Salt-moistened bran stir-frying processing technology of rhizoma alismatis

Abstract

The invention relates to a salt-moistened bran stir-frying processing technology of rhizoma alismatis, which comprises the following steps: slicing rhizoma alismatis, and treating for 40-60 seconds by microwave medium-low fire at the power density of 5-10 W / g; the preparation method comprises the following steps: weighing 2-3g of edible sea salt per 100g of rhizoma alismatis slices, preparing into 15-20g of saline water, uniformly mixing 100g of rhizoma alismatis with the saline water, moistening for 0.5-1 hour at normal temperature, moistening for 1-2 hours at 8-10 DEG C, spreading out, and drying for 1-3 hours under hot air circulation at 50-70 DEG C; mixing wheat bran and yam starch according to the mass ratio of (9-8): (1-2) to obtain a stir-frying medium, scattering the stir-frying medium into a preheated herbal medicine roaster for stir-frying, and adding rhizoma alismatis when green smoke emerges; the stir-frying temperature is 150-180 DEG C, intermittent heating is adopted, real-time regulation and control are performed in cooperation with infrared temperature measurement, automatic turning and stirring are performed, stir-frying is performed for 3-4 min, then the materials are taken out and stacked for 15-30 s, and a stir-frying medium is screened out; and drying for 6-12 hours under the circulation of hot air at 30-40 DEG C, cooling, and screening out chips, thereby obtaining the product. According to the processing technology, the rhizoma alismatis has a better anti-enteritis effect.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine processing, and particularly relates to a processing technology of orientalis rhizome using a salt-moistened bran frying method. Background Art

[0002] Alisma orientalis Alisma orientale (Sam.) Juzep. Or Alisma Alisma plantago-aquatica Linn. The dried tuber of is a traditional diuretic and dampness-injecting medicine. It is cold in nature and sweet in taste. It enters the kidney and bladder meridians. It has the effects of diuresis and swelling, clearing away heat and detoxification, clearing turbidity and reducing lipids. It is often used in the treatment of edema, damp-heat stranguria and hyperlipidemia. According to the literature of various dynasties, its processing methods are mainly salt processing and bran frying. Among them, the Jiangxi characteristic processing technology "salt-moistened bran frying method" uses salt water to soak the medicine downward, supplemented by wheat bran to slowly neutralize the cold nature, and has the effects of tonifying the spleen and replenishing qi and nourishing yin and relieving heat. The current operation method of salt-moistened bran frying method is: (1) Use sodium chloride salt solution to spray or mix the medicinal materials, and steam until the salt is fully penetrated. (2) Bran frying method: preheat the wok to 150-180℃, add wheat bran and fry until smoking, add salt-moistened orientalis, stir-fry quickly over medium heat until the surface is slightly yellow, and sieve out the bran after it is coked. The surface of the medicinal materials is light yellow and the aroma of coke is revealed.

[0003] Modern research has also confirmed that triterpenoid components in Alisma orientalis (such as 23-acetylalusinol B and 23-acetylalusinol C) are its main anti-inflammatory and lipid-lowering active substances. However, the traditional processing technology has the following problems: (1) The parameters lack quantification: the amount of salt water, the amount of wheat bran, the frying temperature and time rely on empirical operations, resulting in fluctuations in the quality of the medicinal materials; (2) The thermal effect during the processing may lead to the transformation or loss of components, thereby affecting the expression of the efficacy. Summary of the invention

[0004] 1. Technical issues to be resolved: In view of the above-mentioned shortcomings and deficiencies of the prior art, the present invention provides a processing technology for Alisma orientalis by frying with salt and bran, which can increase the total content of 23-acetyl alismatol C and 23-alismol B in the processed salt-bran Alisma orientalis slices, and improve the content of volatile components such as olefins, terpenes, esters, organic acid esters, furans, ethylene, etc., thereby improving the quality and anti-inflammatory effect of the salt-bran Alisma orientalis slices, and solve the problems of low standardization and quantitative operation degree of the existing processing technology and large amount of component conversion or loss in the slices.

[0005] (II) Technical solution: In a first aspect, the present invention provides a process for preparing Rhizoma Alismatis by frying with bran and soaking in salt, which comprises the following steps: S1. Microwave treatment: After slicing the rhizome of Alisma, first use a microwave at medium-low heat for 40-60 seconds with a power density of 5-10W / g; S2, Gradient temperature control and salt lubrication: Weigh 2-3g of edible sea salt for every 100g of oriental rhizome slices, add pure water to 2-3g of edible sea salt to prepare 15-20g of brine, mix 100g of oriental rhizome with the brine, first soak at room temperature for 0.5-1h, then continue to soak at 8-10℃ for 1-2h; spread out the oriental rhizome slices that have been soaked thoroughly and dry them under hot air circulation at 50-70℃ for 1-3h; S3, pulse gluten frying: Mix wheat bran and yam powder in a mass ratio of (9-8): (1-2) as a frying medium, sprinkle the frying medium into a preheated frying machine, stir-fry continuously, and add Alisma orientalis when green smoke appears; set the frying temperature to 150-180℃ during the frying process, and use intermittent heating, combined with infrared temperature measurement for real-time automatic stirring, fry for 3-4 minutes, take out, pile and simmer for 15-30 seconds, and sieve out the frying medium; dry for 6-12 hours under hot air circulation at 30-40℃, cool, and sieve out debris to obtain salt bran and Alisma orientalis decoction pieces.

[0006] Preferably, in step S1, the mixture is treated with a 2450 MHz microwave at medium-low heat for 60 seconds with a power density of 10 W / g to prepare for subsequent salting.

[0007] Preferably, in step S2, 2 g of edible sea salt is weighed for every 100 g of Alisma orientalis slices, 2 g of edible sea salt is mixed with pure water to make 15 g of brine, 100 g of Alisma orientalis and 15 g of brine are mixed evenly, firstly soaked at room temperature for 0.5 h, and then continued to soak at 8-10° C. for 2 h.

[0008] Preferably, in step S2, the slices of the oriental rhizome that have been steamed until thoroughly moistened are spread out and dried for 3 hours under hot air circulation at 50°C. Preliminary drying is sufficient (no moisture can be squeezed by hand), and the drying temperature should not be too high, as it is easier to get bran and yam powder on the surface of the oriental rhizome during the heating process so that the bran / yam powder coke product is formed on the surface of the oriental rhizome.

[0009] According to a preferred embodiment of the present invention, in step S3, wheat bran and yam flour are mixed in a mass ratio of 8:2 as a frying medium. The amount of the frying medium accounts for 30-50% of the mass of the dried oriental rhizome slices processed in step S2.

[0010] According to a preferred embodiment of the present invention, in step S3, the frying temperature is set to 180°C during the frying process, and the frying temperature is set to 180°C for 30 seconds → standing for 10 seconds → frying at 180°C for 30 seconds for cyclic intermittent heating, and the infrared temperature measurement is used to automatically stir and stir in real time, and the mixture is taken out after frying for 3 minutes, and the mixture is stuffed for 15-30 seconds, and the frying medium is sieved out; the mixture is dried for 12 hours under a hot air circulation of 30-40°C, cooled, and the debris is sieved out to obtain the mixture. After frying, the frying medium (wheat bran and yam flour) is coked and evenly attached to the surface of the oriental rhizome. When the infrared detects uneven temperature and the local temperature is too high, the stirring speed is accelerated to make the bran coking product more evenly attached to the surface of the oriental rhizome.

[0011] When wheat bran and yam flour are mixed according to (9-8): (1-2) as the frying medium, the sticky component of yam flour (dioscin) can form a coating layer to reduce the release of irritating volatile oils from Rhizoma Alismatis. In addition, the spleen-strengthening components of yam and the diuretic function of Rhizoma Alismatis form a "attack and supplement" effect, which meets the treatment needs of spleen deficiency and dampness, and further achieves the purpose of "alleviating cold and protecting the spleen and stomach".

[0012] In a second aspect, the present invention further provides a salt-brine Alisma orientalis decoction piece, which is prepared by the processing technology of any of the above embodiments.

[0013] (III) Beneficial effects: The salt-moistened bran-frying method of the oriental rhizome of the present invention is adopted. On the basis of the existing salt-moistened bran-frying method, each processing process is further quantified and standardized, and the processing process steps are optimized and improved to shorten the processing process time. Under the premise of achieving the synergistic effect of "salt leads the medicinal properties downward, bran relieves cold and protects the spleen and stomach", the total content of 23-acetyl alismatol C and 23-alismatol B in the decoction pieces can be effectively increased relative to the raw oriental rhizome, and the content of δ-elemene and β-elemene in the volatile components in the decoction pieces is significantly increased, and the content of volatile components such as terpenes, esters, organic acid esters, furans, ethylene, etc. is increased. The oriental rhizome decoction pieces prepared by the present invention have better anti-enteritis effect than the oriental rhizome decoction pieces prepared by the existing processing method.

[0014] Experiments also prove that the moisture content of the salt-bran Alisma orientalis slices prepared by the processing process of the present invention is ≤11%, the total ash content is ≤6%, and the acid-insoluble ash content is ≤0.4%; the alcohol-soluble extract is improved (≥9%), and the slices are golden in color, without burnt spots, and have a significant bran aroma. Its anti-inflammatory (low expression levels of pro-inflammatory factors TNF-α, IL-6, IL-1β, etc.) effect is better than that of the Alisma orientalis slices prepared by the existing processing process. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 : Comparison images between the salt-bran Alisma orientalis decoction pieces (B) prepared according to the method of Example 1 and the raw Alisma orientalis (A).

[0016] Figure 2: The results of HE staining of ileum sections of LPS-induced enteritis mouse model using salt-bran Alisma orientalis decoction pieces prepared with different processing techniques, raw Alisma orientalis, and dexamethasone are presented.

[0017] Figure 3 : The expression of PI3K, Akt, p-Akt, p-IκB and NF-κB p65 proteins in the ileum tissue of LPS-induced enteritis mouse model were compared in salt-bran Alisma orientalis slices prepared with different processing techniques, raw Alisma orientalis and treated with dexamethasone.

[0018] Figure 4 : The growth morphological changes of cultured organoids after administration of lyophilized powder solution of raw extract of Alisma orientalis in the concentration range of 250 to 6400 μg / mL were observed and recorded.

[0019] Figure 5 : The cell viability of organoids treated with lyophilized powder solution of raw extract of Alisma orientalis at concentrations ranging from 250 to 6400 μg / mL was evaluated by CCK-8 detection method.

[0020] Figure 6 : The results of growth morphological observations after administration of salt-bran Alisma orientalis and raw Alisma orientalis with different processing techniques at a concentration of 3200 μg / mL to LPS-induced mouse small intestinal organoids are shown.

[0021] Figure 7 : The cell viability of LPS-induced mouse small intestinal organoids was evaluated by CCK-8 detection method after treatment with 3200 μg / mL of freeze-dried powder solutions of salt-bran Alisma orientalis extracts with different processing techniques and raw Alisma orientalis extracts.

[0022] Figure 8 : The HE staining results of mouse small intestinal organoids induced by LPS were analyzed after the freeze-dried powder solutions of salt-bran Alisma orientalis extract and raw Alisma orientalis extract with different processing techniques were treated with a concentration of 3200μg / mL (HE, ×100).

[0023] Fig. 9 : The effects of 3200 μg / mL of lyophilized powder solutions of salt-bran Alisma orientalis extract and raw Alisma extract with different processing techniques on the levels of inflammatory factors IL-1β, IL-6 and TNF-α in mouse small intestinal organoids induced by LPS were demonstrated. DETAILED DESCRIPTION

[0024] In order to better explain the present invention and facilitate understanding, the present invention is described in detail below through specific implementation modes in conjunction with the accompanying drawings.

[0025] Example 1 The present embodiment provides a process for preparing Rhizoma Alismatis by frying with bran and soaking in salt, comprising the following steps: S1. After processing the Rhizoma Alismatis into 1 cm thick slices, first treat them with 2450 MHz microwave at medium-low heat for 60 seconds with a power density of 10 W / g to prepare for subsequent salting.

[0026] Microwave treatment can generate micro-channels inside the medicinal material, increase the salt penetration efficiency by 50%, and shorten the salt moistening time; at the same time, microwave enzyme inactivation can prevent the enzymatic synthesis of cold-causing components such as alisma acid. Alisma contains endogenous enzymes such as amylase and oxidase. Microwaves at 2450MHz can effectively inactivate enzyme activity and prevent the enzymatic loss of alisma alcohol components. Compared with microwaves of other wavelengths, the retention rate of alisma alcohol can be increased. High-frequency short-time treatment can promote the degradation of cold-causing components (epoxy alisma ene, rhubarb glycoside, tannic acid) and reduce the cold side effects of alisma slices (increasing efficacy and reducing cold). In addition, since the microwave penetration depth is inversely proportional to the frequency, the penetration depth of 2450MHz is about 1~2cm (a slice of alisma with a water content of 15%), which just matches the thickness of the slice of the medicinal material (usually 0.3~0.5cm), avoiding internal overheating. This frequency may have a higher heating efficiency for water molecules, which helps to heat quickly and evenly and promote salt penetration. The microwave treatment time was controlled to exceed 60 seconds to avoid local carbonization and to maintain the structural stability of the polysaccharide components (stomach-protecting substances).

[0027] S2. Weigh 2g of edible sea salt for every 100g of Alisma orientalis slices, add pure water to 2g of edible sea salt to prepare 15g of brine, mix 100g of Alisma orientalis with the brine, first soak at room temperature 25℃ for 0.5h, then continue to soak at 8-10℃ for 2h; spread out the Alisma orientalis slices that have been soaked thoroughly and dry them under 50℃ hot air circulation for 3h.

[0028] Among them, traditional table salt is replaced with deep-sea mineral salt (containing magnesium and potassium ions), and magnesium ions can enhance the diuretic effect (drug property downward). Gradient temperature control salt moistening is adopted, and salt moistening is carried out at room temperature to promote the initial penetration of salt; then moisturizing salt moistening is carried out at low temperature (8-10℃), slowing down the enzymatic reaction and retaining the polysaccharide stomach-protecting ingredients. If it is not pre-treated with microwaves, the whole process of soaking the oriental salt until it is thoroughly soaked will last for 2-8 hours, but after microwave treatment, micropores are formed inside the medicinal material, and it only takes 1.5-3 hours to completely soak the oriental slices until they are thoroughly soaked.

[0029] S3. Mix wheat bran and yam flour in a mass ratio of 8:2 as a frying medium, the frying medium accounts for 50% of the mass of Alisma orientalis, sprinkle the frying medium into a preheated frying machine, stir-fry continuously, and add Alisma orientalis slices when green smoke appears; set the frying temperature to 180°C during the frying process, and adopt a mode of stir-frying at 180°C for 30 seconds → standing for 10 seconds → stir-frying at 180°C for 30 seconds → standing for 10 seconds, cyclic intermittent heating, and automatic stirring in real-time control with infrared temperature measurement. After frying for 3 minutes, take out, pile and suffocate for 20 seconds, and sieve out the frying medium; continue to dry under 30°C hot air circulation for 12 hours, cool, and sieve out debris to obtain salt bran Alisma orientalis slices, such as Figure 1 B is a photo of the salt-bran Alisma orientalis slices processed in Example 1, which are round or oval thick slices. The outer skin is golden yellow or light yellow brown, with occasional burn spots and small protruding root marks. The cut surface is yellowish white with many pores. It has a bran aroma and is slightly salty. Figure 1 A is commercially available raw slices of Alisma orientalis.

[0030] Example 2 The present embodiment provides a process for preparing Rhizoma Alismatis by frying with bran and soaking in salt, comprising the following steps: S1. After processing the Rhizoma Alismatis into 1 cm thick slices, first treat them with 2450 MHz microwave at medium-low heat for 50 seconds with a power density of 8 W / g to prepare for subsequent salting.

[0031] S2. Weigh 3g of edible sea salt for every 100g of Alisma orientalis slices, add pure water to 3g of edible sea salt to prepare 20g of brine, mix 100g of Alisma orientalis with the brine and stir evenly, first soak at room temperature 25℃ for 1h, then continue to soak at 8-10℃ for 1h; spread out the Alisma orientalis slices that have been soaked thoroughly and dry them under 60℃ hot air circulation for 2h.

[0032] S3. Mix wheat bran and yam powder in a mass ratio of 9:1 as a frying medium, with the frying medium accounting for 50% of the mass of Alisma orientalis. Sprinkle the frying medium into a preheated frying machine, stir-fry continuously, and add Alisma orientalis when green smoke appears; set the frying temperature to 180°C during the frying process, and adopt a mode of stir-frying at 180°C for 30 seconds → let stand for 10 seconds → stir-fry at 180°C for 30 seconds → let stand for 10 seconds, cyclically heat intermittently, and automatically stir and stir in real time with infrared temperature measurement. After frying for 3 minutes, take out, pile and simmer for 20 seconds, and sieve out the frying medium; continue to dry under 30°C hot air circulation for 12 hours, let cool, sieve out debris, and obtain the salt bran and Alisma orientalis decoction piece.

[0033] Example 3 The present embodiment provides a process for preparing Rhizoma Alismatis by frying with bran and soaking in salt, comprising the following steps: S1. After processing the oriental rhizome into 1 cm thick slices, first treat it with a 2450 MHz microwave at medium-low heat for 45 seconds with a power density of 5 W / g to prepare for subsequent salting.

[0034] S2. Weigh 2g of edible sea salt for every 100g of Alisma orientalis slices, add pure water to 2g of edible sea salt to prepare 15g of brine, mix 100g of Alisma orientalis with the brine and stir evenly, first soak at room temperature 25℃ for 1h, then continue to soak at 8-10℃ for 2h; spread out the Alisma orientalis slices that have been soaked thoroughly and dry them under 70℃ hot air circulation for 1h.

[0035] S3. Mix wheat bran and yam powder in a mass ratio of 8:2 as a frying medium, with the frying medium accounting for 50% of the mass of Alisma orientalis. Sprinkle the frying medium into a preheated frying machine, stir-fry continuously, and add Alisma orientalis when green smoke appears; set the frying temperature to 170℃ during the frying process, and adopt a mode of stir-frying at 170℃ for 40 seconds → let stand for 10 seconds → stir-fry at 170℃ for 40 seconds → let stand for 10 seconds, cyclically heat intermittently, and automatically stir and stir in real time with infrared temperature measurement. After frying for 3 minutes, take out, pile and simmer for 20 seconds, and sieve out the frying medium; continue to dry under 30℃ hot air circulation for 12 hours, let cool, sieve out debris, and you will get the Zeyan Bunxie decoction piece.

[0036] Comparative Example 1 Comparative Example 1 adopts the traditional salt-moistened bran frying method, and the steps are as follows: (1) Mix 2g of table salt into 15g of brine.

[0037] (2) Spray 100g of Alisma orientalis tablets with 15g of saline and let soak for 4h.

[0038] (3) Preheat the wok to 160°C, add the wheat bran and fry until smoking. Add the salted Alisma orientalis and stir-fry quickly over medium heat until the surface turns slightly yellow. Sieve off the bran after it is charred. Stir-fry for about 8 minutes until the surface of the herb turns light yellow and emits a charred aroma.

[0039] Example 4 15 batches of orientalis from different origins and batches were collected, and the 15 batches of orientalis were processed according to the processing process of Example 1 to obtain 15 batches of salt bran orientalis slice samples, which were recorded as P1, P2, ..., P15 in sequence. The 15 batches of salt bran orientalis slices were tested for the total content of moisture, total ash, acid-insoluble ash, alcohol-soluble extract, and 23-acetyl alismatol B and 23-acetyl alismatol C, and the relevant inspection limits of salt bran orientalis were formulated based on the experimental results of the 15 batches.

[0040] Experimental results: The moisture content of 15 batches of salt-bran Alisma orientalis slices ranged from 4.33% to 7.88%, with an average of 5.66%; the total ash content ranged from 4.38% to 5.59%, with an average of 5.01%; the acid-insoluble ash content ranged from 0.1% to 0.28%, with an average of 0.16%; the alcohol extract content ranged from 9.04% to 19.39%, with an average of 14.46%; the total amount of 23-acetyl alisma alcohol B and 23-acetyl alisma alcohol C ranged from 0.124% to 0.226%, with an average of 0.172%. Table 1 shows the processing effects of 15 batches of salt-bran Alisma orientalis.

[0041] Table 1: Test results of 15 batches of salt bran oriental rhizome slices prepared according to Example 1: .

[0042] With reference to the "Pharmacopoeia of the People's Republic of China 2020 Edition" and "Jiangxi Province Traditional Chinese Medicine Piece Processing Specification" (2008 Edition) Alisma processing inspection content, it is initially formulated that the moisture content of salt bran Alisma slices shall not exceed 11.0%, the total ash content shall not exceed 6.0%, the acid-insoluble ash content shall not exceed 0.4%, the extract content shall not be less than 9.0%, and the total amount of 23-acetyl alisma alcohol B and 23-acetyl alisma alcohol C shall not be less than 0.10%. Measured by this standard, the processing effects of the above 15 batches of salt bran Alisma slices meet the standard. In addition, after these 15 batches of Alisma from different origins and different batches are processed according to the method of Example 1 of the present invention, the final moisture, extract, total ash, acid-insoluble ash, 23-acetyl alisma alcohol B, C total content, etc. are all close to the same, which shows that the processing process of the present invention can achieve a stable processing effect, so that Alisma from different origins and different batches have similar and nearly consistent properties and quality after processing.

[0043] Example 5 The raw Rhizoma Alismatis was processed by the methods of Example 1 and Comparative Example 1, and the volatile components were identified by HS-GC-MS, and the main components were terpenoid compounds.

[0044] Among them, after processing according to the methods of Example 1 and Comparative Example 1, the contents of components such as (+)-limonene, 1,3,3-trimethyl tricyclo (2.2.1.02,6) heptane, Cypera-2,4-diene, eucalyptol and the like were significantly reduced, while the contents of components such as δ-elemene, β-elemene, dianthin, ethyl hexadecanoate and the like were increased. Among them, after processing according to Example 1, the contents of components such as (+)-limonene, 1,3,3-trimethyl tricyclo (2.2.1.02,6) heptane, Cypera-2,4-diene, eucalyptol and the like were reduced to a greater extent than those of the oriental decoction pieces processed in Comparative Example 1, while the contents of components such as δ-elemene, β-elemene, dianthin, ethyl hexadecanoate and the like were increased to a greater extent than those of the oriental decoction pieces processed in Comparative Example 1. See Table 2 and Table 3.

[0045] At the same time, the electronic nose test showed that after the processing of Alisma orientalis by the method of Example 1, the sensor response values ​​related to volatile components such as terpenes, esters, organic acid esters, furans, and ethylene in the electronic nose increased significantly (P<0.05), which is consistent with the detection results of HS-GC-MS. Combined with the traditional processing theory, raw Alisma orientalis is cold in nature and good at clearing heat and removing dampness, while the medicinal properties tend to be mild after processing with salt bran, and the burnt aroma is enhanced, which may be closely related to the transformation and content changes of components such as terpenes and esters. For example, elemene components have anti-inflammatory and lipid-lowering activities, and their increased content may enhance the effects of salt bran Alisma orientalis on tonifying the spleen and replenishing qi and nourishing yin and relieving heat; and the increase of ester substances such as ethyl hexadecanoate may contribute to the formation of bran aroma.

[0046] Table 2: Comparison of volatile components of raw Rhizoma Alismatis and samples processed according to Comparative Example 1 and Example 1 respectively: .

[0047] Note: -. Not detected or matching degree <80%.

[0048] Table 3: Differential components of raw Alismatis orientalis and the Alismatis orientalis slices prepared according to Example 1: .

[0049] In Table 3, the VIP value screening system is a quality control framework for Chinese herbal medicines based on a multi-dimensional quality association model. Its core logic can be summarized as follows: (1) Data layer: integrating spectral (NIRS / HPLC), chemical (content determination), and biological (efficacy verification) data. (2) Algorithm layer: using machine learning models such as PLS-DA (partial least squares discriminant analysis) and random forest to screen key parameters. (3) Decision layer: achieving quality grading through dynamic weight allocation (VIP>1.0 is the key indicator, 0.8≤VIP≤1.0 is the auxiliary indicator).

[0050] Example 5 This example evaluates the intervention effect of salt-bran Alisma orientalis prepared by different processing techniques on lipopolysaccharide (LPS)-induced small intestinal inflammation through in vivo and in vitro experimental systems. The experiment includes the following parts: (1) In vivo test: By inducing small intestinal inflammation in mice with lipopolysaccharide, the improvement effects of the salt-bran Alisma orientalis prepared in Example 1 and the salt-bran Alisma orientalis prepared in Comparative Example 1 on small intestinal inflammation in mice were explored, and their potential mechanisms of action were deeply analyzed.

[0051] Experimental groups and treatments: 80 mice were randomly divided into 8 groups, 10 mice in each group, namely A blank group, B model group, C dexamethasone group (1.0 mg / kg), D salt bran Alisma orientalis slice high dose group (4.0 g / kg) of Example 1, E salt bran Alisma orientalis slice medium dose group (2.0 g / kg) of Example 1, F salt bran Alisma orientalis slice low dose group (1.0 g / kg) of Example 1, G salt bran Alisma orientalis group of comparative example 1, and H raw Alisma orientalis group; all were adaptively raised in a quiet environment at an ambient temperature of 22-25°C for 3 days, and were given free food and water. After 3 days, mice other than the blank group were intraperitoneally injected with 10 mg / kg of lipopolysaccharide suspension according to body weight according to the modeling method of lipopolysaccharide-induced mouse small intestinal inflammation model; then mice in each group were gavaged once a day for 7 consecutive days. The overall state of the mice was observed every day.

[0052] The blank group A and the model group B were gavaged with normal saline at a dose of 2.0 g / kg per unit weight of mice; The remaining groups C to H were infused according to the drug doses noted in the brackets of their respective groups. For example, the intragastric administration of 4.0 g / kg for group D indicated that the decoction boiled with 4 g of salt bran and Alisma orientalis slices per unit body weight of mice was used as the infusion drug.

[0053] ① Effect of Alisma orientalis on ileum morphology in mice with lipopolysaccharide-induced enteritis: The ileum tissues fixed with 4% paraformaldehyde were embedded, sliced, and stained with HE to observe the histopathological changes under a microscope. Three samples were collected in each group, and photographed and analyzed.

[0054] like Figure 2 As shown, the intestinal wall of the ileum tissue of mice in the blank group A was intact, the structure was clear, the villi were neatly arranged, and no inflammatory cell infiltration was observed. In the B model group, some intestinal villi fell off, some crypt structures disappeared, and a large number of inflammatory cells were observed to infiltrate. In the salt bran Alisma orientalis slices of Example 1, the ileum tissue of mice in the medium and high dose groups E, D and dexamethasone group C basically returned to normal, with healthy intestinal walls, clear structures, neatly arranged villi, and disappeared inflammatory cells. The raw Alisma orientalis group H and the salt bran Alisma orientalis group G of comparative example 1 also had certain therapeutic effects, among which the salt bran Alisma orientalis group G of comparative example 1 was better than the raw Alisma orientalis group H, but some inflammatory cells were still observed. This shows that the improvement effect of Alisma orientalis on inflammatory response is enhanced after processing, and the improvement effect of the salt bran Alisma orientalis slices processed in Example 1 is stronger than that of comparative example 1.

[0055] ② Effect of Alisma orientalis on the levels of inflammatory factors in the ileum tissue of mice: According to the operating instructions of the ELISA kit, the ileum tissues of mice in each group were collected, and the contents of IL-1β, IL-6 and TNF-α therein were determined. As shown in Table 4, compared with the blank group, the levels of inflammatory factors TNF-α, IL-6 and IL-1β in the model group mice were significantly increased (P<0.01); compared with the model group, the levels of TNF-α, IL-6 and IL-1β in the ileum tissue of mice in the dexamethasone group, the raw Alisma orientalis group, the salt bran Alisma orientalis group of Comparative Example 1, and the low, medium and high dose groups of the salt bran Alisma orientalis decoction pieces of Example 1 were significantly decreased (P<0.01); the inhibitory effects of different dose groups on the inflammatory factors TNF-α, IL-6 and IL-1β in the ileum tissue of mice were as follows: the high dose group of the salt bran Alisma orientalis decoction pieces of Example 1 > the medium dose group of the salt bran Alisma orientalis decoction pieces of Example 1 > the low dose group of the salt bran Alisma orientalis decoction pieces of Example 1, and the inhibitory effects on the inflammatory factors under the same dose conditions were as follows: the medium dose group of the salt bran Alisma orientalis decoction pieces of Example 1 > the salt bran Alisma orientalis decoction pieces of Comparative Example 1 > the raw Alisma orientalis group.

[0056] Table 4: Effects of Alisma orientalis on the levels of inflammatory factors in mouse ileum tissue: .

[0057] ③ Expression of PI3K, Akt, p-Akt, p-IκB and NF-κB p65 proteins in mouse ileum tissue: See Table 5 and Figure 3 Compared with the blank group, the protein expression levels of PI3K, p-Akt / Akt, p-Akt, p-IκB, and NF-κB p65 in the ileum tissue of mice in the model group were significantly increased (P<0.01); compared with the model group, the protein expression levels of NF-κBp65 in each drug-treated group were significantly decreased (P<0.01); the expression levels of p-Akt, PI3K, and p-IκB in the dexamethasone group and the high and medium dose groups of the salt-bran Alisma orientalis slices in Example 1 were significantly lower than those in the raw Alisma orientalis slices, the low dose group of the salt-bran Alisma orientalis slices in Example 1, and the salt-bran Alisma orientalis group in Comparative Example 1 were significantly lower (P<0.01).

[0058] Table 5: Expression of PI3K, p-Akt / Akt, p-Akt, p-IκB, and NF-κB p6 proteins in mouse ileum tissue : .

[0059] (2) In vitro test: ① Effects of different drug concentrations on small intestinal inflammation organoids: Weigh an appropriate amount of salt-bran Alisma decoction pieces or raw Alisma powder (80 mesh), decoct it into a 1g / mL decoction extract, make the extract into freeze-dried powder, seal and dry for later use. Mice were killed by cervical dislocation after intraperitoneal anesthesia, and 15cm small intestinal tissue was obtained under sterile conditions. The mesentery and adipose tissue were removed by rinsing with 4℃ DPBS. After longitudinally dissecting the intestine to remove the contents, the intestinal villus epithelium was scraped off to prepare 3mm tissue fragments. After low-temperature digestion with 5mM EDTA for 20min, the crypt structure was dissociated by gradient oscillation method, and the crypt suspension was obtained by filtration. After centrifugation enrichment at 300×g, the crypts were inoculated by three-dimensional matrix gel embedding method, and complete culture medium was added after solidification at 37℃ for air-liquid interface culture (5% CO2). The culture medium was changed every 48h in the culture system, and the developmental state of the organoids was monitored by morphology. When the central area became dark, the passage was processed.

[0060] The in vitro intestinal organoid inflammation model was constructed by treating the small intestinal organoids with 200 mg / L LPS. The growth and morphological changes of the small intestinal organoids were observed and photographed under an inverted microscope. For the successfully constructed small intestinal organoid inflammation model, 250 μg / mL-6400 μg / mL extract lyophilized powder solution was used to culture the organoids. Morphological observation and CCK-8 were used to detect cell viability in order to determine the optimal dosage concentration and provide a basis for subsequent drug administration. The experimental results are as follows Figure 4 , Figure 5 As shown (*P<0.05 compared with the blank group; #P<0.05 compared with the model group).

[0061] After being induced by 200mg / L LPS, the small intestinal organoids of mice all showed a significant expansion trend to varying degrees, and the inner cavity was in a black apoptotic state, and the cell viability decreased to varying degrees (P < 0.05). Compared with the model group, after the administration of different concentrations of raw oriental rhizome, the expansion of the small intestinal organoids was alleviated, the inner cavity had a tendency to restore the normal organoid morphology, and the cell viability was restored. Among them, the cell proliferation activity was the most obvious at a concentration of 3200μg / mL (P < 0.05). Figure 4 and Figure 5 , so this dosing concentration (3200 μg / mL) was selected for subsequent experiments.

[0062] ②The effects of different processing techniques on small intestinal inflammation organoids: Raw alisma orientalis, salt bran alisma orientalis of comparative example 1, and freeze-dried powder solutions of salt bran alisma orientalis of example 1 were administered to mouse small intestinal organoids induced by LPS, and the effects of different preparations of alisma orientalis on inflammation of small intestinal organoids were compared by morphological observation and CCK-8 assay for proliferation activity. Figure 6 , Figure 7As shown in the figure, after administration of different processed products, the swelling of small intestinal organoids was relieved, and the inner cavity gradually restored the normal organoid shape. The cell viability of mouse small intestinal organoids induced by LPS decreased significantly, and the cell viability after administration of different processed products of Alisma was far less than that of the blank group (P < 0.05), but the cell viability was above 50%. Compared with the model group, the cell viability of different processed products of Alisma recovered after administration, among which the cell viability of the salt bran Alisma decoction piece group prepared in Example 1 recovered the most significantly (P < 0.05). Figure 7 Compared with the blank group *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001; compared with the model group #P <0.05.

[0063] ③ Effects of different preparations on the morphology of mouse small intestinal inflammation organoids: Mouse intestinal organoids fixed with 4% paraformaldehyde were stained with HE and histopathological changes were observed under a microscope. Two samples were taken from each group and analyzed. Figure 8 As shown in the figure, no inflammatory cell infiltration was observed in the small intestinal organoids of the blank group mice. A large number of inflammatory cells were observed in the model group. Compared with the model group, the degree of inflammation in the small intestinal organoids of mice in each group of Alisma administration was significantly reduced, but some inflammatory cells were still observed. Among them, the effect of the salt-moistened bran-fried Alisma was better, which indicates that the improvement effect of the inflammatory response of the Alisma after the salt-moistened bran-fried method was enhanced. Figure 8 A: blank group; B: model group; D: high-dose group of the salt-bran and Alisma orientalis slices of Example 1; H: raw Alisma orientalis group; G: salt-bran and Alisma orientalis group of comparative example 1.

[0064] ④ Effects of different preparations on inflammatory factors IL-1β, IL-6 and TNF-α in mouse small intestinal organoids: According to the operating instructions of the ELISA kit, the supernatant culture medium of the small intestinal organoids of each group of mice was collected and the contents of IL-1β, IL-6 and TNF-α were determined. Fig. 9As shown, compared with the blank group, the levels of inflammatory factors TNF-α, IL-6 and IL-1β in the model group mice were significantly increased (P<0.0001); the effects of different processed Rhizoma Alismatis on the levels of IL-1β, IL-6 and TNF-α in LPS-induced mouse small intestinal inflammation organoids showed different trends. Compared with the model group, different processed Rhizoma Alismatis had significantly reduced the inflammatory factors IL-1β, IL-6 and TNF-α (P<0.0001). It shows that different processed Rhizoma Alismatis has anti-inflammatory pharmacological effects. Among them, the Rhizoma Alismatis processed by the salt-moistened bran stir-fried decoction preparation method of Example 1 was the most significant, and its reduction in TNF-α, IL-6 and IL-1β levels was the most obvious, followed by the Rhizoma Alismatis processed by the salt-moistened bran stir-fried decoction preparation method of Comparative Example 1. Fig. 9 middle, , compared with the blank group ****P <0.0001; compared with the model group ####P <0.0001.

[0065] In summary, in the in vivo model, salt-bran Alisma significantly alleviated the weight loss, abnormal blood indexes and pathological damage of small intestinal tissues caused by LPS in mice, and reduced the expression levels of proinflammatory factors (TNF-α, IL-6, IL-1β) by inhibiting the PI3K-Akt / NF-κB signaling pathway. The in vitro organoid experiment further verified the anti-inflammatory activity of salt-bran Alisma, which showed a dose-dependent effect consistent with the in vivo experiment by reducing organoid expansion, restoring cell proliferation activity and reducing the release of inflammatory factors. In addition, the experimental results indicate that the anti-inflammatory effect of the salt-bran Alisma prepared in Example 1 is significantly better than that of the raw Alisma and the salt-bran Alisma of Comparative Example 1, indicating that the improvement of the processing technology of the present invention is of key significance to the enhancement of its pharmacological activity.

[0066] The medium and high-dose groups of salt-bran Alisma orientalis slices had the most significant inhibitory effects on p-Akt, p-IκB and NF-κB p65, confirming that the optimization of active ingredients after processing may achieve anti-inflammatory enhancement by targeting and regulating key nodes of signal pathways.

[0067] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that the technical solutions described in the above embodiments may still be modified, or some or all of the technical features therein may be replaced by equivalents. These modifications or replacements, or the technical features in the above embodiments may be combined in the manner described in the embodiments if they do not conflict with each other, and these modifications, replacements or combinations do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A process for preparing Rhizoma Alismatis by frying with bran and soaking in salt, characterized in that: The steps include: S1. Microwave treatment: After slicing the rhizome of Alisma, first use a microwave at medium-low heat for 40-60 seconds with a power density of 5-10W / g; S2, Gradient temperature control and salt lubrication: Weigh 2-3g of edible sea salt for every 100g of oriental rhizome slices, add pure water to 2-3g of edible sea salt to prepare 15-20g of brine, mix 100g of oriental rhizome with the brine, first soak at room temperature for 0.5-1h, then continue to soak at 8-10℃ for 1-2h; spread out the oriental rhizome slices that have been soaked thoroughly and dry them under hot air circulation at 50-70℃ for 1-3h; S3, pulse gluten frying: Mix wheat bran and yam powder in a mass ratio of (9-8): (1-2) as a frying medium, sprinkle the frying medium into a preheated frying machine, stir-fry continuously, and add Alisma orientalis when green smoke appears; set the frying temperature to 150-180℃ during the frying process, and use intermittent heating, combined with infrared temperature measurement for real-time automatic stirring, fry for 3-4 minutes, take out, pile and simmer for 15-30 seconds, and sieve out the frying medium; dry for 6-12 hours under hot air circulation at 30-40℃, cool, and sieve out debris to obtain salt bran and Alisma orientalis decoction pieces.

2. The process for preparing Rhizoma Alismatis by frying with bran and soaking in salt according to claim 1, characterized in that: In step S1, the mixture is treated with a 2450 MHz microwave at medium-low power for 60 seconds with a power density of 10 W / g.

3. The process for preparing Rhizoma Alismatis by frying with bran and soaking in salt according to claim 1, characterized in that: In step S2, 2 g of edible sea salt is weighed for every 100 g of Alisma orientalis slices, 2 g of edible sea salt is mixed with pure water to make 15 g of brine, 100 g of Alisma orientalis and 15 g of brine are mixed evenly, firstly soaked at room temperature for 0.5 h, and then continued to soak at 8-10° C. for 2 h.

4. The process for preparing Rhizoma Alismatis by frying with bran and soaking in salt according to claim 1, characterized in that: In step S2, the slices of the oriental rhizome that have been steamed until thoroughly moistened are spread out and dried under hot air circulation at 50° C. for 3 hours.

5. The process for preparing Rhizoma Alismatis by frying with bran and soaking in salt according to claim 1, characterized in that: In step S3, wheat bran and yam flour are mixed in a mass ratio of 8:2 as a frying medium.

6. The process for preparing Rhizoma Alismatis by frying with bran and soaking in salt according to claim 1, characterized in that: In step S3, the frying temperature is set to 180°C during the frying process, and the frying temperature is set to 180°C, and the frying temperature is adjusted to 180°C for 30 seconds → let stand for 10 seconds → frying at 180°C for 30 seconds for cyclic intermittent heating. The mixture is automatically stirred in real time with infrared temperature measurement. After frying for 3 minutes, the mixture is taken out and simmered for 15-30 seconds, and the frying medium is sieved out.

7. The process for preparing Rhizoma Alismatis by frying with bran and soaking in salt according to claim 6, characterized in that: In step S3, the mixture is dried under hot air circulation at 30-40° C. for 12 h, cooled, and debris is sieved to obtain the salt-bran Alisma orientalis decoction slices.

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