Fructus aurantii fried rhizoma atractylodis macrocephalae and processing technology thereof, and application of fructus aurantii fried rhizoma atractylodis macrocephalae in preparation of medicine for treating spleen deficiency and dampness stagnation type ulcerative colitis

Abstract

The invention discloses a fructus aurantii fried rhizoma atractylodis macrocephalae, which is prepared by moistening and frying rhizoma atractylodis macrocephalae decoction pieces by taking fructus aurantii juice as an auxiliary material. The processing technology comprises the following steps: mixing rhizoma atractylodis macrocephalae decoction pieces and fructus aurantii juice, moistening for 20-30 minutes at room temperature, and frying for 10-20 minutes at the temperature of 120-150 DEG C, so as to obtain the fructus aurantii fried rhizoma atractylodis macrocephalae; the dosage of the fructus aurantii juice is calculated according to fructus aurantii decoction pieces, and the mass of the fructus aurantii decoction pieces is 5-15% of that of the rhizoma atractylodis The invention discloses an extract of rhizoma atractylodis macrocephalae fried with fructus aurantii. The extract is a water extract of rhizoma atractylodis macrocephalae fried with fructus aurantii or freeze-dried powder of the water extract of rhizoma atractylodis macrocephalae fried with fructus aurantii. The invention discloses application of rhizoma atractylodis macrocephalae fried with fructus aurantii or extract of rhizoma atractylodis macrocephalae fried with fructus aurantii in preparation of medicine for treating ulcerative colitis. After the atractylodes macrocephala koidz is fried with the fructus aurantii juice, the content of the effective component atractylenolide is increased compared with bran-fried atractylodes macrocephala koidz decoction pieces, the effects of drying dampness and reducing phlegm are enhanced, and the traditional Chinese medicine composition has a better effect of treating the spleen deficiency dampness stagnation type ulcerative colitis.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine, and relates to stir-fried Atractylodes macrocephala with Fructus Aurantii Immaturus and its processing technology, as well as the application of stir-fried Atractylodes macrocephala with Fructus Aurantii Immaturus or its extract in the preparation of drugs for treating ulcerative colitis of spleen deficiency and dampness entrapment type. Background Art

[0002] Atractylodes macrocephala is the dried rhizome of Atractylodes macrocephala Koidz. of the genus Atractylodes in the Compositae family. The cut slices are irregular thick slices. The outer epidermis is grayish yellow or grayish brown. The cut surface is yellowish white to light brown, scattered with yellowish brown punctate oil chambers, and the xylem has radial texture; the cut surface of the dried one is horny, darker in color or has cracks. There are only eight processing methods of Atractylodes macrocephala included in modern specifications, including stir-frying, stir-frying to char, stir-frying with earth, stir-frying with rice, stir-frying with bran, steaming with earth, soaking in rice-washing water. Only "stir-frying Atractylodes macrocephala with honey and bran" is included in Part I of the current edition of the Chinese Pharmacopoeia, and the regulations on the technology, types of auxiliary materials and dosage of auxiliary materials for various processing methods are not unified in local specifications. Modern pharmacological research shows that the volatile oil in raw Atractylodes macrocephala has the effects of increasing gastrointestinal motility and anti-ulcer; the content of lactone components in stir-fried Atractylodes macrocephala with bran increases, which is beneficial to improving diarrhea due to spleen deficiency; the tannin content in stir-fried Atractylodes macrocephala with earth rises, and the astringent effect is enhanced, which is used for irritable bowel syndrome.

[0003] Since the content of the index components of Atractylodes macrocephala is the key factor determining its medicinal efficacy, therefore, based on the content of the index components, further optimizing parameters such as infiltration time and frying temperature can ensure the scientificity and rationality of the processing technology. Summary of the Invention

[0004] Atractylodes macrocephala can invigorate the spleen and replenish qi, dry dampness and promote diuresis; Fructus Aurantii Immaturus can regulate qi and relieve fullness, promote qi movement and eliminate distension; Atractylodes macrocephala is warm in nature and tonifying, while Fructus Aurantii Immaturus is slightly cold in nature and promoting qi movement. Stir-frying Atractylodes macrocephala with the juice of Fructus Aurantii Immaturus, the two herbs complement and restrict each other, and combine promoting qi movement and tonifying, and can achieve the effects of promoting qi circulation to invigorate the spleen and drying dampness to resolve phlegm. However, there is no application of the stir-fried product of Atractylodes macrocephala with Fructus Aurantii Immaturus in intervening in ulcerative colitis of spleen deficiency and dampness entrapment type.

[0005] The purpose of the present invention is to provide a stir-fried product of Atractylodes macrocephala with Fructus Aurantii Immaturus, and provide an effective guarantee for its reasonable clinical application.

[0006] The first purpose of the present invention is achieved through the following technical solutions:

[0007] A kind of stir-fried Atractylodes macrocephala with Fructus Aurantii Immaturus is obtained by moistening and stir-frying Atractylodes macrocephala cut slices with the juice of Fructus Aurantii Immaturus as the auxiliary material.

[0008] Another object of the present invention is to provide a processing technology for stir-frying Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus, comprising: mixing Atractylodis Macrocephalae Rhizoma slices with Fructus Aurantii Immaturus juice, moistening for 20 - 30 min at room temperature, and then stir-frying at a temperature of 120 - 150 °C for 10 - 20 min to obtain stir-fried Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus; wherein, the dosage of Fructus Aurantii Immaturus juice is based on the mass of Fructus Aurantii Immaturus slices, and the mass of Fructus Aurantii Immaturus slices is 5 - 15% of the mass of Atractylodis Macrocephalae Rhizoma slices.

[0009] Preferably, the dosage of Fructus Aurantii Immaturus juice is based on the mass of Fructus Aurantii Immaturus slices, and the mass of Fructus Aurantii Immaturus slices is 15% of the mass of Atractylodis Macrocephalae Rhizoma slices.

[0010] Preferably, mix Fructus Aurantii Immaturus juice with Atractylodis Macrocephalae Rhizoma slices, moisten for 25 min at room temperature, and then stir-fry at a temperature of 130 - 140 °C for 15 min to obtain stir-fried Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus.

[0011] The Fructus Aurantii Immaturus juice is prepared by the following method: weigh a certain amount of Fructus Aurantii Immaturus slices, use water as the extraction solvent, heat under reflux for extraction 1 - 3 times, each extraction for 0.5 - 1 h, filter after each extraction, combine the decoctions, and concentrate the decoctions under reduced pressure to a concentration of 0.3 g / mL to obtain Fructus Aurantii Immaturus juice.

[0012] Preferably, the Fructus Aurantii Immaturus juice is prepared by the following method: weigh a certain amount of Fructus Aurantii Immaturus slices, use water as the extraction solvent, add water according to the mass ratio of Fructus Aurantii Immaturus slices to water of 1:14 each time, heat under reflux for extraction 3 times in total, each extraction for 1 h, filter after each extraction, combine the decoctions, and concentrate the decoctions under reduced pressure to a concentration of 0.3 g / mL to obtain Fructus Aurantii Immaturus juice.

[0013] Specifically, the Fructus Aurantii Immaturus juice is prepared by the following method: weigh a certain amount of Fructus Aurantii Immaturus slices, add water according to the mass ratio of Fructus Aurantii Immaturus slices to water of 1:14, bring to a boil over high heat, then change to low heat to keep it slightly boiling, reflux and extract for 1 h under slightly boiling state, filter while it is hot with a filter cloth (120 mesh); according to the mass ratio of Fructus Aurantii Immaturus slices to water of 1:14, add water to the medicinal residues and repeat the extraction 2 times, each time bring to a boil over high heat, then change to low heat to keep it slightly boiling, reflux and extract for 1 h under slightly boiling state, filter while it is hot with a filter cloth (120 mesh); combine the three decoctions, and concentrate under reduced pressure to a concentration of 0.3 g / mL to obtain Fructus Aurantii Immaturus juice.

[0014] Another object of the present invention is to provide an extract of stir-fried Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus, which is an aqueous extract of the stir-fried Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus or a freeze-dried powder of the aqueous extract of the stir-fried Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus.

[0015] Preferably, the extract of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii is prepared from the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii as the raw material, adding 10 to 14 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii, soaking for 15 min to 1 h, quickly heating to boiling, decocting or refluxing for 30 min to 1.5 h, pouring out the medicinal liquid, adding water to decoct or reflux the medicinal residues for 2 to 3 times, adding 6 to 14 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii each time for decocting or refluxing, decocting or refluxing for 30 min to 1.5 h each time, combining the medicinal liquids, filtering, and concentrating to 0.1 g / mL; or adding 10 to 14 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii, soaking for 15 min to 1 h, quickly heating to boiling, decocting or refluxing for 30 min to 1.5 h, pouring out the medicinal liquid, adding water to decoct or reflux the medicinal residues for 2 to 3 times, adding 6 to 14 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii each time for decocting or refluxing, decocting or refluxing for 30 min to 1.5 h each time, combining the medicinal liquids, filtering, and freeze-drying to obtain the freeze-dried powder of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii.

[0016] More preferably, the extract of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii is prepared from the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii as the raw material, adding 10 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii, soaking for 1 h, quickly heating to boiling, decocting for 1 h, pouring out the medicinal liquid, adding 8 times the amount of water to the medicinal residues, decocting for 1 h, combining the two medicinal liquids, filtering, and concentrating to 0.1 g / mL; or adding 14 times the weight of water of the crude powder of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii, soaking for 15 min, quickly heating to boiling, refluxing for 1.5 h, pouring out the medicinal liquid, adding 14 times the weight of water of the crude powder of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii to the medicinal residues, refluxing for 1.5 h, pouring out the medicinal liquid, continuing to add 14 times the weight of water of the crude powder of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii to the medicinal residues, refluxing for 1.5 h, combining the three medicinal liquids, filtering, and freeze-drying to obtain the freeze-dried powder of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii.

[0017] The freeze-drying is as follows: first freeze at -80 °C for 48 h, and then perform sublimation analysis with a freeze-dryer.

[0018] Another object of the present invention is to provide the application of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii or the extract of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii in the preparation of a drug for treating ulcerative colitis.

[0019] Preferably, the application is in the drug for treating ulcerative colitis of the spleen-deficiency and dampness-entrapment type.

[0020] Preferably, the application is the application of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii or the extract of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii in the preparation of treating animals with ulcerative colitis of the spleen-deficiency and dampness-entrapment type by exerting the following functions: activating intestinal cell autophagy, repairing the intestinal epithelial barrier, and alleviating intestinal inflammatory damage.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] 1. In view of the problems such as the non-standard determination of the process parameters of the processed products of Atractylodes macrocephala Koidz., the optimal process parameters of Atractylodes macrocephala Koidz. stir-fried with Fructus aurantii were screened out to provide a reference for the research and development of traditional Chinese medicine.

[0023] 2. After Atractylodes macrocephala Koidz. is stir-fried with Fructus aurantii juice, the content of the active ingredient atractylenolide in it is increased compared with that of the Atractylodes macrocephala Koidz. stir-fried with bran, enhancing the efficacy of drying dampness and resolving phlegm.

[0024] 3. The Atractylodes macrocephala Koidz. stir-fried with Fructus aurantii in the present invention tonifies the spleen deficiency and relieves qi stagnation at the same time. The two complement and restrict each other, and are used for abdominal distension and fullness, and phlegm-dampness obstructing the middle energizer. While using raw Atractylodes macrocephala Koidz. alone is more inclined to tonify deficiency, with insufficient qi-promoting effect.

[0025] 4. Pharmacological experiments show that compared with raw Atractylodes macrocephala Koidz., Atractylodes macrocephala Koidz. stir-fried with bran, and Fructus aurantii + Atractylodes macrocephala Koidz., the Atractylodes macrocephala Koidz. stir-fried with Fructus aurantii in the present invention can significantly reduce the disease activity index of animals with ulcerative colitis of spleen deficiency and dampness entrapment type, improve the disease more significantly, and has a stronger ability to repair intestinal inflammatory damage, and has a better effect in treating ulcerative colitis of spleen deficiency and dampness entrapment type. Description of the Drawings

[0026] Figure 1 It is a finished product diagram of Atractylodes macrocephala Koidz. stir-fried with Fructus aurantii.

[0027] Figure 2 It is the HPLC chromatograms of atractylenolide Ⅰ, Ⅱ, and Ⅲ in the test solutions of Atractylodes macrocephala Koidz. stir-fried with Fructus aurantii (A) and Atractylodes macrocephala Koidz. stir-fried with bran (B); among them, peak 1, peak 2, and peak 3 are atractylenolide Ⅲ, Ⅱ, and Ⅰ respectively.

[0028] Figure 3 It is the results of the DAI scores of different combinations of Atractylodes macrocephala Koidz. and Fructus aurantii on rats with ulcerative colitis of spleen deficiency and dampness entrapment type.

[0029] Figure 4 It is the pathological diagram of HE staining of the colon of rats with ulcerative colitis of spleen deficiency and dampness entrapment type with different combinations of Atractylodes macrocephala Koidz. and Fructus aurantii. Detailed Embodiments

[0030] The technical solutions of the present invention will be clearly and completely described below in conjunction with specific embodiments. The described embodiments are only partial examples, not all.

[0031] Example 1

[0032] Preparation of Fructus aurantii Juice

[0033] Weigh a certain amount of Fructus Aurantii Immaturus slices (the dried immature fruits of Citrus aurantium L. of the Rutaceae family and its cultivated varieties). According to the mass ratio of Fructus Aurantii Immaturus slices to water of 1:14, add 14 times the amount of water to the Fructus Aurantii Immaturus slices. Place the mixture in an electric heating mantle, turn on the heating reflux extraction device, first quickly heat it to the boiling state with strong fire. After the liquid medicine boils, start timing, and immediately adjust the fire to gentle fire to keep the liquid medicine in a slightly boiling state, and continuously extract for 1 hour in the slightly boiling state. After the extraction is completed, filter it while it is hot with a 120-mesh filter cloth to obtain the decoction. According to the mass ratio of Fructus Aurantii Immaturus slices to water of 1:14, add 14 times the amount of water to the medicinal residues again, and repeat the extraction 2 times according to the above method. Combine the decoctions obtained from the three extractions, and use a vacuum concentration device for concentration until the concentration of the concentrated liquid reaches 0.3 g / mL, that is, each 1 mL of the medicinal juice contains 0.3 g of Fructus Aurantii Immaturus slices, and then Fructus Aurantii Immaturus juice is obtained.

[0034] Example 2

[0035] Optimization of the processing technology of stir-fried Atractylodes macrocephala with Fructus Aurantii Immaturus

[0036] Atractylodes macrocephala Koidz. slices, Fructus Aurantii Immaturus slices (the dried immature fruits of Citrus aurantium L. of the Rutaceae family and its cultivated varieties); reference substances: naringin, neohesperidin, atractylenolide I, atractylenolide II, atractylenolide III; a frying pan, a high performance liquid chromatograph, an analytical balance, an ultrasonic cleaner, a high-speed refrigerated centrifuge, a freeze dryer; ultrapure water, formic acid, chromatographic grade acetonitrile, chromatographic grade methanol.

[0037] Single factor experiment: Select 4 influencing factors, namely the dosage of auxiliary materials (the Fructus Aurantii Immaturus juice in Example 1, and the dosage of Fructus Aurantii Immaturus juice is calculated based on Fructus Aurantii Immaturus slices), the moistening time, the frying temperature and the frying time. Take the three active ingredients of atractylenolide I, II, and III and the solid content as evaluation indexes to investigate the processing technology of stir-fried Atractylodes macrocephala with Fructus Aurantii Immaturus.

[0038] I: Weigh 5 portions of Atractylodes macrocephala Koidz. slices, each portion is about 50 g, and then weigh 5 portions of Fructus Aurantii Immaturus slices respectively. The masses of the Fructus Aurantii Immaturus slices are successively 5% (2.5 g), 10% (5.0 g), 15% (7.5 g), 20% (10.0 g), and 25% (12.5 g) of the mass of the corresponding Atractylodes macrocephala Koidz. slices. Prepare Fructus Aurantii Immaturus juice according to Example 1. Mix the Fructus Aurantii Immaturus juice and Atractylodes macrocephala Koidz. slices, moisten them at room temperature for 20 min, and then stir-fry them at a temperature of 120 - 130 °C for 10 min.

[0039] II: Weigh 5 portions of Atractylodis Macrocephalae Rhizoma slices, each portion being approximately 50 g, add Fructus Aurantii Immaturus juice (concentration 0.3 g / mL, prepared from Fructus Aurantii Immaturus slices accounting for 15% of the mass of Atractylodis Macrocephalae Rhizoma slices according to Example 1), moisten at room temperature for 20 min, and then stir-fry at 120 - 130 °C for 3 min, 5 min, 10 min, 15 min, and 20 min respectively.

[0040] III: Weigh 5 portions of Atractylodis Macrocephalae Rhizoma slices, each portion being approximately 50 g, add Fructus Aurantii Immaturus juice (concentration 0.3 g / mL, prepared from Fructus Aurantii Immaturus slices accounting for 15% of the mass of Atractylodis Macrocephalae Rhizoma slices according to Example 1), moisten at room temperature for 10 min, 15 min, 20 min, 25 min, and 30 min respectively, and then stir-fry at 120 - 130 °C for 10 min.

[0041] IV: Weigh 5 portions of Atractylodis Macrocephalae Rhizoma slices, each portion being approximately 50 g, add Fructus Aurantii Immaturus juice (concentration 0.3 g / mL, prepared from Fructus Aurantii Immaturus slices accounting for 15% of the mass of Atractylodis Macrocephalae Rhizoma slices according to Example 1), moisten at room temperature for 20 min, and then stir-fry at 100 °C - 120 °C, 120 - 130 °C, 130 - 140 °C, 140 - 150 °C, and 150 - 160 °C for 10 min respectively.

[0042] Preparation of the test solution for processed products of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus: Take 10 g of the coarse powder of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus (passed through No. 2 sieve), add 14 times the amount of water according to the mass ratio of the coarse powder of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus to water of 1:14, soak at room temperature for 15 min, extract by heating under reflux 3 times, each extraction for 1.5 h. After filtration, combine the water decoctions, accurately measure 20 mL, place at -80 °C for freezing for 48 h, then take out and immediately put into a freeze dryer for freeze-drying to obtain the freeze-dried powder of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus. Accurately add 20 mL of methanol for re-dissolution, perform ultrasonic treatment (power 254 W, frequency 40 kHz), centrifuge at 12000 r·min -1 Centrifuge for 10 min, take the supernatant, filter, and obtain the test solution. Detect the content of the index components by HPLC. HPLC detection conditions: Chromatographic column: Agilent ZORBAX SB-C18 chromatographic column (250 mm × 4.6 mm, 5 μm); Mobile phase: 0.1% formic acid water (A) - acetonitrile (B), gradient elution: 0 - 5 min, 20% - 25% B; 5 - 15 min, 25% - 55% B; 15 - 28 min, 55% - 65% B; 28 - 35 min, 65% - 70% B; Detection wavelength: 240 nm (atractylenolide II, III) and 280 nm (naringin, neohesperidin, atractylenolide I); Flow rate 1 mL·min -1 ; Column temperature: 40 °C; Injection volume: 10 μL.

[0043] Use a pipette to measure the decoction of stir-fried Atractylodes macrocephala Koidz. with Fructus Aurantii Immaturus in the "test solution of stir-fried Atractylodes macrocephala Koidz. with Fructus Aurantii Immaturus", and determine the solid content according to the Chinese Pharmacopoeia (2020 Edition).

[0044] The results of the single-factor experiments are shown in Tables 1 to 4. Taking the contents of atractylenolide I, II, III and solids as comprehensive evaluation indexes, the comprehensive scores were calculated by the analytic hierarchy process to determine the importance of the contents of atractylenolide I, II, III and solids. A hierarchical model and a judgment matrix were constructed, in which the weight coefficient of atractylenolide I was 0.1067, the weight coefficients of atractylenolide II and atractylenolide III were 0.4269, and the weight coefficient of the solid content was 0.0395. Combining with the actual production and economic factors, according to the score ranking, the single-factor investigation interval was narrowed, and the investigation factor levels of the orthogonal test for the processing technology of stir-fried Atractylodes macrocephala Koidz. with Fructus Aurantii Immaturus were determined as follows: amount of auxiliary material: 5-15%, moistening time: 20-30 min, frying temperature: 120-150 °C, frying time: 10-20 min.

[0045] Table 1. Results of the investigation on the amount of auxiliary material

[0046]

[0047] Table 2. Results of the investigation on the moistening time

[0048]

[0049] Table 3. Results of the investigation on the frying temperature

[0050]

[0051] Table 4. Results of the investigation on the frying time

[0052]

[0053] According to the results of the single-factor experiments, use the following orthogonal L9(3 4 ) experimental table to design an orthogonal experiment, A frying temperature (120-130 °C, 130-140 °C, 140-150 °C), B frying time (10 min, 15 min, 20 min), C moistening time (20 min, 25 min, 30 min), D amount of auxiliary material (5%, 10%, 15%).

[0054] Table 5. Orthogonal factor level table

[0055]

[0056] The detailed combinations and results of the orthogonal experiment are shown in Tables 6 and 7. According to the results of intuitive analysis and variance analysis, the influence of the four factors on the processing technology of stir-fried Atractylodis Macrocephalae with Fructus Aurantii is as follows: frying time > amount of auxiliary materials > moistening time > frying temperature. The optimal process is selected as follows: take Fructus Aurantii slices according to 15% of the mass of Atractylodis Macrocephalae slices, prepare Fructus Aurantii juice (concentration 0.3 g / mL) according to Example 1, mix Atractylodis Macrocephalae slices and Fructus Aurantii juice, moisten for 25 min at room temperature, and then fry at 130 - 140 °C for 15 min.

[0057] Table 6. Results and Intuitive Analysis Table of Orthogonal Experiment

[0058]

[0059] Table 7. Variance Analysis Table

[0060]

[0061] Example 3

[0062] Preparation of test solution of stir-fried Atractylodis Macrocephalae with bran: Purchase the standard slices of stir-fried Atractylodis Macrocephalae that meet the requirements of the Chinese Pharmacopoeia (2020 Edition) on the market, pulverize and sieve through No. 2 sieve to obtain coarse powder. Weigh about 1 g (0.9917 g) of coarse powder, accurately add 10 mL of methanol to dissolve, perform ultrasonic treatment (power 254 W, frequency 40 kHz) for 30 min, and centrifuge at 12000 r·min -1 for 10 min, take the supernatant, filter, and obtain the test solution of stir-fried Atractylodis Macrocephalae with bran.

[0063] Prepare stir-fried Atractylodis Macrocephalae with Fructus Aurantii according to the optimal process screened in Example 2: Take Fructus Aurantii slices according to 15% of the mass of Atractylodis Macrocephalae slices (i.e., the mass ratio of Fructus Aurantii slices to Atractylodis Macrocephalae slices is 15:100), prepare Fructus Aurantii juice with a concentration of 0.3 g / mL according to Example 1, mix Atractylodis Macrocephalae slices and Fructus Aurantii juice, moisten for 25 min at room temperature, and then fry at 130 - 140 °C for 15 min to obtain stir-fried Atractylodis Macrocephalae with Fructus Aurantii. The finished product is shown in Figure 1 .

[0064] Preparation of test solution of stir-fried Atractylodis Macrocephalae with Fructus Aurantii: Take about 1 g (1.0080 g) of stir-fried Atractylodis Macrocephalae with Fructus Aurantii coarse powder (sieved through No. 2 sieve), accurately add 10 mL of methanol to dissolve, perform ultrasonic treatment (power 254 W, frequency 40 kHz) for 30 min, and centrifuge at 12000 r·min -1 for 10 min, take the supernatant, filter, and obtain the test solution of stir-fried Atractylodis Macrocephalae with Fructus Aurantii.

[0065] Detect according to the HPLC detection conditions in Example 2, and compare the peak areas of atractylenolide Ⅰ, Ⅱ, and Ⅲ in the test solution of stir-fried Atractylodis Macrocephalae with bran and the test solution of stir-fried Atractylodis Macrocephalae with Fructus Aurantii. The results are shown in Table 8 and Figure 2, it was shown that the peak areas of atractylenolide II and atractylenolide III in stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii were significantly higher than those in stir-fried Atractylodes macrocephala Koidz. with bran.

[0066] Table 8. Peak areas of active ingredients in the test solution of stir-fried Atractylodes macrocephala Koidz. with bran and the test solution of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii

[0067]

[0068] Example 4

[0069] Further investigate the therapeutic effect of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii on rats with spleen deficiency and dampness entrapment type ulcerative colitis established by the "internal and external causes + trinitrobenzene sulfonic acid (TNBS)" method.

[0070] Prepare the traditional Chinese medicine decoction:

[0071] ① Preparation of Atractylodes macrocephala Koidz. decoction: Weigh 50 g of Atractylodes macrocephala Koidz. slices, add 10 times the weight of water of the Atractylodes macrocephala Koidz. slices, soak for 1 h, quickly heat to boiling, decoct for 1 h, pour out the medicinal liquid, add 8 times the weight of water of the slices to the medicinal residues, decoct for 1 h, pour out the medicinal liquid, combine the two times of medicinal liquid, filter and concentrate to 0.1 g / mL, store at 4 °C for later use.

[0072] ② Preparation of stir-fried Atractylodes macrocephala Koidz. with bran decoction: Purchase the standard slices of stir-fried Atractylodes macrocephala Koidz. that meet the regulations of the Chinese Pharmacopoeia (2020 Edition) on the market, weigh 50 g of stir-fried Atractylodes macrocephala Koidz. slices, add 10 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. slices, soak for 1 h, quickly heat to boiling, decoct for 1 h, pour out the medicinal liquid, add 8 times the weight of water of the slices to the medicinal residues, decoct for 1 h, pour out the medicinal liquid, combine the two times of medicinal liquid, filter and concentrate to 0.1 g / mL, store at 4 °C for later use.

[0073] ③ Preparation of the decoction of Atractylodes macrocephala Koidz. combined with Fructus aurantii: The mass ratio of raw Atractylodes macrocephala Koidz. to Fructus aurantii is 10:1, with a total of 50 g. Add 10 times the total weight of water of raw Atractylodes macrocephala Koidz. and Fructus aurantii, soak for 1 h, quickly heat to boiling, decoct for 1 h, pour out the medicinal liquid, add 8 times the total weight of water of raw Atractylodes macrocephala Koidz. and Fructus aurantii to the medicinal residues, decoct for 1 h, combine the two times of medicinal liquid, filter and concentrate to 0.1 g / mL, store at 4 °C for later use.

[0074] ④ Preparation of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii decoction: Obtain the stir-fried Atractylodes macrocephala Koidz. slices according to the optimal processing technology in Example 2. Take 50 g of the slices, add 10 times the weight of water of the stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii slices, soak for 1 h, quickly heat to boiling, decoct for 1 h, pour out the medicinal liquid, add 8 times the weight of water of the slices to the medicinal residues, decoct for 1 h, combine the two times of medicinal liquid, filter and concentrate to 0.1 g / mL, store at 4 °C for later use.

[0075] SD male rats, 7 weeks old, were purchased from Spf (Beijing) Biotechnology Co., Ltd., with the license number: SCXK(Beijing) 2019-0010. They were adaptively fed for 1 week under the conditions of 5 rats per cage, room temperature (22±1)°C, humidity (50±10)%, natural light, and free access to food and water. They were randomly divided into a blank group (without modeling and no drug administration), a model group, and a drug administration group (positive drug and 4 traditional Chinese medicine groups). Among them, the model group and the drug administration group were modeled by the TNBS method: on odd days, the rats were only given ice water at 4°C by gavage (2 mL / rat), and on even days, they were supplied with sufficient feed and given lard by gavage (4 mL / rat); from 8:00 to 16:00 every day, the rats were placed in water 2 cm deep to limit their sleep time to 8 hours for 20 consecutive days. After 24 hours of fasting but not water deprivation on the 21st day, the rats were anesthetized, and a polypropylene tube was inserted 8 cm above the anus, and a complex of 0.8 mL of 5% trinitrobenzenesulfonic acid (TNBS) and 50% ethanol mixed at a volume ratio of 12:5 was injected; then the rat's tail was lifted and it was continuously inverted for 1 minute to allow the modeling agent to fully penetrate into the intestinal cavity. After the rats woke up naturally, they were allowed to eat and drink freely. Two rats were randomly sacrificed on the 2nd day after modeling, and the colon tissues were taken to observe the pathological changes to ensure the success of modeling. The success criteria for modeling were the presence of two main symptoms (severe diarrhea and even rectal prolapse, poor appetite) and two concurrent symptoms (weight loss, emaciation, listlessness, limp limbs, dull coat color, huddling together, easy fatigue).

[0076] After successful modeling, the rats were grouped: a model group (without drug intervention), a positive group (given mesalazine 0.2 g / kg), a Atractylodes macrocephala group (given aqueous decoction of Atractylodes macrocephala 1 g / kg), a stir-fried Atractylodes macrocephala group (given aqueous decoction of stir-fried Atractylodes macrocephala 1 g / kg), an Atractylodes macrocephala + Aurantii Fructus Immaturus group (given the compatible aqueous decoction of Atractylodes macrocephala and Aurantii Fructus Immaturus, Atractylodes macrocephala: Aurantii Fructus Immaturus = 10:1, 1 g / kg), and a stir-fried Atractylodes macrocephala with Aurantii Fructus Immaturus group (given aqueous decoction of stir-fried Atractylodes macrocephala with Aurantii Fructus Immaturus 1 g / kg). The drugs in each drug administration group were diluted to the same volume with water and then given by gavage once a day for 14 consecutive days. The rats in the blank group and the model group were given the same volume of water. The general conditions of the rats such as drinking water, eating, and activities were observed every day, and they were weighed and recorded. At the same time, the fecal traits and blood in the stool were observed, and the DAI score was calculated as DAI = (weight loss score + stool trait score + blood in stool score) / 3. The DAI score is shown in Table 9. After 14 days, samples were taken and the colon tissues of the rats were stained with HE.

[0077] Table 9. DAI score table

[0078]

[0079] By Figure 3It can be seen that compared with the blank group, the DAI score of rats in the model group was significantly increased (p < 0.001). Compared with the model group, the DAI scores of rats in the positive group, Atractylodes macrocephala + Aurantii Fructus Immaturus group, and stir-fried Atractylodes macrocephala with Aurantii Fructus Immaturus group were significantly decreased (p < 0.01), and the DAI scores of the Atractylodes macrocephala group and stir-fried Atractylodes macrocephala group were also significantly decreased (p < 0.05). Among them, the stir-fried Atractylodes macrocephala with Aurantii Fructus Immaturus group had the best effect on reducing the DAI score. From Figure 4 It can be seen that the mucosa of the colon tissue of rats in the blank group was normal without damage or inflammation; in the model group, the goblet cells in the mucus layer of the colon tissue of rats were severely damaged, all crypts were lost, and a large number of inflammatory cells infiltrated into the serosa layer, which was in line with the pathological characteristics of ulcerative colitis. The goblet cells in the colon tissue of rats in the Atractylodes macrocephala group and stir-fried Atractylodes macrocephala group were severely damaged, a large number of crypts were distorted or lost, and inflammatory cells infiltrated into the submucosa layer, indicating that they had no obvious effect on the pathological changes of ulcerative colitis. In the Atractylodes macrocephala + Aurantii Fructus Immaturus group, the goblet cells in the colon tissue of rats were moderately damaged, some crypts were distorted, and a small number of inflammatory cells infiltrated but did not reach the submucosa layer, indicating that it had a certain improvement effect. In the stir-fried Atractylodes macrocephala with Aurantii Fructus Immaturus group and the positive drug group, the goblet cells in the colon tissue of rats were slightly damaged, the crypts were closely arranged, occasionally distorted, and scattered inflammatory cells infiltrated but did not reach the submucosa layer, indicating that stir-fried Atractylodes macrocephala with Aurantii Fructus Immaturus had a significant improvement effect on the pathological changes of ulcerative colitis, and its curative effect was similar to that of mesalazine. The above shows that Atractylodes macrocephala processed with Aurantii Fructus Immaturus juice has a more significant effect on improving the disease, a stronger ability to repair intestinal inflammatory damage, and has a better effect on treating ulcerative colitis of spleen deficiency and dampness entrapment type.

Claims

1. Atractylodes macrocephala Koidz. stir-fried with Fructus Aurantii Immaturus, characterized in that: It is obtained by moistening and stir-frying Atractylodis Macrocephalae Rhizoma slices with Fructus Aurantii Immaturus juice as an adjuvant.

2. The processing technology of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii immaturus as claimed in claim 1, characterized in that: It includes: Mix Fructus Aurantii Immaturus juice and Atractylodis Macrocephalae Rhizoma slices, moisten them at room temperature for 20 - 30 min, and then stir-fry at a temperature of 120 - 150 °C for 10 - 20 min to obtain Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus; among them, the dosage of Fructus Aurantii Immaturus juice is calculated based on Fructus Aurantii Immaturus slices, and the mass of Fructus Aurantii Immaturus slices is 5 - 15% of the mass of Atractylodis Macrocephalae Rhizoma slices.

3. The processing technique of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii immaturus according to claim 2, wherein: The dosage of Fructus Aurantii Immaturus juice is calculated based on Fructus Aurantii Immaturus slices, and the mass of Fructus Aurantii Immaturus slices is 15% of the mass of Atractylodis Macrocephalae Rhizoma slices.

4. The processing method of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii immaturus according to claim 2, wherein: Mix Fructus Aurantii Immaturus juice and Atractylodis Macrocephalae Rhizoma slices, moisten them at room temperature for 25 min, and then stir-fry at a temperature of 130 - 140 °C for 15 min to obtain Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus.

5. The processing technology of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii immaturus according to claim 2, characterized in that: The Fructus Aurantii Immaturus juice described above is prepared by the following method: Weigh a certain amount of Fructus Aurantii Immaturus slices, use water as the extraction solvent, heat and reflux for extraction 1 - 3 times, each extraction for 0.5 - 1 h, filter after each extraction, combine the decoctions, and concentrate the decoctions under reduced pressure to a concentration of 0.3 g / mL to obtain Fructus Aurantii Immaturus juice.

6. The processing technique of stir-frying Atractylodis Macrocephalae Rhizoma with Fructus Aurantii Immaturus as claimed in claim 2, wherein: The Fructus Aurantii Immaturus juice described above is prepared by the following method: Weigh a certain amount of Fructus Aurantii Immaturus slices, use water as the extraction solvent, add water according to the mass ratio of Fructus Aurantii Immaturus slices to water of 1:14 each time, heat and reflux for extraction 3 times in total, each extraction for 1 h, filter after each extraction, combine the decoctions, and concentrate the decoctions under reduced pressure to a concentration of 0.3 g / mL to obtain Fructus Aurantii Immaturus juice.

7. An extract of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii, characterized in that: It is the water extract of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus described in claim 1 or the freeze-dried powder of the water extract of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus.

8. The extract of stir-fried Atractylodes macrocephala Koidz. with Fructus aurantii immaturus according to claim 7, characterized in that: Using the Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus described in claim 1 as the raw material, add 10 - 14 times the weight of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus of water, soak for 15 min - 1 h, quickly heat to boiling, decoct or reflux for extraction for 30 min - 1.5 h, pour out the medicinal liquid, add water to the medicinal residues for decoction or reflux extraction 2 - 3 times, each time adding 6 - 14 times the weight of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus of water for decoction or reflux extraction, each time for 30 min - 1.5 h, combine the medicinal liquids, filter, and concentrate to 0.1 g / mL; or add 10 - 14 times the weight of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus of water, soak for 15 min - 1 h, quickly heat to boiling, decoct or reflux for extraction for 30 min - 1.5 h, pour out the medicinal liquid, add water to the medicinal residues for decoction or reflux extraction 2 - 3 times, each time adding 6 - 14 times the weight of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus of water for decoction or reflux extraction, each time for 30 min - 1.5 h, combine the medicinal liquids, filter, and freeze-dry to obtain the freeze-dried powder of Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus.

9. Use of the Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus described in claim 1 or the extract of the Atractylodis Macrocephalae Rhizoma stir-fried with Fructus Aurantii Immaturus described in claim 6 in the preparation of drugs for treating ulcerative colitis.

10. The application according to claim 9, wherein: The ulcerative colitis described above is spleen-deficiency and dampness-entrapment type ulcerative colitis.

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