CN103142935A - Traditional Chinese medicinal composition for treating lung cancer and liver cancer - Google Patents
Traditional Chinese medicinal composition for treating lung cancer and liver cancer Download PDFInfo
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- CN103142935A CN103142935A CN2013100861519A CN201310086151A CN103142935A CN 103142935 A CN103142935 A CN 103142935A CN 2013100861519 A CN2013100861519 A CN 2013100861519A CN 201310086151 A CN201310086151 A CN 201310086151A CN 103142935 A CN103142935 A CN 103142935A
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Abstract
The invention discloses a traditional Chinese medicinal composition for treating lung cancer and liver cancer. The composition consists of rhizoma paridis yunnanensis extract and rhizoma curcumae longae extract according to a mass ratio of 1:(5-25). Experiments prove that the traditional Chinese medicinal composition can obviously inhibit lung cancer cell lines such as LA795 and hepatoma cell lines such as Hep-B3 from growing, the half maximal inhibitory concentrations (IC50) of the lung cancer cell lines and the hepatoma cell lines can respectively reach below 20 micrograms/milliliter; tests on mouse models bearing lung adenocarcinoma and liver cancer tumors show that the lung adenocarcinoma and liver cancer inhibition rates are respectively above 50%, and the spleen index and liver index of mice are increased; and compared with raw medicinal materials which are separately applied, the medicinal composition has high anti-lung cancer and anti-liver cancer activities, an exact effect and a good inhibition effect on lung cancer and liver cancer.
Description
Technical field
The present invention relates to Chinese medicine composition of a kind of anti-hepatocarcinoma and pulmonary carcinoma and preparation method thereof, belong to technical field of Chinese medicines.
Background technology
Rhizoma Paridis is the dry rhizome of liliaceous plant Rhizoma Paridis Paris polyphylla Smith var.yunnanensis (Franch.) Hand.-Mazz. or Rhizoma Paridis Paris polyphylla Smith var.chinensis (Franch.) Hara, has heat-clearing and toxic substances removing, the effect of reducing swelling and alleviating pain, cool liver arresting convulsion, being used for the treatment of the diseases such as furuncle carbuncle, laryngopharynx swelling and pain, venom, traumatic pain, cool breeze tic, is the chief component medicine of Chinese patent medicine YUNNAN BAIYAO, GONGXUENING JIAONANG, jidesheng sheyao tablets etc.Modern pharmacological research show Rhizoma Paridis have cytotoxic activity, antibacterial anti-inflammatory, analgesia, hemostasis, calmness, antiearly pregnancy, spermicidal, to respiratory system and Cardiovascular System etc., evident in efficacy in treatment lung pattern disease, dysfunctional uterine hemorrhage, tuberculous lymphadenitis ulcer, neurodermatitis and surgery inflammation and treatment tumor etc. in clinical practice.There is at present the listing Chinese patent medicine compound recipe that Rhizoma Paridis is used as medicine that the multiple recovery oral liquid of gold, softening the hard mass oral liquid, building lotus capsule, GANFULE PIAN etc. are arranged.In recent years, the bibliographical information of Rhizoma Paridis and antitumor effective ingredient Rhizoma Paridis total saponins thereof is increasing.CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2008,33(16): 2057-2060 discloses cell toxicant spectrum and the structure activity study of Rhizoma Paridis saponin to 10 kinds of tumor cell lines; Huang Xian schools etc. are at Chinese herbal medicine magazine 2009,40(3): 483-489 has delivered the progress of Paris Linnaeus(Paris L.) medicinal plants saponins chemical composition and source of students approach thereof; Chen Zhihong etc. have investigated Rhizoma Paridis total saponins to human lung cancer cell A549's inhibited proliferation and the impact of cell cycle; Zhu Lili etc. suppress SGC-7901 cell proliferation and apoptosis-induced being studied to Rhizoma Paridis saponin; Lin Yunhua research finds that Rhizoma Paridis and oxaliplatin may have the effect of certain angiogenesis inhibitor in experiment in external, body, and both use in conjunction may have synergistic function.The discovery Rhizoma Paridis total saponins such as Jia Ke can significantly suppress the growth of MGC-803 cell and meta-dose-dependence when being, and can block cell in the S phase, the rising of cell death inducing rate; Can significantly suppress EphA2, survivin protein expression, raise simultaneously the Caspase-3 protein expression, and then its mechanism of action of reaching a conclusion may and promote that with the expression of lowering EphA2 and survivin the expression of Caspase-3 is relevant.Chinese patent 200610052223.8 discloses the preparation method of Rhizoma Paridis total saponins, and 200810052092.2 also disclose pharmaceutical preparation take Rhizoma Paridis total saponins as effective ingredient and preparation method thereof.
The Rhizoma Curcumae Longae beginning is stated from Tang Materia Medica, be the dry rhizome of zingiberaceous plant Rhizoma Curcumae Longae Curcuma longa L., its nature and flavor acrid, bitter, warm is returned spleen, Liver Channel, has the removing blood stasis circulation of qi promoting, the effect of inducing menstruation to relieve menalgia can be used for the twinge of the breast side of body, obstruction of qi in the chest and cardialgia, dysmenorrhea amenorrhea mass in the abdomen, rheumatism shoulder arm pain, the treatment of the diseases such as tumbling and swelling.Rhizoma Curcumae Longae contains the number of chemical composition, and main component is curcumin chemical compounds: curcumin, bisdemethoxycurcumin, demethoxycurcumin, dihydro curcumin etc.; Sesquiterpenoids: the new ketone of Rhizoma Curcumae Longae, Rhizoma Curcumae Longae keto-alcohol A, B etc.; Volatile oil (4.2%): turmerone, curcumene, curcumenol etc.The pharmacological action of the effective ingredient such as curcumin is extensive, the effects such as its antiinflammatory, antioxidation, atherosclerosis, antidepressant, antitumor have generally been approved, wherein antitumor action after proposing in 1985 first, existing a large amount of experimentatioies confirms, mechanism of action mainly comprises inducing apoptosis of tumour cell, suppress angiogenesis, the retardance cell cycle progression is arranged, the aspects such as reversing multiple medicine resistance of tumor cells.Huang Dongsheng etc. study discovery, and curcumin can strengthen the expression of caspase-8, thereby start exogenous apoptosis pathway, induce the human lung carcinoma cell apoptosis.The report curcumins such as Fang Yue have the effect of the digestive system tumors such as anti-esophageal carcinoma, gastric cancer, hepatocarcinoma, colon cancer.Chinese patent CN1931353A discloses curcumin and has extracted and the pharmaceutical composition preparation method.
At present, the patent of the pharmaceutical composition of Rhizoma Paridis saponin and astragalus polysaccharides (CN101152408A) has been seen in report, and not yet finds to have simultaneously anti-hepatocarcinoma and pulmonary carcinoma effect and mix by Rhizoma Paridis extract and Rhizoma Curcumae Longae extract the Chinese medicine composition that forms.
Summary of the invention
The object of the present invention is to provide the Chinese medicine composition of a kind of anti-hepatocarcinoma and pulmonary carcinoma.
Technical scheme of the present invention is summarized as follows:
The Chinese medicine composition of a kind of anti-hepatocarcinoma and pulmonary carcinoma, the Rhizoma Paridis extract and the Rhizoma Curcumae Longae extract that are 1:5-25 by mass ratio form.
The mass ratio of described Rhizoma Paridis extract and Rhizoma Curcumae Longae extract is preferably 1:10.
Advantage of the present invention:
Prove through external mtt assay activity test, Chinese medicine composition of the present invention can significantly suppress the growth of the hepatoma cell strains such as the lung cancer cell lines such as LA795 and Hep-B3, IC
50Can reach respectively below 20 μ g/mL; Adenocarcinoma of lung and the test of hepatocarcinoma bearing mouse model are shown, tumour inhibiting rate improves mouse spleen index and liver index respectively all more than 50%, compares with independent application crude drug, the activity of the anti-hepatocarcinoma of this pharmaceutical composition and pulmonary carcinoma is higher and effect is clear and definite, and hepatocarcinoma, pulmonary carcinoma are had good inhibitory action.
The specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment.
The following examples are in order to enable those skilled in the art to understand better the present invention, but the present invention are not imposed any restrictions.
Rhizoma Paridis extract: be commercially available Rhizoma Paridis extract (specification 5:1 or 10:1 or 20:1).
The preparation of Rhizoma Curcumae Longae extract:
Rhizoma Curcumae Longae medical material or decoction pieces are ground into fritter, in proportion, the mass concentration that 1g Rhizoma Curcumae Longae fritter is soaked in 7ml is in 1.3% NaOH aqueous solution, soaked 1 hour, in 90 ℃ of reflux, extract, 1 hour, filter, and carry out again reflux, extract, 2 times by the 1st reflux, extract, operating condition, filter, merge 3 times extracting solution, filtrate is 1.0 through being evaporated to 25 ℃ of lower relative densities, be that 1.7% hydrochloric acid-alcoholic solution is regulated pH to neutral with mass concentration, add dehydrated alcohol to the concentration of dehydrated alcohol to account for 65% of cumulative volume, mix, standing 24 hours, filtration is precipitated, the precipitation lyophilization obtains Rhizoma Curcumae Longae extract.
Embodiment 1
The preparation of the Chinese medicine composition of a kind of anti-hepatocarcinoma and pulmonary carcinoma is in mass ratio with the Rhizoma Paridis extract (specification 10:1) of 1 part and the Rhizoma Curcumae Longae extract mixing of 10 parts.
Embodiment 2
The preparation of the Chinese medicine composition of a kind of anti-hepatocarcinoma and pulmonary carcinoma is in mass ratio with the Rhizoma Paridis extract (specification 20:1) of 1 part and the Rhizoma Curcumae Longae extract mixing of 5 parts.
Embodiment 3
The preparation of the Chinese medicine composition of a kind of anti-hepatocarcinoma and pulmonary carcinoma is in mass ratio with the Rhizoma Paridis extract (specification 5:1) of 1 part and the Rhizoma Curcumae Longae extract mixing of 25 parts.
The test of pesticide effectiveness of Chinese medicine composition of the present invention is as follows:
One. the anti-tumor activity test method
1. anti tumor activity in vitro test
1.1 cell strain and cultivation
The mouse pulmonary adenocarcinoma cell line LA 795 strain is incubated at and contains 10% inactivated fetal bovine serum, the 100U/mL penicillin, and in the RPMn640 culture medium of 100 μ g/mL streptomycins, in 37 ℃, 5%CO
2Cultivate under incubator and saturated humidity condition, went down to posterity once in 3-4 days.
Human lung adenocarcinoma A549 cell strain is incubated at and contains 10% inactivated fetal bovine serum, the 100U/mL penicillin, and in the DMEM culture medium of 100 μ g/mL streptomycins, in 37 ℃, 5%CO
2Cultivate under incubator and saturated humidity condition, went down to posterity once in 3～4 days.
The BEL-7402 hepatoma cell strain is incubated at and contains 10% inactivated fetal bovine serum, the 100U/mL penicillin, and in the RPMn640 culture medium of 100 μ g/mL streptomycins, in 37 ℃, 5%CO
2Cultivate under incubator and saturated humidity condition, went down to posterity once in 3-4 days.
The Hep-B3 hepatoma cell strain is incubated at and contains 10% inactivated fetal bovine serum, the 100U/mL penicillin, and in the DMEM culture medium of 100 μ g/mL streptomycins, in 37 ℃, 5%CO
2Cultivate under incubator and saturated humidity condition, went down to posterity once in 3～4 days.
1.2 Antitumor Activity of Drugs is measured
Mtt assay: it is 3 * 10 that the exponential phase cell is mixed with concentration after with trypsinization
4The cell suspension of individual/mL is inoculated in 96 hole ELISA Plate, and every hole adds 200 μ L.Change after 24h and do not contain serum free culture system liquid and make cell synchronization growth, every hole 200 μ L.Added in the hole in the 3rd day to be subjected to reagent, namely add respectively Rhizoma Paridis extract aqueous solution (being called for short the Rhizoma Paridis group), Rhizoma Curcumae Longae extract aqueous solution (being called for short the Rhizoma Curcumae Longae group) and press this drug composition aqueous solution (being called for short compositions group embodiment 1 prepares) of embodiment 1 preparation; Separately establish matched group: 0.1% dimethyl sulfoxide (DMSO), every hole add 200 μ L.Be subjected to every group of reagent to establish respectively 0.01,0.1,1,10,100,1,000 six of μ g/mL concentration, each concentration is established 8 parallel holes, and after cultivating 24h under 37 ℃, every hole adds serum-free without the freshly prepared 0.5mg/mL MTT100 of phenol red culture fluid μ L, continue to cultivate 4h, then, every hole adds 100 μ L DMSO dissolving MTT formazan granules, after microoscillator vibration mixing, measure optical density value (OD) on microplate reader, the experiment triplicate is averaged.Process tumor cell as matched group take solvent control, calculate suppression ratio by the OD value, formula is: inhibitory rate of cell growth=(matched group OD value-experimental group OD value)/matched group OD * 100%, and take drug level as abscissa, the OD value is vertical coordinate, draws cell growth curve, and tries to achieve IC
50(half suppression ratio), IC
50=inhibitory rate of cell growth is 50% drug level.The results are shown in Table 1.
Table 1 compositions Anticancer Activity in vitro result of the test
2. anti-tumor in vivo activity test
2.1HepA liver cancer model experiment
2.1.1 animal model
(female, the pharmaceutical composition that 18-20g) gives embodiment 1 preparation carries out the test of pesticide effectiveness in mice HepA liver cancer model body with 80 (be divided at random 8 groups, 10 every group, each group name claims to see 2.1.2) kunming mices.
Get the full tumor-bearing mice of ascites after inoculation HepA hepatoma carcinoma cell, the sterilization abdominal part, extracting milky ascites in superclean bench is the tumor source, the normal saline washing, ascites 4mL, the ratio of ascites and normal saline is 1:1, normal saline 4mL, be made into cell suspension, extract 0.1mL with asepsis injector, be inoculated in respectively 80 mice oxters.Whole operating under aseptic condition carried out.
2.1.2 animal grouping and administration
Inoculate and begin administration, every day 1 time next day: (1) model group gavages normal saline, every 0.2mL; (2) chemotherapy group: press the 20mg/kgbw intraperitoneal injection of cyclophosphamide; (3) Rhizoma Paridis high dose group: by 5g(crude drug amount)/kgbw gavages; (4) Rhizoma Paridis low dose group: by 2.5g(crude drug amount)/kgbw gavages; (5) Rhizoma Curcumae Longae high dose group: by 5g(crude drug amount)/kgbw gavages; (6) Rhizoma Curcumae Longae low dose group: by 2.5g(crude drug amount)/kgbw gavages; (7) pharmaceutical composition high dose group: by 5g(crude drug amount)/kgbw gavages; (8) pharmaceutical composition low dose group: by 2.5g(crude drug amount)/kgbw gavages; Two weeks, next day, mice was put to death in the cervical region dislocation after administration finishes, and dissected and got its tumor tissue, won liver, and spleen is also weighed.
Survey tumour inhibiting rate: (i) tumour inhibiting rate (%)=(the average tumor of the average tumor weight/normal saline of 1-administration group group is heavy) * 100%, win liver, spleen is also weighed.Liver refers to=(liver weight/body weight) * 1000, and spleen refers to=(spleen weight/body weight) * 1000.
The results are shown in Table 2 and table 3.
Table 2 compositions is to HepA hepatocarcinoma tumor-bearing mice tumour inhibiting rate result of the test
Table 3 compositions to HepA hepatocarcinoma tumor-bearing mice immune organ weight index relatively
Annotate: compare * P＜0.05, * * P＜0.01 with model group
2.2LA795 adenocarcinoma of lung metastasis model experiment
2.2.1 animal model
(female, the pharmaceutical composition that 18-20g) gives embodiment 1 preparation carries out the test of pesticide effectiveness in mice LA795 adenocarcinoma of lung metastasis model body with 80 (be divided at random 8 groups, 10 every group, each group name claims to see 2.2.2) T739 mices.
Get 6-8 days good T739 mice with tumor of tumor growth after inoculation LA795 lung adenocarcinoma cell, break cervical vertebra and put to death mice, peel off tumor under aseptic condition, select fresh shredding without downright bad tumor tissue to put in the glass grinding device, adding appropriate normal saline grinds gently and is prepared into tumor cell suspension, add normal saline dilution, counting after the cell sieve filters, being deployed into cell concentration is the cell suspension of 1,000 ten thousand/milliliter, and it is subcutaneous that every of 0.2mL/ is inoculated in healthy mice right fore axillary fossa.
2.2.2 animal grouping and administration
Inoculate and begin administration, every day 1 time next day: (1) model group gavages normal saline, every 0.2mL; (2) chemotherapy group: press the 20mg/kgbw intraperitoneal injection of cyclophosphamide; (3) Rhizoma Paridis high dose group: by 5g(crude drug amount)/kg gavages Rhizoma Paridis extract 0.2mL; (4) Rhizoma Paridis low dose group: by 2.5g(crude drug amount)/kg gavages Rhizoma Paridis extract 0.2mL; (5) Rhizoma Curcumae Longae high dose group: by 5g(crude drug amount)/kgbw gavages; (6) Rhizoma Curcumae Longae low dose group: by 2.5g(crude drug amount)/kgbw gavages; (7) pharmaceutical composition high dose group: by 5g(crude drug amount)/kgbw gavages; (8) pharmaceutical composition low dose group: 2.5g(crude drug amount)/kgbw gavages; Two weeks, next day, mice was put to death in the cervical region dislocation after administration finishes, and dissected and got its tumor tissue, won liver, spleen, thymus, kidney, the heart, brain, lung, stomach, ileum, colon, liver wherein, spleen is weighed respectively.The tissue that takes off is stored in 10% neutral formalin, and does paraffin section, carry out conventional H E dyeing, wherein tumor tissue is also done TUNEL dyeing, observation of cell apoptosis.
Survey tumour inhibiting rate: (i) tumour inhibiting rate (%)=(the average tumor of the average tumor weight/normal saline of 1-administration group group is heavy) * 100%, win liver, spleen is also weighed.Liver refers to=(liver weight/body weight) * 1000, and spleen refers to=(spleen weight/body weight) * 1000.The results are shown in Table 4, table 5 and table 6.
Table 4 compositions is to T739 lung cancer metastasis mice tumour inhibiting rate result of the test
Table 5 compositions to T739 lung cancer metastasis mouse immune organ weight index relatively
Annotate: compare * P＜0.05, * * P＜0.01 with model group
Table 6T739 lung cancer metastasis mice is respectively organized HE dyeing and tumor tissue TUNEL coloration result
2.3 Hepatoma Induced By Diethylnitrosamine experiment
2.3.1 animal model and administration
90 rats are divided into 9 groups at random, and except 20 of B groups, all the other respectively organize 10.The A treated animal is freely drunk water, is eaten food, and the B treated animal is mixed with 1% aqueous solution with diethylnitrosamine (DEN), every Mus gavage regularly weekly, and weekly dose press 70mg/kg and is calculated, continuous 15 weeks of gavage.C to H treated animal awards each Chinese medicine: (1) normal group (A group) simultaneously except giving DEN by above-mentioned dosage method; (2) model group (B group); (3) Rhizoma Paridis high dose group (C group): by 5g(crude drug amount)/kgbw gavages; (4) Rhizoma Paridis low dose group (D group): by 2.5g(crude drug amount)/kgbw gavages; (5) Rhizoma Curcumae Longae high dose group (E group): by 5g(crude drug amount)/kgbw gavages; (6) Rhizoma Curcumae Longae low dose group (F group): by 2.5g(crude drug amount)/kgbw gavages; (7) pharmaceutical composition high dose group (G group): by 5g(crude drug amount)/kgbw gavages; (8) pharmaceutical composition low dose group (H group): by 2.5g(crude drug amount)/kgbw gavages.
2.3.2 detection method and index
Above-mentioned each treated animal and the 15th week put to death, get blood system and detect alanine aminotransferase (ALT), aspartate amino transferase (AST), the gamma glutamyl transpeptidase (liver function index such as γ-GT), hepatic tissue fixing 24h in 10% formalin from serum and automatic biochemistry analyzer.Row dehydration after fixing, transparent, waxdip, routine paraffin wax embedding, the preparation section adopts the HE staining to observe histology's variation of liver under light microscopic.The results are shown in Table 7 and table 8.
Table 7 is respectively organized Liver Function and is detected the index result relatively
Annotate: compare * P＜0.05, * * P＜0.01 with normal group; Compare with model group,
ΔP＜0.05,
The Δ ΔP＜0.01
Table 8 is respectively organized the liver tissues of rats pathological examination relatively
With top identical method, the compositions of embodiment 2 and embodiment 3 preparations is tested, experimental result shows, the compositions of embodiment 2 and embodiment 3 also has above-mentioned similar effect.
Three, conclusion
The Chinese medicine composition preparation method of anti-hepatocarcinoma provided by the invention and pulmonary carcinoma is simple, and through external MTT evidence, this pharmaceutical composition can significantly suppress the hepatoma cell strain growths such as the lung cancer cell line such as LA795 and Hep-B3, IC
50Reach below 20 μ g/mL; Adenocarcinoma of lung and the test of hepatocarcinoma bearing mouse model show, after compatibility, tumour inhibiting rate significantly improves (more than 50%), and mouse spleen index and liver index also increase to some extent, compare with independent use, this pharmaceutical composition activity is higher and effect is clear and definite, and hepatocarcinoma, pulmonary carcinoma are had good inhibitory action.
Claims (2)
1. the Chinese medicine composition of an anti-hepatocarcinoma and pulmonary carcinoma, is characterized in that, the Rhizoma Paridis extract and the Rhizoma Curcumae Longae extract that are 1:5-25 by mass ratio form.
2. the Chinese medicine composition of a kind of anti-hepatocarcinoma according to claim 1 and pulmonary carcinoma, the mass ratio that it is characterized in that described Rhizoma Paridis extract and Rhizoma Curcumae Longae extract is 1:10.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103705697A (en) * | 2013-12-27 | 2014-04-09 | 南阳医学高等专科学校 | Pain-relieving powder for navel therapy for liver cancer |
| CN103933472A (en) * | 2014-04-04 | 2014-07-23 | 天津大学 | Traditional Chinese medicine composition for resisting liver cancer and lung cancer |
| CN104623215A (en) * | 2015-01-30 | 2015-05-20 | 天津大学 | Anti-tumor medicine composition |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102225180A (en) * | 2011-06-17 | 2011-10-26 | 天津大学 | Traditional Chinese medicine composition having anticancer effect and preparation method thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102225180A (en) * | 2011-06-17 | 2011-10-26 | 天津大学 | Traditional Chinese medicine composition having anticancer effect and preparation method thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103705697A (en) * | 2013-12-27 | 2014-04-09 | 南阳医学高等专科学校 | Pain-relieving powder for navel therapy for liver cancer |
| CN103933472A (en) * | 2014-04-04 | 2014-07-23 | 天津大学 | Traditional Chinese medicine composition for resisting liver cancer and lung cancer |
| CN103933472B (en) * | 2014-04-04 | 2016-10-05 | 天津大学 | A kind of Traditional Chinese medicinal composition for treating lung cancer and liver cancer |
| CN104623215A (en) * | 2015-01-30 | 2015-05-20 | 天津大学 | Anti-tumor medicine composition |
| CN104623215B (en) * | 2015-01-30 | 2018-05-04 | 天津大学 | A kind of antitumor medicine composition |
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Application publication date: 20130612 |