CN103202875B - Application of Shen-Gui pills in preparation of drugs for tumor inhibition and combined chemotherapy drugs - Google Patents
Application of Shen-Gui pills in preparation of drugs for tumor inhibition and combined chemotherapy drugs Download PDFInfo
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- CN103202875B CN103202875B CN201310085846.5A CN201310085846A CN103202875B CN 103202875 B CN103202875 B CN 103202875B CN 201310085846 A CN201310085846 A CN 201310085846A CN 103202875 B CN103202875 B CN 103202875B
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Abstract
The invention discloses application of Shen-Gui pills in preparation of drugs for tumor inhibition and combined chemotherapy drugs. The application has the advantages that effective tumor inhibition rate of the Shen-Gui pills is 30%, effective body weight of cis-platinum chemotherapy tumor-bearing mouse is increased, living states of the tumor-bearing mouse are improved, reduction of neutrophile granulocyte and monocytes caused by cis-platinum chemotherapy is reduced, reduction of spleen and thymus indexes caused by cis-platinum chemotherapy can be resisted by the Shen-Gui pills, the spleen and thymus indexes can be increased, important immune organs of the mouse are protected to some extent, concentration of HIF-1alpha (hypoxia inducible factor-1 alpha) can be reduced, hypoxia resistance of tumor cells is weakened, death of the tumor cells is accelerated, activity of LDH (lactate dehydrogenase) is lowered, damages of LDH on normal cells are reduced, and toxic action of the cis-platinum chemotherapy can be alleviated.
Description
Technical field
The present invention relates to the application of Chinese medicine, be specifically related to join and return the application of ball in the medicine preparing inhibition tumor cell and combined chemotherapy medicine.
Background technology
Ginseng is returned ball head to see " curing mirror at all times " and is rolled up nine.By Radix Sophorae Flavescentis (only end) 4 liang, Radix Angelicae Sinensis (only end) 2 liang of compositing formulas.There is the merit of clearing away heat and cooling blood.In multitype database, search for ginseng returns ball at present, only obtains " Technical Condition of Semi-bionic Extraction of uniform Design preferred DANGGUI KUSHEN WAN side medicine " one section of article, has no and returns ball to be used for the clinical of oncotherapy or Experimental report ginseng.Research proves that Radix Angelicae Sinensis polysaccharide and sulfuric ester thereof have direct facilitation, Radix Angelicae Sinensis polysaccharide energy enhancing non-specific immunity function to activating T cell propagation, and has inhibitory action to humoral immune function.Sulfation Radix Angelicae Sinensis polysaccharide has remarkable anti-tumor activity in vivo, and Radix Angelicae Sinensis polysaccharide can induce DC to break up with ripe, stimulates T cell propagation and anti-tumor activity.Radix Angelicae Sinensis polysaccharide has certain tumor-inhibiting action to sarcoma S-180 tumor-bearing mice and obviously promotes the increase of the Thymus and Spleen index of tumor-bearing mice.Oxymatrine and cyclophosphamide share and can improve tumour inhibiting rate, and the leukocyte count minimizing cause high dose cyclophosphamide and hepatic injury have certain mitigation.Matrine can transfer the immunologic function of body, produces Suppressive effect, in chemotherapy of tumors adjuvant drug, has application prospect.Matrine has certain inhibitory action for the growth of Mice Bearing Lewis Lung Cancer primary tumor, and can significantly suppress tumor tissue blood capillary to be formed, may be relevant with protein expression with its Tumor suppression VEGF mRNA.Matrine is external in vivo all shows stronger antitumor action and apoptosis-induced effect to mice H22 hepatoma carcinoma cell, and in apoptosis-promoting effect and its rise cell, Bax expresses, and suppresses Bc-l 2 to express relevant.
Ginseng returns ball to have effect of heat clearing and damp drying, cures mainly and matches with tumor pathogenesis.Modern study proves that Radix Angelicae Sinensis polysaccharide and oxymatrine all have antitumor action.Tolerance hypoxia ability and the metabolism of the generation of malignant tumor and transfer and tumor cell are relevant.HIF-1 α is the adaptation response of histiocyte to hypoxia, is the responsive controlling elements independent of blood oxygen concentration, can the expression of active cell intracellular metabolite waste scavenging gene and VEGF, promotes removing and the angiogenesis of tumor tissues metabolite.HIF-1 α can become the potential target spot of oncotherapy, and medicine, by lowering the removing of HIF-1 α inhibition tumor cell intracellular metabolite product, promotes death of neoplastic cells.LDH isozyme is the important indicator of diagnosis conscience injury of kidney, is discharged into extracellular after cell membrane damage.Cohort study shows that LDH is high, poor prognosis.Traditional cancer therapy drug to the normal cell of body also damaging action, causes the degradation of the several functions such as immunity of organism while killing malignant cell, the reduction of index and spleen index and thymus index, leukopenia.Therefore, research has tumor-inhibiting action and the Chinese medicine that chemotherapeutics has an efficacy enhancing and toxicity reducing effect is become gradually to the focus of Chinese medicine study.
Summary of the invention
Object of the present invention is exactly for above-mentioned defect of the prior art, provides ginseng and returns the application of ball in the medicine preparing inhibition tumor cell and combined chemotherapy medicine.
To achieve these goals, technical scheme provided by the invention is: ginseng returns the application of ball in the medicine preparing inhibition tumor cell.
Second object of the present invention there is provided ginseng and returns the application of ball in the combined chemotherapy medicine preparing inhibition tumor cell, and the ginseng that consists of of described combined chemotherapy medicine returns ball and cisplatin.
Further, above-mentioned application, described ginseng is returned in ball, according to weight ratio Radix Angelicae Sinensis: Radix Sophorae Flavescentis is 1:2.
Further, above-mentioned application, described ginseng returns the form of being used as medicine of ball to be decoction.
Further, above-mentioned application, described tumor cell is H22 hepatoma carcinoma cell.
Ginseng returns the application of ball in the medicine preparing inhibition tumor cell, and described ginseng returns the consumption of ball to be every kg body weight per day 9g, and application method is gavage, and administration time is 14 days.
Ginseng returns the application of ball in the combined chemotherapy medicine preparing inhibition tumor cell, and in described combined chemotherapy medicine, ginseng returns the consumption of ball to be every kg body weight per day 9g, and application method is gavage, and administration time is 14 days; The consumption of cisplatin is every kg body weight per day 2.0mg, and application method is lumbar injection, and administration time is 14 days.
Beneficial effect of the present invention is: ginseng provided by the invention returns the application of ball in the medicine preparing inhibition tumor cell and combined chemotherapy medicine, and ginseng returns ball to have effective tumour inhibiting rate (>30%); Increase effective weight of cisplatin chemotherapy tumor-bearing mice; Improve the survival condition of tumor-bearing mice; Neutrophilic granulocyte caused by cisplatin chemotherapy and mononuclear cell are lowered and has rising effect; Ginseng returns ball the index and spleen index that causes of antagonism cisplatin chemotherapy and thymus index to reduce, and has the effect raising index and spleen index and thymus index, has certain protective effect to the important immune organ of mice; The concentration level of HIF-1 α can be reduced, slacken the ability of tumor cell tolerance hypoxia, promote death of neoplastic cells; Reduce the activity of LDH, reduce LDH to Normocellular damage, thus have Attenuation to cisplatin chemotherapy.
Experimental verification:
1 material:
1.1 animals:
SPF level kunming mice 100, public 45, female 55, quality 20-22g, thered is provided by Gansu Chinese of Traditional Chinese Medicine SPF level animal experimental center, the animal quality certification number: SCXK (sweet) 2011-0001, SPF level animal facility use certificate number: SYXK (sweet) 2011-0001.
1.2 cell strains:
The strain of H22 hepatocarcinoma tumor is by Gansu Chinese of Traditional Chinese Medicine's combination of Chinese and Western medicine institute Secondary Culture.
1.3 medicines and reagent:
CO
2incubator (NAPCO France), RPMI1640 (American I nvitro company), cisplatin injections, 10mg/ props up, Shandong Zhiyue Co., Ltd (lot number: 201002CF); HIF1 α test kit, Yuan Ye bio tech ltd, Shanghai (lot number: 20121001A); LDH test kit, Bioengineering Research Institute's (lot number: 20121011) is built up in Nanjing.Ginseng returns ball to be self-control decoction (every ml medicinal liquid contains 0.45g crude drug, Radix Angelicae Sinensis 0.15g/ml, Radix Sophorae Flavescentis 0.30g/ml).
1.4 instruments:
SW-CJ-IF super-clean bench (Chinese Suzhou Decontamination Equipment Plant), electronic balance (LD100001, Long Teng Electronics Co., Ltd.), Abacus Junior vet 5 animal serum analyser (blue bridge medical treatment), microplate reader (680 types, BD-RAD company of the U.S.), UV1101 spectrophotometry instrument (Shanghai Tian Mei science company limited).
2 methods:
2.1 ginsengs return the preparation of Fish ball soup agent:
By angelica sinensis 15g, Radix Sophorae Flavescentis decoction pieces 30g is placed in potter's clay medicine pot, add 500ml deionized water and soak 30min, after boiling, slow fire is boiled medicine 1h, is poured into by medicinal liquid in beaker, medicinal residues are still stayed in medicine pot, after medicine pot temperature declines, then add 500ml deionized water and soak 30min, after boiling, slow fire is boiled medicine 1h, after being mixed by twice fried liquid, in water-bath, heating in water bath is concentrated to 100ml, autoclaving.Its outward appearance is in dark brown brown, sticky moderate, transparent in flint glass graduated cylinder, and drug level is that 0.45g/mL, SPF level experiment normal temperature laboratory preserves 3 days by raw medicine calculation.
2.2 animal model preparations:
Aseptically, in SPF level laboratory, get 100 KM mices, female 55, public 45, after the 4d that conforms under laboratory condition, separate each 8 of male and female by random digits table, be set to Normal group.Get the ascitic type H22 mouse ascites gone down to posterity 7 days, being adjusted to cell concentration with normal saline is 2 × 10
6individual mL
-1, by 84 mices with 75% ethanol disinfection, the H22 hepatoma carcinoma cell suspension 0.2mL for preparing of subcutaneous injection according to a conventional method, be inoculated in the right front axil of kunming mice subcutaneous, cell counting is about 4 × 10
5individual, with occur skin mound for subcutaneous vaccination position correct.Observe the tumor growth situation at mouse inoculation position in 5-7 days, inoculation position is visible or touch tuberosity for inoculate successfully.
2.3 animal pattern grouping and process:
Within 8th day, do not eliminate tumor good or inoculate unsuccessful mice, random choose inoculates successful mice, and male and female is each, and 32 are divided into model control group, cisplatin group at random, ginseng returns ball group and ginseng to return ball to add cisplatin group, often organize 16.Weigh after grouping, model control group is with normal saline 20mLkg
-1d
-1gavage, cisplatin group is with 2.0mgkg
-1d
-1lumbar injection 0.2ml, ginseng returns ball group with 9gkg
-1d
-1gavage 0.4ml ginseng returns pill liquid, and ginseng returns ball to add cisplatin group group with 9gkg
-1d
-1gavage ginseng returns pill liquid, lumbar injection 2.0mgkg
-1d
-1cisplatin 0.2ml, successive administration 14 days.Observe mouse survival state, as one of index evaluating drug toxicity; After drug withdrawal, next day takes weight, eyeball get blood examination have a blood test conventional and centrifugal after get serum and put less than-20 DEG C preservations.Peel off tumor soma, weigh respectively after winning liver,spleen,kidney, thymus.Calculate index and the availability indexes of tumour inhibiting rate, tumor, liver, kidney, spleen, thymus etc., and the changes of body mass before and after medication.Weight during weight during changes of body mass=execution-divided into groups.Mice dead before putting to death, does not participate in the calculating of meansigma methods.Tumour inhibiting rate (%)=(model control group average tumor quality-experimental group average tumor quality) average tumor quality of/model control group.Shoot formation=organ mass (mg)/weight (g).Effective shoot formation=organ mass/(weight-tumor quality).Weight when weight-tumor during effective changes of body mass=execution weigh-has been divided into groups.
2.4 evaluating drug effects:
The Pharmacological Evaluation of Herbal Medicine standard of the treatment malignant tumor formulated according to health ministry bureau of drug policy & administration:
(1) control group mice tumor average must not lower than 1g;
(2) tumor control rate is more than 30%, and comparing with matched group has significant difference;
(3) administration treated animal average body Quality Down (with own control) must not more than 15%;
(4) animal dead number is no more than 20%.
2.5 drug combination curative effects:
Q value is calculated: q=E according to WebbShi mark product method
aB/ [E
a+ (1-E
a) E
b.E in formula
afor A medicine (ginseng returns ball or ginseng to return ball) tumour inhibiting rate, E
bfor B medicine (cisplatin) tumour inhibiting rate, E
aBfor the tumour inhibiting rate that drug combination group two medicine share, q=0.85-1.15 is that two medicine effects are added, and q>1.25 is that two medicine effects strengthen (or collaborative), and q<0.85 is two medicine antagonisms.
2.6 statistical dispositions:
Experimental data all with (
± s) represent, adopt SPSS 17.0 statistical software to carry out statistical procedures, after homogeneity test of variance, compare between group and check with t.
2 results:
2.1 modeling success rates:
Tumor tuberosity is all there is, average diameter 3mm, modeling success rate 100% in 37 male Mus when inoculation 7 days.39 are had to occur tumor tuberosity, average diameter 2.3mm in 47 female Mus, modeling success rate 83%.Repeatedly preliminary experiment finds that the tumor cognition of indivedual beans size is died away or do not grow, and makes modeling success rate bigger than normal than actual value, and therefore this research is many for the theoretical quantity of mice number ratio of modeling.
The general status of 2.2 tumor-bearing mices:
Inoculate successfully, tumor block is long-pending to be increased gradually, and outward appearance is spherical in shape, hemispherical or mountain range shape.Initial stage can be movable, mostly hardening gradually afterwards.Tumor-bearing mice figure well-balanced (except tumor body bump pad) before grouping, agile, flexibly, fur is smooth in reaction, and feed water inlet is normal; Divide into groups after 4 days, along with the continuation of experimental period extends, there is lethargy in cisplatin group mice at first, diet activity reduces, and assembles one, and fur one-tenth is seized and tarnished, the phenomenons such as hypogastric region is unholiness, urine flavescence, it is more late than cisplatin group that ginseng returns ball to add the appearance of above-mentioned phenomenon in cisplatin group.In model group, the appearance of above-mentioned phenomenon again a little later, and in Radix Angelicae Sinensis Bulbus Fritillariae Uninbracteatae Radix Sophorae Flavescentis group ball, the appearance of above-mentioned phenomenon the latest.From diet, activity and chroma of hair, ginseng returns ball group mouse survival quality and survival condition all apparently higher than cisplatin group.General status compares and shows that cisplatin integral status is the poorest.As shown in table 1ly to compare for respectively organizing mice general status during inoculation the 21st day.
Table 1
。
The mensuration of 2.2 tumor weights, tumour inhibiting rate:
Ginseng return ball, cisplatin, ginseng to return ball combination with cisplatin all to have tumor-inhibiting action, as shown in table 2 be each group of tumor weight and tumour inhibiting rate comparison (
± s).Cisplatin group dead mouse 3, time dead, weight is respectively 15.4g, 15.8g, 14.6g.3 dead mouses a few days ago curling oneself up, without wish state.Dead mice does not participate in list statistics.According to
, illustrate that ginseng returns ball and cisplatin combined application to there is summation action.
Table 2
。
Compare with model group, ★ P < 0.01.
2.3 tumor-bearing mice treatment forebody-afterbody quality comparation:
As shown in table 3ly to compare for Murine Hepatoma22 tumor-bearing mice changes of body mass (
± s).
Table 3
。
Weight during first administration is compared, ★ P > 0.05 between each group; Compare with model group, * * P > 0.05; Compare with model group, * P < 0.01; Ball group is returned to compare with ginseng, * * * P < 0.01.
2.4 tumor-bearing mice shoot formation compare:
As shown in table 4 be Murine Hepatoma22 tumor-bearing mice liver index, renal index, index and spleen index, thymus index (
± s).
Table 4
。
Note: compare with cisplatin group, ★ P < 0.01; ★ ★ P < 0.01.
The effective shoot formation of 2.5 tumor-bearing mice compares (
± s):
。As shown in table 5 for the effective liver index of Murine Hepatoma22 tumor-bearing mice, renal index, index and spleen index, thymus index compare (
± s).
Table 5
。
Note: compare with cisplatin group, ★ P < 0.01; ★ ★ P < 0.01.
2.6 tumor-bearing mice cytometries compare:
Numeration of leukocyte reflects inflammation and the bone marrow depression situation of mice body, in 2ml centrifuge tube, drip heparin-saline 80 microlitre, then add whole blood 20 microlitre, detect in the conventional instrument of animal blood, Murine Hepatoma22 tumor-bearing mice cytometry (n=10
± s) statistical result is as shown in table 6.
Table 6
。
Note: compare with cisplatin group, * P>0.05; ▲, ★ P < 0.05.
2.7 tumor-bearing mice serum HIF1 α concentration and LDH activity:
In strict accordance with the operation of test kit description, the OD value obtained is converted into the concentration value of HIF-1 α and the energy value of LDH.Each group of horizontal comparative result of mice serum HIF-1 α, LDH (
± s) as shown in table 7.
Table 7
。
Compare with model group, * P<0.05; Compare with cisplatin group, ★ P<0.01.
Detailed description of the invention
embodiment 1:
Ginseng returns the application of ball in the medicine preparing inhibition tumor cell, and ginseng returns ball according to weight ratio Radix Angelicae Sinensis: Radix Sophorae Flavescentis is 1:2, and ginseng returns ball form of being used as medicine to be decoction, tumor cell is H22 hepatoma carcinoma cell, ginseng returns the consumption of ball to be every kg body weight per day 9g, and application method is gavage, and administration time is 14 days.
embodiment 2:
Ginseng returns the application of ball in the combined chemotherapy medicine preparing inhibition tumor cell, the ginseng that consists of of combined chemotherapy medicine returns ball and cisplatin, ginseng returns ball according to weight ratio Radix Angelicae Sinensis: Radix Sophorae Flavescentis is 1:2, ginseng returns ball form of being used as medicine to be decoction, tumor cell is H22 hepatoma carcinoma cell, ginseng returns the consumption of ball to be every kg body weight per day 9g, and application method is gavage, and administration time is 14 days; The consumption of cisplatin is every kg body weight per day 2.0mg, and application method is lumbar injection, and administration time is 14 days.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (2)
1. ginseng returns the application of ball in the medicine preparing inhibition tumor cell, and it is characterized in that, described ginseng is returned in ball, according to weight ratio Radix Angelicae Sinensis: Radix Sophorae Flavescentis is 1:2; Described ginseng returns the form of being used as medicine of ball to be decoction; Described tumor cell is H22 hepatoma carcinoma cell; Described ginseng returns the consumption of ball to be every kg body weight per day 9g, and application method is gavage, and administration time is 14 days.
2. ginseng returns the application of ball in the combined chemotherapy medicine preparing inhibition tumor cell, and it is characterized in that, the ginseng that consists of of described combined chemotherapy medicine returns ball and cisplatin, and described ginseng is returned in ball, according to weight ratio Radix Angelicae Sinensis: Radix Sophorae Flavescentis is 1:2; Described ginseng returns the form of being used as medicine of ball to be decoction; Described tumor cell is H22 hepatoma carcinoma cell; In described combined chemotherapy medicine, ginseng returns the consumption of ball to be every kg body weight per day 9g, and application method is gavage, and administration time is 14 days; The consumption of cisplatin is every kg body weight per day 2.0mg, and application method is lumbar injection, and administration time is 14 days.
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| Title |
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