阅读说明：本技术 黄芪-蝉拟青霉发酵菌质及其用途 (Astragalus root-paecilomyces cicadae fermentation mycoplasm and application thereof ) 是由 张加余 代龙 王少平 耿子凯 王喻淇 于 2019-11-15 设计创作，主要内容包括：本发明公开了一种黄芪-蝉拟青霉发酵菌质及其用途。所述发酵菌质以黄芪药材粉末作为发酵底物,蝉拟青霉作为发酵菌种,通过固体发酵制备得到。本发明的黄芪-蝉拟青霉发酵菌质具有显著的治疗高尿酸血症和高甘油三酯血症的作用。(The invention discloses an astragalus-paecilomyces cicadae fermentation mycoplasm and application thereof. The fermentation mycoplasm is prepared by taking astragalus medicinal material powder as a fermentation substrate and paecilomyces cicadae as a fermentation strain through solid fermentation. The astragalus-paecilomyces cicadae fermentation mycoplasm has obvious effect of treating hyperuricemia and hypertriglyceridemia.)

1. The radix astragali-paecilomyces cicadae fermentation mycoplasm is prepared by taking radix astragali medicinal material powder as a fermentation substrate and paecilomyces cicadae as a fermentation strain through a solid fermentation way.

2. The use according to claim 1, wherein the astragalus-paecilomyces cicadae zymophyte is used for preparing a medicament for treating hyperuricemia complicated with hypertriglyceridemia.

3. The use according to claim 1 or 2, wherein the astragalus-paecilomyces cicadae zymophyte is prepared by the following method:

(1) adding water into the astragalus medicinal powder, uniformly mixing, and sterilizing to obtain an astragalus solid culture medium;

(2) and (2) inoculating the paecilomyces cicadae into the astragalus membranaceus solid culture medium, and culturing for 7-30 days under the conditions that the culture temperature is 24-30 ℃ and the relative humidity is 50-90% to obtain the astragalus membranaceus-paecilomyces cicadae fermentation mycoplasm.

4. The use of claim 3, wherein in the step (1), the granularity of the astragalus root medicinal material powder is 6-20 meshes; the weight ratio of the astragalus medicinal material powder to water is 1-5: 1.

5. The use according to claim 3, wherein in the step (2), the Paecilomyces cicadae is obtained by subjecting a Paecilomyces cicadae strain to activation culture.

6. The use according to claim 5, wherein the activated culture method of the Paecilomyces cicadae strain in the step (2) comprises the following steps: inoculating Paecilomyces cicadae strain on Potato Dextrose Agar (PDA) slant culture medium, placing in a constant temperature and humidity box with 27 deg.C and relative humidity of 80%, activating and culturing for 5 days to obtain activated Paecilomyces cicadae; then, the activated paecilomyces cicadae is picked by an inoculating loop and placed in a potato glucose liquid culture medium at the temperature of 25 ℃ and the temperature of 140 r.min^-1Culturing for 7 days under the condition of a shaking table to obtain the paecilomyces cicadae and a seed solution.

7. The use as claimed in claim 6, wherein in the step (2), the inoculation amount of the paecilomyces cicadae is 5-20 wt% of the weight of the astragalus root medicinal material powder.

8. The use as claimed in claim 7, wherein, in the step (2), the paecilomyces cicadae seed liquid is also inoculated in the astragalus membranaceus solid culture medium.

9. The use according to claim 3, wherein in the step (2), the culture temperature is 26-28 ℃, the relative humidity is 70-90%, and the culture time is 20-24 days.

10. An astragalus-paecilomyces cicadae zymophyte prepared by the preparation method according to any one of claims 3 to 9.

Technical Field

The invention relates to a traditional Chinese medicine fermentation mycoplasm and application thereof, in particular to astragalus-paecilomyces cicadae fermentation mycoplasm and application thereof in preparing a medicine for treating hyperuricemia and/or hypertriglyceridemia.

Background

The traditional Chinese medicine fermentation is characterized in that the original performance of the medicine is changed, new effects are enhanced or generated, and the medicine variety is expanded through the fermentation process under certain environmental conditions (such as temperature, humidity, air, moisture and the like) by means of the action of enzyme and microorganism so as to adapt to the requirements of clinical medication. Traditional Chinese medicine fermentation is a microbial fermentation technology, and refers to a method for foaming a medicine to generate a yellowish-white mildew coat by virtue of the catalytic decomposition action of microorganisms and enzymes under certain temperature and humidity conditions after the medicine is cleaned or treated. Because the variety and the number of strains participating in fermentation in traditional Chinese medicine fermentation are influenced by geographical environment and seasonal variation, and the fermentation process is judged and controlled by human subjective experience, the safety, the effectiveness, the stability and the controllability of the fermented Chinese medicine product are difficult to ensure.

The traditional Chinese medicine fermentation is gradually changed from the traditional natural fermentation to the modern traditional Chinese medicine fermentation. The latter can monitor the fermentation process in real time, has stronger pertinence and is more beneficial to the deep research on the fermentation mechanism of the traditional Chinese medicine. Meanwhile, the novel fermentation technology utilizes traditional Chinese medicinal materials as medicinal substrates to perform solid or liquid fermentation by applying certain fungi, and the chemical components and the content of the medicinal materials are changed by various enzymes and metabolites generated by the fungi in the fermentation process, so that the function of modifying the original medicinal materials is realized. The novel bidirectional fermentation has the advantages of low cost and easy availability of equipment, wide raw material source, simple experimental operation, simple fermentation method, high economic benefit, short production period and the like, and can realize large-scale industrialized production in practical application.

Radix astragali is a common traditional Chinese medicine and has the effects of tonifying qi and invigorating yang, consolidating superficial resistance and arresting sweating, inducing diuresis and relieving swelling, and the like. Regarding modern fermentation research of astragalus, quantan and the like (anti-hyperuricemia activity of astragalus residue fermentation product, Jiangsu agricultural science [ J ], vol.41, No. 9, page 288-. Zhao Chongyan and the like (establishment of a astragalus-paecilomyces cicadae bidirectional fermentation system and component research, world traditional Chinese medicine, 13 th 12 th of 12 months in 2018, 3195 page 3198) establish a bidirectional fermentation system of an astragalus-paecilomyces cicadae medicinal material, and find that after fermentation, the contents of polysaccharide and total saponin in the mycoplasm of the astragalus medicinal material are reduced, and the content of flavone is increased.

At present, no research report on the pharmacological activity of the astragalus-paecilomyces cicadae zymophyte is found.

Disclosure of Invention

The invention aims to provide astragalus-paecilomyces cicadae fermentation mycoplasm and a novel pharmaceutical application thereof.

The purpose of the invention is realized by the following technical scheme.

The invention provides application of astragalus-paecilomyces cicadae fermentation mycoplasm in preparing a medicine for treating hyperuricemia and/or hypertriglyceridemia, wherein the astragalus-paecilomyces cicadae fermentation mycoplasm is prepared by taking astragalus medicinal material powder as a fermentation substrate and paecilomyces cicadae as a fermentation strain through a solid fermentation way.

In the invention, the astragalus-paecilomyces cicadae fermentation mycoplasm is preferably used for preparing a medicine for treating hyperuricemia complicated with hypertriglyceridemia.

In the invention, preferably, the astragalus-paecilomyces cicadae fermentation mycoplasm is prepared by the following method:

(1) adding water into the astragalus medicinal powder, uniformly mixing, and sterilizing to obtain an astragalus solid culture medium;

(2) and (2) inoculating the paecilomyces cicadae into the astragalus membranaceus solid culture medium, and culturing for 7-30 days under the conditions that the culture temperature is 24-30 ℃ and the relative humidity is 50-90% to obtain the astragalus membranaceus-paecilomyces cicadae fermentation mycoplasm.

In the invention, in the step (1), the granularity of the astragalus root medicinal material powder is preferably 6-20 meshes, more preferably 8-14 meshes, and further preferably 8 meshes. The granularity of the astragalus medicinal material powder is favorable for the growth of paecilomyces cicadae.

In the invention, preferably, in the step (1), the weight ratio of the astragalus membranaceus medicinal material powder to water is 1-5: 1, more preferably 1.5-3: 1, and still more preferably 2-2.5: 1. By adopting the proportion, the inoculated paecilomyces cicadae and the astragalus medicinal material powder are in full contact, and a better fermentation effect is obtained. Particularly, the weight ratio of the astragalus membranaceus medicinal material powder to water is 2-2.5: 1, the obtained astragalus membranaceus solid culture medium is more suitable for growth of paecilomyces cicadae, and the growth rate of the paecilomyces cicadae and the yield of astragalus membranaceus-paecilomyces cicadae fermentation mycoplasm are higher.

In the step (1), the sterilization operation may employ conventional sterilization conditions, and is not particularly limited. According to one embodiment of the present invention, the sterilization operation is sterilization in an autoclave at 90 ℃ for 60 min.

In the present invention, preferably, in step (2), the paecilomyces cicadae is obtained by subjecting a paecilomyces cicadae strain to activation culture. The paecilomyces cicadae strain is a known strain, and is preferably a strain with the number of cfcc81169 preserved by China forestry microorganism preservation center. The activation culture method may be any activation method conventionally used in the art, and is not limited. According to one embodiment of the present invention, the activation culture method is: inoculating Paecilomyces cicadae strain on Potato Dextrose Agar (PDA) slant culture medium, placing in a constant temperature and humidity box with 27 deg.C and relative humidity of 80%, activating and culturing for 5 days to obtain activated Paecilomyces cicadae; then, the activated paecilomyces cicadae is picked by an inoculating loop and placed in a potato glucose liquid culture medium at the temperature of 25 ℃ and the temperature of 140 r.min^-1Culturing for 7 days under the condition of a shaking table to obtain the paecilomyces cicadae and the paecilomyces cicadae seed liquid. The obtained paecilomyces cicadae can be directly inoculated in the astragalus membranaceus solid culture medium.

In the invention, in the step (2), the inoculation amount of the paecilomyces cicadae is preferably 5-20 wt%, more preferably 8-15 wt%, and still more preferably 8-12 wt% of the astragalus membranaceus medicinal material powder. By adopting the inoculation amount, the growth rate of the paecilomyces cicadae and the yield of the astragalus-paecilomyces cicadae fermentation mycoplasm are higher, and the using amount of the paecilomyces cicadae is saved.

In the invention, preferably, in the step (2), the culture temperature is 25-29 ℃, and more preferably 26-28 ℃; the relative humidity is 60-90%, and more preferably 70-90%. The culture time is preferably 14 to 24 days, and more preferably 20 to 24 days. According to a preferred embodiment of the invention, the culture temperature is 26-28 ℃, the relative humidity is 70-90%, and the culture time is 20-24 days. The culture condition can achieve a better fermentation effect. The cultivation can be carried out in a constant temperature and humidity incubator.

In the present invention, preferably, in the step (2), the paecilomyces cicadae seed liquid is also inoculated in the astragalus membranaceus solid medium.

The invention also provides the astragalus-paecilomyces cicadae fermentation mycoplasm prepared by the preparation method. The content of the astragalus-paecilomyces cicadae fermentation mycoplasm polysaccharide is obviously higher than that of the astragalus medicinal material.

Compared with the astragalus root medicinal material, the astragalus root-paecilomyces cicadae fermentation mycoplasm has the advantages that the polysaccharide content is obviously improved, and the astragalus root-paecilomyces cicadae fermentation mycoplasm has the obvious effect of treating hyperuricemia and/or hypertriglyceridemia.

Drawings

FIG. 1 is a graph comparing the levels of UA in serum from hyperuricemia rats.

FIG. 2 is a graph showing a comparison of serum BUN levels in hyperuricemia rats.

FIG. 3 is a graph comparing serum CRE levels in hyperuricemia rats.

FIG. 4 is a graph comparing the levels of TG in hyperuricemia rats.

Note: indicated very significant difference (P) compared to normal group<0.001) indicating significant difference (P)<0.01); in comparison to the set of models,⁺⁺⁺shows a very significant difference (P)<0.001)，⁺⁺Indicates significant difference (P)<0.01)，⁺Indicates that there is a statistical difference (P)<0.05)。

Detailed Description

The invention provides a new pharmaceutical application of astragalus-paecilomyces cicadae fermentation mycoplasm, and the following embodiment of the invention is specifically explained.

The following further illustrates embodiments of the invention by way of specific examples.

In the following examples, the Paecilomyces cicadae strain is provided by China forestry microbiological Collection center, with the number cfcc 81169. The paecilomyces cicadae strain is inoculated on a Potato Dextrose Agar (PDA) slant culture medium and is placed in a constant temperature and humidity box with the temperature of 27 ℃ and the relative humidity of 80 percent for activation culture for 5 days to obtain the activated paecilomyces cicadae. The activated paecilomyces cicadae is picked by an inoculating loop and placed in a potato glucose liquid culture medium at the temperature of 25 ℃ and the temperature of 140 r.min^-1Culturing for 7 days under the condition of a shaking table to obtain the paecilomyces cicadae and the paecilomyces cicadae seed liquid.

Reagent: potassium oxonate (Potasisonate) was purchased from Sigma, USA; benzbromarone tablets (Narcarin) were purchased from Excella GmbH, germany. The calibration solution, the quality control solution and the detection kit are all purchased from Beijing Lidemann Biochemical technology GmbH. Triglyceride assay kit, batch number: 112293K. Uric acid assay kit, batch number: 00275. urea nitrogen assay kit, batch No.: 01049; creatinine assay kit, lot No.: 04210.

animals: sprague Dawley (SD) rats, male, with a body mass of 220-250 g, purchased from Beijing Wintolite laboratory animal technology, Inc., with a license number of SCXK (Jing) 2016-.

The instrument comprises the following steps: the CX-4Pro type full-automatic biochemical analyzer is a product of Beckman company in America; an electronic analytical balance (one hundred thousand) model R200D is a product of Sartorius, germany; the Millipore Synergy UV type ultra-pure water machine is a product of Millipore company in America; the BXM-30R high-pressure steam sterilization pot is a product of laboratory instruments and equipment Limited of the Xian apparatus; the UV-2000 ultraviolet visible spectrophotometer is a product of Beijing Rayleigh analysis instruments company; the FW-100 high-speed pulverizer is a product of Tensted instruments, Inc. of Tianjin; the LHS-80 HC-II type constant temperature and humidity incubator is a product of Shanghai-constant technology limited company.

The fermentation culture of the following examples 1-2 and comparative examples 1-3 is performed in parallel, and the used radix astragali material, the amount thereof, and the operation method of the culture process are completely the same.