A pancreatic enzymolysate and its preparation method and application

Abstract

This invention relates to the field of pancreatic extract technology, and discloses a pancreatic enzymatic hydrolysate, its preparation method, and its application, comprising the following steps: using porcine pancreas as raw material, 10% of the total mass of porcine pancreas is used to prepare a fermentation substrate; the fermentation substrate and the remaining porcine pancreas are enzymatically hydrolyzed, followed by solid-liquid separation and drying to obtain the pancreatic enzymatic hydrolysate; a fermenting agent is added during the preparation of the fermentation substrate, wherein the fermenting agent includes Pediococcus pentosaceus and Lactobacillus sakei. The pancreatic extract of this invention can alleviate inflammation.

Description

Technical Field

[0001] The present invention relates to the technical field of pancreatic extracts, and in particular to a pancreatic enzymolysate and a preparation method and application thereof. Background Art

[0002] The inflammatory response is a protective physiological reaction of the body to infection, injury or irritation. It is part of the immune system and is designed to eliminate the cause of the inflammation and begin repairing damaged tissue. The inflammatory response can be acute or chronic, varying in duration and nature.

[0003] As a by-product of animal husbandry, pig and cattle pancreas have high utilization value. The pancreas can secrete insulin to balance the blood sugar level in the body and enzymes such as protease and amylase that decompose carbohydrates, proteins and fats. The insulin produced domestically is derived from pig and cattle pancreas. Due to the different sources of insulin extraction, the composition of the obtained insulin is different. The structural difference between pig insulin and human insulin is the smallest, and only one amino acid is different. In the prior art, some researchers have separated and purified insulin from pig pancreas through modern separation and purification technology, and it is well known that different extracts will be obtained by treating the pancreas in different ways. Based on this background, from the perspective of turning waste into treasure, it is obvious to study the application value and social benefits of pig pancreas. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a pancreatic enzymatic hydrolysate and a preparation method and application thereof. The pancreatic extract of the present invention can relieve inflammation.

[0005] The present invention provides a method for preparing pancreatic enzymatic hydrolysate, comprising the following steps:

[0006] Using pig pancreas as raw material, 10% of the total mass of the pig pancreas is used to prepare a fermentation substrate, and the fermentation substrate and the remaining pig pancreas are enzymatically hydrolyzed, solid-liquid separated, and dried to obtain a pancreatic enzymolyzate.

[0007] Furthermore, the specific process of preparing the fermentation substrate includes: washing and crushing the pig pancreas, adding the fermentation agent, and fermenting at a temperature of 35° C.-37° C. for 11 h-13 h.

[0008] Furthermore, the fermentation agent includes Pediococcus pentosaceus and Lactobacillus sakei.

[0009] Furthermore, the Pediococcus pentosaceus is Pediococcus pentosaceus TS289313 (Testo Biotech).

[0010] Furthermore, the sake Lactobacillus is sake Lactobacillus TS344651 (Testo Biotechnology).

[0011] Furthermore, the method for preparing the starter includes: inoculating the bacterial powder of Pediococcus pentosaceus and the bacterial powder of Lactobacillus sakei into a primary activated seed culture medium for activation culture, and then inoculating into a secondary activated seed culture medium for culture to obtain the starter.

[0012] Furthermore, the formula of the primary activated seed culture medium and the secondary activated seed culture medium includes: 10 g / L peptone, 10 g / L beef extract, 5 g / L yeast extract, 20 g / L glucose, 5 g / L sodium acetate, 2 g / L diamine hydrogen citrate, 1 mL / L Tween-80, 2 g / L dipotassium hydrogen phosphate, 0.2 g / L magnesium sulfate heptahydrate, and 0.05 g / L manganese sulfate heptahydrate, and the pH value is adjusted to 5.5-5.9.

[0013] Those skilled in the art should understand that deionized water is used when preparing the culture medium.

[0014] Furthermore, the activation culture and culture conditions are: temperature of 36°C-38°C, and culture time of 23h-25h.

[0015] Furthermore, the mass ratio of the bacterial powder of Pediococcus pentosaceus to the bacterial powder of Lactobacillus sakei is 1:1.

[0016] Furthermore, when inoculated into the primary activated seed culture medium, the mass concentration of the bacterial powder of Pediococcus pentosaceus in the primary activated seed culture medium is 1 g / 250 mL.

[0017] Furthermore, the inoculation amount in the secondary activated seed culture medium is 1% (v / v).

[0018] Furthermore, the mass ratio of the added fermentation agent in the crushed pig pancreas is 1%.

[0019] Furthermore, the enzymolysis also includes sterilization, and the specific process includes: washing the remaining pig pancreas, adding water, grinding it into a paste, and then heating the paste to sterilize it.

[0020] Furthermore, the specific heating process includes: heating to 100°C, keeping at 100°C for 25min-35min, with a heating rate of 100°C / 3h; and then cooling to 35°C-50°C.

[0021] Furthermore, the mass ratio of the remaining porcine pancreas to water is 1:4.

[0022] Furthermore, the specific process of enzymatically hydrolyzing the fermentation substrate and the remaining porcine pancreas includes: mixing the fermentation substrate and the sterilized substance, then adding papain, and maintaining at 35°C-50°C for 4h-6h, and performing inactivation treatment after enzymatic hydrolysis.

[0023] Furthermore, during the enzymatic hydrolysis process, in order to promote sufficient enzymatic hydrolysis, those skilled in the art may stir the solution during the enzymatic hydrolysis process.

[0024] Furthermore, the mass ratio between the fermentation substrate and the sterilized substance is 1:45.

[0025] Furthermore, the mass ratio of the added mass of the papain to the fermentation substrate is 0.1%-0.3%.

[0026] Furthermore, the specific process of the inactivation treatment includes: adjusting the pH value of the environment after enzymatic hydrolysis to 6, and then heating to 80°C-100°C and maintaining it for 15min-25min.

[0027] Furthermore, the specific process of the solid-liquid separation includes: after solid-liquid separation, cooling to between 35°C and 50°C, keeping the temperature constant for 3h-5h, adding citric acid to the filtrate after solid-liquid separation, heating the filtrate to 70°C-80°C, and then continuing to heat the filtrate to 90°C-100°C and keeping it for 1.5h-2.5h, passing the filtrate through an 80-mesh sieve, and spray drying to obtain pancreatic enzymatic hydrolysate.

[0028] Further, the filtrate is stirred while being heated to 70°C-80°C, and the stirring is stopped when the temperature reaches 70°C-80°C.

[0029] Furthermore, the mass concentration of the citric acid in the filtrate after solid-liquid separation is 1.5%.

[0030] The present invention also provides pancreatic hydrolysate obtained by the pancreatic hydrolysate preparation method.

[0031] The present invention also provides the use of the pancreatic enzymolysate in preparing a medicine for alleviating inflammation.

[0032] The invention also provides application of the pancreatic enzymolysate in food.

[0033] The embodiments of the present invention have the following technical effects:

[0034] 1. The method for preparing pancreatic hydrolysate of the present invention is simple and easy to implement. In the method of the present invention, biomacromolecule nucleic acid is hydrolyzed into small molecule nucleic acid degradation products, which is conducive to the absorption of the prepared pancreatic hydrolysate.

[0035] 2. In the present invention, the porcine pancreas is used as a fermentation substrate, and Pediococcus pentosaceus and Lactobacillus sakei are used as fermentation agents to ferment the fermentation substrate, which can not only hydrolyze biological macromolecular nucleic acids into small molecular nucleic acid degradation products, which is beneficial to the absorption of the prepared pancreatic hydrolysate, but also the pancreatic hydrolysate obtained by the method of the present invention can alleviate and treat lipopolysaccharide-induced systemic inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0037] Figure 1 The weight and body temperature changes of the rats in the protection group provided by the present invention.

[0038] Figure 2 The weight and body temperature changes of the rats in the protection group provided by the present invention.

[0039] Figure 3 The present invention relates to changes in body weight and body temperature of rats after intragastric administration of the pancreatic enzymolysate provided by the present invention.

[0040] Figure 4 The present invention relates to changes in body weight and body temperature of rats after intragastric administration of the pancreatic enzymolysate provided by the present invention.

[0041] Figure 5 It is the percentage of neutrophils in the blood routine test of rats provided by the embodiment of the present invention.

[0042] Figure 6 The present invention is the detection of inflammatory factor IL-6 in rat blood.

[0043] Figure 7 The present invention is the detection of the inflammatory factor IgA in the blood of rats.

[0044] Figure 8 The present invention is the detection of the inflammatory factor IL-1β in the blood of rats.

[0045] Fig. 9 The present invention is the detection of inflammatory factor IgG in rat blood.

[0046] Fig.10 The present invention is the detection of the inflammatory factor CRP in the blood of rats.

[0047] Fig.11 The present invention is the detection of the inflammatory factor hs-CRP in the blood of rats.

[0048] Fig.12 The present invention is the detection of inflammatory factor TNF-α in rat blood.

[0049] Fig.13 The HE staining (H&E) of the intestine provided by the present invention, wherein Fig.13 Middle a is HE staining of the intestine of the normal group. Fig.13 Middle b is HE staining of the intestine of the model group. Fig.13 Middle c is HE staining of the intestine of the positive drug group. Fig.13 Middle d is HE staining of the intestine of the normal dose treatment group. Fig.13 Middle e is HE staining of the intestine in the high-dose treatment group. Fig.13 Middle f is HE staining of the intestine in the ultra-high-dose treatment group. Fig.13 Figure a1 is an enlarged view of Figure a. Fig.13 Figure b1 is an enlarged view of Figure b. Fig.13 In the figure c1 is an enlarged view of figure c. Fig.13 Figure d1 is an enlarged view of Figure d. Fig.13 Figure e1 is an enlarged view of Figure e. Fig.13 Figure f1 is an enlarged view of Figure f. Fig.13 Figure a2 is an enlarged view of Figure a. Fig.13 Figure b2 is an enlarged view of Figure b. Fig.13 Figure c2 is an enlarged view of Figure c. Fig.13 Figure d2 is an enlarged view of Figure d. Fig.13 Figure e2 is an enlarged view of Figure e. Fig.13 Figure f2 is an enlarged view of Figure f. DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0051] In a first aspect, some embodiments of the present invention provide a method for preparing pancreatic hydrolysate, comprising the following steps:

[0052] Using pig pancreas as raw material, 10% of the total mass of the pig pancreas is used to prepare a fermentation substrate, and the fermentation substrate and the remaining pig pancreas are enzymatically hydrolyzed, solid-liquid separated, and dried to obtain a pancreatic enzymolyzate.

[0053] In some embodiments, the specific process of preparing the fermentation substrate includes: washing and crushing the pig pancreas, adding the fermentation agent, and fermenting at a temperature of 35° C.-37° C. for 11 h-13 h.

[0054] In some embodiments, the fermentation agent includes Pediococcus pentosaceus and Lactobacillus sakei.

[0055] In some embodiments, the method for preparing the starter includes: inoculating the bacterial powder of Pediococcus pentosaceus and the bacterial powder of Lactobacillus sakei into a primary activated seed culture medium for activation culture, and then inoculating into a secondary activated seed culture medium for culture to obtain the starter.

[0056] In some embodiments, the formula of the primary activated seed culture medium and the secondary activated seed culture medium includes: 10 g / L peptone, 10 g / L beef extract, 5 g / L yeast extract, 20 g / L glucose, 5 g / L sodium acetate, 2 g / L diamine hydrogen citrate, 1 mL / L Tween-80, 2 g / L dipotassium hydrogen phosphate, 0.2 g / L magnesium sulfate heptahydrate, and 0.05 g / L manganese sulfate heptahydrate, and the pH value is adjusted to 5.5-5.9.

[0057] In some embodiments, the activation culture and culture conditions are: temperature of 36° C.-38° C., and culture time of 23 h-25 h.

[0058] In some embodiments, the mass ratio of the bacterial powder of Pediococcus pentosaceus to the bacterial powder of Lactobacillus sakei is 1:1.

[0059] In some embodiments, when inoculated into the primary activated seed culture medium, the mass concentration of the bacterial powder of Pediococcus pentosaceus in the primary activated seed culture medium is 1 g / 250 mL.

[0060] In some embodiments, the inoculation amount of the secondary activated seed culture medium is 1% (v / v).

[0061] In some embodiments, the mass ratio of the added fermentation agent in the mashed porcine pancreas is 1%.

[0062] In some embodiments, the enzymatic hydrolysis further includes sterilization, and the specific process includes: washing the remaining pig pancreas, adding water, grinding it into a paste, and then heating the paste to sterilize it.

[0063] In some embodiments, the specific heating process includes: heating to 100°C, keeping at 100°C for 25min-35min, with a heating rate of 100°C / 3h; and then cooling to 35°C-50°C.

[0064] In some embodiments, the mass ratio of the remaining porcine pancreas to water is 1:4.

[0065] In some embodiments, the specific process of enzymatically hydrolyzing the fermentation substrate and the remaining porcine pancreas includes: mixing the fermentation substrate and the sterilized substance, then adding papain, and maintaining at 35°C-50°C for 4h-6h, and performing inactivation treatment after enzymatic hydrolysis.

[0066] In some embodiments, the mass ratio between the fermentation substrate and the sterilized substance is 1:45.

[0067] In some embodiments, the mass ratio of the added mass of papain to the fermentation substrate is 0.1%-0.3%.

[0068] In some embodiments, the specific process of the inactivation treatment includes: adjusting the pH value of the environment after enzymatic hydrolysis to 6, and then heating to 80° C.-100° C. and maintaining it for 15 min-25 min.

[0069] In some embodiments, the specific process of solid-liquid separation includes: after solid-liquid separation, cooling to between 35°C and 50°C, keeping the temperature constant for 3h-5h, adding citric acid to the filtrate after solid-liquid separation, heating the filtrate to 70°C-80°C, and then continuing to heat the filtrate to 90°C-100°C and maintaining it for 1.5h-2.5h, passing the filtrate through an 80-mesh sieve, and spray drying to obtain pancreatic enzymatic hydrolysate.

[0070] In some embodiments, the filtrate is stirred while being heated to 70°C-80°C, and the stirring is stopped when the temperature reaches 70°C-80°C.

[0071] In some embodiments, the mass concentration of citric acid in the filtrate after solid-liquid separation is 1.5%.

[0072] In a second aspect, some embodiments of the present invention also provide pancreatic hydrolysate obtained by the pancreatic hydrolysate preparation method.

[0073] In a third aspect, some embodiments of the present invention also provide use of the pancreatic hydrolysate in the preparation of a drug for alleviating inflammation.

[0074] In a fourth aspect, some embodiments of the present invention also provide the use of the pancreatic hydrolysate in food.

[0075] The following describes it in conjunction with specific embodiments.

[0076] Example 1

[0077] (1) Take 1 kg of fresh porcine pancreas, wash and mince it into a paste, and obtain porcine pancreatic pulp as a fermentation substrate. Add a starter culture to the porcine pancreatic pulp, and culture it in a constant temperature incubator at a temperature of about 36°C for about 12 h. The starter culture consists of Pediococcus pentosaceus and Lactobacillus sakei, and the mass ratio of the starter culture to the porcine pancreatic pulp is 1%.

[0078] (2) Preparation process of starter: Inoculate the powder of Pediococcus pentosaceus and the powder of Lactobacillus sake into the primary activated seed culture medium for activation culture at 37°C for 24 hours; then inoculate into the secondary activated seed culture medium (inoculation amount is 1% (v / v)), culture at 37°C for 24 hours to obtain the starter. Among them, the mass ratio of the powder of Pediococcus pentosaceus and the powder of Lactobacillus sake is 1:1, and the mass concentration of the powder of Pediococcus pentosaceus in the primary activated seed culture medium is 1g / 250mL. Among them, Pediococcus pentosaceus TS289313 (Testo Biotechnology) and Lactobacillus sake TS344651 (Testo Biotechnology) are used.

[0079] (3) Take 9 kg of fresh pig pancreas, add 4 times the weight of water to make it easier to crush. Heat to 100°C and keep warm for 20 minutes. Lower the temperature to 50°C, add 1 kg of pig pancreatic pulp cultured in the constant temperature incubator in (1), stir evenly, keep at 35-50°C for 5 hours, and keep stirring. After the enzymatic hydrolysis is completed, adjust the pH of the solution to 6, heat to 100°C and keep for 20 minutes, and filter to obtain the enzymatic hydrolysis clear liquid. After keeping at 48°C for 4 hours, pour the filtered liquid into the heating tank and add citric acid. The mass concentration of citric acid in the filtrate after solid-liquid separation is 1.5%; start stirring and gradually increase the temperature to 75°C, stop stirring, gradually increase the temperature to 100°C, keep the temperature for 2 hours, remove all impurities in the liquid, and pass the liquid with impurities through an 80-mesh sieve for use. Finally, spray dry the liquid to obtain the extract.

[0080] The pancreatic hydrolysate obtained in Example 1 of the present invention was used:

[0081] (1) The pancreatic hydrolysate obtained in Example 1 was experimentally verified to have a protective effect on a lipopolysaccharide (LPS)-induced rat systemic inflammation model.

[0082] In order to study the protective effect of pancreatic hydrolysate on rats with LPS-induced systemic inflammation, pancreatic hydrolysate was prepared into capsules and used in rats with systemic inflammation, and the changes of related factors were observed.

[0083] 1) Experimental materials:

[0084] Sixty healthy male Wistar rats weighing 200±20g were raised in a laboratory at room temperature of (22±2)℃.

[0085] 2) Animal grouping:

[0086] After one week of adaptive feeding of healthy rats, the rectal temperature of the rats was measured once, and 60 rats with normal body temperature were randomly selected and divided into normal dose protection group (P1), high dose protection group (P2), ultra-high dose protection group (P3), protection normal group, protection model group, and protection positive drug group, with 10 rats in each group. According to the normal human body daily intake of 1.8g pancreatic enzymatic hydrolysate, the adult body weight is about 60kg, so the daily intake of adults is about 1.8g / 60kg≈0.03g / kg, but there is a difference in the metabolic rate between rats and humans, and the conversion factor is 6.3. Therefore, according to the rat body weight, the daily intake of pancreatic enzyme extract should be 0.03g / kg×6.3=0.189g / kg≈0.19g / kg. The normal dose protection group was set with an intake of 0.19 g / kg (rat body weight), the high dose protection group was set with an intake of 3 times, and the ultra-high dose protection group was set with an intake of 5 times. The normal diet of the protection normal group was not intervened, the normal diet of the protection model group was not intervened after modeling, and the positive drug group was fed with 0.0015 g / kg hydrocortisone acetate solution once a day before modeling. The rats were gavaged twice a day. Therefore, the single gavage dose was finally determined to be 0.19 g / kg (P1), 0.57 g / kg (P2), 0.95 g / kg (P3), and the positive drug group was protected according to the rat body weight. In addition to the normal diet, the P1 group, P2 group, and P3 group took pancreatic enzymatic hydrolysate according to the dose for one week in advance, and gavage twice a day.

[0087] Three days before modeling, the rectal temperature of all rats was measured once a day, and the measurement was performed at the same time and place as much as possible. Rats with temperature fluctuations (∆T>1.0℃) and single body temperatures above 38℃ were eliminated. Finally, 8 rats with qualified temperatures were selected from the normal dose protection group (P1), high dose protection group (P2), and ultra-high dose protection group (P3).

[0088] After one week of drug administration, rats were allowed to eat and drink freely before the start of the animal model establishment experiment. On the day of the experiment, the rectal temperature of the rats was measured once every 0.5 hours before the injection of LPS at a concentration of 2 mg / kg. The measurement was performed 3 times, and the average value was taken as the basal body temperature of the rats. The body temperature of the rats was measured every 0.5 hours after the injection, and the general condition of the rats was observed. The model was successfully established when the rats showed typical manifestations of systemic inflammatory response syndrome such as fever, increased heart rate, and shortness of breath.

[0089] 3) Sample preparation:

[0090] The rats were first subjected to adaptive feeding for one week, and then gavage with pancreatic enzymatic hydrolysate capsules for one week before modeling experiments. At the end of the experiment (a total of 15 days from adaptive feeding to the end of the experiment), all rats were killed and serum was collected. Some tissues were fixed in 4% paraformaldehyde, and the rest were stored at -80°C.

[0091] 4) Determination method:

[0092] Serum was collected for routine blood tests, and Luye Bio's ELISA kit was used to detect high-sensitivity C-reactive protein (hs-CRP), C-reactive protein (CRP), immunoglobulin A (IgA), immunoglobulin G (IgG), interleukin-6IL-6, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the serum.

[0093] 5) Data processing statistics:

[0094] Data are expressed as mean ± SEM. Data were analyzed using Prism 8 (GraphPad Software, USA), and statistical significance was determined by two-tailed unpaired Student's t-test. Differences with p values ​​< 0.05 were considered statistically significant.

[0095] (2) Experimental verification of the therapeutic effect of pancreatic hydrolysate on LPS-induced systemic inflammation model in rats:

[0096] In order to study the therapeutic effect of pancreatic hydrolysate on rats with LPS-induced systemic inflammation, the pancreatic hydrolysate was made into capsules and used in rats with systemic inflammation, and the changes of related factors were observed.

[0097] 1) Experimental materials:

[0098] 55 healthy male Wistar rats, weighing 200±20 g, were raised in a laboratory at room temperature of (22±2)°C.

[0099] 2) Animal grouping:

[0100] 48 rats with qualified temperature were screened and randomly divided into 6 groups, 8 in each group, namely normal group (N), model group (M), positive drug group (PC), normal dose treatment group (T1), high dose treatment group (T2), ultra-high dose treatment group (T3) and pancreatic enzyme extract treatment group. Establishment of animal model Before the experiment, rats were free to eat and drink water. On the day of the experiment, the rectal temperature of rats was measured once every 0.5 hours before LPS injection. The average value was taken as the basal body temperature of rats. In the experiment, except for the blank control group, each group was intraperitoneally injected with LPS at 2 mg / kg according to the weight of rats. The body temperature of rats was measured every 0.5 hours after injection. The body temperature of rats was measured 3 times and the general condition of rats was observed. The model was successfully established when rats showed typical manifestations of systemic inflammatory response syndrome such as fever, increased heart rate and shortness of breath. The groups were divided into 8 rats in each group. The single gavage dose of the treatment group was 0.19 g / kg (normal dose treatment group), 0.57 g / kg (high dose treatment group), and 0.95 g / kg (ultra-high dose treatment group), and the mice were gavaged twice a day. The normal group (N) had a normal diet without other interventions. The model group (M) had a normal diet after modeling without other interventions. The positive drug group (PC) was fed 0.0015 g / kg hydrocortisone acetate solution once a day after modeling. The mice were gavaged on the day of modeling and on the 2nd, 3rd, and 4th days. The mice were killed on the fifth day.

[0101] 3) Sample preparation:

[0102] The rats were first modeled, and then fed with drugs. At the end of the experiment (a total of 15 days from the beginning of feeding to the experimental results), all rats were killed and serum was collected. Some tissues were fixed in 4% paraformaldehyde, and the rest were stored at -80°C.

[0103] 4) Determination method:

[0104] Serum was collected for routine blood tests, and Luye Bio's ELISA kit was used to detect high-sensitivity C-reactive protein (hs-CRP), C-reactive protein (CRP), immunoglobulin A (IgA), immunoglobulin G (IgG), interleukin-6IL-6, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the serum.

[0105] 5) Data processing statistics:

[0106] Data are expressed as mean ± SEM. Data were analyzed using Prism 8 (GraphPad Software, USA), and statistical significance was determined by two-tailed unpaired Student's t-test. Differences with p values ​​< 0.05 were considered statistically significant.

[0107] Results and Analysis:

[0108] After oral administration of pancreatic enzymatic hydrolysate, the rats had higher body weight and lower body temperature compared with the protection model group, and the overall condition was well improved. Figure 1-Figure 2 Compared with the protection model group, the number of neutrophils in the blood routine test was reduced, and the effect of the ultra-high dose protection group (P3) was the most obvious. Figure 5 As shown. The data of ELISA test showed that the protection group had a good protection effect. Compared with the model group (M), the high-sensitivity C-reactive protein test (hs-CRP), C-reactive protein test (CRP), immunoglobulin A (IgA), immunoglobulin G (IgG), interleukin-6IL-6, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were all reduced. Among them, the ultra-high dose protection group (P3) had the most obvious effect. Figure 6-Figure 12 As shown, it can be seen that the pancreatic hydrolysate obtained by the present invention has a protective effect in systemic inflammation.

[0109] After intragastric administration of pancreatic enzymatic hydrolysate, the rats had higher body weight and lower body temperature compared with the model group (M), and the overall condition was well improved. Figure 3-Figure 4 Compared with the model group (M), the number of neutrophils in the blood routine test was reduced, and the ultra-high dose treatment group (T3) had the best treatment effect. Figure 5 Compared with the model group (M), the data of ELISA test showed that the protection group had a good protective effect, and the levels of high-sensitivity C-reactive protein (hs-CRP), C-reactive protein (CRP), immunoglobulin A (IgA), immunoglobulin G (IgG), interleukin-6IL-6, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were all reduced. Among them, the ultra-high-dose treatment group (T3) had the best treatment effect. Figure 6-Figure 12 As shown. H&E staining results showed that after LPS modeling, the mouse muscle layer was partially broken and detached, the intestinal glands in the lamina propria were irregularly arranged and severely dilated, and the intestinal mucosal epithelium was incomplete. Compared with the model group (M), the normal dose treatment group (T1), the high dose treatment group (T2), and the ultra-high dose treatment group (T3) showed no significant changes in colon morphology compared with the model group (M). The intestinal wall of the high dose treatment group (T2) and the ultra-high dose treatment group (T3) was slightly thicker, as shown in Figure 2. Fig.13 It can be seen that the pancreatic enzymatic hydrolysate obtained by the present invention has the effect of treating inflammation.

[0110] In summary, the method of the present invention can promote the hydrolysis of biological macromolecular nucleic acids into small molecule nucleic acid degradation products, and the generated small molecule nucleic acid degradation products have the effect of protecting and treating systemic inflammation.

[0111] It should be noted that the terms used in the present invention are only for describing specific embodiments, rather than limiting the scope of the present application. As shown in the present specification, unless the context clearly indicates an exception, the words "one", "a", "a kind of" and / or "the" do not specifically refer to the singular, but may also include the plural. The terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that the process, method or device including a series of elements includes not only those elements, but also includes other elements not explicitly listed, or also includes elements inherent to such process, method or device. In the absence of more restrictions, the elements defined by the sentence "include one..." do not exclude the presence of other identical elements in the process, method or device including the elements.

[0112] It should also be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. Unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", etc. should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be an indirect connection through an intermediate medium, or it can be a connection between the two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0113] 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 aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing pancreatic enzymatic hydrolysate, characterized in that: The steps include: Using pig pancreas as raw material, washing and crushing 10% of the total mass of the pig pancreas, adding a fermentation agent, fermenting at a temperature of 35°C-37°C for 11h-13h, enzymolysis, solid-liquid separation, and drying to obtain pancreatic enzymolysate; The mass ratio of the fermentation agent in the crushed pig pancreas is 1%; The fermentation agent is Pediococcus pentosaceus and Lactobacillus sakei; The enzymolysis also includes sterilization, and the specific process includes: washing 90% of the total mass of the pig pancreas of the raw material, adding water, grinding into a uniform paste, and then heating the paste to sterilize; The specific process of the enzymatic hydrolysis includes: mixing the fermented product and the sterilized material, then adding papain, and maintaining at 35°C-50°C for 4h-6h, and performing inactivation treatment after enzymatic hydrolysis; The specific process of the solid-liquid separation includes: after solid-liquid separation, cooling to between 35°C and 50°C, keeping the temperature constant for 3h-5h, adding citric acid to the filtrate after solid-liquid separation, heating the filtrate to 70°C-80°C, and then continuing to heat the filtrate to 90°C-100°C and keeping it for 1.5h-2.5h, passing the filtrate through an 80-mesh sieve, and spray drying to obtain pancreatic enzymatic hydrolysate.

2. The preparation method according to claim 1, characterized in that: The specific heating process includes: heating to 100°C, keeping at 100°C for 25min-35min, and then cooling to 35°C-50°C; the heating rate is 100°C / 3h; The mass ratio of 90% of the total mass of the raw materials to water is 1:

4.

3. The preparation method according to claim 1, characterized in that: The mass ratio between the fermented product and the sterilized substance is 1:45; The mass ratio of the added mass of the papain to the fermented product is 0.1%-0.3%.

4. The preparation method according to claim 1, characterized in that: The specific process of the inactivation treatment includes: adjusting the pH value of the environment after enzymatic hydrolysis to 6, and then heating to 80° C.-100° C. and maintaining it for 15 min-25 min.

5. The preparation method according to claim 1, characterized in that: The filtrate is stirred during the process of heating to 70°C-80°C, and the stirring is stopped when the temperature reaches 70°C-80°C; The mass concentration of the citric acid in the filtrate after solid-liquid separation is 1.5%.

6. The pancreatic hydrolysate obtained by the method for preparing pancreatic hydrolysate according to any one of claims 1 to 5.

7. Use of the pancreatic enzymatic hydrolysate according to claim 6 in the preparation of a drug for alleviating inflammation.

8. Use of the pancreatic hydrolysate according to claim 6 in food.

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