CN103623143B - A kind of Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof - Google Patents
A kind of Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof Download PDFInfo
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Abstract
The invention discloses a kind of Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof, this RP-HPLC is made up of more than 100 tastes such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Crocis.Found by pharmacological evaluation, the life span of RP-HPLC energy significant prolongation Leukemia Model mice reaches more than 3 times, makes model mice T cell subset proportions recover normal, IL-2, the levels of cytokine secretion such as IL-6, IFN-γ go up, and recover the immunosuppressant that tumor causes.RP-HPLC can also improve P
388the activity of natural killer cell (NKC), interleukin-2 (IL-2) after leukemia mouse chemotherapy, the immunosuppressant that chemotherapy is caused recovers gradually, thus to the immunosuppressant caused by leukemic treatment and immunity function restructuring, control chemotherapy of tumors.
Description
Technical field
The present invention relates to a kind of Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof, belong to field of medicaments.
Background technology
Leukemia is a class hematopoietic stem cell malignant clone disease.Clonal leukaemia breeds accumulation in a large number because of mechanism such as proliferation out of control, dysdifferentiation, apoptosis are obstructed in bone marrow and other hemopoietic tissue, and infiltrates its hetero-organization and organ, and normal hematopoiesis is suppressed simultaneously.Clinical visible anemia in various degree, hemorrhage, infectious fever and liver, spleen, lymphadenectasis and skeleton pain.It is reported, the leukemic sickness rate in China each department accounts for the 6th in various tumor.Because leukaemia comes from blood system, there is again the feature of tumor cell, so people are called leukemia traditionally.Leukemic distribution is global, accounts for about 5% of cancer total incidence, fall ill with child and youth in the majority.China leukaemic is about 3-4 people/100,000 populations, and male is more than women.In the mortality rate of each age group malignant tumor of China, leukemia accounts for the 6th (male) and the 8th (women), in the crowd of child and less than 35 years old, then account for the 1st.
Tibetan of RP-HPLC system proved recipe, Cheng Fang is in eightth century of Christian era, and be hide king, the Panchen, Dalai and high official and noble lord's medicine dedicated the earliest, go through the applicating history of more than 1,000 year, nowadays, RP-HPLC records in " Chinese Pharmacopoeia " (2010 editions).RP-HPLC is made up of more than 100 taste medicines such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Crocis, and function is heat-clearing and toxic substances removing, mediation nourishing.For " grand, red bar, Baconic " each disease, old gastroenteritis, ulcer, " wooden cloth " is sick, atrophic gastritis, various nosotoxicosis; Syphilis, leprosy, outmoded calentura, anthrax, furuncle pain, yellow fluid reducing, suppurates.Modern pharmacodynamic study result shows, this product has antiinflammatory, suppresses ulcer, promotes the effects such as ulcer healing.Acute toxicity test, long term toxicity test result shows, this product is without overt toxicity.Clinical practice result shows, this product treatment digestive system disease, as aspect display good efficacy such as various acute or chronic gastritis, chronic atrophic gastritis, peptic ulcer, liver cirrhosis, the untoward reaction of chemotherapy metenteron, and having good health-care effect, is one of Tibetan medicine's health care medicine.
The aspects such as the function of RP-HPLC cures mainly, document, patent all do not find that it can be used for treating leukemic research.
Summary of the invention
The object of this invention is to provide a kind of Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof.
The present invention is achieved through the following technical solutions:
Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof, described RP-HPLC records kind RP-HPLC for " Chinese name republic pharmacopeia " version first 573 pages in 2010.
RP-HPLC preparation of the present invention is by RP-HPLC crude drug formula, the preparation that the extract adopting 30-80% concentration ethanol to extract acquisition is made.
RP-HPLC preparation of the present invention is by RP-HPLC crude drug formula, adopts the preparation that water extraction acquisition extract is made.
The application by improving in NKC, IL-2, IL-6, IFN-gamma activity treatment leukemia medicament prepared by RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof, especially RP-HPLC and preparation thereof.
Described RP-HPLC and preparation thereof and the chemical drugs or Chinese medicine or natural drug of being applied as is preparing the application for the treatment of in leukemia medicament.
Described be applied as medicine that RP-HPLC and preparation thereof and chemical drugs or Chinese medicine or natural drug form preparing and treat application in leukemia medicament by improving NKC, IL-2, IL-6, IFN-gamma activity.
RP-HPLC preparation of the present invention, is characterized in that, by RP-HPLC crude drug formula, is prepared into capsule, tablet, granule, powder or oral liquid.
For enabling above-mentioned dosage form realize, need be add pharmaceutically acceptable adjuvant in these dosage forms of preparation, such as: filler, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, antiseptic etc., filler comprises: starch, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc., disintegrating agent comprises: starch, pregelatinized Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose etc., lubricant comprises: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc., binding agent comprises starch slurry, polyvinylpyrrolidone, hydroxypropyl emthylcellulose etc., sweeting agent comprises: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc., correctives comprises: sweeting agent and various essence, and antiseptic comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and salt thereof, benzalkonium bromide, acetic acid chloroethene is fixed, Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods.
Beneficial effect: in the application for the treatment of field of RP-HPLC, people know the effect of its heat-clearing and toxic substances removing, mediation nourishing, and know it for " grand, red bar, Baconic " each disease, old gastroenteritis, ulcer, " wooden cloth " is sick, atrophic gastritis, various nosotoxicosis; Syphilis, leprosy, outmoded calentura, anthrax, furuncle pain, yellow fluid reducing, the treatment of suppuration etc.
Instant invention overcomes the technology prejudice of above-mentioned application aspect, found by pharmacological evaluation, the life span of RP-HPLC energy significant prolongation Leukemia Model mice reaches more than 3 times, model mice T cell subset proportions is made to recover normal, IL-2, the levels of cytokine secretion such as IL-6, IFN-γ go up, and recover the immunosuppressant that tumor causes.RP-HPLC can also improve the activity of natural killer cell (NKC) after P388 leukemia mouse chemotherapy, interleukin-2 (IL-2), the immunosuppressant that chemotherapy is caused recovers gradually, thus to leukemic treatment and immunity function restructuring, there is positive meaning for the immunosuppressant caused by control chemotherapy of tumors.
Detailed description of the invention
Following embodiment is for illustrating the preparation of RP-HPLC of the present invention and preparation thereof, but it can not form any restriction to scope of the present invention.
The preparation of embodiment 1 RP-HPLC
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded:
Get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci and be processed into pill, the heavy 1g of every ball.
RP-HPLC prepared by said method is commercially available RP-HPLC (Jin He Tibetan medicine limited company produces, the accurate word Z63020078 of traditional Chinese medicines).
The preparation of embodiment 2 RP-HPLC sheet
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded:
Get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci and be processed into tablet.
The preparation of embodiment 3 RP-HPLC capsule
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded:
Get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci and be processed into capsule.
The preparation of embodiment 4 RP-HPLC granule
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded:
Get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci and be processed into granule.
Embodiment 5 RP-HPLC tablet
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded; get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci; add 30% concentration ethanol to extract; reclaim ethanol; concentrate drying; extract is ground into fine powder, and conveniently formulation method makes RP-HPLC tablet.
Embodiment 6 RP-HPLC granule
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded; get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci; add 80% concentration ethanol to extract; reclaim ethanol; concentrate drying; extract is ground into fine powder, and conveniently formulation method obtains RP-HPLC granule, subpackage and get final product.
Embodiment 7 RP-HPLC capsule
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded; get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci; extracting in water; filter; concentrate drying; extract is ground into fine powder, and conveniently formulation method makes RP-HPLC capsule.
Embodiment 8 RP-HPLC oral liquid
By " Chinese Pharmacopoeia " (2010 editions) RP-HPLC formula recorded; get the flavour of a drug such as Margarita, Cinnabaris, Lignum Santali Albi, Lignum Dalbergiae Odoriferae, Lignum Aquilariae Resinatum, Fructus Chebulae, Calculus Bovis, artificial Moschus, Stigma Croci, extracting in water, filter; concentrated, conveniently formulation method makes RP-HPLC oral liquid.
Embodiment 9 RP-HPLC tablet
(add 30% concentration ethanol and extract, reclaim ethanol, concentrate drying, extract is ground into fine powder, and conveniently formulation method makes RP-HPLC tablet for the accurate word Z63020078 of traditional Chinese medicines, product batch number: 20120922) to get RP-HPLC.
Embodiment 10 RP-HPLC granule
(add 80% concentration ethanol and extract, reclaim ethanol, concentrate drying, extract is ground into fine powder, and conveniently formulation method obtains RP-HPLC granule, subpackage and get final product for the accurate word Z63020078 of traditional Chinese medicines, product batch number: 20120922) to get RP-HPLC.
Embodiment 11 RP-HPLC capsule
(extracting in water, filter, concentrate drying, extract is ground into fine powder, and conveniently formulation method makes RP-HPLC capsule for the accurate word Z63020078 of traditional Chinese medicines, product batch number: 20120922) to get RP-HPLC.
Embodiment 12 RP-HPLC oral liquid
(extracting in water, filters, and concentrated, conveniently formulation method makes oral liquid for the accurate word Z63020078 of traditional Chinese medicines, product batch number: 20120922) to get RP-HPLC.
Below by experimental example, RP-HPLC and the drug effect of preparation in treatment leukemia thereof are described.
Experimental example 1: RP-HPLC is to the test of pesticide effectiveness of L615 leukemia mouse
1, modeling method: aseptically get L
615leukemia mouse spleen makes 25% normal saline cell suspension, gives the L that grows up
615mouse hypodermic inoculation, all there is leukemia in result, transplants continuously, set up stable L with morbidity mice spleen oncocyte at Syngenic mice
615leukemia Model.
L
615the foundation of leukemia cell line: remove L
615the spleen of leukemia mouse, shreds spleen in Hanks liquid, makes leukaemia free out, by collected by centrifugation free cell, then uses RPMI1640 culture fluid (containing 10% calf serum) to be diluted to 8*10
^6the Cell sap of/ml, cultivates at 37 DEG C.Cultivate 20d and find there is adherent fusiform cells in culture, around it, have the round cell island of propagation.When separately being cultivated by cells different for two kinds of forms, spindle cell can normal adherent growth, and suspension cell growth is slow; Again by two kinds of cells Mixed culture again, two kinds of Growth of Cells are vigorous, after 45d, by the round cell of suspension growth single culture again, pass to for 12 generations always, and Growth of Cells is good, thus sets up this cell strain.
2, the RP-HPLC treatment leukemia test of pesticide effectiveness.
Material: Kunming mouse (Shandong Province's Experimental Animal Center).RP-HPLC (Jin He Tibetan medicine limited company, lot number: 20110809), RP-HPLC-1, RP-HPLC-2, RP-HPLC-3(is respectively by the embodiment of the present invention 9, embodiment 10, the RP-HPLC sheet that embodiment 11 method prepares, RP-HPLC granule, RP-HPLC capsule, lot number is respectively: 20120920, 20120921, 20120922), cyclophosphamide (Hengrui Medicine Co., Ltd., Jiangsu Prov.), standard RhIL-2 (Beijing Shuanglu Pharmaceutical Co., Ltd.), school of life and health sciences cellular resources center, Yackl and IL-2 dependent cells strain CTLL(Chinese Academy of Sciences Shanghai), RPMII640 cell culture fluid and hyclone (Huamei Bio-Engrg Co.), tetramethyl azo azoles salt (MTT), dodecyl sodium sulfate and canavaline, dimethyl formamide is (Sigma Products), sample injector, CO
2incubator, ELX-800 microplate reader (Bio-Tek company of the U.S.), the 96 flat Tissue Culture Plates in hole, Epics flow cytometer (Beckman company of the U.S.).
Method: modeling as stated above.Experiment grouping first group is Normal group, 20 mices, after conventional raising 1w, and intraperitoneal injection of saline 0.2ml/d, normal saline gavage 0.3ml, each 1 time of upper and lower noon.2nd group is model group, after conventional raising 1w, and inoculation L
615leukaemia, the same Normal group of inoculation post processing.3rd group is Western medicine group, and 20 conventional raisings, after one week, inoculate L
615leukaemia, after inoculation the 3rd day starts ip cyclophosphamide, 200mg/kg, 1 time/d.4th group of-7 groups is respectively RP-HPLC group, RP-HPLC-1 group, RP-HPLC-2 group, RP-HPLC-3 group, get respectively 20 conventional raise 1d after, inoculation L
615leukaemia, gives RP-HPLC, RP-HPLC-1, RP-HPLC-2, RP-HPLC-3 gavage on the 3rd day after inoculation respectively, and each 0.3ml(content of dispersion conversion is 300mg to medical material), each 1 time of upper and lower noon.Above each treated animal often group get 10 in the 5th day put to death, get spleen and carry out related immune Indexs measure, remaining each animal carries out observation life cycle.
Observation index: 1. observe life cycle.2. T cell Subsets.3. IL-2, IL-6, IFN-gamma activity measures.
Statistical method: data with
represent, adopt MicrosoftExcel to carry out t inspection.
Result:
Life cycle: RP-HPLC respectively organize life cycle comparatively model group and Western medicine group obviously extend, statistics there were significant differences P<0.01, in table 1.
Table 1 each experimental group compares life cycle
| Group | Number of elements | Life span (d) |
| Model group | 10 | 6.50±0.57 |
| Western medicine group | 10 | 17.43±1.34 * |
| RP-HPLC group | 10 | 28.00±3.14 *△ |
| RP-HPLC-1 group | 10 | 25.23±3.62 *△ |
| RP-HPLC-2 group | 10 | 27.45±3.28 *△ |
| RP-HPLC-3 group | 10 | 26.48±3.78 *△ |
Note: * is P<0.01 compared with model group, △ is P<0.01 compared with Western medicine group.
T cell subgroup: RP-HPLC respectively organizes CD4
+cell comparatively model group and Western medicine group high.CD8
+leukopenia, CD4/CD8 ratio recovers normal.In table 2.
Table 2 respectively group T cell subgroup change
| Group | Number of elements | CD3 + | CD4 + | CD8 + | CD4/CD8 |
| Normal group | 10 | 67.56±2.18 | 35.28±2.13 | 24.01±1.26 | 1.47±0.15 |
| Model group | 10 | 52.75±2.34 * | 22.33±2.18 * | 29.56±1.32* | 0.76±0.08 * |
| Western medicine group | 10 | 42.45±2.02 △ | 16.32±1.98 △ | 33.78±0.63 △ | 0.48±0.07 △ |
| RP-HPLC group | 10 | 64.37±2.10 ▲△ | 33.21±1.90 ▲△ | 25.42±1.43 ▲△ | 1.31±0.07 ▲△ |
| RP-HPLC-1 group | 10 | 60.63±2.12 ▲△ | 30.03±1.87 ▲△ | 26.04±1.53 ▲△ | 1.15±0.06 ▲△ |
| RP-HPLC-2 group | 10 | 61.45±2.34 ▲△ | 31.20±1.88 ▲△ | 24.35±1.62 ▲△ | 1.28±0.06 ▲△ |
| RP-HPLC-3 group | 10 | 62.60±2.32 ▲△ | 32.36±1.78 ▲△ | 24.76±1.48 ▲△ | 1.31±0.05 ▲△ |
Note: * is P<0.01 compared with Normal group, △ is P<0.01 compared with model group, ▲ compared with Western medicine group P<0.01.
IL-2, IL-6, IFN-gamma activity: after Western medicine group leukemia mouse chemotherapy, it secretes IL-2, IL-6, the level of IFN-γ significantly declines, and RP-HPLC respectively organizes IL-2, the secretion level of IL-6, IFN-γ obviously increases, both have significant difference (P<0.01), in table 3 at contrast.
Table 3 respectively organizes IL-2, and IL-6, IFN-gamma activity changes
| Group | Number of elements | IL-2 | IL-6 | IFN-γ |
| Normal group | 10 | 22.38±2.14 | 45.53±5.55 | 10.68±1.55 |
| Model group | 10 | 8.88±2.23 * | 15.48±4.13 * | 5.87±2.11 * |
| Western medicine group | 10 | 6.08±1.76 △ | 10.36±3.86 △ | 4.02±1.24 △ |
| RP-HPLC group | 10 | 14.26±2.11 ▲ | 28.44±1.90 ▲△ | 8.02±1.19 ▲△ |
| RP-HPLC-1 group | 10 | 15.42±2.09 ▲ | 25.67±2.06 ▲△ | 8.11±1.16 ▲△ |
| RP-HPLC-2 group | 10 | 12.12±1.98 ▲ | 23.14±1.97 ▲△ | 8.23±1.14 ▲△ |
| RP-HPLC-3 group | 10 | 13.44±1.88 ▲ | 26.55±1.99 ▲△ | 7.98±1.21 ▲△ |
Note: * is P<0.01 compared with Normal group, △ is P<0.01 compared with model group, ▲ compared with Western medicine group P<0.01.
The whether normal of immunologic function be one of key factor of tumor development, and immunologic function is that the ratio of T cell subgroup also plays an important role in stable regulation immunologic function by performed by the cytokine secreted by immunocyte.T cell subset proportions imbalance after leukemia mouse chemotherapy, IL-2, the levels of cytokine secretion such as IL-6, IFN-γ decline, and by after the treatment of use RP-HPLC, T cell subset proportions recovers normal, the levels of cytokine secretion such as IL-2, IL-6, IFN-γ go up, illustrate that RP-HPLC is by regulatory T-cell subset proportions and promote that the secretion of these cytokines plays its immunoregulation effect, has certain effect to leukemic therapy apparatus immunity function restructuring.
Experimental example 2: RP-HPLC is to the pharmacodynamics test of P388 leukemia mouse
1. modeling method: get ascitic type P
388leukemia mouse, de-cervical vertebra is put to death, and abdominal part is sterilized, suction ascites, and add normal saline mixing, it is (1-1.5) * 10 that microscope lowers intact cells concentration
^7individual/ml, the inoculum density of every Mus 0.2ml(Kunming mouse and dosage) carry out lumbar injection.
2. RP-HPLC is to P
388the impact of natural killer cell (NKC), interleukin-2 (IL-2) activity after leukemia mouse chemotherapy.
Material: Kunming kind one-level mice (Shandong Province's Experimental Animal Center), 50, male and female half and half, body weight 18-22g.RP-HPLC (Jin He Tibetan medicine limited company, lot number: 20110809), the RP-HPLC sheet that RP-HPLC-1(is prepared by the embodiment of the present invention 9 method, lot number is: 20120920), cyclophosphamide (Hengrui Medicine Co., Ltd., Jiangsu Prov.), standard RhIL-2 (Beijing Shuanglu Pharmaceutical Co., Ltd.), school of life and health sciences cellular resources center, Yackl and IL-2 dependent cells strain CTLL(Chinese Academy of Sciences Shanghai), RPMII640 cell culture fluid and hyclone (Huamei Bio-Engrg Co.); Tetramethyl azo azoles salt (MTT), dodecyl sodium sulfate and canavaline, dimethyl formamide is (Sigma Products); Sample injector, CO2 incubator, ELX-800 microplate reader (Bio-Tek company of the U.S.); The 96 flat Tissue Culture Plates in hole, Epics flow cytometer (Beckman company of the U.S.).
Modeling and group technology: get 40 mices, inoculation P
388the strain of Lymphocytic leukemia tumor (inoculation method: get ascitic type P388 leukemia mouse, de-cervical vertebra is put to death, and abdominal part is sterilized, suction ascites, adds normal saline mixing, and it is 1.5*107/ml that microscope lowers intact cells concentration, every mice right oxter injection 0.2ml), be divided into 4 groups at random.Another 10 is normal blank group.
Processing method: 1. normal blank group: with 0.4ml/20g normal saline ig; 2. P
388leukemia Model group: with 0.4ml/20g normal saline ig; 3. chemotherapy group: with 0.4ml/20g normal saline ig; 4. chemotherapy adds RP-HPLC group: respectively with 50mg extract in 0.4ml normal saline ig; 5. chemotherapy adds RP-HPLC-1 group: be dissolved in 0.4ml normal saline ig with 100mg extract.3. the continuous ig14d of each group above, give simultaneously, 4., 5. organize ip cyclophosphamide 35mg/kg/d, be used in conjunction 6d.In the 15th day, de-cervical vertebra was put to death, and aseptically got spleen NKC, IL-2 active.
Detection method: NKC Activity determination is with reference to Jiang Zhonghua method: the external test [J] of the NK cell words property of lepromatous leprosy animal model. the assorted poison of China Immunology, 1985,4 (1): 238; IL-2 activity test method is with reference to Qian Yukun method: practical immunology new technique [M]. Beijing: Beijing combined publication society of China Concord Medical Science University of Ya Ke university .1994.49.
Statistical method: adopt many group means to compare, namely first do variance analysis, remake T inspection.
Result shows, it is active that RP-HPLC can improve NKC and IL-2.In table 4
Table 4 is group NKC and IL-2 expression activitiy respectively
| Group | n | NKC activity (%) | IL-2 activity (Iu/ml) |
| Normal group | 8 | 44.38±3.17 | 17.66±2.98 |
| Model control group | 8 | 35.77±3.47 * | 13.56±2.11 * |
| Chemotherapy group | 8 | 27.76±3.68 **△△ | 10.98±2.22 **△△ |
| Chemotherapy adds RP-HPLC group | 8 | 34.68±3.45 ** | 13.25±2.56 ** |
| Chemotherapy adds RP-HPLC-1 group | 8 | 39.45±3.57 ** | 15.86±2.98 ## |
Note: compare with normal group, * P<0.05, * * P<0.01; Compare with model group, △ P<0.05, △ △ P<0.01; Compare with chemotherapy group, #P<0.05, ##P<0.01.
NKC is one of three major types lymphocyte, it is a group broad-spectrum anti-tumor cell, a large amount of cytokine can be secreted rapidly as interferon-γ (IFN-γ), tumor necrosis factor-alpha (TNF-α), GM-CSF (GMCSF), IL-2 etc., regulate phagocyte and T, B cell function.IL-2 is main, the most strong SCIF in body, can induce or promote the activity of various kinds of cell toxic cell, as NKC, cytotoxic T lymphocyte (CTL), Lymphatic circulation (LAK) and tumor infiltrating lymphocyte (TIL) etc., these cells are most important in the immune surveillance and antineoplastic immune of body.
Above-mentioned experimental result shows, P388 leukemia mouse self and immunity function after chemotherapy are suppressed, NKC and IL-2 activity all has reduction in various degree, after Treated with Chemotherapy with Cyclophosphamide, NKC and the IL-2 activity of P388 leukemia mouse is starkly lower than normal group and model group, after giving RP-HPLC treatment, NKC and IL-2 all has rebound significantly.Experimental result shows, RP-HPLC has enhancing IL-2 and NKC active function, has certain protective effect to the immunosuppressant of P388 leukemia mouse after chemotherapy.Infer its mechanism of action may with enhancing Exploration of Red Cell Immuno-adhesive Function, impel erythrocyte to discharge NKC activity factor, regulate NKC-IFN-IL-2 immunological network relevant.The above-mentioned effect of RP-HPLC, for the immunosuppressant caused by control chemotherapy of tumors, effectively ensures completing of chemotherapy, has positive meaning.
Claims (4)
1. a Tibetan medicine RP-HPLC and the application of preparation in preparation treatment leukemia medicament thereof, it is characterized in that, described Tibetan medicine RP-HPLC preparation is by RP-HPLC crude drug formula, the preparation adopting 30-80% concentration ethanol to extract or adopt the extract of water extraction acquisition to make.
2. Tibetan medicine RP-HPLC according to claim 1 and the application of preparation in preparation treatment leukemia medicament thereof, it is characterized in that, described Tibetan medicine RP-HPLC and the preparation thereof of being applied as improves NKC, IL-2, IL-6, IFN-gamma activity in preparation and treats application in leukemic medicine.
3. Tibetan medicine RP-HPLC according to claim 1 and the application of preparation in preparation treatment leukemia medicament thereof, it is characterized in that, described RP-HPLC and preparation thereof and the chemical drugs or Chinese medicine or natural drug of being applied as is preparing the application for the treatment of in leukemia medicament.
4. the Tibetan medicine RP-HPLC according to any one of claim 1 ~ 3 and the application of preparation in preparation treatment leukemia medicament thereof, it is characterized in that, described Tibetan medicine RP-HPLC preparation is by RP-HPLC crude drug formula, add pharmaceutically acceptable adjuvant, the capsule be prepared into according to a conventional method, tablet, granule, powder or oral liquid.
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