CN103877435B - The application in immunity function after chemotherapy of the positive eliminating mass side is supported in spleen invigorating - Google Patents
The application in immunity function after chemotherapy of the positive eliminating mass side is supported in spleen invigorating Download PDFInfo
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- CN103877435B CN103877435B CN201410055831.9A CN201410055831A CN103877435B CN 103877435 B CN103877435 B CN 103877435B CN 201410055831 A CN201410055831 A CN 201410055831A CN 103877435 B CN103877435 B CN 103877435B
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Abstract
The invention discloses a kind of spleen invigorating and support the application in preparing the medicine for reducing damnification of immunity function after chemotherapy of the positive eliminating mass side.This application is a kind of new application that positive eliminating mass side is supported in spleen invigorating, positive eliminating mass is supported in spleen invigorating can improve index and spleen index, thymus index, Peritoneal Macrophage Phagocytosis, NK cytoactive and T cell multiplication capacity, reducing T-reg cell proportion, therefore positive eliminating mass side is supported in spleen invigorating the good effect alleviating the immunologic injury that chemotherapy causes.Reduction clearly is had particularly in Treated with Chemotherapy with Cyclophosphamide damnification of immunity function after aspect.
Description
Technical field
The present invention relates to the new opplication of a kind of compound composite medicament, be specifically related to a kind of spleen invigorating and support the application in damnification of immunity function after chemotherapy of the positive eliminating mass side.
Background technology
Gastric cancer is one of malignant tumor common in world wide, is the 4th malignant tumor occurred frequently, and mortality rate is then in second, and the life and health of the mankind is caused great threat.2002, whole world incidence gastric cancer number was 930,000, and death is then more than 700,000.China is the district occurred frequently of gastric cancer, and morbidity accounts for the 2/5 of the whole world.Annual morbidity about 400,000, death about 300,000, number of the infected accounts for the 41% of whole world incidence gastric cancer number, and its prevention and control situation is in severe state.In China, the patients with gastric cancer of about 40% is late period when medical, and relapse and metastasis will occur after accepting radical operation in the gastric cancer of other about 60%.Data show according to the study: the annual rate of depositing of late gastric cancer is 8%, and median survival interval is 7～9 months.For this some patients, how to extend its life cycle by reasonably treatment, improve life quality, be emphasis and difficult point, the major fields that great diseases prevention and treatment are worked by Ye Shi China of current cancer research.
Positive eliminating mass side is supported in current spleen invigorating has good therapeutical effect to the patient of vital QI being weakened and pathogen being violent after chemotherapy.We, to prove that the party has the multiple digestive tract cell proliferation of suppression by experiment in vivo and vitro, induce stomach cancer cell early apoptosis, regulate the effect of gastric cancer cell cycle.But spleen invigorating is supported the impact on immunity function after chemotherapy of the positive eliminating mass side and was not studied.
Summary of the invention
The technical problem to be solved is in that to provide a kind of spleen invigorating to support the application in preparing the medicine for reducing damnification of immunity function after chemotherapy of the positive eliminating mass side.
In order to solve above-mentioned technical problem, the technical solution adopted in the present invention is as follows: the application in preparing the medicine for reducing damnification of immunity function after chemotherapy of the positive eliminating mass side is supported in a kind of spleen invigorating.
The present invention improves further, and the application in preparing the medicine for reducing the damnification of immunity function after chemotherapy that cyclophosphamide (CTX) is induced of the positive eliminating mass side is supported in described spleen invigorating.
In the present invention, described spleen invigorating is supported positive eliminating mass side and is included following crude drug: Radix Codonopsis 15g, Rhizoma Atractylodis Macrocephalae (parched) 10g, Poria 10g, Semen Coicis 20g, Radix Angelicae Sinensis 10g, Rhizoma dioscoreae 15g, Radix Aucklandiae 10g, Radix Paeoniae Alba 10g, Pericarpium Citri Reticulatae 6g, Rhizoma Smilacis Chinensis 30g, Herba Salviae Chinensis 30g, Radix Glycyrrhizae Preparata 3g.
Beneficial effect: the new opplication that the application in preparing the medicine for reducing damnification of immunity function after chemotherapy of the positive eliminating mass side is this prescription is supported in spleen invigorating of the present invention, positive eliminating mass is supported in spleen invigorating can improve index and spleen index, thymus index, Peritoneal Macrophage Phagocytosis, NK cytoactive and T cell multiplication capacity, reducing T-reg cell proportion, therefore positive eliminating mass side is supported in spleen invigorating the good effect alleviating the immunologic injury that chemotherapy causes.Reduction clearly is had particularly in Treated with Chemotherapy with Cyclophosphamide damnification of immunity function after aspect.
Accompanying drawing explanation
Fig. 1 is that spleen invigorating of the present invention supports positive eliminating mass side to the impact of Mouse Weight after CTX chemotherapy.
Detailed description of the invention
The application in preparing the medicine for reducing damnification of immunity function after chemotherapy of the positive eliminating mass side is supported in spleen invigorating of the present invention.Wherein, spleen invigorating is supported positive eliminating mass side and is included following crude drug: Radix Codonopsis 15g, Rhizoma Atractylodis Macrocephalae (parched) 10g, Poria 10g, Semen Coicis 20g, Radix Angelicae Sinensis 10g, Rhizoma dioscoreae 15g, Radix Aucklandiae 10g, Radix Paeoniae Alba 10g, Pericarpium Citri Reticulatae 6g, Rhizoma Smilacis Chinensis 30g, Herba Salviae Chinensis 30g, Radix Glycyrrhizae Preparata 3g.
Due to the chemotherapeutics that CTX is that bone marrow injury is the most serious.And immunocyte is many differentiates from bone marrow, so alternatively CTX is the chemotherapeutic that damnification of immunity function is the most serious.
Below in conjunction with experimental example, beneficial effects of the present invention is described in further detail.
Experimental example:
1 material and instrument
1.1 animals and feeding environment:
BALB/c mouse (Shanghai Si Laike animal center, the quality certification number: SCXK (Shanghai) 2007-0005).21, male and female half and half, 10-12 week old, body weight 18-20g, often group 7；SPF environment is raised, indoor regularly ultra-vioket radiation, the equal strict sterilization of mouse cage, bedding and padding, drinking-water.
1.2 medicines are bought and preparation:
Positive eliminating mass side is supported in spleen invigorating: the party includes following crude drug: Radix Codonopsis 15g, Rhizoma Atractylodis Macrocephalae (parched) 10g, Poria 10g, Semen Coicis 20g, Radix Angelicae Sinensis 10g, Rhizoma dioscoreae 15g, Radix Aucklandiae 10g, Radix Paeoniae Alba 10g, Pericarpium Citri Reticulatae 6g, Rhizoma Smilacis Chinensis 30g, Herba Salviae Chinensis 30g, Radix Glycyrrhizae Preparata 3g.
First adding crude drug gross weight 10 times amount distilled water toward crude drug and soak 30min, after boiling, little fire boils 30min again, after pouring out liquid, then adds the distilled water of crude drug gross weight 5 times, then boils 30min, is merged by the liquid of twice, and water-bath is concentrated into 2.5g mL-1, 4 DEG C of Refrigerator stores.
Cyclophosphamide (CTX): Hengrui Medicine Co., Ltd., Jiangsu Prov., lot number: 12042425.
1.3 reagent and instrument:
RPMI-1640 complete medium (Kai Ji company), enzyme micro-plate reader (Beijing Pu Lang Technew SA product), CO2Incubator (Shanghai Fuma Experiment Equipment Co., Ltd.), superclean bench (SuZhou Antai Air Tech Co., Ltd.), inverted microscope (south of the River, Nanjing Shui Xin Optical Co., Ltd).
2 experimental techniques
2.1 packets and administration
Be randomly divided into blank group, group (positive eliminating mass side+CTX group is supported in spleen invigorating) combined by CTX group, Chinese medicine.Testing 1d to rise, blank group and CTX group gavage NS, Chinese medicine is combined and is organized gavage spleen invigorating and support positive eliminating mass side, 50g kg-1, 1/ day, continuous 10 days.Group combined by gavage 3d, CTX group and Chinese medicine, disposable celiac injection CTX200mg kg-1, the NS of blank group lumbar injection equivalent.11st day, eye socket took blood, put to death mice.
2.2 spleens, thymus index
After sacrifice, aseptic take off spleen, thymus, weigh and calculate organ index.
Organ index=organ weights (g)/the weight of animals (kg)
The mensuration of 2.3 Peritoneal Macrophage Phagocytosis
Testing the 8th day, mouse web portion of sterilizing, lumbar injection mass concentration is 4.5% soluble starch (starch configures with normal saline, autoclaving), 2mL/ mice.11st day, after putting to death mice, cut off mouse skin, expose peritoneum.Inject cold PBS liquid (phosphate buffer) 10mL of 4 degrees Celsius with syringe along ventrimeson, gently rub abdominal part 5min, pumpback abdominal cavity liquid.So repeat to rinse abdominal cavity 2 times.Merging the peritoneal lavage fluid collected, 4 DEG C, 250g, 10min is centrifuged, and abandons supernatant.Wash twice with cold RPMI-1640 culture fluid, regulate cell concentration to 2 × 106/ mL, 100 μ l/ holes add in 96 well culture plates, and each sample sets 3 multiple holes.37 DEG C, 5%CO2Cultivating 2h in incubator, the warm PBS liquid of 37 degrees Celsius is washed and the non-attached cell of sucking-off, obtains macrophage monolayer.It is 0.03% neutral red solution 100 μ l that every hole adds mass concentration, puts CO2Incubator cultivates 1h, the dimethyl diaminophenazine chloride that the warm PBS liquid flushing of 37 degrees Celsius is not swallowed 3 times.Every hole adds cell pyrolysis liquid (Kai Ji company, article No.: KGP601) 100 μ l, 4 DEG C of refrigerator overnight, and microplate reader measures 570nm place OD value.
2.4NK cytoactive detection
2.4.1 prepared by spleen bar cell suspension: after sacrifice, aseptic spleen of winning, and rejects fatty tissue.Under cold PBS liquid 4 degrees Celsius instils, 200 order steel meshes grind and filter, and 1500rmp is centrifuged, 5 minutes.Add erythrocyte cracked liquid (Kai Ji company, article No.: KGP11100).
Resuspended 3 minutes, lysed erythrocyte, PBS stopped, and washs 2 times, obtains spleen bar cell suspension.
2.4.2 with the RPMI-1640 culture fluid containing mass concentration 10% calf serum, spleen bar cell suspension is adjusted to 1 × 107/ mL action effect cell.Taking L929 l cell (Kai Ji company, article No.: the KG087) mass concentration of Secondary Culture is 0.25% pancreatin to adjust L929 cell concentration be 2 × 105/ mL is as target cell, and on 96 orifice plates, every hole adds target cell suspension 100 μ l, 37 DEG C, 5%CO2Cultivating 90min so that it is adherent one-tenth cell monolayer, then the every hole of experimental port adds 100 μ l effector lymphocyte's suspensions, often the multiple hole of group 4.Target cell control wells adds the RPMI-1640 culture fluid 100 μ l containing mass concentration 10% calf serum.96 orifice plates are placed on 37 DEG C, 5%CO2Hatch 20h, remove supernatant, fill each hole with normal saline gently, 3 times repeatedly, to remove effector lymphocyte and the target cell come off by killing, filter paper blots moisture in hole, every hole adds the dimethyl diaminophenazine chloride dye liquor 100 μ l that mass concentration is 0.1%, hatches 30min, discards dye liquor, normal saline washes 3 times, every hole adds 100 μ l cell pyrolysis liquids, makes the target cell lysis containing dimethyl diaminophenazine chloride, shakes up, microplate reader 450nm surveys OD value, calculates killing rate by below equation:
Killing rate (%)=(target cell OD value-experimental group OD value)/target cell OD value × 100%
The mensuration of 2.5T lymphopoiesis ability
The conventional splenocyte suspension that makes, tune splenocyte suspension concentration is to 5 × 106/ L, 100 μ l/ holes add 96 well culture plates, then are separately added into ConA (con A) 100 μ l/ hole, make the final concentration of 5 μ g mL of ConA-1.37 DEG C, volumetric concentration be 5%CO2Incubator takes out after placing 56h, and every hole sucking-off 10 μ l adds 0.5g L-1MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide bromine salt) 10 μ l, put 4h in incubator.Taking out culture plate, abandon supernatant, every hole adds DMSO (dimethyl sulfoxide) 100 μ l, shakes 30 seconds, stands 20min, and microplate reader surveys absorbance (OD) value of 570nm.T lymphopoiesis ability is represented with sample OD value.
3. statistical procedures
Adopt SPSS13.0 statistical software, all data row variance analyses, compare employing ANOVA inspection between group, with P < 0.05 for there being significant difference.
4. experimental result
The observation of 4.1 ordinary circumstances
Within after cyclophosphamide group mouse chemotherapy the 2nd day, occurring losing weight, feeding activities declines, hence it is evident that diarrhoea.And positive eliminating mass side (group combined by Chinese medicine) is supported in coupling spleen invigorating, losing weight less, feeding activities has no significant effect, without substantially diarrhoea.In Table 1, Fig. 1.
Table 1 spleen invigorating supports positive eliminating mass side to the impact of Mouse Weight after CTX chemotherapy
(*: compared with cyclophosphamide group, P < 0.05；△: compared with blank group, P < 0.05)
The impact on chemotherapy mouse thymus index and spleen index of the positive eliminating mass side is supported in 4.2 spleen invigorating
Cyclophosphamide group mouse spleen index, thymus index are all decreased obviously, significant difference (P < 0.05) compared with blank group, and associating spleen invigorating support positive eliminating mass side treatment after, both of which significantly rises, significant difference (P < 0.05) compared with cyclophosphamide group.Result is in Table 2.
Table 2 spleen invigorating supports positive eliminating mass side to the impact of mouse thymus index, spleen index after CTX chemotherapy
(*: compared with cyclophosphamide group, P < 0.05；△: compared with blank group, P < 0.05)
The impact on chemotherapy Phagocytosis By The Peritoneal Macrophages In Mice of the positive eliminating mass side is supported in 4.3 spleen invigorating
As shown in Table 3, cyclophosphamide group Phagocytosis By The Peritoneal Macrophages In Mice is decreased obviously, and has significant difference (P < 0.05) with blank group.After positive eliminating mass side is supported in coupling spleen invigorating, phagocytic function rises, and blank group zero difference (P>0.05) has notable difference (P<0.05) with cyclophosphamide group.
The impact on chemotherapy Phagocytosis By The Peritoneal Macrophages In Mice of the positive eliminating mass side is supported in table 3 spleen invigorating
(*: compared with cyclophosphamide group, P < 0.05；△: compared with blank group, P < 0.05)
Positive eliminating mass side is supported in 4.4 spleen invigorating to be affected chemotherapy NK cells in mice killing rate
Compared with alone chemotherapy, after positive eliminating mass side is supported in coupling spleen invigorating, NK cell killing rate substantially increases, and difference statistically significant (P < 0.05), in Table 3.
The impact on chemotherapy mouse T lymphocyte multiplication capacity of the positive eliminating mass side is supported in 4.5 spleen invigorating
Mice is after CTX chemotherapy, and the lymphocytic breeder reaction of T reduces, significant difference notable (P < 0.05).After positive eliminating mass side is supported in associating coupling spleen invigorating, promoting that the lymphocytic multiplication capacity of T substantially rises, difference statistically significant (P < 0.05), in Table 3.
Claims (2)
1. the application in preparing the medicine for reducing damnification of immunity function after chemotherapy of the positive eliminating mass side is supported in a spleen invigorating；
Wherein, described spleen invigorating is supported positive eliminating mass side and is prepared from by following crude drug: Radix Codonopsis 15g, Rhizoma Atractylodis Macrocephalae (parched) 10g, Poria 10g, Semen Coicis 20g, Radix Angelicae Sinensis 10g, Rhizoma dioscoreae 15g, Radix Aucklandiae 10g, Radix Paeoniae Alba 10g, Pericarpium Citri Reticulatae 6g, Rhizoma Smilacis Chinensis 30g, Herba Salviae Chinensis 30g, Radix Glycyrrhizae Preparata 3g.
2. application according to claim 1, it is characterised in that the application in the medicine of the damnification of immunity function after chemotherapy prepared for reducing cyclophosphamide induction of the positive eliminating mass side is supported in described spleen invigorating.
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| CN1645364A (en) * | 2004-08-18 | 2005-07-27 | 范启康 | Applying method for realizing computer Chinese medicinal disease and prescription index by network technology |
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| CN1645364A (en) * | 2004-08-18 | 2005-07-27 | 范启康 | Applying method for realizing computer Chinese medicinal disease and prescription index by network technology |
Non-Patent Citations (3)
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| 刘沈林治疗消化道肿瘤的经验;舒鹏等;《江苏中医药》;20121231;第44卷(第12期);13-15 * |
| 胃癌术后中西医结合治疗的临床研究;崔琳;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20070515(第5期);E058-2 * |
| 胃癌术后的中医药治疗进展;李鹰等;《中医药临床杂志》;20120430;第24卷(第4期);372-374 * |
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