CN110840924A - Application of rhizoma atractylodis stem and leaf extract - Google Patents
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- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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Abstract
The invention belongs to the field of traditional Chinese medicine processing, and particularly relates to an application of an atractylodes rhizome stem leaf extract. The application of the atractylodes rhizome stem and leaf extract comprises the steps of crushing atractylodes rhizome stem and leaf, extracting with a solvent, standing, performing rotary evaporation and the like. The invention realizes the effective utilization of the byproduct in the industrialization process of the traditional Chinese medicine resources, effectively extends the economic industrial chain of the resources and generates good social, economic and ecological benefits.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicine processing, and particularly relates to an application of an atractylodes rhizome stem leaf extract.
Background
Chinese herbal medicine is the material basis and resource guarantee supporting the development of the animal remedy and feed additive industry in our country, it has a long history to be used for prevention and cure of poultry and livestock diseases, in recent years, the industry cluster taking Chinese medicine and natural medicine resources as raw materials develops rapidly, the non-medicinal part and the output of by-products produced in the process of producing medicinal materials and deep processing industrialization are up to hundred million tons, these materials are all not utilized effectively, not only cause the enormous waste of resources, but also cause the serious pollution of the ecological environment.
Atractylodes Lancea (academic name: Atractylodes Lancea (Thunb.) DC.) belongs to Compositae family Atractylodes perennial herb. The rhizome of atractylodes is used as a medicine for activating spleen, is bitter, warm, pungent and strong in property and has the effects of eliminating dampness, eliminating turbidity and relieving pain, and the rhizome of atractylodes contains 5 to 9 percent of volatile oil as a main medicinal component. The non-medicinal parts of the rhizoma atractylodis, such as stems and leaves, are often discarded, and meanwhile, the medicinal components in the stems and leaves of the rhizoma atractylodis are low in content and uncertain in effect, which causes great difficulty in further utilizing the stems and leaves of the rhizoma atractylodis.
Disclosure of Invention
The invention aims to provide application of rhizoma atractylodis stems and leaves, in particular to application in the aspects of inhibiting pathogenic bacteria, resisting oxidation and inhibiting inflammatory factors.
The application of the atractylodes rhizome stem and leaf extract in inhibiting pathogenic bacteria is disclosed, wherein the pathogenic bacteria are escherichia coli CVCC195, escherichia coli CVCC 1515, salmonella CVCC 3377, salmonella ATCC14028 and/or staphylococcus aureus ATCC 43300.
The rhizoma atractylodis stem and leaf extract is applied to antioxidation.
The application of the atractylodes rhizome stem and leaf extract in inhibiting the inflammatory factors is disclosed by the invention, wherein the inflammatory factors comprise IL-1 β and/or IL-6 in an aqueous extract, and the inflammatory factors inhibited by alcohol extraction comprise L-6 and/or TNF- α.
According to the application of the rhizoma atractylodis extract, the rhizoma atractylodis stem and leaf extract is obtained by a method comprising the following steps:
(1) pulverizing stem and leaf of rhizoma Atractylodis to obtain medicinal powder;
(2) adding an extraction solvent into the medicinal powder, and performing ultrasonic extraction, wherein the mass volume ratio of the medicinal powder to the extraction solvent is 1: 40, obtaining an extract by ultrasonic power of 500W, temperature of 50 ℃ and time of 30 min;
(3) standing the extract obtained in the step (2), taking the upper layer solution, centrifuging for 10min at 5000rpm, and taking the supernatant;
(4) and (4) performing rotary evaporation on the supernatant obtained in the step (3), and freeze-drying the supernatant into powder.
According to the application of the rhizoma atractylodis extract, the extraction solvent is water or 95% ethanol solution.
The invention has the beneficial effects that:
the invention extracts the effective components in the stems and leaves of the rhizoma atractylodis to obtain the extract of the stems and leaves of the rhizoma atractylodis, the water/alcohol extract of the stems and leaves of the rhizoma atractylodis has obvious inhibiting effect on pathogenic bacteria such as escherichia coli CVCC195, escherichia coli CVCC 1515, salmonella CVCC 3377, salmonella ATCC14028 and/or staphylococcus aureus ATCC43300, the water/alcohol extract of the stems and leaves of the rhizoma atractylodis has good antioxidation, the water extract of the stems and leaves of the rhizoma atractylodis has good inhibiting effect on inflammatory factors IL-1 β and/or IL-6, and the alcohol extract of the stems and leaves of the rhizoma atractylodis has good inhibiting effect on inflammatory factors L-6 and/or TNF- α.
Detailed Description
Example 1 preparation of aqueous extract of leaves and stems of Atractylodes lancea
(1) Crushing the stems and leaves of the rhizoma atractylodis by a miniature plant crusher, and sieving the crushed stems and leaves of the rhizoma atractylodis by a 100-mesh analysis sieve for later use;
(2) weighing 20g of rhizoma atractylodis stem leaf coarse powder in a conical flask containing 800mL of distilled water, wherein the ultrasonic condition is as follows: performing ultrasonic extraction with ultrasonic power of 500W and temperature of 50 ℃ for 30 min;
(3) standing for 10min after ultrasonic extraction, pouring the upper layer extracting solution into a centrifuge tube, and centrifuging for 10min at 5000 rpm;
(4) and (4) rotatably evaporating the centrifuged supernatant to dryness, freeze-drying the supernatant into powder, and placing the powder into a refrigerator at the temperature of minus 80 ℃ for later use.
Example 2 preparation of alcoholic extract of atractylodes rhizome stem and leaf
(1) Crushing the stems and leaves of the rhizoma atractylodis by a miniature plant crusher, and sieving the crushed stems and leaves of the rhizoma atractylodis by a 100-mesh analysis sieve for later use;
(2) weighing 20g of rhizoma atractylodis stem leaf coarse powder in an erlenmeyer flask containing 800mL of 95% alcohol solution, and performing ultrasonic treatment under the following conditions: performing ultrasonic extraction with ultrasonic power of 500W and temperature of 50 ℃ for 30 min;
(3) standing for 10min after ultrasonic extraction, pouring the upper layer extracting solution into a centrifuge tube, and centrifuging for 10min at 5000 rpm;
(4) and (4) rotatably evaporating the centrifuged supernatant to dryness, freeze-drying the supernatant into powder, and placing the powder into a refrigerator at the temperature of minus 80 ℃ for later use.
The alcohol extracts of the stem and leaf of rhizoma atractylodis, the root and hair of rhizoma atractylodis, the seed coat of astragalus mongholicus, the leaf of astragalus mongholicus, the peel of trichosanthes kirilowii maxim and the ganoderma lucidum fungus stick are prepared by the same method.
Example 3 examination of the efficacy of the extract of the stems and leaves of Atractylodes lancea
3.1 detection of bacteriostatic effect of Atractylodes lancea stem and leaf extract
Extracts of atractylodes rhizome stem and leaf, atractylodes rhizome root hair, astragalus membranaceus seed coat, astragalus membranaceus leaf, astragalus membranaceus seed coat, trichosanthes kirilowii Maxim peel, ganoderma lucidum stick and dictamnus alba fruit peel were prepared as described in examples 1 and 2.
Plate coating method: pulverizing the by-products of Chinese herbal medicine processing, and sieving with 100 mesh sieve. Then, 0.6g of rhizoma atractylodis extract freeze-dried powder which is sieved by a 100-mesh sieve is added into 60mL of MHA culture medium, stirred and uniformly mixed, sterilized for 15min at 121 ℃, adjusted to pH 7.2 +/-0.2, and subpackaged into three drug-adding flat plates. A drug-adding flat plate (the drug concentration is 10mg/mL) is manufactured 24h before the experiment,
taking the pathogenic bacteria liquid in the exponential phase, adjusting the concentration of the bacteria to be 1 multiplied by 108cfu, sucking 50 mu L of the bacteria liquid, uniformly coating the bacteria liquid on three parallel drug adding plates and a control plate, and culturing for 24h at 37 ℃.
TABLE 1 comparison of bacteriostatic ability of Chinese medicine processing waste
As can be seen from table 1, at a concentration of 10mg/mL, the bacteriostatic ability of each waste of traditional Chinese medicine processing is different, for example, the stem and leaf of atractylodes has significant bacteriostatic effect on 5 strains of pathogenic bacteria except s.aureus ATCC 25923; the rhizoma atractylodis root hair is from rhizoma atractylodis as well as the stem and leaf of the rhizoma atractylodis, but only has good inhibition effect on S.aureus ATCC 43300; the ganoderma lucidum fungus stick, the astragalus membranaceus leaves and the astragalus membranaceus seed coat can completely inhibit the growth of S.aureus ATCC 43300; the astragalus membranaceus leaves and the astragalus membranaceus seed coats can also obviously inhibit the growth of S.enterica CVCC 3377.
3.2 detecting the total antioxidant capacity of the rhizoma atractylodis stem and leaf extract
Extracts of atractylodes rhizome stem and leaf, atractylodes rhizome root hair, astragalus membranaceus seed coat, astragalus membranaceus leaf, astragalus membranaceus seed coat, trichosanthes kirilowii Maxim peel were prepared as described in examples 1 and 2.
FeSO provided by 27.8mg kit is weighed4·7H2O, dissolved and diluted to 1mL, and the concentration is 100 mM. Taking a proper amount of 100mM FeSO4The solution was diluted to 0.15, 0.3, 0.6, 0.9, 1.2 and 1.5 mM. Typically, the standard may be formulated using distilled water or a sample formulation solution.
a: to each assay well of a 96-well plate, 180. mu.L of FRAP working solution (TPTZ dilution 150. mu.L + TPTZ solution 15. mu.L + assay buffer 15. mu.L) was added.
b: add 5. mu.L of distilled water to the blank control well; adding 5 mu L of FeSO4 standard solution with various concentrations into the detection holes of the standard curve; mu.L of each sample (200. mu.g/mL) or 0.8mM Trolox was added to the sample wells as a positive control. Mix gently.
c: a593 was determined after incubation at 37 ℃ for 3-5 min. If the A593 measurement is difficult, the measurement can also be carried out in the 585-605nm range.
d: the Total Antioxidant Capacity (TAC) of the sample was calculated from the standard curve. If the absorbance measured by the sample is outside the range of the standard curve, the sample is diluted properly and then measured.
TABLE 2 Total antioxidant capacity of Chinese medicine processing waste
As can be seen from Table 2, the oxidation resistance of the Chinese medicinal processing waste is different under the concentration of 200 mug/mL, wherein the total oxidation resistance of the rhizoma atractylodis stem and leaf extract is higher than that of the rhizoma atractylodis root hair; the antioxidant capacity of the water extract of the stem and leaf of the rhizoma atractylodis is stronger than that of the alcohol extract thereof.
3.3 testing the anti-inflammatory ability of the extract of the stems and leaves of Atractylodes lancea
(1) Preparation of pharmaceutical solutions
Extracts of atractylodes rhizome stem and leaf, atractylodes rhizome root hair, astragalus membranaceus seed coat, astragalus membranaceus leaf, astragalus membranaceus seed coat, trichosanthes kirilowii Maxim peel, ganoderma lucidum stick and dictamnus alba fruit peel were prepared as described in examples 1 and 2.
Weighing water extracts and alcohol extract freeze-dried powders of dictamnus pericarp, rhizoma atractylodis stem leaves, rhizoma atractylodis root hair, astragalus membranaceus seed coat, astragalus membranaceus leaf, pachyrhizus peel and ganoderma lucidum stick 10mg, dissolving with 1mL of serum-free culture medium in a super clean bench, and passing through a membrane for later use;
(2) grouping and dosing
Inoculating the cell suspension in logarithmic growth phase into 96-well plate at cell density of 1.5 × 106each/mL of the cells is 100 muL per well, after 24 hours of adhesion, a blank control group, an LPS group (1mg/mL), LPS + cortex dictamni/rhizoma atractylodis stem leaves/rhizoma atractylodis root hair/astragalus membranaceus diagonal seed coat/astragalus membranaceus diagonal leaves/trichosanthes kirilowii maxim peel/ganoderma lucidum stick (100 mug/mL) group are arranged, 6 repeats are arranged in each group, corresponding drugs are added into each administration group for pretreatment for 2 hours, a basal medium with the same volume is added into the blank control group and the LPS group, after 2 hours, 1mg/mL of LPS is added into the other groups except the blank control group for stimulation for 24 hours, cell supernatants are collected, the NO content is detected by adopting a Griess method, and the levels of cell factors IL-6 and IL-1 β - α are detected by an ELISA method.
TABLE 3 anti-inflammatory action of aqueous extract of waste from Chinese medicine processing
As shown in Table 3, the concentration of each of the processed Chinese medicinal wastes was different from that of the LPS group, for example, the root hairs of Atractylodes lancea were capable of simultaneously inhibiting the production of IL-1 β, IL-6 and TNF- α inflammatory factors, the stem leaves of Atractylodes lancea were capable of simultaneously inhibiting the production of IL-1 β and IL-6 inflammatory factors, the seed coats of Astragalus membranaceus with oblique stems were capable of simultaneously inhibiting the production of IL-6 and TNF- α inflammatory factors, and the pericarpium cucumis melo and the stick of Ganoderma lucidum were capable of simultaneously inhibiting the production of NO, IL-1 β and IL-6 inflammatory factors.
TABLE 4 anti-inflammatory action of alcoholic extract of Chinese medicinal processing waste
As shown in Table 4, the concentration of 100. mu.g/mL is different from that of the traditional Chinese medicine processing waste, for example, compared with LPS group, the pericarp of dictamnus alba, the root hair of rhizoma atractylodis, the seed coat of astragalus membranaceus with oblique stem and the stick of ganoderma lucidum can simultaneously and obviously inhibit the generation of IL-1 β, IL-6 and TNF- α inflammatory factors, the stem and leaf of rhizoma atractylodis can inhibit the generation of NO, IL-6 and TNF- α inflammatory factors, and the peel of pachyrhizus up can simultaneously inhibit the generation of IL-6 and TNF- α inflammatory factors.
Claims (10)
1. The rhizoma atractylodis stem and leaf extract is used for inhibiting pathogenic bacteria, wherein the pathogenic bacteria are escherichia coli CVCC195, escherichia coli CVCC 1515, salmonella CVCC 3377, salmonella ATCC14028 and/or staphylococcus aureus ATCC 43300.
2. The use as claimed in claim 1, wherein the extract of the stem and leaf of atractylodes lancea is obtained by a method comprising the steps of:
(1) pulverizing stem and leaf of rhizoma Atractylodis to obtain medicinal powder;
(2) adding an extraction solvent into the medicinal powder, and performing ultrasonic extraction, wherein the mass volume ratio of the medicinal powder to the extraction solvent is 1: 40, obtaining an extract by ultrasonic power of 500W, temperature of 50 ℃ and time of 30 min;
(3) standing the extract obtained in the step (2), taking the upper layer solution, centrifuging for 10min at 5000rpm, and taking the supernatant;
(4) and (4) performing rotary evaporation on the supernatant obtained in the step (3), and freeze-drying the supernatant into powder.
3. The use according to claim 2, wherein in step (2), the extraction solvent is water or a 95% ethanol solution.
4. The rhizoma Atractylodis stem and leaf extract can be used for resisting oxidation.
5. The use as claimed in claim 4, wherein the extract of the stem and leaf of Atractylodes lancea is obtained by a method comprising the steps of:
(1) pulverizing stem and leaf of rhizoma Atractylodis to obtain medicinal powder;
(2) adding an extraction solvent into the medicinal powder, and performing ultrasonic extraction, wherein the mass volume ratio of the medicinal powder to the extraction solvent is 1: 40, obtaining an extract by ultrasonic power of 500W, temperature of 50 ℃ and time of 30 min;
(3) standing the extract obtained in the step (2), taking the upper layer solution, centrifuging for 10min at 5000rpm, and taking the supernatant;
(4) and (4) performing rotary evaporation on the supernatant obtained in the step (3), and freeze-drying the supernatant into powder.
6. The use according to claim 5, wherein in step (2), the extraction solvent is water or a 95% ethanol solution.
7. Application of rhizoma Atractylodis stem and leaf water extract in inhibiting inflammatory factors, wherein the inflammatory factors include IL-1 β and/or IL-6.
8. The use as claimed in claim 7, wherein the aqueous extract of stem and leaf of atractylodes lancea is obtained by a method comprising the steps of:
(1) pulverizing stem and leaf of rhizoma Atractylodis to obtain medicinal powder;
(2) adding water into the medicinal powder, and performing ultrasonic extraction, wherein the mass volume ratio of the medicinal powder to the water is 1: 40, obtaining an extract by ultrasonic power of 500W, temperature of 50 ℃ and time of 30 min;
(3) standing the extract obtained in the step (2), taking the upper layer solution, centrifuging for 10min at 5000rpm, and taking the supernatant;
(4) and (4) performing rotary evaporation on the supernatant obtained in the step (3), and freeze-drying the supernatant into powder.
9. The application of the alcohol extract of the stem and leaf of the atractylodes lancea in inhibiting inflammatory factors is disclosed, wherein the inflammatory factors comprise L-6 and/or TNF- α.
10. The use as claimed in claim 9, wherein the alcohol extract of the stem and leaf of atractylodes lancea is obtained by a method comprising the steps of:
(1) pulverizing stem and leaf of rhizoma Atractylodis to obtain medicinal powder;
(2) adding the medicinal powder into a 95% ethanol solution, and performing ultrasonic extraction, wherein the mass volume ratio of the medicinal powder to the 95% ethanol solution is 1: 40, obtaining an extract by ultrasonic power of 500W, temperature of 50 ℃ and time of 30 min;
(3) standing the extract obtained in the step (2), taking the upper layer solution, centrifuging for 10min at 5000rpm, and taking the supernatant;
(4) and (4) performing rotary evaporation on the supernatant obtained in the step (3), and freeze-drying the supernatant into powder.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111743845A (en) * | 2020-07-17 | 2020-10-09 | 广州本心生物医药科技有限公司 | Traditional Chinese medicine extract no-clean hand sanitizer and preparation method thereof |
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| CN114152705B (en) * | 2021-12-06 | 2023-11-17 | 中国农业科学院饲料研究所 | HPLC fingerprint quality evaluation method for rhizoma atractylodis stem and leaf |
| CN115895963A (en) * | 2022-11-29 | 2023-04-04 | 内蒙古九禾农业科技发展有限公司 | Preparation method of rhizoma atractylodis stem and leaf extract and application of rhizoma atractylodis stem and leaf extract in promoting proliferation of lactobacillus plantarum |
| CN115895963B (en) * | 2022-11-29 | 2024-05-14 | 内蒙古九禾农业科技发展有限公司 | Preparation method of rhizoma atractylodis stem and leaf extract and application of rhizoma atractylodis stem and leaf extract in promoting proliferation of lactobacillus plantarum |
| CN116942720A (en) * | 2023-09-21 | 2023-10-27 | 畜科生物工程有限公司 | Application of rhizoma Atractylodis extract in inhibiting bacterial biofilm and composition for inhibiting bacterial biofilm formation |
| CN116942720B (en) * | 2023-09-21 | 2023-12-15 | 畜科生物工程有限公司 | Application of rhizoma Atractylodis extract in inhibiting bacterial biofilm and composition for inhibiting bacterial biofilm formation |
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