Ganoderma lucidum, radix salviae miltiorrhizae and radix salviae miltiorrhizae traditional Chinese medicine composition and preparation method thereof

Abstract

The invention discloses a radix linderae, radix salviae miltiorrhizae and radix salviae miltiorrhizae traditional Chinese medicine composition and a preparation method thereof, and relates to the field of food / health-care products. The ganoderma lucidum, ginseng and salvia miltiorrhiza traditional Chinese medicine composition is prepared from the following components: 22.5 to 45 parts of wall-broken ganoderma lucidum spore powder, 26 to 45.5 parts of gynostemma pentaphylla, 21.6 to 48 parts of salvia miltiorrhiza, 6 to 18 parts of ginseng and 6.5 to 19.5 parts of safflower carthamus. By precisely proportioning natural medicinal materials such as wall-broken ganoderma lucidum spore powder, fiveleaf gynostemma herb, red sage root, ginseng and safflower, the problem of insufficient single drug effect or improper formula proportion in the prior art is solved. Through the specific proportion setting, the synergistic effect of the medicine effects of the two is effectively promoted, the immunity can be enhanced, the blood circulation can be improved, and the fatigue can be resisted. Meanwhile, the comprehensive curative effect of the product can be further improved through reasonable matching of the radix salviae miltiorrhizae and the flos carthami. All the raw materials are optimally combined, so that the stability and the absorption rate of the medicine effect are improved, the comprehensive health-care effect of the product is enhanced, and the health requirements of different people can be met.

Description

Technical Field

[0001] The present invention relates to the field of food / health products, and particularly to a Ling Shen Dan traditional Chinese medicine composition and a preparation method thereof. Background Art

[0002] In recent years, with the improvement of people's health awareness, the demand for natural plant drugs and health products has gradually increased. Natural plant herbs such as Ganoderma lucidum, Gynostemma pentaphyllum, Salvia miltiorrhiza, Panax ginseng, and Carthamus tinctorius are widely used to improve health, enhance immunity, promote blood circulation, and improve cardiovascular and cerebrovascular functions. Especially in the field of traditional Chinese medicine, herbs such as Ganoderma lucidum and Panax ginseng are considered to have various pharmacological effects, such as enhancing immunity, anti-fatigue, and anti-aging functions.

[0003] Although existing related health products are relatively abundant in the market, most products do not fully exert the synergistic effects of various herbs, and the proportion and combination method of the drug formula are often not scientifically and reasonably formulated, resulting in unstable or insufficient efficacy of the products. For example, the patent with the application number 201811551693.8 provides a Gynostemma pentaphyllum and Panax ginseng functional health food capsule and a preparation method thereof. Especially in traditional formulas, the proportion between herbs may not be able to maximize the drug efficacy, and there may be problems such as incomplete drug efficacy, poor absorption, or slow effects. Therefore, optimizing the herb formula and proportion and enhancing the comprehensive benefits of products have become important topics in current health product research.

[0004] Currently, although health products such as Ganoderma lucidum and Panax ginseng on the market have been recognized by some consumers, they generally have the following problems: First, many formulas are too single in the selection of herbs or overly rely on a certain ingredient, failing to achieve the synergistic effects of multiple components and multiple targets; Second, the dosage form or the way of processing herbs in some products fails to fully release the active ingredients, resulting in a low absorption rate; Finally, traditional Chinese medicine formulas often do not consider the optimal proportional relationship between herbs, resulting in unsatisfactory drug efficacy.

[0005] Therefore, there is an urgent need for an innovative product that can optimize the compatibility ratio of various herbs and exert the synergistic effects of each component to improve the drug efficacy and meet the needs of consumers for comprehensive health care effects. Summary of the Invention

[0006] One object of the present invention is to provide a Ling Shen Dan traditional Chinese medicine composition, and another object is to provide a preparation method of the Ling Shen Dan traditional Chinese medicine composition. The Ling Shen Dan traditional Chinese medicine composition includes Ganoderma lucidum spore powder, Panax ginseng, Carthamus tinctorius, Gynostemma pentaphyllum, and Salvia miltiorrhiza. The combination of each raw material can achieve strengthening the healthy qi and consolidating the constitution, replenishing qi and generating essence, and balancing the yin and yang qi and blood of the human body's zang-fu organs, so that the healthy qi of the human body is taken into account and the immunity is comprehensively enhanced, and it is easy to be absorbed by the human body.

[0007] To achieve the above objects, the present invention adopts the following technical solutions: A Ganoderma lucidum ginseng pill traditional Chinese medicine composition, comprising the following components: 22.5 - 45 parts of broken Ganoderma lucidum spore powder, 26 - 45.5 parts of Gynostemma pentaphyllum, 21.6 - 48 parts of Salvia miltiorrhiza, 6 - 18 parts of ginseng, and 6.5 - 19.5 parts of safflower.

[0008] Among them, the ratio of broken Ganoderma lucidum spore powder to ginseng is (5:4) - (15:2).

[0009] Preferably, it further comprises 0.6 - 2 parts of soluble starch, 0.75 - 7.5 parts of silicon dioxide, and 0.75 - 7.5 parts of enteric-coated film coating premix.

[0010] As a preference, a Ganoderma lucidum ginseng pill traditional Chinese medicine composition, comprising the following components: 25 parts of broken Ganoderma lucidum spore powder, 35 parts of Gynostemma pentaphyllum, 20 parts of Salvia miltiorrhiza, 8 parts of ginseng, 8 parts of safflower, 1 part of soluble starch, 1 part of silicon dioxide, and 1 part of enteric-coated film coating premix.

[0011] The salvianolic acid B in the Ganoderma lucidum ginseng pill traditional Chinese medicine composition ≥ 960 mg / 100 g, total saponins ≥ 2.5 g / 100 g, ganoderic acid A ≥ 25 mg / 100 g. The present invention also adopts another technical solution: A preparation method of a Ganoderma lucidum ginseng pill traditional Chinese medicine composition, comprising the following steps: S1: Extraction of each raw material: The broken Ganoderma lucidum spore powder is pretreated and extracted with water. Gynostemma pentaphyllum, Salvia miltiorrhiza, ginseng, and safflower are respectively coarsely crushed, extracted with water, filtered, concentrated, spray-dried, and sieved through a 60-mesh sieve to prepare the extract for standby; S2: Granulation: The above-mentioned extract, soluble starch, and 1 / 2 prescription amount of silicon dioxide are put into a fluidized bed granulator. After mixing evenly, purified water is used as the wetting agent, and water is sprayed in to boil and granulate, and dried until the water content ≤ 5.0%, and granulated through a 20-mesh sieve; S3: Add the remaining 1 / 2 prescription amount of silicon dioxide, mix for 8 - 15 minutes, and press tablets; S4: The enteric-coated film coating premix is added to purified water and stirred evenly to prepare a suspension; the plain tablets are coated, and the inlet air temperature is controlled at 60°C - 80°C to keep the core temperature of the tablets at 37 - 42°C.

[0012] In the extraction of each raw material in S1, combined extraction can also be adopted: After Gynostemma pentaphyllum, Salvia miltiorrhiza, ginseng, and safflower are respectively coarsely crushed, the broken Ganoderma lucidum spore powder, Gynostemma pentaphyllum, Salvia miltiorrhiza, ginseng, and safflower are weighed according to the ratio, added with 8 - 20 times the amount of water, and mixed and extracted 2 - 3 times under the conditions of 90°C - 100°C for 1 - 2 hours each time, filtered, concentrated, spray-dried, and processed through a 60-mesh sieve at an inlet air temperature of 180°C - 205°C to prepare the extract for standby.

[0013] In S1, add 8 - 20 times the amount of water, heat at 90°C - 100°C, and extract 2 - 3 times, each time for 0.5 - 2 hours.

[0014] In S1, the inlet air temperature for spray drying is 180°C - 205°C.

[0015] In S2, the weight of the water sprayed is 35 - 65% of the weight of the material, and the inlet air temperature is 65°C - 85°C.

[0016] In S4, the concentration of the suspension is 10 - 15%.

[0017] The dosage form of the Lingcan Dan traditional Chinese medicine composition can be dosage forms such as capsules, tablets, and granules.

[0018] Ganoderma lucidum spore powder is the tiny oval reproductive cells ejected from the gills of Ganoderma lucidum during its growth and maturity period. Each spore is only 46 micrometers and contains many genetic materials of Ganoderma lucidum. It mainly contains nutrients such as ganoderic triterpenoids and ganoderma polysaccharides, and has the effects of enhancing immunity, beautifying the skin, etc. The polysaccharides and ganoderic triterpenoid compounds in Ganoderma lucidum spore powder can significantly improve the function of the body's immune system. By promoting the activities of immune cells such as macrophages, T cells, and B cells, the disease resistance ability is enhanced. After Ganoderma lucidum spore powder is broken wall, it is more suitable for direct absorption by the human stomach and intestines. Broken wall Ganoderma lucidum spore powder has the effects of improving immunity, protecting the liver and detoxifying, regulating blood sugar and blood lipids, calming the nerves and helping sleep, protecting cardiovascular health, delaying aging, anti - inflammation and antibacterial.

[0019] Ginseng contains rich active ingredients such as ginsenosides, polysaccharides, amino acids, vitamins, and minerals. The polysaccharides and saponin compounds in ginseng can promote the activities of immune cells and enhance the body's immune defense function against infection and diseases. In addition, ginseng also has the effect of promoting blood circulation, which helps to improve the transport ability of oxygen and nutrients, improve microcirculation, and is beneficial to cardiovascular health.

[0020] Gynostemma pentaphyllum is a herbaceous climbing plant of the Cucurbitaceae family. Gynostemma pentaphyllum tastes bitter and cold, and has the effects of clearing heat and detoxifying, relieving cough and clearing lung phlegm, nourishing the heart and calming the nerves, replenishing qi and generating essence, tonifying qi and strengthening the spleen, and tonifying the kidney and calming the nerves. It can be used to lower blood pressure, lower blood lipids, protect the liver, promote sleep, and treat gastroenteritis, tracheitis, pharyngolaryngitis, and is used in the anti - cancer clinical treatment of various cancers.

[0021] The main active ingredient of Salvia miltiorrhiza is tanshinone compounds, which have the effects of antioxidant, anti - inflammation, and improving blood circulation. Salvia miltiorrhiza dilates blood vessels, improves microcirculation, and promotes blood flow, thereby enhancing the transportation and distribution of immune cells. In addition, Salvia miltiorrhiza has an anti - inflammatory effect, which can reduce the over - reaction of the immune system or chronic inflammation, so that the immune system is in a more balanced state.

[0022] Safflower has the effects of promoting blood circulation to remove blood stasis and dredging meridians to relieve pain, and its main components are safflower flavonoids. Safflower can improve blood circulation, promote the flow of immune cells, enhance the distribution of immune cells in the blood, and promote immune responses. At the same time, safflower also has antioxidant and anti-inflammatory effects, which can reduce the generation of free radicals in the body, thereby reducing the burden on the immune system and enhancing immune function.

[0023] Ganoderma lucidum spore powder and ginseng can significantly improve the activities of T cells and B cells, and enhance immune surveillance and immune responses. Gynostemma pentaphyllum further enhances immune function by promoting the proliferation of immune cells. Salvia miltiorrhiza and safflower improve the microcirculation and blood flow, thereby enhancing the transport efficiency of immune cells and the timeliness of immune responses. Ganoderma lucidum spore powder, ginseng, and gynostemma pentaphyllum all have antioxidant effects, which can scavenge free radicals in the body, reduce oxidative stress, and maintain the normal function of the immune system. Safflower also contains rich antioxidant components, which can synergistically reduce oxidative damage and protect immune cells. The anti-inflammatory effects of salvia miltiorrhiza and safflower help regulate the overreaction of the immune system, avoid immune disorders or chronic inflammatory states, provide a good immune environment for other medicinal materials, and improve the overall immunity; they can also improve blood circulation, promote the flow of immune cells, and make immune responses more rapid and comprehensive. The components of ginseng and gynostemma pentaphyllum help improve the overall metabolic function of the body and increase the anti-fatigue ability of the body, so that the immune system can work effectively without external pressure.

[0024] Therefore, the present invention has the following beneficial effects: By precisely proportioning natural medicinal materials such as Ganoderma lucidum spore powder, gynostemma pentaphyllum, salvia miltiorrhiza, ginseng, and safflower, the problems of insufficient single drug efficacy or improper formula ratio in the prior art are overcome. In particular, the synergy between Ganoderma lucidum spore powder and ginseng, through specific ratio settings, effectively promotes the synergistic effect of the two drugs, which can not only enhance immunity but also improve blood circulation and anti-fatigue. At the same time, the reasonable combination of other medicinal materials such as salvia miltiorrhiza and safflower can further improve the comprehensive curative effect of this product. Through this optimized combination, the present invention not only improves the stability and absorption rate of drug efficacy but also enhances the comprehensive health care effect of the product, and can meet the health needs of different populations. Detailed implementation mode

[0025] Example 1 A traditional Chinese medicine composition of Lingcan Dan comprises the following components: 25 parts of Ganoderma lucidum spore powder, 35 parts of gynostemma pentaphyllum, 20 parts of salvia miltiorrhiza, 8 parts of ginseng, 8 parts of safflower, 1 part of soluble starch, 1 part of silicon dioxide, and 1 part of gastric-soluble film coating premix.

[0026] Example 2 The traditional Chinese medicine composition of Lingcan Dan in this example is the same as that in Example 1, and the preparation method adopts separate extraction.

[0027] A method for preparing a Lingshendan Chinese medicine composition comprises the following steps: S1: Extraction of raw materials: Extraction of broken spore powder of Ganoderma lucidum: add 14 times and 12 times the amount of water respectively, extract twice at 90°C, each time for 2 hours, filter, concentrate and spray dry after extraction, the air inlet temperature is 200°C; pass through 60 mesh sieve, package and other main processes to make extract, and set aside; Gynostemma pentaphyllum extraction: Gynostemma pentaphyllum is roughly crushed, and extracted twice with 15 times and 10 times the amount of water at 90°C, each time for 1 hour. After extraction, it is filtered, concentrated, and spray-dried, and the air inlet temperature is 200°C; the extract is processed through a 60-mesh sieve and packaged, and is set aside; Salvia miltiorrhiza extraction: Salvia miltiorrhiza is roughly crushed, and extracted twice with 14 times and 10 times the amount of water at 95°C, each time for 1 hour. After extraction, it is filtered, concentrated, spray-dried, and the air inlet temperature is 205°C; the extract is made by the main process of sieving through a 60-mesh sieve and packaging, and is set aside; Ginseng extraction: Ginseng is roughly crushed, and extracted twice with 12 times and 12 times the amount of water at 90°C, each time for 1.5 hours. After extraction, it is filtered, concentrated, spray-dried, and the air inlet temperature is 195°C; the extract is made by the main process of sieving through a 60-mesh sieve and packaging, and is set aside; Safflower extraction: safflower is roughly crushed, and extracted twice with 16 times and 13 times the amount of water at 100°C, each time for 0.5 hours. After extraction, it is filtered, concentrated, spray-dried, and the air inlet temperature is 180°C; the extract is made by the main process of sieving through a 60-mesh sieve and packaging, and is set aside; S2: Granulation: After sieving and mixing the extracts, soluble starch and 1 / 2 of the prescription amount of silicon dioxide, put them into a one-step granulator. After the materials are evenly mixed, spray water for boiling granulation. The material temperature is controlled at about 60°C, and the weight of the sprayed water is 65.6% of the material weight. After further drying, take out and sieve, the moisture content is 4.7%, and granulate through a 20-mesh sieve; S3: Add the remaining 1 / 2 amount of silicon dioxide, mix well, and press into tablets, mix for 10 minutes, and press into tablets; S4: Add purified water to the film coating premix to prepare a 15% coating solution for coating. The coating volume is 1.0 g / tablet.

[0028] The test results of each index are shown in Table 1.

[0029] Table 1 Results of various indicators of the Lingshendan Chinese medicine composition prepared in Example 2.

[0030] Example 3 The Ganoderma lucidum ginseng pill traditional Chinese medicine composition of this embodiment is the Ganoderma lucidum ginseng pill traditional Chinese medicine composition in Embodiment 1, and the preparation method adopts combined extraction.

[0031] A preparation method of a Ganoderma lucidum ginseng pill traditional Chinese medicine composition comprises the following steps: S1: Extraction: After Gynostemma pentaphyllum, Salvia miltiorrhiza, Panax ginseng, and Carthamus tinctorius are roughly crushed respectively, take Ganoderma lucidum spore powder, Gynostemma pentaphyllum, Salvia miltiorrhiza, Panax ginseng, and Carthamus tinctorius according to the ratio for mixed extraction. Add 15 times the amount of water and 12 times the amount of water respectively, and extract at 100 °C for 2 times, 1 hour each time. After extraction, filter, concentrate, and spray-dry, with the inlet air temperature at 200 °C; pass through a 60-mesh sieve, package and other main processes to process and make an extract for standby.

[0032] S2: Granulation: After the extract, soluble starch, and 1 / 2 of the prescription amount of silicon dioxide are sieved and mixed, put them into a one-step granulator. After the materials are mixed evenly, spray water to boil and granulate, control the material temperature at about 60 °C, and the weight of the sprayed water is 46.8% of the weight of the materials. After continuous drying, take out and sieve, with the moisture content of 5.1%, and screen and size with a 20-mesh sieve.

[0033] S3: Add the remaining 1 / 2 of the prescription amount of silicon dioxide, mix evenly and then press tablets, mix for 10 minutes and then press tablets.

[0034] S4: Prepare a 15% coating solution by mixing a film coating premix with purified water, and conduct coating, with the coated tablets being 1.0 g / tablet.

[0035] The test results of each index are shown in Table 2.

[0036] Table 2 Results of each index of the Ganoderma lucidum ginseng pill traditional Chinese medicine composition prepared in Example 3.

[0037] As can be seen from Table 1 and Table 2, in the preparation of the Ganoderma lucidum ginseng pill traditional Chinese medicine composition in Example 1, the results of each index of the Ganoderma lucidum ginseng pill traditional Chinese medicine composition prepared by separate extraction and combined extraction of each component are not very different and there is no obvious difference. Therefore, it shows that in the preparation process of the Ganoderma lucidum ginseng pill traditional Chinese medicine composition of the present invention, separate extraction and combined extraction of each component are both acceptable, and both can achieve good physical stability and a high content of active ingredients, and can exert the effects of enhancing immunity, antioxidation, anti-inflammation, anti-fatigue, etc.

[0038] Efficacy evaluation I. Sample treatment In this test, the test sample prepared in Example 2 was crushed into powder as the test substance, and before the test, it was formulated into each dose with deionized water as the solvent and mixed evenly for testing.

[0039] II. Experimental animals and detection conditions The license number for the use of experimental animals is SYXK(Zhe)2023-0042. The ICR mice used in the experiment were provided by Zhejiang Vital River Laboratory Animal Technology Co., Ltd. The production license number for experimental animals is SYXK(Zhe)2024-0001, SPF grade, female, weighing 20±2g, and the quality certificate number: 20240313Abzz0619060571. The feed was provided by Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd., and the production license number: Su Feed License (2019)01008. The detection environmental conditions were barrier environment, temperature 22-25°C, relative humidity 40-70%. The animals were acclimatized in the animal house environment for 4 days before the experiment.

[0040] III. Dose Design The experiment was set up with three dose groups and a negative control group (deionized water). The recommended human dose of this finished product is 6.0 g / 60 kg / day. The low, medium, and high doses were 0.5, 1.0, and 3.0 g / kg body weight respectively, which are equivalent to 5, 10, and 30 times the recommended human dose. For the three dose groups, 5, 10, and 30 g of the sample powder were weighed respectively and added with deionized water to 100 mL to prepare test substances with concentrations of 0.05, 0.10, and 0.30 g / mL. The test substances were administered by gavage, and the gavage volume was calculated as 0.1 mL / 10 g body weight. The negative control group was treated the same except for being given deionized water by gavage.

[0041] IV. Experimental Methods and Results 1. ConA-induced Mouse Splenic Lymphocyte Transformation Test Each group was given the test substance by gavage once a day for 32 consecutive days. On the 33rd day of the experiment, the spleen of each mouse was aseptically removed and placed in a small petri dish containing an appropriate amount of sterile Hanks solution. It was gently torn with forceps to make a single cell suspension, filtered through a 200-mesh sieve, washed, counted, and finally the cell concentration was adjusted to 3×10 6 cells / mL with RPMI1640 complete culture medium. The cell suspension was added to two wells of a 24-well culture plate, 1 mL per well. In one well, 25 μL of ConA solution (equivalent to 2.5 μg / mL) was added, and the other well was used as a control. The cells were cultured in an incubator at 37°C and 5% CO2 for 72 hours. Four hours before the end of the culture, 0.7 mL of the supernatant was gently aspirated from each well, 0.7 mL of RPMI1640 culture medium without calf serum was added, and at the same time 50 μL / well of MTT (5 mg / mL) was added, and the culture was continued for 4 hours. After the culture ended, 1 mL of acidic isopropanol was added to each well, and it was pipetted and mixed well to completely dissolve the purple crystals. The optical density value (OD) was measured at a wavelength of 570 nm. Finally, the optical density value of the well with ConA minus the optical density value of the well without ConA was used to represent the proliferation ability of lymphocytes. The results are shown in Table 3.

[0042] Table 3 Effects of the test samples on the proliferation ability of ConA-induced mouse splenic lymphocytes (x-±s). Group Number of animals (pcs) Optical density difference <![CDATA[Negative control (H2O)]]> 10 0.276 ± 0.076 Low-dose group 10 0.278±0.046 Medium-dose group 10 0.284±0.046 High-dose group 10 0.284±0.089

[0043] As can be seen from Table 3, compared with the negative control group, the optical density differences of the drug administration groups (low, medium, and high dose groups) increased, and the proliferation ability of lymphocytes increased, indicating that the Lingcan Dan traditional Chinese medicine composition of the present invention helps the proliferation of mouse lymphocytes, can effectively enhance the immune function, and improve immunity.

[0044] 2. Induction of DTH in mice by sheep red blood cells (SRBC) (plantar thickness method) Each group was intragastrically administered the test substance once a day for 30 consecutive days. On the 26th day of the experiment, each mouse was intraperitoneally injected with 0.2 mL of 2% (v / v) packed sheep red blood cell (SRBC) suspension for immunization. Four days after immunization, the thickness of the left hind paw was measured, and then 20% (v / v) SRBC was subcutaneously injected at the measurement site, 20 μL per mouse (about 1×10⁸ SRBC). The thickness of the left hind paw was measured 24 hours after injection, and the same site was measured three times and the average value was taken. The results are shown in Table 4.

[0045] Table 4 Effects of the test samples on DTH induced by sheep red blood cells in mice (x-±s). Group Number of animals (pcs) Difference in thickness of left hind paw sole (mm) <![CDATA[Negative control (H2O)]]> 10 0.50±0.10 Low-dose group 10 0.52±0.10 Medium-dose group 10 0.65±0.12 High-dose group 10 0.72±0.15

[0046] As can be seen from Table 3, compared with the negative control group, the thickness differences of the left hind paws of the mice in the drug administration groups (low, medium, and high dose groups) increased, and the differences in the low and high dose groups were significant, which could significantly enhance the DTH response of the mice, thus showing the effect of enhancing the cellular immune function of the mice.

[0047] 3. Determination of mouse serum hemolysin (hemagglutination method) Each group was intragastrically administered the test substance once a day for 30 consecutive days. On the 26th day of the experiment, each mouse was intraperitoneally injected with 0.2 mL of 2% (v / v) packed sheep red blood cell (SRBC) suspension for immunization. Five days later, blood was taken and centrifuged to collect serum. The serum was serially diluted with physiological saline and incubated in a 37°C incubator for 3 hours. The degree of hemagglutination was observed and the antibody titer was calculated. The results are shown in Table 5.

[0048] Table 5 Effects of the test samples on mouse serum hemolysin (x-±s). Group Number of animals (pcs) Anti-volume value <![CDATA[Negative control (H2O)]]> 10 213.4±30.4 Low-dose group 10 231.9±23.3 Medium-dose group 10 248.8±34.3 High-dose group 10 240.4±36.8

[0049] As can be seen from Table 5, the antibody titer values of each group were higher than those of the negative control group, and the antibody titer value of the medium dose group was the highest. This indicates that the Lingcan Dan traditional Chinese medicine composition may have a significant promoting effect on the production of mouse serum hemolysin, especially at the medium dose, thus indicating that the Lingcan Dan traditional Chinese medicine composition of the present invention has a significant enhancing effect on the humoral immune response.

[0050] 4. Detection of antibody-producing cells (Jerne modified slide method) Each group was given the test substance by gavage once a day for 30 consecutive days. On the 26th day of the experiment, each mouse was intraperitoneally injected with 0.2 mL of 2% (v / v) packed sheep red blood cell (SRBC) suspension for immunization. Five days later, the spleen of each mouse was aseptically removed and placed in a small Petri dish containing an appropriate amount of sterile Hanks solution. It was gently torn with forceps to make a single-cell suspension, filtered through a 200-mesh sieve, washed, counted, and finally the cell concentration was adjusted to 5×106 cells / mL with RPMI 1640 complete culture medium. After the surface medium (1 g of agarose added to distilled water to 100 mL) was heated and dissolved, it was kept warm in a 45°C water bath, mixed with an equal volume of Hank's solution with a pH of 7.2 - 7.4 and a double concentration, and aliquoted into small test tubes, 0.5 mL per tube. Then, 50 μL of 10% SRBC (w / v, prepared with SA buffer) and 20 μL of spleen cell suspension (5×106 cells / mL) were added to the tube, quickly mixed, and poured onto a slide coated with a thin layer of agarose to make parallel slides. After the agar solidified, the slides were horizontally buckled on a slide rack and placed in a carbon dioxide incubator for incubation for 1.5 hours. Then, the complement diluted with SA buffer (1:8) was added to the groove of the slide rack, and incubation was continued for 1.5 hours. The number of hemolytic plaques was counted. The results are shown in Table 6.

[0051] Table 6 Effects of the test samples on antibody-producing cells in mice (x-±s). Group Number of animals (pcs) Number of hemolytic plaques (pcs / 105 spleen cells) Negative control (H2O) 10 46±9 Low-dose group 10 57±12 Medium-dose group 10 52±12 High-dose group 10 45±13

[0052] As can be seen from Table 6, compared with the negative control group, the number of hemolytic plaques in the low-dose group was the highest. The number of hemolytic plaques in the medium-dose group and high-dose group was slightly lower than that in the low-dose group, but still higher than that in the negative control group, indicating that at a low dose, the Lingcan Dan traditional Chinese medicine composition has a certain promoting effect on antibody-producing cells and has an enhanced immune function.

[0053] 5. Phagocytosis of chicken red blood cells by mouse peritoneal macrophages (semi-in vivo method) Each group was given the test substance by gavage once a day for 32 consecutive days. Thirty minutes before the animals were sacrificed, each mouse was intraperitoneally injected with 1 mL of 20% (v / v) chicken red blood cell suspension. After sacrifice, 2 mL of normal saline was injected. The peritoneal fluid was taken for dropping on a slide, incubated in a 37°C incubator for 30 minutes, fixed, stained, examined under a microscope, and 100 macrophages were counted to calculate the phagocytosis rate and phagocytosis index. The results are shown in Table 7.

[0054] Table 7 Effects of the test samples on phagocytosis of chicken red blood cells by mouse peritoneal macrophages (x-±s). Group Number of animals (pcs) Phagocytosis rate (%) Phagocytosis index <![CDATA[Negative control (H2O)]]> 10 20.2±4.0 0.40±0.11 Low-dose group 10 21.1±4.7 0.45±0.17 Medium-dose group 10 29.1±11.1 0.50±0.16 High-dose group 10 30.4±9.9 0.53±0.15

[0055] As shown in Table 7, compared with the negative control group, the phagocytosis rate and phagocytosis index of each dose group increased, and the effect was better with the increase of the dose, indicating that the Lingcan Dan traditional Chinese medicine composition of the present invention can enhance the phagocytosis ability of mouse peritoneal macrophages, significantly improve the phagocytosis rate, and thus enhance the non-specific immune ability of the body.

[0056] 6. Mouse carbon clearance test Each group was given the test substance by gavage once a day for 29 consecutive days. On the 29th day of the experiment, Indian ink diluted 6 times with normal saline was injected into the tail vein of each mouse at a rate of 0.1 mL per 10 g body weight. Immediately after the ink injection, the timing started. At 2 and 10 minutes after the ink injection, 20 μL of blood was taken from the medial canthus venous plexus and added to 2 mL of 0.1% Na2CO3 solution. The optical density value (OD) was measured at a wavelength of 600 nm using a 721 spectrophotometer, with the Na2CO3 solution as the blank control. After the second blood sampling, the mice were immediately sacrificed, and the liver and spleen were taken, blotted dry on the surface of the organs with filter paper, and weighed. The phagocytosis index was calculated. The results are shown in Table 8.

[0057] Table 8 Effects of the test samples on the phagocytosis index in the mouse carbon clearance test (x-±s). Group Number of animals (pcs) Phagocytosis index <![CDATA[Negative control (H2O)]]> 10 6.53±0.55 Low-dose group 10 6.93±0.64 Medium-dose group 10 7.12±0.82 High-dose group 10 7.06±0.60

[0058] As shown in Table 8, the phagocytosis index of each dose group increased compared with the negative control group, especially the phagocytosis index of the medium dose group was the highest. It shows that under the experimental conditions of this experiment, the Lingcan Dan traditional Chinese medicine composition has a significant effect on the phagocytic function of mouse macrophages, effectively enhances the phagocytic activity of macrophages, and enhances the non-specific immune ability.

[0059] 7. Determination of NK cell activity (LDH assay method) Each group was given the test substance by gavage once a day for 33 consecutive days. On the 33rd day of the experiment, the spleen of each mouse was aseptically taken and placed in a small petri dish containing an appropriate amount of sterile Hanks solution. It was gently torn with forceps to make a single cell suspension, filtered through a 200-mesh sieve, washed, counted, and finally the cell concentration was adjusted to 2×10 7cells / mL. The YAC-1 cells (target cells) were subcultured 24 hours before the experiment. Before use, they were washed 3 times with Hanks solution, and the cell concentration was adjusted to 4×105 cells / mL with RPMI1640 complete culture medium. Take 100 μL each of the target cells and spleen cell suspension (effector cells) (effector-to-target ratio 50:1), and add them to a U-shaped 96-well culture plate. Add 100 μL each of the target cells and culture medium to the natural release wells of the target cells, and add 100 μL each of the target cells and 2.5% Triton to the maximum release wells of the target cells. Incubate in a 37°C, 5% CO2 incubator for 4 hours, centrifuge, and aspirate 100 μL of the supernatant from each well into a flat-bottom 96-well culture plate. At the same time, add 100 μL of LDH substrate solution, react for 8 minutes, add 30 μL of 1 mol / L HCl to each well, and measure the optical density (OD) value at a wavelength of 490 nm to calculate the NK cell activity. The results are shown in Table 9.

[0060] Table 9 Effects of the test samples on the NK cell activity of mice (x-±s). Group Number of animals (pcs) NK cell activity (%) <![CDATA[Negative control (H2O)]]> 10 45.9±2.8 Low-dose group 10 46.7 ± 5.3 Medium-dose group 10 42.1±6.1 High-dose group 10 42.8±6.3

[0061] As shown in Table 9, compared with the negative control group, the NK cell activity value in the low-dose group was higher than that in the negative control group, and the effect on NK cell activity was significant. Therefore, it is indicated that the Lingcan Dan traditional Chinese medicine composition of the present invention can effectively enhance immunity.

[0062] 8. Organ / body weight ratio Each group was given the test substance by gavage once a day for 29 consecutive days. On the 29th day of the experiment, the animals were sacrificed, and their thymus and spleen were taken and weighed to calculate the thymus / body weight and spleen / body weight ratios. The results are shown in Table 10.

[0063] Table 10 Effects of the test samples on the organ / body weight ratio of mice (x-±s). Group Number of animals (pcs) Thymus / body weight (mg / g) Spleen / body weight (mg / g) <![CDATA[Negative control (H2O)]]> 10 2.04±0.27 4.22±0.64 Low-dose group 10 2.38±0.32 4.41±0.60 Medium-dose group 10 2.10±0.40 4.59±1.17 High-dose group 10 2.10±0.40 4.34±0.85

[0064] As shown in Table 10, compared with the negative control group, the thymus / body weight and spleen / body weight ratios of mice in each dose group increased, and the effect in the low-dose group was the most significant, indicating that the Lingcan Dan traditional Chinese medicine composition of the present invention helps to maintain or enhance the function of the immune system and has the effect of regulating and enhancing the function of immune cells.

[0065] Therefore, in the Lingcan Dan traditional Chinese medicine composition of the present invention, Ganoderma lucidum spores, ginseng, safflower, Gynostemma pentaphyllum, and Salvia miltiorrhiza are used. The combination of each raw material can achieve the effects of strengthening the healthy qi and consolidating the root, replenishing qi and generating essence, balancing the yin, yang, qi, and blood of the human body's zang-fu organs, so that the healthy qi of the human body is taken into account, the immunity is comprehensively enhanced, blood circulation is improved and fatigue resistance is achieved, and it is easy to be absorbed by the human body.

Claims

1. A Lingshendan Chinese medicine composition, characterized in that The invention comprises the following components: 22.5-45 parts of broken wall ganoderma spore powder, 26-45.5 parts of gynostemma pentaphyllum, 21.6-48 parts of salvia miltiorrhiza, 6-18 parts of ginseng, and 6.5-19.5 parts of safflower; Among them, the ratio of broken wall Ganoderma lucidum spore powder and ginseng is (5:4)-(15:2).

2. The Lingshendan Chinese medicine composition according to claim 1, characterized in that The invention also comprises 0.6-2 parts of soluble starch, 0.75-7.5 parts of silicon dioxide and 0.75-7.5 parts of gastric soluble film coating premix.

3. The Lingshendan Chinese medicine composition according to any one of claims 1-2, characterized in that: The invention comprises the following components: 25 parts of broken wall ganoderma spore powder, 35 parts of gynostemma pentaphyllum, 20 parts of salvia miltiorrhiza, 8 parts of ginseng, 8 parts of safflower, 1 part of soluble starch, 1 part of silicon dioxide and 1 part of gastric soluble film coating premix.

4. The Lingshendan Chinese medicine composition according to claim 1, characterized in that Its content of salvianolic acid B ≥ 960 mg / 100 g, total saponins ≥ 2.5 g / 100 g, and ganoderic acid A ≥ 25 mg / 100 g.

5. A method for preparing a Lingshendan Chinese medicine composition, characterized in that: The following steps are involved: S1: Extraction of raw materials: pre-treatment and water extraction of broken spore powder of Ganoderma lucidum, respectively, coarsely crush Gynostemma pentaphyllum, Salvia miltiorrhiza, Ginseng, and Carthamus tinctorius, respectively, extract with water, filter, concentrate, spray dry, and pass through a 60-mesh sieve to prepare extracts for use; S2: Granulation: Put the above extract, soluble starch and 1 / 2 of the prescription amount of silicon dioxide into a fluidized bed granulator, mix them evenly, use purified water as a wetting agent, spray water for boiling granulation, dry until the moisture content is ≤5.0%, and the granules are 20 mesh; S3: Add the remaining 1 / 2 amount of silicon dioxide, mix well, and press into tablets, mix for 8-15 minutes, and press into tablets; S4: Add purified water to the gastric-soluble film coating premix and stir thoroughly to prepare a suspension; coat the plain tablets, control the inlet air temperature at 60°C-80°C, and keep the tablet core temperature at 37-42°C.

6. The method for preparing the Lingshendan Chinese medicine composition according to claim 5, characterized in that: The mixed extraction in S1 is added with 8-20 times of water, the temperature is 90° C.-100° C., and the extraction is performed 2-3 times, each time for 0.5-2 hours.

7. The method for preparing the Lingshendan Chinese medicine composition according to claim 6, characterized in that: The inlet air temperature of the spray drying in S1 is 180°C-205°C.

8. The method for preparing the Lingshendan Chinese medicine composition according to claim 5, characterized in that: The weight of water sprayed into S2 is 35-65% of the weight of the material, and the inlet air temperature is 65°C-85°C.

9. The method for preparing the Lingshendan Chinese medicine composition according to claim 5, characterized in that: The concentration of the suspension in S4 is 10-15%.

10. The method for preparing the Lingshendan Chinese medicine composition according to claim 5, characterized in that: The extraction of the raw materials in S1 can also be carried out by combined extraction: after Gynostemma pentaphyllum, Salvia miltiorrhiza, Ginseng, and Carthamus tinctorius are coarsely chopped separately, broken wall Ganoderma lucidum spore powder, Gynostemma pentaphyllum, Salvia miltiorrhiza, Ginseng, and Carthamus tinctorius are weighed according to the ratio, 8 to 20 times of water are added, mixed and extracted at 90° C.-100° C. for 2 to 3 times, each time for 1 to 2 hours, filtered, concentrated, spray-dried, and processed into an extract at an inlet air temperature of 180° C.-205° C. and sieved through a 60-mesh sieve for standby use.

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