Preparation method and application of dendrobe salt roasted product

Abstract

The invention discloses a preparation method and application of a dendrobe salt roasted product, and belongs to the technical field of traditional Chinese medicines. The preparation method comprises the following steps: drying fresh dendrobium huoshanense strips serving as a raw material, adding saline water for moistening until the core of the dendrobium huoshanense is completely penetrated, and frying to obtain the salt roasted dendrobium huoshanense product. According to the preparation method, a single factor is combined with a Box-Behnken response surface method, the contents of main medicinal components including total polysaccharides, total flavonoids, alcohol-soluble extracts and water-soluble extracts of dendrobium huoshanense are taken as evaluation indexes, comprehensive scoring is carried out by combining an entropy weight method, and the preparation method of the dendrobium huoshanense salt roasted product is obtained through optimization. The quality of the dendrobium huoshanense salt honey-fried product obtained by the method is obviously improved, the total polysaccharide content and the alcohol-soluble extract content of the dendrobium huoshanense salt honey-fried product exceed 12.82% and 51.25% in the specification of pharmacopeia respectively, and the effect of improving hyperthyroidism yin deficiency is also obviously improved.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine, and in particular to a preparation method and application of a salt-roasted dendrobium product. Background Art

[0002] Dendrobium has a long history of use and a variety of processing methods. However, due to its scarcity and extremely high economic value, it is over-exploited and its resources are on the verge of extinction. Many traditional methods of processing Dendrobium are close to being lost, and the application of traditional efficacy is also missing. The current 2020 edition of the "Chinese Pharmacopoeia" does not include any processed Dendrobium products, and the market lacks deep-processed products processed by ancient methods. After investigation, the ancient books on the salt-roasted processing method of Dendrobium are simple. The Materia Medica records that "stir-fried Dendrobium with salt water" can "tonify the kidney and clear kidney fire". The ancient and modern medical records contain salt-water cultivation of Dendrobium. The Dendrobium in Volume 3 Guipi Decoction needs to be fried in salt water before being used as medicine, but the specific usage is not indicated in the ancient books, and there is a lack of regulations on specific processing process parameters. By verifying the salt-roasting methods of other medicinal materials, it was found that the ancient books did not indicate the specific usage and dosage of salt-roasted medicinal materials, and most of them were simply fried in salt water. Moreover, there are few studies on the preparation of Huoshan Dendrobium, a local medicinal material.

[0003] Therefore, conducting research on the ancient method of salt-roasting of Dendrobium is of great significance to fully exert the important effects of Huoshan Dendrobium. Summary of the invention

[0004] The purpose of the present invention is to provide a preparation method and application of salt-baked dendrobium products to solve the problems existing in the above-mentioned prior art. The quality of the salt-baked dendrobium products obtained by the preparation method of the present invention is significantly improved, and its total polysaccharide content and alcohol-soluble extract content exceed the pharmacopoeia requirements by 12.82% and 51.25%, respectively, and the effect of improving hyperthyroidism and yin deficiency is also significantly improved.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a method for preparing a salt-roasted Huoshan dendrobium, comprising the following steps:

[0007] The fresh strips of Huoshan Dendrobium are dried as raw materials, added with salt water to moisten them until they are completely cooked, and then fried to obtain the salt-roasted Huoshan Dendrobium.

[0008] Optionally, the fresh strips are three-year-old fresh strips of Dendrobium huoshanense.

[0009] Optionally, the drying temperature is 60° C. and the drying time is 25 hours.

[0010] Optionally, when adding salt water for moistening, based on the mass of the dried Dendrobium huoshanense strips and NaCl, salt water containing 4.6 g NaCl is added for every 100 g of the dried Dendrobium huoshanense strips.

[0011] Optionally, the ratio of the mass of the dried Dendrobium huoshanense strips to the volume of the added brine is 1 g:10 mL.

[0012] Optionally, the steaming time is 18 hours.

[0013] Optionally, the frying temperature is 100° C. and the frying time is 8 minutes.

[0014] The present invention also provides a Huoshan dendrobium salt-roasted product obtained by the preparation method.

[0015] The present invention also provides application of the Huoshan Dendrobium salt-roasted product in preparing a medicine for improving the Yin deficiency syndrome of hyperthyroidism.

[0016] The present invention also provides the use of the Huoshan Dendrobium salt-roasted product in the preparation of a medicine for improving body disorders caused by hyperthyroidism Yin deficiency syndrome.

[0017] The present invention discloses the following technical effects:

[0018] The present invention adopts a single factor combined with the Box-Behnken response surface method, takes the total polysaccharide, total flavonoids, alcohol-soluble extract and water-soluble extract contents of the main medicinal components of Huoshan Dendrobium as evaluation indicators, combines the entropy weight method for comprehensive scoring, and optimizes the preparation method of a Huoshan Dendrobium salt-fried product. The quality of the Huoshan Dendrobium salt-fried product obtained by the method is significantly improved, and the total polysaccharide content and the alcohol-soluble extract content exceed the pharmacopoeia regulations by 12.82% and 51.25%, respectively. The Huoshan Dendrobium salt-fried product obtained by the preparation process of the present invention is also significantly improved in improving hyperthyroidism and yin deficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 The pictures of samples obtained by different salt-roasting methods in Example 1, wherein A is the salt-roasted product A of Dendrobium officinale from Huoshan obtained by method 1 (first add salt water to moisten and then stir-fry); B is the salt-roasted product B of Dendrobium officinale from Huoshan obtained by method 2 (first stir-fry to dry and then spray salt water while stir-frying);

[0021] Figure 2 It is the comprehensive score of Huoshan Dendrobium salt-roasted products calculated by entropy weight method under different salt addition amounts;

[0022] Figure 3 It is the comprehensive score of Huoshan Dendrobium salt-fried products at different frying temperatures calculated by entropy weight method;

[0023] Figure 4 It is the comprehensive score of Huoshan Dendrobium salt-fried products at different frying times calculated by entropy weight method;

[0024] Figure 5 It is a three-dimensional response surface diagram of the quadratic term model of the independent variables of salt addition, frying temperature and frying time and the dependent variable comprehensive score, among which a is the influence diagram of salt addition and frying temperature on the comprehensive score; b is the influence diagram of frying temperature and frying time on the comprehensive score; c is the influence diagram of salt addition and frying time on the comprehensive score;

[0025] Figure 6 The results of organ index experiments of thymus (A), spleen (B), kidney (C), and liver (D) of mice in different groups;

[0026] Figure 7 The experimental results of cyclic nucleotide levels of mice in different groups are shown in Figure 1, where A is the level of cyclic guanosine monophosphate (cGMP); B is the level of cyclic adenosine monophosphate (cAMP); C is the ratio of cyclic adenosine monophosphate / cyclic guanosine monophosphate (cAMP / cGMP);

[0027] Figure 8 The results of the thyroid function test of mice in different groups are shown in Figure 1, where A is the T3 content level; B is the T4 content level;

[0028] Fig. 9 The experimental results of the serum indicators corticosterone (CORT) and creatinine (CRE) levels of mice in different groups, where A is the CORT level; B is the CRE level;

[0029] Fig.10 The results of ATPase activity in renal tissue of mice in different groups, where A is Na + -K + -ATPase activity, B is Ca 2+ -Mg 2+ -ATPase activity;

[0030] Figure 7-10 In the table, * indicates p<0.05; ** indicates p<0.01; *** indicates p<0.001; **** indicates p<0.0001. DETAILED DESCRIPTION

[0031] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0032] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0033] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0034] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0035] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0036] Example 1 Preparation method and process optimization of Huoshan Dendrobium salt-roasted product

[0037] 1. Preparation method of Huoshan Dendrobium salt-roasted

[0038] Pick three-year-old fresh strips of Huoshan Dendrobium, wash and remove impurities, and then put them into a forced air drying oven at 60℃ for 25h to obtain the dried Huoshan Dendrobium. Take 10g of the dried Huoshan Dendrobium, add 10mL of a mixed salt solution according to a material-liquid ratio of 1:1, and the amount of salt added is 2% (that is, 2gNaCl is added to 100g of Huoshan Dendrobium medicinal materials). After being steamed for 18h, take out and stir-fry (a concave induction cooker is used for frying). The frying temperature is 80℃ and the frying time is 8min. After frying, take out and cool, sieve out the debris, and the Huoshan Dendrobium salt-roasted product is obtained.

[0039] 2. Process optimization of the preparation method of Huoshan Dendrobium salt-roasted

[0040] In order to optimize the salt-roasting process, single-factor experiments were conducted to investigate the effects of salt-roasting method, drying time, steaming time, material-liquid ratio, salt addition amount, frying temperature and frying time on the quality of salt-roasted Dendrobium officinale from Huoshan. The contents of total polysaccharides, total flavonoids and water-soluble / alcohol-soluble extracts of Dendrobium officinale were used as evaluation indicators, and a comprehensive score was given in combination with the entropy weight method. The response surface methodology was used to optimize the optimal processing technology, and process verification was carried out.

[0041] 2.1 Entropy weight method to determine the weight of each indicator

[0042] The entropy weight method is an objective value assignment method. It measures the effect of each indicator on the comprehensive indicator based on the dispersion of each indicator, such as polysaccharide, alcohol-soluble extract, and water-soluble extract, so as to determine the weight of each indicator. The calculation is as follows:

[0043] First, we establish an initial matrix consisting of i samples and j indicators (1):

[0044]

[0045] Then, the indicator values ​​are dimensionlessly standardized according to algorithm (2):

[0046]

[0047] According to algorithm (3), the weight of the i sample values ​​under this indicator is calculated:

[0048]

[0049] Calculate the index entropy value according to algorithm (4):

[0050]

[0051] Calculate the information entropy redundancy according to algorithm (5):

[0052] d j =1-e j (5)

[0053] Finally, the weight values ​​of each indicator are calculated according to algorithm (6):

[0054]

[0055] The final calculated weights of total polysaccharides, total flavonoids, alcohol-soluble extracts and water-soluble extracts were 0.35, 0.37, 0.144 and 0.135, respectively. Therefore, the comprehensive score (Y) = total polysaccharide content × 0.35 + total flavonoids content × 0.37 + alcohol-soluble extract content × 0.144 + water-soluble extract content × 0.135.

[0056] 2.2 Single factor investigation

[0057] 2.2.1 Salt roasting method

[0058] In order to explore the salt-roasting method suitable for Huoshan Dendrobium, the samples were prepared by first adding salt water to moisten and then frying and roasting, and by spraying salt water while frying. The specific operations are as follows:

[0059] Method 1 (add salt water to moisten first and then stir-fry and roast): Same as “1. Preparation method of Huoshan Dendrobium Salt-Roasted Product”, to obtain Huoshan Dendrobium Salt-Roasted Product A.

[0060] Method 2 (fry until dry and then spray with salt water while frying): pick three-year-old fresh strips of Huoshan Dendrobium, wash and remove impurities, weigh 10g of fresh strips, put them in a frying container (a concave induction cooker is used for frying), the frying temperature is 80°C, fry until the Dendrobium is slightly yellow, slightly dry and loose in texture, then add salt water and fry, the process of adding salt water is to spray and add 2mL of salt water every 30s, the amount of salt added is 2% (that is, 2gNaCl is added to 100g of Huoshan Dendrobium medicinal material), the total amount of salt water is 10mL, take out and cool after frying, sieve out the debris, and obtain Huoshan Dendrobium salt-roasted product B.

[0061] The salt-roasted products of Dendrobium huoshanense obtained by two methods are as follows Figure 1 As shown in the figure, since the surface of the fresh Huoshan Dendrobium is dense, the flesh is thick, and the polysaccharide content is high, it is not easy to fry dry. Method 2 greatly prolongs the entire frying time. In addition, salt water is sprayed while frying. The salt water evaporates and crystallizes instantly when it meets the hot pan, and cannot be evenly covered on the medicinal materials, nor can it be completely absorbed by the medicinal materials, which easily causes the Dendrobium to become mushy when fried.

[0062] The samples prepared by two different salt-roasting methods were tested separately, and the contents of total polysaccharides, total flavonoids, and water-soluble / alcohol-soluble extracts were used as evaluation indicators. The results are shown in Table 1.

[0063] Table 1 Investigation results of different salt-roasting methods

[0064]

[0065] Judging from the comprehensive scoring results, the method of first steaming with salt water and then frying is better than spraying salt water while frying. Considering the difficulty of the frying process, the method of first steaming with salt water and then frying is chosen.

[0066] 2.2.2 Pretreatment of medicinal materials - Investigation of drying time

[0067] The method of first adding salt water to moisten the herbs and then stir-frying and roasting means that the fresh Huoshan Dendrobium strips need to be dried first to moisten them. In order not to destroy the chemical composition of the medicinal materials and to shorten the drying time as much as possible, the drying temperature is selected to be 60°C. After the medicinal materials are sent into the oven, the status of the Dendrobium is observed and recorded every 1 hour. The herbs are taken out when the whole plant is easy to break. It has been found that the fresh Dendrobium strips meet the experimental standards after 25 hours of drying.

[0068] Table 2 Investigation results of salt roasting method

[0069]

[0070] 2.2.3 Material-liquid ratio

[0071] The amount of brine used during steaming was investigated, with the main criteria being whether the medicinal materials were completely steamed and the brine was completely absorbed, and the difficulty of frying. The results showed that when the amount of Dendrobium was 10 g and the amount of brine was 10 mL, the material-liquid ratio was 1:1, which was the best (Table 3).

[0072] Table 3 Investigation results of material-liquid ratio

[0073]

[0074] Note: In different material-liquid ratios, the amount of salt used is 2% of the amount of medicinal materials, that is, different amounts of brine contain 0.2g NaCl.

[0075] 2.2.4 Stuffing time

[0076] Put 10g of dried Dendrobium huoshanense into a beaker, add 10mL of saline (containing a total of 0.2g NaCl), and observe the degree of penetration every two hours. When the Dendrobium huoshanense is completely penetrated and the weight of the medicinal material does not increase significantly, record the steaming time at this time.

[0077] The results showed that after 18 hours of steaming, the core of the Huoshan Dendrobium was completely penetrated and the weight no longer increased significantly, so the steaming time was determined to be 18 hours (Table 4).

[0078] Table 4 Investigation results of the moistening time

[0079]

[0080] 2.2.5 Salt Amount

[0081] Take 4 portions of 10 g of Huoshan Dendrobium medicinal materials, add 10 mL of saline mixed solution to each portion, with the salt amounts of 2%, 4%, 6%, and 8% respectively (i.e., add 2g, 4g, 6g, and 8g NaCl to 100g of Huoshan Dendrobium medicinal materials), mix well, stir frequently for 18 hours, then take out and stir-fry (use a concave induction cooker for stir-frying), the frying temperature is 80°C, the frying time is 8 minutes, take out and cool, sieve out the debris, and the Huoshan Dendrobium salt-roasted product is obtained.

[0082] Each sample was ground into powder and mixed by a ball mill and then passed through a No. 3 sieve. The sample powder was taken to determine the contents of total polysaccharides, total flavonoids, water-soluble extracts and alcohol-soluble extracts in turn, and the comprehensive score Y value was calculated by the entropy weight method. The results are as follows: Figure 2 shown.

[0083] 2.2.6 Stir-frying temperature

[0084] Take 4 portions of 10 g of Huoshan Dendrobium medicinal materials, add 10 mL of saline mixed solution to each portion, with the salt amount being 2% (i.e., add 2 g NaCl to 100 g of Huoshan Dendrobium medicinal materials), mix well and simmer for 18 hours, then take out and stir-fry (use a concave induction cooker for stir-frying), select 60°C, 80°C, 100°C, and 120°C for 8 minutes, take out and let cool, sieve out the debris, and obtain the Huoshan Dendrobium salt-roasted product.

[0085] Each sample was ground into powder and mixed by a ball mill and then passed through a No. 3 sieve. The sample powder was taken to determine the contents of total polysaccharides, total flavonoids, water-soluble extracts and alcohol-soluble extracts in turn, and the comprehensive score Y value was calculated by the entropy weight method. The results are as follows: Figure 3 shown.

[0086] 2.2.7 Stir-frying time

[0087] Take 4 portions of 10 g of Huoshan Dendrobium medicinal materials, add 10 mL of saline mixed solution to each portion, with the salt amount added being 2% (i.e., add 2 g NaCl to 100 g of Huoshan Dendrobium medicinal materials), mix well and stew for 18 hours, then take out and stir-fry (use a concave induction cooker for stir-frying), select a stir-frying temperature of 80°C, and the stir-frying times are 4 min, 8 min, 12 min, and 16 min respectively. Take out and let cool, sieve out the debris, and the Huoshan Dendrobium salt-roasted product is obtained.

[0088] Each sample was ground into powder and mixed by a ball mill and then passed through a No. 3 sieve. The sample powder was taken to determine the contents of total polysaccharides, total flavonoids, water-soluble extracts and alcohol-soluble extracts in turn, and the comprehensive score Y value was calculated by the entropy weight method. The results are as follows: Figure 4 shown.

[0089] 3. Response surface optimization process

[0090] The results of the previous single-factor investigation experiment showed that when other factors were consistent, the salt addition amount was 6%, the frying temperature was 100℃, and the frying time was 12min, and the comprehensive score of the salt-roasted Huoshan Dendrobium reached the highest level. Therefore, 6%, 100℃, and 12min were selected as the center points of the salt addition amount, frying temperature, and frying time in the response surface, respectively.

[0091] The response surface method of design expert 13 software was used to rationally design a 3-factor 3-level experiment. The salt addition amount (%), frying temperature (℃), and frying time (min) were used as the investigation factors, and each factor was selected into 3 levels: low, medium, and high, which were -1, 0, and 1, respectively. The factor and level design are shown in Table 5.

[0092] The Box-Behnken response surface method was used to design and arrange a total of 17 groups of experiments. The specific arrangements and results of the experiments are shown in Table 6.

[0093] Table 5. Response surface test factors and levels of Huoshan Dendrobium salt roasting process

[0094]

[0095] Table 6 Response surface experiment arrangement and results

[0096]

[0097]

[0098] Design expert 13 was used to perform polynomial regression analysis on the data in Table 6, and the quadratic regression equation of the independent variables salt addition (A), frying temperature (B), frying time (C) and the dependent variable comprehensive score (Y) was obtained as follows: Y = 13.52 + 0.4078 × A-2.36 × B-0.9573 × C + 0.3981 × AB + 0.8231 × AC-1.27 × BC-0.3185 × A 2 -2.59×B 2 +0.3218×C 2 . The results of variance analysis are shown in Table 7. The model p-value is <0.01, and the p-value of the lack of fit term is >0.05, indicating that the quadratic term model can better reflect the functional relationship between the three factors and the comprehensive score value, and can be used for the process prediction and optimization of salt-roasted Huoshan Dendrobium. The p-values ​​of the single factors are all less than 0.05, and the degree of influence on the process of salt-roasted Huoshan Dendrobium is B>C>A. It is known that the frying temperature has the greatest influence on the process of salt-roasted Huoshan Dendrobium. The p-values ​​of AC and BC items are <0.01, indicating that there is an interaction between the amount of salt added (A) and the frying time (C), and between the frying temperature (B) and the frying time (C).

[0099] Table 7 Response surface variance analysis results

[0100]

[0101]

[0102] 4. Process prediction

[0103] When any one of the three factors, namely, the amount of salt, frying temperature and frying time, is fixed, the influence of the other two factors on the comprehensive score is investigated. The three-dimensional response surface diagram constructed according to the obtained model is shown in the figure below. Figure 5 The maximum value of the comprehensive score corresponds to the highest point of the response surface, and the center point of the contour line is the optimal value of the process. The best salt-roasting process obtained by software fitting is 4.6% salt addition, 96℃ frying temperature, and 8min frying time. The predicted comprehensive score under this condition is 14.99.

[0104] Combined with the parameters predicted by the software and the applicability of actual operation, three samples of Dendrobium huoshanense were taken and processed according to the process of adding salt 4.6%, frying temperature 100℃, and frying time 8min. The results show that the contents of total polysaccharides, total flavonoids, alcohol-soluble extracts, and water-soluble extracts are shown in Table 8. The average comprehensive score is 14.92, and the RSD is 0.5%, which is close to the predicted value of the software fitting, indicating that the salt-roasting process of Dendrobium huoshanense optimized by Box-Behnken response surface method has good stability and good application value.

[0105] Table 8 Process verification results

[0106]

[0107] According to the 2020 edition of the "Chinese Pharmacopoeia", the alcohol-soluble extract of Huoshan Dendrobium shall not be less than 8%, and the total polysaccharide content shall not be less than 17%. Table 8 shows that the average contents of total polysaccharides, total flavonoids, alcohol-soluble extracts and water-soluble extracts of the salt-roasted Huoshan Dendrobium obtained after the process optimization in this embodiment are 19.18%, 1.35%, 12.1% and 44.18%, respectively. The total polysaccharide content and alcohol-soluble extract content exceed the pharmacopoeia requirements by 12.82% and 51.25%, respectively.

[0108] Example 2 Effect of Huoshan Dendrobium Salt Products on the Improvement of Body Disorders Caused by Hyperthyroidism Yin Deficiency Syndrome

[0109] 1. Experimental Methods

[0110] Huoshan Dendrobium samples: dried Huoshan Dendrobium strips, Huoshan Dendrobium buds, and Huoshan Dendrobium salt-roasted products (prepared using the optimized process of Example 1).

[0111] Since Huoshan Dendrobium is mostly used as medicine in the form of decoction and oral administration, the present embodiment uses the water extract of Dendrobium to administer the medicine. Take 4g of the powder of Huoshan Dendrobium sample (powdered and passed through a No. 3 sieve), add distilled water at a solid-liquid ratio of 1g:50mL, heat and reflux at 100°C for 3h, retain the supernatant, evaporate and concentrate, then freeze-dry, and dilute to the corresponding concentration when used.

[0112] ICR male mice were randomly divided into blank control group, model group, positive drug Liuwei Dihuang Wan group, high, medium and low dose groups of Huoshan Dendrobium salt roasted products, high, medium and low dose groups of Huoshan Dendrobium dried strips, and high, medium and low dose groups of Huoshan Dendrobium Fengdou. The body weight, temperature and total food intake of each group of mice were recorded every 5 days. After the end of the drug administration, the organ index, serum index and renal tissue ATP metabolic enzyme of each group of mice were detected.

[0113] 1. Modeling method

[0114] Every afternoon, except for the blank control group, which was given an equal amount of normal saline, the other groups of mice were given thyroxine suspension solution (160 mg / kg / d) once a day for 15 days to prepare the hyperthyroidism Yin deficiency model. During the modeling period, the fur color, food intake, autonomous activity and mental state of each group of mice were observed, and the changes in body weight, temperature and food intake of each group of mice were recorded. When the hair turned yellow and dull, the urine was yellow and the stool was dry, the reaction was active and irritable, the food intake was reduced, the body temperature was increased, and the body weight was reduced, the model was successful.

[0115] 2. Grouping and Dosing

[0116] A total of 130 SPF male ICR mice (30-35g, 5 weeks old) were selected for the experiment. After 7 days of adaptive feeding, they were randomly divided into 12 groups: blank control group, model group, positive medicine Liuwei Dihuang Pills group (10g / kg / d), Huoshan Dendrobium Salt-roasted / Dried Strips / Fengdou high (3.4g / kg / d), medium (1.7g / kg / d), and low (0.8g / kg / d) dose groups.

[0117] At the beginning of the experiment, except for the blank control group, the other groups were gavaged with thyroxine suspension (160 mg / kg) daily to induce the hyperthyroidism Yin deficiency model. The modeling period was 15 days, and the modeling criteria were dry and yellow hair, restlessness and irritability, reduced food intake, increased body temperature and decreased body weight. The drug intervention started on the 16th day and lasted for 15 consecutive days. At the end of the experiment, the organ index, serum index and renal ATPase activity of mice in each group were measured.

[0118] 3. Detection indicators

[0119] 3.1 Organ index

[0120] After the administration, the mice were killed, and the thymus, spleen, kidney, and liver of the mice were immediately taken and weighed, rinsed with 0.9% sodium chloride solution, blotted dry with filter paper, and the organ index was calculated. The organ index was calculated according to the following formula: organ index (mg / g) = organ wet weight (mg) / mouse final body weight (g).

[0121] 3.2 Serum detection indicators

[0122] After the administration, the eyeballs were removed to collect blood, and the serum was allowed to stand at room temperature for 2 hours, centrifuged, and the serum was separated. At the same time, the mice in each group were killed by cervical dislocation. The blood was centrifuged at 3000r / min for 15 minutes, and the blood supernatant was collected.

[0123] ELISA was used to detect triiodothyronine (T3), thyroxine (T4), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), and the cAMP / cGMP ratio was calculated. As well as corticosterone (CORT) and creatinine (CRE) levels.

[0124] 3.3 ATPase detection in renal tissue

[0125] After removing the eyeballs of mice and taking blood, the kidneys were quickly taken out, washed with saline, and frozen in liquid nitrogen for later use. 0.5 g of kidney was taken out, rinsed with ice saline, dried with filter paper, cut into pieces, added with 4.5 mL of saline, homogenized mechanically in an ice water bath, centrifuged at 3000 r / min for 10 min, and the supernatant (i.e., 10% of the homogenized supernatant) was taken out and diluted to 0.4% with saline. At the same time, the tissue protein was determined with Coomassie brilliant blue reagent, and the Na was determined using an ultra-micro ATPase test kit. + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase activity.

[0126] 4. Statistical methods

[0127] The experimental data were analyzed using SPSS statistical software, and the data were normal distribution and were expressed as mean ± standard deviation. The comparison among the groups was performed with one-way ANOVA. When the variances were equal, the least significant difference (LSD) method was used for statistical analysis. When the variances were unequal, the Dunnett's method was used for statistical analysis. P < 0.05 was considered statistically significant.

[0128] 2. Experimental Results

[0129] 1. Organ index

[0130] After the administration, the thymus, spleen, kidney, and liver of the mice were weighed and their organ indexes were calculated. Figure 6 As shown, the results showed that compared with the blank group, the thymus, spleen and kidney indexes of the mice in the model group were significantly reduced (p<0.0001); compared with the model group, the thymus indexes of the mice in each drug administration group were significantly increased (p<0.05); except for the medium-dose group of dried strips, the spleen indexes of the mice in the other drug administration groups were significantly increased (p<0.05); compared with the model group, except for the high-dose and low-dose groups of dried strips and the low-dose group of maple thrush, the kidney indexes of the mice in the other drug administration groups were significantly increased (p<0.05); compared with the dried strips and maple thrush groups, the Huoshan Dendrobium salt-fried group could significantly improve the thymus, spleen and kidney indexes of the mice (p<0.001). The changes in the liver organ indexes of each group were not significant. The results of the organ indexes of each group showed that Huoshan Dendrobium can improve the decrease of the thymus, spleen and kidney organ indexes caused by hyperthyroidism Yin deficiency syndrome and improve the immune function of mice. The improvement of the organ indexes of Huoshan Dendrobium after salt-fried is more significant.

[0131] 2. Serum indicators

[0132] 2.1 Serum cyclic nucleotide levels

[0133] Cyclic nucleotide levels are often used to evaluate the body's Yin deficiency state. In medicine, a decrease in cyclic guanosine monophosphate (cGMP) levels, an increase in cyclic adenosine monophosphate (cAMP) levels, and an increase in the ratio of cyclic adenosine monophosphate to cyclic guanosine monophosphate (cAMP / cGMP) are used as indicators of Yin deficiency in the body. Figure 7 ) Compared with the blank group, the cyclic guanosine monophosphate level of the model group mice was significantly reduced (p<0.0001), and the cyclic adenosine monophosphate level and the cyclic adenosine monophosphate / cyclic guanosine monophosphate ratio were significantly increased (p<0.0001), proving that the model of hyperthyroidism Yin deficiency in mice was successfully established. Compared with the model group, the serum cyclic adenosine monophosphate level and the cyclic adenosine monophosphate / cyclic guanosine monophosphate ratio of the mice in each drug administration group were significantly reduced (p<0.05); except for the low-dose dry strip and low-dose maple dou drug administration groups, the cyclic guanosine monophosphate level of the mice in the other groups was significantly increased (p<0.05); the salt-fried product drug administration groups of each dose could significantly reduce the cyclic adenosine monophosphate level and the cyclic adenosine monophosphate / cyclic guanosine monophosphate ratio (p<0.0001), increase the cyclic guanosine monophosphate level (p<0.0001), and improve the body's cyclic nucleotide level (p<0.0001). Experiments have shown that Huoshan Dendrobium can regulate the disordered state of cyclic nucleotides in the body under Yin deficiency state, and the salt-roasted Huoshan Dendrobium can improve the cyclic nucleotide levels of Yin deficiency mice more significantly.

[0134] 2.2 Serum T3 and T4 levels

[0135] Modern research shows that hyperthyroidism Yin deficiency syndrome can cause elevated T3 and T4 levels. Experimental results show that ( Figure 8 ), compared with the blank group, the T3 and T4 levels in the model group were significantly increased (p<0.0001), indicating that the model of hyperthyroidism Yin deficiency in mice was successfully established; compared with the model group, except for the low-dose dry strip group, the T3 and T4 levels in each group were significantly reduced (p<0.05); among them, the salt-roasted product groups at all doses could significantly reduce the T3 and T4 levels (p<0.0001), and the T4 levels of mice in the salt-roasted product groups at all doses were significantly reduced (p<0.05) when compared with the salt-roasted product groups at the same dose and the dry strip and maple pod groups. The experimental results show that Huoshan Dendrobium can regulate the level of thyroid hormones, reduce the content of T3 and T4 in serum, and the regulatory effect is stronger after salt-roasting.

[0136] 2.3 Serum CORT and CRE levels

[0137] Serum corticosterone and creatinine levels were tested and the results showed that ( Fig. 9), compared with the blank group, the serum corticosterone and creatinine levels of the mice in the model group were extremely significantly increased (p<0.0001); compared with the model group, the serum corticosterone and creatinine levels of the mice in each drug-treated group were significantly decreased (p<0.05); among them, the salt-roasted product groups at each dose could extremely significantly reduce the serum corticosterone and creatinine levels (p<0.0001), and compared with the salt-roasted product groups at the same dose and the dried strips and maple thrush groups, the serum corticosterone levels of the mice in the salt-roasted product groups at each dose were significantly lower than those in the dried strips and maple thrush groups (p<0.01); the creatinine level in the salt-roasted product medium dose group was significantly lower than that in the maple thrush group (p<0.05), and the creatinine level in the salt-roasted product low dose group was significantly lower than that in the dried strips and maple thrush groups (p<0.05). The experimental results show that Huoshan Dendrobium can effectively regulate the hormone levels of the hypothalamus-pituitary-adrenal axis and renal function indicators in mice with hyperthyroidism and Yin deficiency syndrome, improve hyperthyroidism and Yin deficiency syndrome, and the regulatory effect of Huoshan Dendrobium after salt roasting is stronger.

[0138] 3. Renal tissue ATPase

[0139] Detection of ATPase content in mouse kidney tissue. Experimental results ( Fig.10 ) showed that compared with the blank group, the Na + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase activity was significantly increased (p<0.0001); compared with the model group, the Na in the kidney tissue of mice in each drug group + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase activity was significantly reduced (p<0.05), and the salt-fried product in each dose group could significantly reduce the Na + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase activity (p<0.0001). The results showed that Huoshan Dendrobium could inhibit the ATPase activity of renal tissue in mice with hyperthyroidism and Yin deficiency syndrome. + -K + -ATPase and Ca 2+ -Mg 2+ -ATPase activity inhibition is stronger and can effectively improve the symptoms of Yin deficiency and internal heat in mice with hyperthyroidism and Yin deficiency syndrome.

[0140] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A method for preparing Huoshan Dendrobium salt-roasted product, characterized in that: The following steps are involved: The fresh strips of Huoshan Dendrobium are dried as raw materials, added with salt water to moisten them until they are completely cooked, and then fried to obtain the salt-roasted Huoshan Dendrobium.

2. The preparation method according to claim 1, characterized in that: The fresh strips are three-year-old Huoshan Dendrobium fresh strips.

3. The preparation method according to claim 1, characterized in that: The drying temperature is 60° C. and the drying time is 25 h.

4. The preparation method according to claim 1, characterized in that: When adding salt water for moistening, based on the mass of the dried Huoshan Dendrobium officinale strips and NaCl, salt water containing 4.6 g NaCl is added for every 100 g of the dried Huoshan Dendrobium officinale strips.

5. The preparation method according to claim 4, characterized in that: The ratio of the mass of the dried strips of Dendrobium huoshanense to the volume of the added salt water is 1 g:10 mL.

6. The preparation method according to claim 4, characterized in that: The time of moistening is 18 hours.

7. The preparation method according to claim 4, characterized in that: The frying temperature is 100° C. and the frying time is 8 minutes.

8. The salt-roasted Dendrobium huoshanense obtained by the preparation method according to any one of claims 1 to 7.

9. Use of the salt-roasted Huoshan Dendrobium as claimed in claim 8 in the preparation of a medicine for improving hyperthyroidism Yin deficiency syndrome.

10. Use of the salt-roasted Huoshan Dendrobium as claimed in claim 8 in the preparation of a medicine for improving body disorders caused by hyperthyroidism Yin deficiency syndrome.

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