to separate MDR reversal agents, it is relatively low that the MDR reversal agents of natural origin is generally of toxicity, the feature that human tolerance is good, but these researchs also rest on laboratory stage mostly, there is no the biggest progress at present.Therefore, there is no at present can be used for clinic MDR reversal agents.
Ramulus Cinnamomi Poria pill is Zhang Zhongjing " Medical Treasures of the Golden Chamber " contained classical prescription, and former side, for disease, is made up of Ramulus Cinnamomi, Poria, Cortex Moutan, Radix Paeoniae Rubra, Semen Persicae equivalent, the applicating history of existing more than one thousand years, has the advantages that safety is high, toxicity is low.Modern pharmacological research shows, Ramulus Cinnamomi Poria pill decocting liquid has faint tumor-inhibiting action, it is impossible to use clinically as antitumor drug.Have now been found that the composition containing reversion MDR effect in Poria and Radix Paeoniae Rubra, but not yet have whether document report Ramulus Cinnamomi Poria pill can play the effect of reversing tumor MDR as a complete classical prescription.
Summary of the invention
The technical problem to be solved is to provide a kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, this Chinese medicine composition meets the organic conception of tcm prescription compatibility, can be preferably from Mutiple Targets, multipath, too many levels reversing tumor MDR, have the advantages that safety is high, curative effect is lasting, treatment cost is low, thus reach to improve chemotherapy effect, change
Kind patient's prognosis, and produce economic and social benefit.
For achieving the above object, the technical scheme is that: a kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, be made up of following weight proportioning raw material: Ramulus Cinnamomi 9-12 part, Poria 10-15 part, Cortex Moutan 9-12 part, Radix Paeoniae Rubra 10-15 part, Semen Persicae 9-12 part.
Further, the described Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, it is made up of following weight proportioning raw material: Ramulus Cinnamomi 9 parts, 15 parts of Poria, Cortex Moutan 9 parts, Radix Paeoniae Rubra 15 parts, 9 parts of Semen Persicae.
The present invention utilizes the multiple pharmacologically active that Ramulus Cinnamomi Poria pill is had, by changing the proportion of composing of the former side of Ramulus Cinnamomi Poria pill, become a kind of Chinese medicine composition based on Ramulus Cinnamomi Poria pill, prepare the concentrated solution of this Chinese medicine composition, and prepare Contained Serum according to this concentrated solution, it is experimentally confirmed this concentrated solution and Contained Serum can reverse the drug resistance of ovarian cancer cell of height drug resistance effectively, have the advantages that safety is high, toxicity is low, curative effect is lasting, treatment cost is low, can effectively reverse the drug resistance of the ovarian cancer cell of height drug resistance.
Detailed description of the invention
The present invention reverses the Chinese medicine composition of oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, is made up of following weight parts proportioning raw material: Ramulus Cinnamomi 9-12 part, Poria 10-15 part, Cortex Moutan 9-12 part, Radix Paeoniae Rubra 10-15 part, Semen Persicae 9-12 part.
Embodiment 1
The present invention reverses the Chinese medicine composition of oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, and the weight of stating of its each component raw material may is that Ramulus Cinnamomi 9 parts, 10 parts of Poria, Cortex Moutan 9 parts, Radix Paeoniae Rubra 10 parts, 9 parts of Semen Persicae.
Embodiment 2
The present invention reverses the Chinese medicine composition of oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, the stating weight and can also be of its each component raw material: Ramulus Cinnamomi 10 parts, 13 parts of Poria, Cortex Moutan 10 parts, Radix Paeoniae Rubra 13 parts, 10 parts of Semen Persicae.
Embodiment 3
The present invention reverses the Chinese medicine composition of oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, the stating weight and can also be of its each component raw material: Ramulus Cinnamomi 12 parts, 15 parts of Poria, Cortex Moutan 12 parts, Radix Paeoniae Rubra 15 parts, 12 parts of Semen Persicae.
Embodiment 4
The present invention reverses the Chinese medicine composition of oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill, its each component raw material state weight it may also is that Ramulus Cinnamomi 9 parts, 15 parts of Poria, Cortex Moutan 9 parts, Radix Paeoniae Rubra 15 parts, 9 parts of Semen Persicae.
Experimental example 1:
Experimental technique:
Ramulus Cinnamomi, Poria, Cortex Moutan, Radix Paeoniae Rubra, Semen Persicae is taken by described proportioning, Cortex Moutan steam distillation, collect distillate, it is standby point to take volatile ingredient；Cortex Moutan medicinal residues to without alcohol taste, are evaporated to appropriate with Ramulus Cinnamomi, Radix Paeoniae Rubra, Semen Persicae and Poria 90% ethanol extraction secondary, united extraction liquid, recovery ethanol；Ramulus Cinnamomi, Poria, Cortex Moutan, Radix Paeoniae Rubra, Semen Persicae and Cortex Moutan medicinal residues add water decoction secondary, filter, merging filtrate, be evaporated to appropriate, merge mixing with above-mentioned concentrated solution and described volatile ingredient, reach the standard (hereinafter referred to as concentrated solution) containing crude drug 1.5 g in 1 ml medicinal liquid.
Cell line SKOV3 is human ovarian cancer sensitive cells strain, and SKOV3/DDP is the drug-resistant cell strain induced with cisplatin (DDP).The concentrated solution prepared according to the method described above, prepare Ramulus Cinnamomi Poria pill Contained Serum (hereinafter referred to as Contained Serum) after being administered to Wistar rat oral gavage, use CCK-8(Cell Counting Kit-8) test kit detection Contained Serum combination with cisplatin/paclitaxel half-inhibition concentration (IC in human ovarian cancer sensitive cells strain SKOV3 and the cell of resistance to cisplatin SKOV3/DDP thereof respectively50), calculate drug resistance multiple and drug resistance reversal fold.Use Flow Cytometry Assay through the fluorescent dyes rhodamine 123(Rhodamine123 of drug resistance P-gp glycoprotein transport) accumulation in above-mentioned sensitive strain, persister cell, and after Contained Serum processes, the change of persister intracellular Rhodamine 123 accumulation.After using Western blot and RT-PCR technology detection SKOV3/DDP cell Contained Serum to process, P-gp expresses and encoding geneMDR1The expression change of mRNA, specifies Contained Serum and expresses P-gp and function and encoding gene thereofMDR1Effect.
Result: SKOV3/DDP The mensuration of cells resistance multiple
The cisplatin IC to SKOV3/DDP cell50Value is: 14.79 ± 0.41 mgL-1(n=3), the cisplatin IC to its sensitive strain cell SKOV350Value is: 3.58 ± 0.12 mgL-1(n=3), SKOV3/DDP cell is 4.13 times to the drug resistance multiple of cisplatin.
The SKOV3/DDP cell IC to paclitaxel50Value is: 6.86 ± 0.22 mgL-1(n=3), its sensitive strain cell SKOV3 IC to paclitaxel50Value is: 1.78 ± 0.31 mgL-1(n=3), SKOV3/DDP cell is 3.85 times to the drug resistance multiple of paclitaxel.
Contained Serum pair SKOV3/DDP Cell and SKOV3 The cytotoxicity of cell:
Concentrated solution is with 16 g kg-1·d-1(high), 8 g kg-1·d-1In (), 4 g kg-1·d-1(low) dosage gives rat oral gavage respectively, prepare the Contained Serum of high, medium and low three kinds of concentration, it is respectively (20.4 ± 4.0) %, (11.5 ± 1.1) % and (4.7 ± 1.4) % to the suppression ratio of SKOV3 cell, the suppression ratio of SKOV3/DDP cell is respectively (14.9 ± 2.4) %, (6.3 ± 2.0) % and (3.3 ± 0.7) %, wherein low concentration Contained Serum is below 5% to the suppression ratio of two kinds of cells, the most acellular toxic action.Middle and high concentration Contained Serum is more weak to the cytotoxicity of two kinds of cells.
Contained Serum pair SKOV3/DDP The drug resistance inversion effect of cell:
IC to SKOV3 cell after the low concentration Contained Serum of acellular toxic action and Cisplatin50Value is: (3.22 ± 0.08) mgL-1(n=3), the IC to SKOV3/DDP cell50Value is: (4.39 ± 0.17) mgL-1(n=3).Low concentration Contained Serum is to being 3.04 times to the reversal index of SKOV3/DDP cell
Contained Serum pair SKOV3/DDP Cell rhodamine 123 The impact of accumulation:
Along with Contained Serum drug concentration increases, the accumulation of intracellular Rhodamine 123 increases, and its fluorescence intensity gradually strengthens, and illustrates that the P-gp function of SKOV3/DDP drug-resistant cell strain is suppressed.
Contained Serum pair SKOV3/DDP Cell P-gp The impact expressed:
Sensitive cells strain SKOV3 expresses without P-gp, and the P-gp of drug-resistant cell strain SKOV3/DDP expresses substantially to be increased, but along with the increase of Contained Serum drug level, the P-gp of SKOV3/DDP cell expresses and is gradually reduced, illustrates that Contained Serum is inhibited to P-gp.
Contained Serum pair SKOV3/DDP Cell MDR1 mRNA The impact expressed:
Drug-resistant cell strain SKOV3/DDP'sMDR1Mrna expression is higher, but along with the increase of Contained Serum drug level, SKOV3/DDP cellMDR1Mrna expression is gradually reduced, and illustrates that Contained Serum is to coding P-gp'sMDR1Inhibited.
Experimental result is pointed out:
Contained Serum prepared by Chinese medicine composition based on Ramulus Cinnamomi Poria pill has to a certain degree cytotoxicity when higher concentration to ovarian cancer drug-resistant strain SKOV3/DDP and sensitive strain cell SKOV3, but under acellular poison mass action, can effectively reverse the SKOV3/DDP cell drug resistance to cisplatin.This Contained Serum can suppress the function of the white P-gp of pump medicated egg of persister SKOV3/DDP, simultaneously suppression P-gp and the expression of encoding gene MDR1 mRNA thereof, thus plays the effect of its reversing tumor MDR.
Experimental example 2:
Taking 6 week old female BAl BIc/c nude mice, in omoplate district, mice both sides, subcutaneous symmetric position injects 1 × 10 respectively7~1×108Individual SKOV3/DDP cell, sets up subcutaneous implantation tumor model.When tumor growth is to about 3 cm, puts to death mice, take tumor tissues.Separately take 6 week old female BAl BIc/c nude mice row implantation in situ: under aseptic condition, choose fresh subcutaneous tumors tissue and immerse rinsing in physiological saline solution, tumor tissue is cut into 1 mm3Little bar is stand-by；Will Mus be transplanted barbital sodium general anesthesia, make abdominal part center longitudinal incision, expose side ovary, cut off ovary peplos, strip tumor tissue is sewed up Ovarian surface, sew up peplos, successively close abdomen, set up BALB/c nude mice drug resistance Transplanted tumor model (hereinafter referred to as resistant models) in situ.Additionally use same method, set up nude mice sensitivity Transplanted tumor model (hereinafter referred to as sensitive model) in situ with SKOV3 cell.
Above two Orthotopic implantation in nude mice tumor model is respectively divided into 6 groups, respectively 6 g kg-1·d-1(high), 3 g kg-1·d-1In (), 1.5 g kg-1·d-1(low) dosage concentrated solution group (Chinese drug-treated group；Gastric infusion), cisplatin/paclitaxel group (Western medicine chemotherapy group；Intravenous administration, cisplatin dose 2.8 mg/kg, dose of paclitaxel 6.0 mg/kg), low dosage concentrated solution and Western medicine chemotherapy combination group and blank group, often 10 drug resistance Orthotopic Transplantation Model animals of group.After each treated animal is administered 10 days, puts to death, take ovarian carcinoma, weigh, calculate tumour inhibiting rate, investigate Ramulus Cinnamomi Poria pill concentrated solution and reverse the effect of ovarian cancer drug-resistant in vivo.The computing formula of tumour inhibiting rate is:
Experimental result:
In sensitive model animal, high, medium and low dosage concentrated solution group, Western medicine chemotherapy group, low dosage concentrated solution are respectively 31.6%, 18.5%, 6.1%, 66.8%, 86.5% with the tumour inhibiting rate of Western medicine chemotherapy combination group.
In resistant models animal, high, medium and low dosage concentrated solution group, Western medicine chemotherapy group, low dosage concentrated solution are respectively 29.6%, 16.5%, 4.1%, 46.8%, 82.5% with the tumour inhibiting rate of Western medicine chemotherapy combination group.
Experimental result is pointed out: in two kinds of models, increase along with concentrated solution dosage, its tumor-inhibiting action strengthens, even if being high dose concentrated solution, its tumor-inhibiting action is the most weak, and the tumour inhibiting rate of various dose concentrated solution is through statistical analysis, no difference of science of statistics (P > 0.05) in two kinds of models, concentrated solution tumor-inhibiting action zero difference in two kinds of models is described.Western medicine chemotherapy group tumor killing effect in resistant models animal is weaker than sensitive model animal, and through statistical analysis, difference has significance (P < 0.05), illustrates that resistant models animal creates drug resistance to Western medicine chemotherapy, makes chemotherapy effect decline.
In order to reduce the concentrated solution impact on tumor suppression, we choose low dosage concentrated solution and Western medicine chemotherapy without tumor-inhibiting action substantially and are combined, after we have found that Chinese medicine and western medicine combination, in resistant models animal, its tumour inhibiting rate has been returned to 82.5% by 46.8% before being combined, and illustrates that concentrated solution can be with the reversing drug resistance animal pattern multidrug resistance to chemotherapeutics.
Above-mentioned experiment shows: the Chinese medicine composition based on Ramulus Cinnamomi Poria pill that the present invention provides can reverse the oophoroma multidrug resistance model multidrug resistance to chemotherapeutics.Being found by cell experiment, this Chinese medicine composition is by the suppression function of P-gp and expression and encoding gene thereofMDR1The expression of mRNA, plays its effect reversing oophoroma multidrug resistance.

Claims (1)

1. one kind based on Ramulus Cinnamomi Poria pill reverse oophoroma multidrug resistance Chinese medicine composition, it is characterised in that: be by The raw material of following weight portion is made: Ramulus Cinnamomi 9 parts, 15 parts of Poria, Cortex Moutan 9 parts, Radix Paeoniae Rubra 15 parts, Fructus Persicae Core 9 parts.
CN201510036927.5A 2015-01-26 2015-01-26 A kind of Chinese medicine composition reversing oophoroma multidrug resistance based on Ramulus Cinnamomi Poria pill Active CN104523902B (en)

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