CN1425463A - Reinforced lucid ganoderma capable of immune preventing and immune curing cancer and its preparing method - Google Patents
Reinforced lucid ganoderma capable of immune preventing and immune curing cancer and its preparing method Download PDFInfo
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- CN1425463A CN1425463A CN 02151784 CN02151784A CN1425463A CN 1425463 A CN1425463 A CN 1425463A CN 02151784 CN02151784 CN 02151784 CN 02151784 A CN02151784 A CN 02151784A CN 1425463 A CN1425463 A CN 1425463A
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Abstract
The reinforced glossy ganodermka for immunological prevention and treatment of cancer consists of fermented glossy ganoderma powder in 1-3 weight portions and glossy ganoderma polysaccharide extracted from fermented glossy ganoderma powder in 1-2 weight portions. On the basis of glossy ganoderma effective components, some specific effective components are added selectively to strengthen the medicinal effect. Animal experiment and pathological experiment show that the present invention has obvious effects of immunological prevention and treatment on cancer.
Description
Affiliated technical field
But the present invention relates to the reinforcement Ganoderma of a kind of immunoprophylaxis and immunotherapy of cancer, the invention still further relates to the preparation method of this reinforcement Ganoderma.
Background technology
Past is because the extremely difficult acquisition of Ganoderma resource, add the restriction of scientific level, research to Ganoderma rests on the empirical accumulation of treatment more, over nearest two, 30 years, along with the successful Application of cultivation of glossy ganoderma or other production technologies and modern age modern science and technology such as medicine and pharmacology, biochemistry development, Ganoderma is proved to be once more that to be applied to treat multiple disease effective, has also obtained very big progress about effective component of glossy ganoderma and pharmacological research thereof.
Studies have shown that Ganoderma has inhibitory action to the tumor of experimental animal and human body, can dwindle tumor to a certain extent and weigh, slight illness is eased.In addition, in clinical practice, prove that also Ganoderma to prolonging the life cycle of cancer patient, alleviates patient's misery, there is certain or significant curative effect the quality of life of improving patient.The effect that use not only can be strengthened treating is united in Ganoderma and chemotherapy, radiotherapy, and can reduce the side effect of chemotherapy or radiotherapy significantly.Most relevant scientific literatures are thought: Ganoderma is to the treatment for cancer effect; not owing to there is certain composition can the direct killing cancerous cell in the Ganoderma; but (comprising: the immunity of cellular level and molecular level) by having transferred the immunity of self in the human body; can promote the level of killing and wounding by the immune system after this enhancing again to cancerous cell; in addition; under the situation of radiotherapy or chemotherapy, can protect the immune system of self again., existing Ganderma lucidum strain can make the big phase of product quality footpath different, adds place of production difference, the ecological condition difference of growth promoter, and the difference of the condition of cultivation causes the change of medicinal ingredient again.In addition; the mode difference of processing; as method commonly used in the family is to boil with other food to boil 1-4 hour; reasons such as its effective ingredient more can't be protected; add different cancerous cell kind differences; its toxicity also is not quite similar, thereby it is lasting or not ideal enough to cause therapeutic effect often to be difficult to.
It is exactly will be with Chinese medicine simple " Western medicineization " that existing many people (particularly Wai Guo medical producer) understand traditional Chinese medical science modernization, and its single certain chemical constituent of promptly purifying is understood fully its structural formula and understood fully its pharmacology and toxicity, comprises its pharmacokinetics etc.But the effect of Chinese medicine (natural drug) is a kind of effect background of effective composite parts, though to different diseases, the composition that has is main effective ingredient, has plenty of less important active ingredient, in addition, may also exist complementary effect between the various compositions of medicine; Event is above-mentioned simply can not to bring ideal therapeutic effect with Ganoderma " Western medicineization ".
Make eye bright
But the object of the present invention is to provide the reinforcement Ganoderma of a kind of immunoprophylaxis and immunotherapy of cancer, under stable with Ganoderma, abundant, the background of multiple active ingredient, increase specific active ingredient selectively, to strengthen drug action as the basis.
Another purpose of purpose of the present invention is to provide the preparation method of this reinforcement Ganoderma.
The object of the present invention is achieved like this: but the reinforcement Ganoderma of immunoprophylaxis and immunotherapy of cancer is to be mixed by the ganoderan that 1～3 weight portion fermented Ganoderma Lucidum powder and 1～2 weight portion extract from fermented Ganoderma Lucidum powder.
Synthetic the present invention strengthens the preparation method of Ganoderma with said components, may further comprise the steps successively:
(1) by protoplast ultraviolet mutagenesis method selection-breeding Ganoderma novel bacterial, and by 10 tons of extensive ganoderma lucidum liquid deep layer fermenting process of jar fermentation makes the Ganoderma wet mycelium to the Ganoderma novel bacterial of selection-breeding, the Ganoderma wet mycelium is ground into the ganoderma lucidum powder of granular size about 80 orders at 50 ℃～60 ℃ temperature dryings;
(2) get above-mentioned ganoderma lucidum powder, add 5～6 times of distilled water, 98-100 ℃ was extracted 5 hours, the yarn coarse filtration, and 3000 rev/mins are centrifugal, discard residue, get clear liquid; Add 3～4 times of 95% ethanol precipitation again in clear liquid, 3000 rev/mins centrifugal, discards clear liquid, gets precipitate; With the precipitate lyophilization, get the ganoderma lucidum crude polysaccharide powder;
(3) with the leaching process in the multiple step 2 of above-mentioned ganoderma lucidum crude polysaccharide grain weight more than three times, purity is 40%～100% ganoderan powder;
(4) the ganoderan powder of getting gained in the ganoderma lucidum powder of gained in the step 1 and the step 3 promptly gets and strengthens Ganoderma by 1～2 part of mix homogeneously of 1～3 part of ratio of described part by weight.
Use the present invention to strengthen Ganoderma prevented and treated tumor to laboratory animal experiment.To the feeding of laboratory animal, the present invention is strengthened Ganoderma be made into powder-type for convenience, detailed process is: add a small amount of 0.1% tween 80 mixing and mill in strengthening Ganoderma, add 0.5% sodium cellulosate solution gradually, through abundant porphyrize, be mixed into suspension, even to quantitative.Also the present invention can be strengthened Ganoderma and be made into the decocting dosage form, detailed process is: get ganoderma lucidum powder 1～3g and mix, add the 100ml normal saline with the ganoderan 1～2g of extraction, after soaking at room temperature is spent the night, be heated to 80 ℃ 15 minutes, get filtrate making; Each each experimental mouse administration volume is 0.3ml water preparation/10g body weight.Concrete experiment is as follows:
One, medication
(a) laboratory animal is selected the cleaning level kunming mice that the period of development is identical, body constitution is tried one's best identical for use, mice body weight 18～22kg, and other male mice of homogeneity is all used in every group of experiment, and each every group of experimental mouse is 10.
(b) before laboratory animal inoculation cancerous cell, experimental drug powder-type dosage is divided into basic, normal, high three kinds (being 1g/kg, 2g/kg, 3g/kg), and every day, gastric infusion was 1 time, continuous 10 days.Water decoction gastric infusion every day 1 time, administration volume are 0.3ml water preparation/10g body weight, continuous 10 days.
(c) by above-mentioned dosage and mode, behind the mouse inoculation cancerous cell, continue to strengthen Ganoderma with booster immunization power, examine the effect of strengthening Ganoderma after 10 days to the prevention and the treatment of cancer.
Two, connect the cancer method
Adopt the H22 hepatoma carcinoma cell as the cancer source cell, get the mice with tumor that contains eugonic H22 hepatoma carcinoma cell respectively and take off vertebra execution, with iodine tincture and the sterilization of ethanol skin of abdomen, extract the ascites that contains oncocyte then, the number of contained cancerous cell in every milliliter of the accurate Calculation, injection normal saline diluting cells to the concentration of reuse doses is 9.6 * 10
4/ ml, getting the above-mentioned cancerous cell of 0.2ml respectively, to be inoculated in the right axil of each experimental mouse subcutaneous, gastric infusion 10 days continuously again, anatomize the result of its immunoprophylaxis of research and immunization therapy next day after the drug withdrawal.
Three, experimental result
By above-mentioned medication with connect the cancer method, through three repeated trials (establish three physiology saline simultaneously and irritate the stomach contrast), the result proves: strengthening Ganoderma has the effect of prophylaxis of cancer preferably, each symptom phenomenon of not observing long cancer in the cancer mice body that connects that all has more than 20%, three times average its prevention cancer rate reaches 22.5%, the effect cancer suppressing ratio of three average its immunization therapies reaches 65.58%, specifically sees Table one, table two, table three.
Table one is strengthened Ganoderma prevention and cancer resistant effect (test for the first time)
| Group and sample | Dosage mg/kg/day | Pre-protection against rodents number | Prevention percent | The tumor weight (X ± SD) | Gross tumor volume (cm2) | Suppression ratio % |
| Normal saline matched group high-quality is strengthened the Ganoderma group | 0.5ml ? ? 1g/kg/d | 0 ? ? 2 | 0 ? ? 20％ | 1.30±0.58 ? ? 0.38±0.33 * | 2.16±0.42 ? ? 0.85± 0.21 * | ? ? ??68.59％ |
*P＜0.05
Table two is strengthened Ganoderma prevention and cancer resistant effect (test for the second time)
| Group and sample | Dosage mg/kg/day | Pre-protection against rodents number | Prevention percent | The tumor weight (X ± SD) | Gross tumor volume (cm2) | Suppression ratio % |
| Normal saline matched group high-quality is strengthened the Ganoderma suspensoid | 0.5ml ? ? 2g/kg/d | 0 ? ? 2 | 0 ? ? 20％ | 1.26±0.347 ? ? ?0.64±0.21 * | ?2.39±0.73 ? ? 0.64±0.21 ????** | ? ? 63.49％ |
*P＜0.05?????
**P＜0.01
Table three different dosage form is strengthened Ganoderma prevention and cancer resistant effect (test for the third time)
| Group and sample | Dosage mg/kg/d | Pre-protection against rodents number | Prevention percent | The tumor weight (* ± SD) | Gross tumor volume (cm2) | Suppression ratio % |
| Physiological saline control group high-quality is strengthened the glossy ganoderma supensoid agent and is strengthened high-quality glossy ganoderma water decoction | ??0.5ml ? ? ?1g/kg/day ? ? ?1g/kg/day | ????0 ? ? ????3 ? ? ????2 | ????0 ? ? ????30％ ? ? ????20％ | 1.19± 0.53 *? ? 0.43±0.30 ? ? 0.40± ? 0.25 * | 1.67±0.23 ? ? 0.82± 0.11 **? ? 0.75± ? 0.21 * | ? ? ? ??63.86％ ? ? ??66.％38 |
*P＜0.05
*P＜0.01 four, pathology experimental result
1, strengthen the deterioration degree that Ganoderma can significantly suppress tumor, obviously reduce tumor cell canceration degree and improve the pathological changes level of tumor cell, show following some:
(a) significantly reduce the deterioration degree of tumor
P
53Be important tumor-inhibiting factor, P
53Gene mutation be one of occurrence cause of many malignant tumor, normal P
53The albumen half-life is very short, and the sudden change back half-life obviously prolongs, and is easily detected by immuning tissue's method.From the visible matched group tumor P of table four
53Positive rate is along with inoculation cancerous cell time lengthening, this ratio still can constantly increase, suppression ratio at inoculation its grade malignancy after 10 days can reach 119.48 ± 7.23%, but after strengthening the Ganoderma administration and inoculating 20 days, the suppression ratio of its deterioration degree still can be retained in 57.83 ± 4.43% levels, in view of the above, strengthen the deterioration degree that Ganoderma can obviously reduce tumor cell, just can significantly suppress the ability of the infiltration and the transfer of tumor cell.
Table top four Ganoderma is to P
53Due to the influence of tumor deterioration degree
| Group and sample | Positive cell rate before the administration | Inoculate and contrast positive cell rate after 10 days | Administration is positive cell rate after 10 days | The grade malignancy suppression ratio |
| Matched group is strengthened the Ganoderma group | ??63.34±5.43 ??58.98±12.05 | ??76.68±10.54 ???????- | ???????- ??34.69±1.24 | ??119.48±7.23 *??57.83±4.43 ** |
*P＜0.05?????
**P＜0.001
(b) strengthen the activity that Ganoderma can obviously reduce tumor-marker enzyme in tumor tissues and the interior serum of its body, demonstration can obviously reduce the degree of canceration
From table five, six as seen: (γ-BT) and the activity of gsh proglumide-S-transferring enzyme (GST) indicate that once more the reinforcement Ganoderma of this experiment can obviously reduce tumor cell canceration degree and can promote cancerous cell to reverse the possible probability of differentiation again to normal cell with strengthening two kinds of tumor-marker enzyme γ-glutamine transpeptidases that can obviously reduce after the Ganoderma prevention administration tumor cell and the serum thereof.
Table five is strengthened Ganoderma to the active influence of r-GT in serum and the tumor tissues
| Group | Dosage | Serum r-GT activity (IV/g) | Tumor tissues r-GT activity (IV/g) |
| Matched group is strengthened the Ganoderma group | ????0.5ml ??1000mg/kg/day | ????5.85±0.74 ????1.69±0.63 * | ????72.5±3.562 ????48.42±3.49 * |
*P＜0.05
Table six is strengthened Ganoderma to the active influence of GST in serum and the tumor tissues
| Group | Dosage | Serum r-GT activity (IV/g) | Tumor tissues r-GT activity (IV/g) |
| Matched group is strengthened the Ganoderma group | ????0.5ml ??1000mg/kg/day | ????6.23±0.16 ????2.08±0.74 * | ????54.39±6.36 ????46.53±2.08 ** |
*P＜0.05????
**P＜0.01
2, prevention of reinforcement Ganoderma and treatment administration obviously improve the pathological changes level of tumor tissue cell
The tumor of matched group and administration group is taken out, be cut into film-making, significant difference is arranged with the histologic characteristics in the tumor of visible matched group of microscopic examination and reinforcement Ganoderma administration group.From Fig. 1 as seen, organizing of matched group tumor is frangible, and be nonelastic, irregularity boundary, and middle downright bad, the cavity is big.From Fig. 2 as seen, after with the administration of reinforcement Ganoderma, in the tumor tissues that it obviously dwindles, tissue slice is more flexible, and is non-friable, the border is clearly demarcated, middle nothing is downright bad, and the cavity is little or do not have the cavity, and the part cell has the nucleus scrambling to increase, membranolysis, showing wherein has phenomena of apoptosis to take place.This proves first: strengthen Ganoderma and can not only obviously suppress tumor growth, and can improve the inner cytopathic degree of tumor tissues.
3, strengthen Ganoderma and can obviously increase body autoimmune ability, show in the following areas:
(a) strengthen Ganoderma and can obviously improve cytophagous phagocytic activity
Utilize sodium thioglycollate to stimulate Turnover of Mouse Peritoneal Macrophages, can make its activation and obtain responsibility and the macrophage of acquisition tool phagocytic function, from table seven as seen: through the effect of variable concentrations Ganoderma extracting solution, as seen it is engulfed the dimethyl diaminophenazine chloride ability and can improve 10～45% respectively than matched group.
Table seven is strengthened Ganoderma to the phagocytotic influence of phagocyte
| Concentration of treatment (ug/ml) | Phagocyte is engulfed dimethyl diaminophenazine chloride (OD) ability | The P value |
| Contrast Ganoderma extracting solution 500 Ganoderma extracting solution 1000 | ????0.443±0.025 ????0.646±0.073 ????0.583±0.064 | ????＞0.05 ????＜0.001 |
(b) strengthen Ganoderma and can significantly improve the structure of T lymph subgroup, thereby can strengthen intravital immunity
The ratio of CD4/CD8 (being T lymph accessory cell/T lymph poison cell) can be used as an important indicator of reflection cell immunocompetent.From table eight as seen; can significantly increase the percentage rate of T lymph accessory cell with strengthening the Ganoderma prevention, and can reduce the percentage rate of T lymph poison cell simultaneously, thereby can reverse the undesired ratio of CD4/CD8 cell; make it near normalization, can strengthen the immunocompetence of cell thus.
Table top eight Ganoderma is to the influence of CD4/CD8 ratio
| Give Ganoderma preceding to behind the Ganoderma |
| ????CD4???????35.8±5.6?????43.7±6.3 **????CD8???????36.3±3.2?????35.7±.0.33 ????CD4/CD8???0.98±0.13????1.22±0.14 * |
*P＜0.001??????
**P＜0.001
(c) but strengthen the generation that Ganoderma obvious stimulation bone marrow hematogenesis is organized medium-sized lymphocyte and colony thereof
From Fig. 3,4 as seen, after high-quality is strengthened the Ganoderma administration, but the growth of obvious stimulation bone marrow hematogenesis tissue, the as seen generation of lymphocyte colonies in a large number, thus can effectively promote the immunocompetence of body.
(d) use the transfer ability that the reinforcement Ganoderma can obviously reduce cancerous cell, prolong the life cycle of suffering from the cancer body
Mus body to the inoculation cancerous cell is dissected and observation analysis, discovery is in matched group, there is visible tumor tissues to be transferred to lung and liver in the mice more than 80%, but in strengthening Ganoderma administration group, all do not find in lung and liver, to have the tumor tissues of visible transfer.In addition, the matched group of one winding kind cancerous cell and the mice of reinforcement Ganoderma administration group are continued to observe its dead speed and survival period, discovery began to have the death of Mus body in back 13 days in inoculation, and the individual death frequency is very fast in matched group, and through Ganoderma administration group, individual death was just arranged, and the individual death frequency is slower after 16 days, the discrete life cycle that still can keep normal mouse, this shows, strengthen Ganoderma and can prolong the survival period that connects the cancer mice to some extent.
In sum, the present invention strengthens Ganoderma all has significant effect to the prevention of cancer and treatment.
Description of drawings
Fig. 1 is a physiology saline control group tumor tissue section;
Fig. 2 is for strengthening Ganoderma group tumor tissue section;
Fig. 3 is a physiology saline control group mouse femur tumor tissue section;
Fig. 4 is for strengthening Ganoderma group mouse femur tumor tissue section.
The specific embodiment
Employing is at " microorganism journal " 2001,41 (2): the 89-93 page or leaf, exercise question is a disclosed protoplast ultraviolet mutagenesis method selection-breeding Ganoderma novel bacterial in " research of protoplast ultraviolet mutation breeding Ganoderma novel bacterial " literary composition, and adopt the above-mentioned Ganoderma novel bacterial fermentation of the extensive ganoderma lucidum liquid deep layer fermenting process of the disclosed 10 tons of jars of Chinese invention patent ZL981131085 to make the Ganoderma wet mycelium to selection-breeding, the detailed process of preparation Ganoderma wet mycelium is: the Ganoderma novel bacterial of selection-breeding is inserted in ten 500ml (filling shake-flask culture base 100ml) triangular flask, cultivated 96 hours on 27 ℃ of following shaking tables, then per two 500ml being shaken bottle strain pours in the triangular flask of a 5000ml (filling 1000ml shake-flask culture base), obtain 5 bottles of 5000ml altogether and shake the bottle kind, shaking bottle with 5 bottles plants on 25 ℃ of following shaking tables and to cultivate 48 hours, shaking bottle with 5 bottles then plants in the seed tank of the 100L (filling the 50L fermentation medium) that transfers, transfer pH in 6.0, keeping a jar temperature is 26 ℃, tank pressure is 20kpa, stir speed (S.S.) is 130 rev/mins, ventilation is 1: 0.4, ferment after 48 hours, be transferred in three tons of fermentation tanks (filling 1 ton of fermentation medium), transfer pH in 6.0, keeping a jar temperature is 28 ℃, tank pressure is 40kpa, stir speed (S.S.) is 140 rev/mins, ventilation is 1: 0.4, ferment after 48 hours, be transferred in 15 tons of fermentation tanks (filling 10 tons of fermentation medium), transfer pH to 6.0, keep 27 ℃ of jar temperature, tank pressure is 30kpa, stir speed (S.S.) is 150 rev/mins, ventilation is 1: 0.55, ferments after 100 hours the fermentation liquid very thickness that becomes, microscopy, finding has a large amount of Ganoderma myceliums in the fermentation liquid, and a small amount of mycelia begins autolyze, puts jar immediately, filter pressing obtains wet mycelium.The Ganoderma wet mycelium with 55 ℃ of temperature dryings, is ground into the ganoderma lucidum powder of granular size about 80 orders.
Get above-mentioned ganoderma lucidum powder, add 5 times of distilled water, 98 ℃ were extracted 5 hours, the yarn coarse filtration, and 3000 rev/mins are centrifugal, discard residue, get clear liquid; Add 3 times of 95% ethanol precipitation again in clear liquid, 3000 rev/mins centrifugal, discards clear liquid, gets precipitate; With the precipitate lyophilization, get the ganoderma lucidum crude polysaccharide powder; Again the ganoderma lucidum crude polysaccharide grain weight is answered the said extracted process five times, get purity and be about 60% ganoderan powder.
1: 1 by weight or 3: 2 or 2: 1 ratios are got ganoderma lucidum powder and ganoderan powder mix homogeneously, promptly get and strengthen Ganoderma.
Claims (2)
- But 1, the reinforcement Ganoderma of a kind of immunoprophylaxis and immunotherapy of cancer is characterized in that being mixed by the ganoderan that 1～3 weight portion fermented Ganoderma Lucidum powder and 1～2 weight portion extract from fermented Ganoderma Lucidum powder.
- But 2, the preparation method of the reinforcement Ganoderma of described immunoprophylaxis of claim 1 and immunotherapy of cancer is characterized in that may further comprise the steps successively:(1) by protoplast ultraviolet mutagenesis method selection-breeding Ganoderma novel bacterial, and by 10 tons of extensive ganoderma lucidum liquid deep layer fermenting process of jar fermentation makes the Ganoderma wet mycelium to the Ganoderma novel bacterial of selection-breeding, the Ganoderma wet mycelium is ground into the ganoderma lucidum powder of granular size about 80 orders at 50 ℃～60 ℃ temperature dryings;(2) get above-mentioned ganoderma lucidum powder, add 5～6 times of distilled water, 98-100 ℃ was extracted 5 hours, the yarn coarse filtration, and 3000 rev/mins are centrifugal, discard residue, get clear liquid; Add 3～4 times of 95% ethanol precipitation again in clear liquid, 3000 rev/mins centrifugal, discards clear liquid, gets precipitate; With the precipitate lyophilization, get the ganoderma lucidum crude polysaccharide powder;(3) with the leaching process in the multiple step 2 of above-mentioned ganoderma lucidum crude polysaccharide grain weight more than three times, purity is 40%～100% ganoderan powder;(4) the ganoderan powder of getting gained in the ganoderma lucidum powder of gained in the step 1 and the step 3 promptly gets and strengthens Ganoderma by 1～2 part of mix homogeneously of 1～3 part of ratio of described part by weight.
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| CN 03106578 CN1284542C (en) | 2002-12-31 | 2003-03-03 | Strengthened ganoderma lucidum for possible immunoprophylaxis and immunotherapy of cancers and related diseases as well as its preparing method |
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| CN103285048A (en) * | 2012-02-27 | 2013-09-11 | 德阳仙鹤生物技术有限公司 | Lucid ganoderma composition, preparation with effect of improving immune function and preparation method of preparation |
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| CN103285048A (en) * | 2012-02-27 | 2013-09-11 | 德阳仙鹤生物技术有限公司 | Lucid ganoderma composition, preparation with effect of improving immune function and preparation method of preparation |
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