Application of mixture of formic acid and calcium lignosulphonate in treatment of chronic colitis

Abstract

The invention discloses application of a mixture of formic acid and calcium lignosulphonate in treatment of chronic colitis, and belongs to the field of biological medicine. On the premise of not influencing the application of formic acid, the adverse effect of formic acid on human health is reduced by adopting a mixture of formic acid and calcium lignosulphonate. Through animal experiments, the relieving effect of lignoformic acid on mice with chronic colitis is further investigated in combination with pathological characteristics, oxidation resistance, inflammatory factor level change and other aspects. Experimental results show that the ligninecarboxylic acid can remarkably improve the tissue structure of the colon, promote secretion of anti-inflammatory factors and improve the oxidation resistance of the intestinal tract, so that the ligninecarboxylic acid can be used as a good treatment product for relieving colitis.

Description

Technical Field

[0001] The invention relates to application of a mixture of formic acid and calcium lignin sulfonate in treating chronic colitis, belonging to the technical field of biomedicine. Background Art

[0002] Chronic colitis is a digestive system disease characterized by long-term or recurrent inflammation of the colon, with clinical manifestations including abdominal pain, diarrhea, mucus and blood in the stool. This disease is common in young and middle-aged people, and is more common in areas with a high incidence of digestive system diseases. With the changes in modern lifestyles and differences in eating habits, the incidence of chronic colitis has increased year by year and has become one of the important public health issues worldwide.

[0003] The pathogenesis of chronic colitis is complex, involving many factors such as immune system disorders, intestinal microbiome imbalance, genetic susceptibility, and environmental factors. Although there are currently a variety of treatments, including drug therapy, surgical treatment, etc., these treatments still face many challenges. Traditional drug treatments (such as steroids, immunosuppressants, antibiotics, etc.) often have side effects caused by long-term use, such as immune suppression, gastrointestinal adverse reactions, etc. In addition, existing treatments have failed to completely solve the fundamental problems of chronic colitis, and the recurrence rate of patients is high, the treatment cycle is long, and the quality of life of patients has a great impact. Therefore, the development of new drugs with fewer side effects and that can effectively control the disease has become an urgent issue to be solved. Summary of the invention

[0004] Lignosulfonate has been proven to have hypoglycemic and antibacterial properties in mice. LA is a high molecular weight, large volume, and sterically hindered substance on the aromatic ring. These characteristics lead to low LA reactivity. Chemical reactions will only occur under special conditions, such as in a strong acid environment. The special negative macromolecular spatial structure of LA has a strong adsorption effect on positive molecules such as small molecule organic acids. Therefore, after LA is mixed with FA, LA can be used as a carrier of FA to maintain a relatively stable state of the mixture. This can make lignosulfonic acid have a coating effect similar to that of a lipid outer membrane, which can slowly release the coated organic acid, maintain its antibacterial properties, and prolong its action time in the intestine.

[0005] In view of the existing situation, the purpose of the present invention is to provide a new type of formic acid product for use in the treatment of chronic colitis, and to study the protective effect of lignin formic acid on the intestine by comparing it with 5-aminosalicylic acid. The lignin formic acid provided by the present invention can reduce the harm of formic acid to the human body and alleviate the intestinal damage of chronic colitis without affecting the application of formic acid, thereby achieving the purpose of treating chronic colitis.

[0006] The lignin formic acid of the present invention is a mixture of formic acid and calcium lignin sulfonate. Calcium lignin sulfonate is a water-soluble derivative obtained by sulfonation reaction of lignin, and has high bioavailability. Formic acid, as an organic acid, has good antibacterial and anti-inflammatory effects. Lignin formic acid is a mixture of calcium lignin sulfonate and formic acid.

[0007] Compared with single lignin or formic acid, lignin formic acid combines the advantages of both, not only enhancing its anti-inflammatory and antioxidant activities, but also improving its solubility and bioavailability. Studies have shown that lignin formic acid has the effects of inhibiting intestinal inflammation, regulating immune function, and repairing intestinal barriers, making it a potential new drug for the treatment of digestive system diseases such as chronic colitis.

[0008] Technical solution:

[0009] The technical solution of the present invention aims to provide a new type of drug lignin formic acid for treating chronic colitis, and play a therapeutic role on chronic colitis by regulating the intestinal barrier, antioxidant capacity and inhibiting intestinal inflammation.

[0010] In one embodiment of the present invention, the preparation method of lignin formic acid is a mixture of formic acid and calcium lignin sulfonate in a volume ratio of 1: (2-3.5). Preferably, the mixture is composed of formic acid and calcium lignin sulfonate in a volume ratio of 1: 3. More preferably, the mass concentration of formic acid in the formic acid aqueous solution is greater than or equal to 70%, and the mass concentration of calcium lignin sulfonate in the calcium lignin sulfonate aqueous solution is 90%-95%.

[0011] Furthermore, the concentration of the formic acid is 75%, and the concentration of the calcium lignin sulfonate is 95%.

[0012] The invention provides application of lignin formic acid in preparing a preparation for alleviating, preventing and / or treating DSS-induced colitis.

[0013] The present invention provides the use of lignin formic acid in preparing a product for alleviating, preventing and / or treating chronic colitis, characterized in that the product contains an effective dose of lignin formic acid, and the product is a food, a medicine, a dietary nutritional supplement or a health product.

[0014] In one embodiment of the present invention, the preparation method of lignin formic acid is a mixture of formic acid and calcium lignin sulfonate in a volume ratio of 1: (2-3.5). Preferably, the mixture is composed of formic acid and calcium lignin sulfonate in a volume ratio of 1: 3. More preferably, the mass concentration of formic acid in the formic acid aqueous solution is greater than or equal to 70%, and the mass concentration of calcium lignin sulfonate in the calcium lignin sulfonate aqueous solution is 90%-95%.

[0015] Furthermore, the concentration of the formic acid is 75%, and the concentration of the calcium lignin sulfonate is 95%.

[0016] In one embodiment of the present invention, the product is used in at least one of (a) to (d):

[0017] (a) Relieve weight loss and bloody stools, reduce disease activity index, relieve colon shortening, and reduce colon tissue damage;

[0018] (b) reduce colon inflammation;

[0019] (c) Regulate the antioxidant capacity of the intestine.

[0020] In one embodiment of the present invention, the reduction of colon inflammation includes inhibiting colon inflammatory factors, promoting the expression of anti-inflammatory factors, and inhibiting the activity of colitis-related enzymes.

[0021] In one embodiment of the present invention, the inhibiting the expression of inflammatory factors includes reducing the levels of IL 1β and TNFα.

[0022] In one embodiment of the present invention, the promoting the expression of anti-inflammatory factors includes increasing the content of anti-inflammatory cytokines IL4 and IL-10.

[0023] In one embodiment of the present invention, the regulating the activity of antioxidant-related enzymes comprises increasing the concentrations of antioxidant-related enzymes CAT, GSH-PX, SOD and T-AOC.

[0024] In one embodiment of the present invention, the regulating the activity of antioxidant-related enzymes comprises reducing the concentrations of MDA and MPO.

[0025] In one embodiment of the present invention, the product is a medicine, a food, a health product, a veterinary drug or a feed additive.

[0026] In one embodiment of the present invention, when the product is a medicine, veterinary drug or feed additive, the effective dose is not less than 1 g / kg.

[0027] In one embodiment of the present invention, the effective dose is 1 to 4 g / kg.

[0028] In one embodiment of the present invention, the drug further comprises a pharmaceutically acceptable excipient; the pharmaceutically acceptable excipient refers to any diluent, adjuvant and / or carrier that can be used in the pharmaceutical field.

[0029] In one embodiment of the present invention, the veterinary drug or feed additive further contains conventional auxiliary materials.

[0030] In one embodiment of the present invention, the conventional auxiliary materials include one or more of fillers, flavoring agents, binders, disintegrants, lubricants, and antacids.

[0031] In one embodiment of the present invention, the food includes, but is not limited to, cereal products, vegetable products, fruit products, fermented foods, meat products, egg products, dairy products, drinks, and snacks;

[0032] In one embodiment of the present invention, the food further comprises special dietary food;

[0033] In one embodiment of the present invention, the health care product further contains acceptable excipients.

[0034] In one embodiment of the present invention, the dosage form of the drug includes any one of drops, oral liquid, tablets, capsules, granules, films, gels, powders, emulsions, pills or solutions;

[0035] In one embodiment of the present invention, the drug further contains pharmaceutically acceptable excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field;

[0036] In one embodiment of the present invention, the auxiliary materials include one or more of the following: adhesives such as cellulose derivatives, alginate, gelatin and polyvinyl pyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol, etc.; fillers such as starch and sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid and glycerol fatty acid esters; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, etc.; lubricants such as hydrogenated vegetable oil, talc and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, cellulose acetate; in addition, other auxiliary materials such as flavoring agents and sweeteners can be added to the composition.

[0037] The present invention also provides the use of the calcium lignin sulfonate in preparing a product for reducing the side effects of formic acid in treating chronic colitis. The use is to prepare a product for treating chronic colitis by mixing formic acid and calcium lignin sulfonate.

[0038] Preferably, the mixture is composed of formic acid and calcium lignin sulfonate in a volume ratio of 1:(2-3.5). Preferably, the mixture is composed of formic acid and calcium lignin sulfonate in a volume ratio of 1:3. Further preferably, the mass concentration of formic acid in the formic acid aqueous solution is greater than or equal to 70%, and the mass concentration of calcium lignin sulfonate in the calcium lignin sulfonate aqueous solution is 90%-95%.

[0039] Furthermore, the concentration of the formic acid is 75%, and the concentration of the calcium lignin sulfonate is 95%.

[0040] In one embodiment of the present invention, the product is a food, a medicine, a dietary supplement or a health product.

[0041] In one embodiment of the present invention, the food includes, but is not limited to, cereal products, vegetable products, fruit products, fermented foods, meat products, egg products, dairy products, drinks, and snacks;

[0042] In one embodiment of the present invention, the food further comprises special dietary food;

[0043] In one embodiment of the present invention, the health care product further contains acceptable excipients.

[0044] In one embodiment of the present invention, the dosage form of the drug includes any one of drops, oral liquid, tablets, capsules, granules, films, gels, powders, emulsions, pills or solutions.

[0045] In one embodiment of the present invention, the drug further contains pharmaceutically acceptable excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field.

[0046] In one embodiment of the present invention, the auxiliary materials include one or more of the following: adhesives such as cellulose derivatives, alginate, gelatin and polyvinyl pyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol, etc.; fillers such as starch and sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid and glycerol fatty acid esters; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red, etc.; lubricants such as hydrogenated vegetable oil, talc and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, cellulose acetate; in addition, other auxiliary materials such as flavoring agents and sweeteners can be added to the composition.

[0047] Beneficial Effects

[0048] (1) The lignin formic acid product of the present invention provides for the first time a new type of formic acid product, which is an effective solution for treating chronic colitis. Through anti-inflammatory, anti-oxidative and regulating and repairing intestinal barrier functions, lignin formic acid can significantly relieve colitis symptoms, reduce oxidative damage, and repair the intestinal barrier. Animal experiments have shown that lignin formic acid significantly improves the clinical symptoms of colitis caused by colitis, such as weight loss, bloody stools, inflammatory cell infiltration and abdominal pain, and in animal experiments, the disease activity index (DAI) is significantly reduced, indicating that it can effectively reduce the severity of colitis and promote pathological recovery.

[0049] (2) The lignin formic acid product of the present invention not only has anti-inflammatory effects, but also improves the therapeutic effect through multiple effects such as anti-oxidation and enhancing intestinal barrier function. Its anti-inflammatory effect effectively reduces the inflammatory response in the colon by inhibiting the secretion of inflammatory factors; at the same time, the antioxidant properties of lignin formic acid effectively reduce oxidative stress and repair oxidative damage caused by chronic inflammation.

[0050] (3) The lignin formic acid product of the present invention improves the intestinal microenvironment and promotes the repair and regeneration of intestinal epithelial cells by regulating the intestinal barrier function. The experimental results show that lignin formic acid can repair the damaged intestinal barrier and enhance the intestinal resistance to external harmful substances, thereby achieving the effect of treating enteritis and preventing further aggravation of colitis.

[0051] (4) Compared with existing colitis treatments, lignin formic acid has a wider range of adaptability and higher clinical application potential. Its multiple mechanisms of action enable it to alleviate the symptoms of colitis while effectively avoiding the side effects of long-term drug use, reducing the probability of recurrence, and having good safety and tolerability.

[0052] (5) Lignin formic acid has a good long-term effect. Experimental results show that it can continuously improve colitis symptoms within a certain treatment cycle and effectively prevent the recurrence of inflammation. Compared with existing drugs that have significant short-term efficacy but are prone to recurrence, lignin formic acid has a more prominent advantage in long-term efficacy.

[0053] (6) Since lignin formic acid can effectively reduce the recurrence and aggravation of colitis, it reduces the cost of long-term drug treatment for patients. The widespread application of this technology is expected to significantly reduce the medical expenses of patients with colitis, while also bringing greater social benefits by improving the quality of life of patients.

[0054] (7) As a new type of natural product, lignin formic acid has the advantages of natural origin, high safety, and low toxicity and side effects compared to existing chemical drugs. This innovative technology not only fills the gap in the treatment of colitis, but also provides new ideas and directions for the treatment of other inflammatory diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] In order to more clearly illustrate the technical solution of this embodiment, the following briefly introduces the drawings required in the description of this embodiment. Obviously, the drawings described below are some embodiments of the present application, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0056] Figure 1 Lignin formic acid improved DSS-induced chronic colitis in mice; (a) Schematic diagram of DSS-induced chronic colitis in C57BL / 6J mice; (b) Weight changes of mice in each group during the entire treatment period (n=10); (c) Disease Activity Index (DAI) score of mice in each group; (d) Representative images of colon length; (e) Colon length statistics. Data are expressed as SEM±mean. The same letters indicate no statistically significant difference between groups (P>0.05), and different letters indicate significant difference between groups (P<0.05).

[0057] Figure 2 H&E staining of colon tissue in mice with chronic colitis treated with lignin formic acid.

[0058] Figure 3 The results of lignin formic acid regulating inflammatory cytokines are shown in Figure 2; (a) serum TNF-α; (b) serum IL-1β; (c) serum IL-4; (d) serum IL-10. Data are expressed as SEM±mean. The same letters indicate no statistically significant difference between the groups (P>0.05), and different letters indicate significant difference between the groups (P<0.05).

[0059] Figure 4 The effect of lignin formic acid on the antioxidant level of mouse colon tissue. (a) CAT activity in colon tissue; (b) GSH-PX activity in colon tissue; (c) MDA activity in colon tissue; (d) SOD activity in colon tissue; (e) T-AOC activity in colon tissue; (f) MPO activity in colon tissue. The same letters indicate no significant difference between the groups (P>0.05), and different letters indicate significant difference between the groups (P<0.05). DETAILED DESCRIPTION

[0060] The preparation method of lignin formic acid (LFA) involved in the following examples is:

[0061] Lignin formic acid is prepared by mixing a formic acid aqueous solution and a calcium lignin sulfonate aqueous solution in a volume ratio of 1:3, wherein the mass concentration of formic acid in the formic acid aqueous solution is 75%, and the mass concentration of calcium lignin sulfonate in the calcium lignin sulfonate aqueous solution is 95%. The mixture of the two is then freeze-dried, and the sample is quickly frozen to -40°C to -80°C to ensure that the water is converted into solid ice. Then, the frozen sample is placed in a freeze dryer, and the temperature is set to -40°C to -50°C, the vacuum degree is 10Pa, and the time is 72h.

[0062] The mouse feed involved in the following examples was purchased from: Jiangsu Collaborative Medicine Bioengineering Co., Ltd., and the ingredients are shown in Table 1 below:

[0063] Table 1: Basic feed composition and nutritional level (air-dried basis)

[0064]

[0065] The premix provided the following per kg of the diet:Fe 66.5mg, Zn 88mg, Mn 110mg, I 0.7mg, Se 0.288mg, Cu 8.8mg, VA 1000IU, VB13mg, VB29mg, VB69mg, VB 12 0.025mg, VD3 3500IU, VE 30mg, VK3 5mg, choline 800mg, nicotinic acid 45mg, folic acid 1.2mg.

[0066] The 5-ASA (5-aminosalicylic acid) involved in the following examples was purchased from Shanghai MacLean Biochemical Technology Co., Ltd., CAS: 89-57-6.

[0067] Example 1: Effect of lignin formic acid on epidemiological data of DSS-induced chronic colitis mice

[0068] 1. Animal experiments

[0069] C57BL / 6J mice (male, 21–24 g, 6 weeks old) were purchased from Weitong Lihua Laboratory Animal Technology Co., Ltd. (Beijing, China). All mice were housed under specific pathogen-free conditions with controlled temperature (22°C) and humidity and a 12-h light / dark cycle. The animal experiments were reviewed and approved by the Animal Experiment Ethics Committee of Jiangnan University (JN.No20230915c0881213

[399] ) and were performed in accordance with the animal experiment protocol.

[0070] The mice were randomly divided into 6 groups (n=10): blank group, model group, positive control group, low-dose LFA (1g / kg) group, medium-dose LFA (2g / kg) group, and high-dose LFA (4g / kg) group. The chronic colitis model was used, and 2% (w / v) dextran sulfate sodium (DSS) was administered for 3 consecutive cycles (each cycle was 7 days), and then DSS was replaced with normal sterile drinking water for 2 cycles (each cycle was 14 days). The experiment lasted for 49 days ( Figure 1 a). To avoid adverse effects on mice due to improper intragastric administration and long experimental time, all drugs were added to the mouse feed.

[0071] The specific process is as follows:

[0072] I) Blank group (Con group): from day 0 to day 7, normal diet; from day 7 to day 56, normal drinking water and normal feed were fed every day;

[0073] II) Model group (Mod group): 0-7 days, normal diet; 7-14 days, drinking 2% DSS aqueous solution; 15-28 days, normal drinking water; 29-35 days, drinking 2% DSS aqueous solution; 36-49 days, normal drinking water; 50-56 days, drinking 2% DSS aqueous solution; 7-56 days, fed normal feed every day;

[0074] III) Positive control group (5-ASA group): on days 0-7, normal diet; on days 7-14, drinking 2% DSS aqueous solution; on days 15-28, normal drinking water; on days 29-35, drinking 2% DSS aqueous solution; on days 36-49, normal drinking water; on days 50-56, drinking 2% DSS aqueous solution; on days 7-56, feeding feed supplemented with 200 mg / kg of 5-ASA every day;

[0075] IV) Low-dose LFA (1 g / kg) group (F1 group): 0-7 days, normal diet; 7-14 days, drinking 2% DSS aqueous solution; 15-28 days, normal drinking water; 29-35 days, drinking 2% DSS aqueous solution; 36-49 days, normal drinking water; 50-56 days, drinking 2% DSS aqueous solution; 7-56 days, feed supplemented with 1 g / kg LFA every day;

[0076] V) LFA medium dose (2g / kg) group (F2 group): 0-7 days, normal diet; 7-14 days, drinking 2% DSS aqueous solution; 15-28 days, normal drinking water; 29-35 days, drinking 2% DSS aqueous solution; 36-49 days, normal drinking water; 50-56 days, drinking 2% DSS aqueous solution; 7-56 days, feed supplemented with 2g / kg LFA every day;

[0077] VI) LFA high dose (4 g / kg) group (F3 group): 0-7 days, normal diet; 7-14 days, drinking 2% DSS aqueous solution; 15-28 days, normal drinking water; 29-35 days, drinking 2% DSS aqueous solution; 36-49 days, normal drinking water; 50-56 days, drinking 2% DSS aqueous solution; 7-56 days, fed feed supplemented with 4 g / kg LFA every day.

[0078] After inducing chronic colitis, the body weight, bloody stools and stool consistency of each mouse were recorded every two days to assess the disease activity index (DAI) of the mouse. At the end of the experiment, the colon segment was removed and the length from the ileocecal part to the anal verge was measured.

[0079] 2. Experimental results

[0080] The experimental results are as follows Figure 1 As shown, the results show:

[0081] (1) By Figure 1 As shown in b: After the experiment, the body weight of mice in the model group decreased significantly compared with the blank group, and the body weight decreased by 23.23% during the modeling period. The positive control group (5-ASA group) reduced the degree of weight loss of mice after intervention, and the body weight decreased by about 18.8%. The low-dose LFA (1g / kg) group (F1 group) reduced the degree of weight loss of mice after intervention, and the body weight decreased by about 20.14%. The medium-dose LFA (2g / kg) group (F2 group) reduced the degree of weight loss of mice after intervention, and the body weight decreased by about 18.75%. The high-dose LFA (4g / kg) group (F3 group) reduced the degree of weight loss of mice after intervention, and the body weight decreased by about 15.08%.

[0082] It can be seen that compared with the Mod group, lignin formic acid can effectively slow down the weight loss of mice ( Figure 1 b);

[0083] (2) By Figure 1 It can be seen from the figure c that after the experiment, compared with the model group, the positive control group (5-ASA group) reduced the degree of increase in the DAI index of mice after intervention, and the DAI index decreased by about 33.33%. The low-dose LFA (1g / kg) group (F1 group) reduced the degree of increase in the DAI index of mice after intervention, and the DAI index decreased by about 12.22%. The medium-dose LFA (2g / kg) group (F2 group) reduced the degree of increase in the DAI index of mice after intervention, and the DAI index decreased by about 28.9%. The high-dose LFA (4g / kg) group (F3 group) reduced the degree of increase in the DAI index of mice after intervention, and the DAI index decreased by about 37.8%.

[0084] It can be seen that compared with the Mod group, lignin formic acid can effectively reduce the DAI index ( Figure 1 c)

[0085] (3) From d to e in the figure, we can see that the colon length of the blank group mice was 7.8 cm, and the colon length of the mice was shortened to 4.5 cm after DSS modeling. After intervention with the positive control group (5-ASA group), the colon length of the mice recovered to 5.75 cm; after intervention with the low-dose LFA (1 g / kg) group (F1 group), the colon length of the mice recovered to 5.36 cm; after intervention with the medium-dose LFA (2 g / kg) group (F2 group), the colon length of the mice recovered to 5.77 cm; after intervention with the high-dose LFA (4 g / kg) group (F3 group), the colon length of the mice recovered to 6.5 cm;

[0086] It can be seen that compared with the Mod group, lignin formic acid can effectively and significantly slow down the shortening of the colon ( Figure 1 de).

[0087] (4) At the end of the experiment, the colon segments were removed and the colon tissue was analyzed. The colon samples were immersed in 4% paraformaldehyde and embedded in paraffin. The paraffin-embedded colon tissue was sliced ​​(thickness 4 μm) and stained with hematoxylin and eosin (H&E).

[0088] The results are as follows Figure 2 As shown, the Mod group showed a significant increase in colon mucosal inflammatory cell infiltration, crypt destruction, and tissue edema compared with the Con group. However, the administration of lignin formic acid improved colon damage. This included improved colon tissue architecture, more intact mucosal epithelium, and reduced inflammatory cell infiltration. At the same time, the improvement in tissue architecture became more obvious as the dose of lignin formic acid increased.

[0089] Example 2: Effect of lignin formic acid on the secretion of pro-inflammatory and anti-inflammatory factors

[0090] The treatment of each group of mice was the same as in Example 1.

[0091] At the end of the animal experiment, blood was collected from the mouse orbits and placed in a 1.5 mL Eppendorf tube. The tube was allowed to stand for 1 hour and centrifuged at 3000 g for 10 min at 4 °C to collect the supernatant. The levels of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4), and interleukin-10 (IL-10) in serum were measured using a commercial Elisa kit (Xiamen Huijia Biotechnology Co., Ltd.). The results are shown in Table 1. Figure 3 shown.

[0092] Depend on Figure 3It can be seen from a that after the experiment, the TNF-α content in the serum of mice in the blank group was: 45.06ng / L. The TNF-α content in the serum of mice in the model group was: 113.1ng / L. After intervention in the positive control group (5-ASA group), the TNF-α content in the serum of mice was: 86.82ng / L. After intervention in the low-dose LFA (1g / kg) group (F1 group), the TNF-α content in the serum of mice was: 91.24ng / L. After intervention in the medium-dose LFA (2g / kg) group (F2 group), the TNF-α content in the serum of mice was: 82.41ng / L. After intervention in the high-dose LFA (4g / kg) group (F3 group), the TNF-α content in the serum of mice was: 70.45ng / L.

[0093] Depend on Figure 3 As shown in b, after the experiment, the IL-1β content in the serum of the blank group mice was 4.88ng / L, and the IL-1β content in the serum of the model group mice was 14.64ng / L. After the intervention of the positive control group (5-ASA group), the IL-1β content in the serum of the mice was 9.06ng / L. After the intervention of the low-dose LFA (1g / kg) group (F1 group), the IL-1β content in the serum of the mice was 9.48ng / L. After the intervention of the medium-dose LFA (2g / kg) group (F2 group), the IL-1β content in the serum of the mice was 9.83ng / L. After the intervention of the high-dose LFA (4g / kg) group (F3 group), the IL-1β content in the serum of the mice was 8.63ng / L.

[0094] Depend on Figure 3 As shown in c, after the experiment, the IL-4 content in the serum of the blank group mice was 53.80pg / ml, and the IL-4 content in the serum of the model group mice was 31.07pg / ml. After the intervention of the positive control group (5-ASA group), the IL-4 content in the serum of the mice was 45.37pg / ml. After the intervention of the low-dose LFA (1g / kg) group (F1 group), the IL-4 content in the serum of the mice was 32.64pg / ml. After the intervention of the medium-dose LFA (2g / kg) group (F2 group), the IL-4 content in the serum of the mice was 35.09pg / ml. After the intervention of the high-dose LFA (4g / kg) group (F3 group), the IL-4 content in the serum of the mice was 49.91pg / ml.

[0095] Depend on Figure 3It can be seen from the d that after the experiment, the IL-10 content in the serum of the blank group mice was 38.37pg / ml, and the IL-10 content in the serum of the model group mice was 18.96pg / ml. After the intervention of the positive control group (5-ASA group), the IL-10 content in the serum of the mice was 26.52pg / ml. After the intervention of the low-dose LFA (1g / kg) group (F1 group), the IL-10 content in the serum of the mice was 22.08pg / ml. After the intervention of the medium-dose LFA (2g / kg) group (F2 group), the IL-10 content in the serum of the mice was 27.31pg / ml. After the intervention of the high-dose LFA (4g / kg) group (F3 group), the IL-10 content in the serum of the mice was 32.51pg / ml.

[0096] The results showed that lignin formic acid-treated DSS-induced mice could effectively reduce the secretion of pro-inflammatory factors TNF-α and IL-1β in the serum, while promoting the secretion of anti-inflammatory factors IL-4 and IL-10.

[0097] Example 3: Effect of lignin formic acid on the antioxidant level of mouse colon tissue

[0098] The treatment of each group of mice was the same as in Example 1.

[0099] After the animal experiment, the mice were anesthetized and killed, and the mouse colon tissue was removed and quickly frozen in liquid nitrogen. Then, the tissue was cut into small pieces, an appropriate amount of PBS buffer was added, and a mechanical homogenizer was used for homogenization, ensuring that the temperature was kept low during the homogenization process. After the homogenization, the sample was centrifuged (10000-12000rpm, 4°C, 10-20 minutes), and the supernatant was collected for subsequent determination. A commercial kit was used to quantitatively measure the levels of representative oxidoreductases in colon tissue (Nanjing Jiancheng Bioengineering Institute, Nanjing, China): catalase (CAT), reduced glutathione (GSH-PX), malondialdehyde (MDA), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) as well as the inflammatory marker myeloperoxidase (MPO). The results are shown in Figure 4 shown.

[0100] Depend on Figure 4From a in the figure, it can be seen that after the experiment, the content of catalase (CAT) in the colon tissue of mice in the blank group was 50.87U / mg prot, and the content of catalase (CAT) in the colon tissue of mice in the model group was 17.42U / mg prot. After intervention in the positive control group (5-ASA group), the content of catalase (CAT) in the colon tissue of mice was 26.33U / mg prot, after intervention in the low-dose LFA (1g / kg) group (F1 group), the content of catalase (CAT) in the colon tissue of mice was 20.18U / mg prot, after intervention in the medium-dose LFA (2g / kg) group (F2 group), the content of catalase (CAT) in the colon tissue of mice was 22.76U / mg prot, and after intervention in the high-dose LFA (4g / kg) group (F3 group), the content of catalase (CAT) in the colon tissue of mice was 38.64U / mg prot.

[0101] Depend on Figure 4 As can be seen from b, after the experiment, the contents of reduced glutathione (GSH-PX) in the colon tissue of mice in the blank group, model group, positive control group (5-ASA group), low-dose LFA (1g / kg) group (F1 group), medium-dose LFA (2g / kg) group (F2 group), and high-dose LFA (4g / kg) group (F3 group) were 440.76U / mg prot, 143.89U / mg prot, 251.67U / mg prot, 236.14U / mg prot, 251.22U / mg prot, and 335.32U / mg prot, respectively.

[0102] Depend on Figure 4 As can be seen from c, after the experiment, the malondialdehyde (MDA) contents in the colon tissue of mice in the blank group, model group, positive control group (5-ASA group), low-dose LFA (1g / kg) group (F1 group), medium-dose LFA (2g / kg) group (F2 group), and high-dose LFA (4g / kg) group (F3 group) were 0.51U / mg prot, 2.77U / mg prot, 1.58U / mg prot, 1.82U / mg prot, 1.28U / mg prot, and 0.98U / mg prot, respectively.

[0103] Depend on Figure 4It can be seen from d that after the experiment, the superoxide dismutase (SOD) contents in the colon tissue of mice in the blank group, model group, positive control group (5-ASA group), low-dose LFA (1g / kg) group (F1 group), medium-dose LFA (2g / kg) group (F2 group), and high-dose LFA (4g / kg) group (F3 group) were 13.05U / mg prot, 5.82U / mg prot, 9.42U / mgprot, 6.89U / mg prot, 8.50U / mg prot, and 9.65U / mg prot, respectively.

[0104] Depend on Figure 4 It can be seen from e that after the experiment, the total antioxidant capacity (T-AOC) contents in the colon tissue of mice in the blank group, model group, positive control group (5-ASA group), low-dose LFA (1g / kg) group (F1 group), medium-dose LFA (2g / kg) group (F2 group), and high-dose LFA (4g / kg) group (F3 group) were 0.160mM, 0.078mM, 0.111mM, 0.089mM, 0.099mM, and 0.127mM, respectively.

[0105] Depend on Figure 4 As can be seen from f, after the experiment, the myeloperoxidase (MPO) contents in the colon tissue of mice in the blank group, model group, positive control group (5-ASA group), low-dose LFA (1g / kg) group (F1 group), medium-dose LFA (2g / kg) group (F2 group), and high-dose LFA (4g / kg) group (F3 group) were 0.55pg / mg prot, 1.81pg / mg prot, 0.61pg / mgprot, 1.39pg / mg prot, 1.38pg / mg prot, and 0.72pg / mg prot, respectively.

[0106] It can be seen that compared with the Con group, the degree of oxidative stress in the Mod group increased (P<0.05). After the administration of lignin formic acid, the activities of antioxidant enzymes such as CAT, GSH-PX, SOD and T-AOC in colon tissue increased significantly (P<0.05), while the MDA level decreased significantly (P<0.05). As an inflammatory marker, the MPO level in the Mod group increased significantly ( Figure 4 ), while all three intervention groups could significantly reduce MPO levels, indicating that lignin formic acid has great potential in treating chronic colitis in mice.

[0107] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. Use of lignin formic acid in preparing a product for relieving, preventing and / or treating chronic colitis, characterized in that: The product contains an effective dose of lignin formic acid and is a food, a medicine, a dietary nutritional supplement or a health product.

2. The use according to claim 1, characterized in that: The preparation method of the lignin formic acid is to prepare a mixture of formic acid and calcium lignin sulfonate in a volume ratio of 1: (2-3.5); Preferably, the effective dosage of the lignin formic acid in the product is not less than 1 g / kg; further preferably, the effective dosage is 1 to 4 g / kg; Preferably, the product is used for at least one of (a) to (c): (a) Relieve weight loss and bloody stools, reduce disease activity index, relieve colon shortening, and reduce colon tissue damage; (b) reduce colon inflammation; (c) regulating intestinal antioxidant capacity; Preferably, the reducing of colon inflammation comprises inhibiting colon inflammatory factors, promoting the expression of anti-inflammatory factors, and inhibiting the activity of colitis-related enzymes; Preferably, the inhibiting the expression of inflammatory factors comprises reducing the content of IL 1β and TNFα; Preferably, the promoting the expression of anti-inflammatory factors comprises increasing the content of anti-inflammatory cytokines IL-4 and IL-10; Preferably, the regulating the activity of antioxidant-related enzymes includes increasing the concentrations of antioxidant-related enzymes CAT, GSH-PX, SOD and T-AOC, and decreasing the concentrations of MDA and MPO.

3. The use according to claim 1 or 2, characterized in that: The food includes, but is not limited to, cereal products, vegetable products, fruit products, fermented foods, meat products, egg products, dairy products, beverages, and snacks; Preferably, the food also includes special dietary food; Preferably, the health care product further contains acceptable excipients.

4. The use according to any one of claims 1 to 3, characterized in that: The dosage form of the drug includes any one of drops, oral liquid, tablets, capsules, granules, films, gels, powders, emulsions, pills or solutions; Preferably, the drug further contains pharmaceutically acceptable excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field; Preferably, the auxiliary materials include one or more of the following: adhesives such as cellulose derivatives, alginate, gelatin and polyvinyl pyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol; fillers such as starch, sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid and glycerol fatty acid esters; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red; lubricants such as hydrogenated vegetable oil, talc and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, cellulose acetate; in addition, other auxiliary agents such as flavoring agents and sweeteners can be added to the composition.

5. Use of calcium lignin sulfonate in the preparation of a product for reducing the side effects of formic acid in the treatment of chronic colitis, characterized in that: The application is to prepare a product for treating chronic colitis by mixing formic acid and calcium lignin sulfonate in a volume ratio of 1:(2-3.5); Preferably, the effective dosage of the mixture in the product is not less than 1 g / kg; More preferably, the effective dose is 1 to 4 g / kg.

6. The use according to claim 5, characterized in that: The product is a food, medicine, dietary supplement or health product.

7. The use according to claim 6, characterized in that: The food includes, but is not limited to, cereal products, vegetable products, fruit products, fermented foods, meat products, egg products, dairy products, beverages, and snacks; Preferably, the food also includes special dietary food; Preferably, the health care product further contains acceptable excipients.

8. The use according to claim 6 or 7, characterized in that: The dosage form of the drug includes any one of drops, oral liquid, tablets, capsules, granules, films, gels, powders, emulsions, pills or solutions.

9. The use according to claim 8, characterized in that: The medicine also contains pharmaceutically acceptable excipients; the pharmaceutical excipients refer to conventional drug carriers in the pharmaceutical field.

10. The use according to claim 9, characterized in that: The auxiliary materials include one or more of the following: adhesives such as cellulose derivatives, alginate, gelatin and polyvinyl pyrrolidone; diluents such as starch, pregelatinized starch, dextrin, sucrose, lactose, mannitol; fillers such as starch and sucrose; wetting agents such as glycerol; disintegrants such as sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone and dry starch; absorption promoters such as quaternary ammonium compounds; surfactants such as polysorbate, sorbitan fatty acid and glycerol fatty acid esters; colorants such as titanium dioxide, sunset yellow, methylene blue, medicinal iron oxide red; lubricants such as hydrogenated vegetable oil, talc and polyethylene glycol; coating materials such as acrylic resin, hydroxypropyl methylcellulose, povidone, cellulose acetate; in addition, other auxiliary agents such as flavoring agents and sweeteners can be added to the composition.

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