Equine serum cholinesterase activator and application thereof

Abstract

The present invention belongs to the technical field of serum cholinesterase, and specifically relates to an equine serum cholinesterase activator and its application. The formula of the equine serum cholinesterase activator is as follows: 5-10g of nano-silicon dioxide, 0.05-0.1g of sodium hexametaphosphate, and an inorganic liquid fixed to 1L, wherein the particle size of the nano-silicon dioxide is 20-60nm, and the pH of the equine serum cholinesterase activator is neutral or alkaline. The activator of the present invention can significantly increase the activity of cholinesterase, especially the activity of acetylcholinesterase.

Description

Technical Field

[0001] The invention belongs to the technical field of serum cholinesterase, and in particular relates to an equine serum cholinesterase activator and application thereof. Background Art

[0002] Cholinesterase is an important enzyme in serum, mainly used to diagnose liver diseases and organophosphorus poisoning. Cholinesterase is produced by the liver and secreted into the blood. It can reflect the ability of the liver parenchyma to synthesize protein. It is roughly parallel to the decrease in serum albumin, but can more sensitively reflect changes in the condition. When the condition improves, cholinesterase will rise rapidly. Common types are acetylcholinesterase and pseudocholinesterase. Acetylcholine is a neurotransmitter released by cholinergic nerve endings, mainly present in sympathetic nerves, parasympathetic nerves, and motor nerves. When nerve endings are stimulated, acetylcholine is released to play a role in neuromuscular excitation transmission. If acetylcholine is hydrolyzed by cholinesterase, it loses its original efficacy. If the action of cholinesterase is inhibited, acetylcholine will accumulate in excess, causing manifestations similar to organophosphorus poisoning or excessive excitation of the sphenopalatine nerve.

[0003] From the perspective of mechanism of action, the reaction catalyzed by cholinesterase generally includes several processes including substrate binding, catalytic reaction and product release. Substrate binding refers to the binding of acetylcholine or other choline compounds as substrates to the active site of cholinesterase. The active site of cholinesterase usually has a hydrophobic pocket. The substrate molecule binds to the pocket through the choline part, while the acetyl part is exposed to the outside. Catalytic reaction refers to the hydrolysis reaction of the substrate catalyzed by cholinesterase through its amino acid residues. Product release refers to the release of the products such as choline from the active site of cholinesterase after the reaction is completed, while cholinesterase can continue to participate in the reaction.

[0004] Horse serum cholinesterase is a serum cholinesterase of equine animals and is also a commercially available product, such as the horse serum cholinesterase sold by Shanghai Yansheng Industrial Co., Ltd. Early studies have shown that horse serum cholinesterase has been widely used as an analytical reagent in the detection of organophosphorus compounds, such as "He Yongbin. Purification and determination of horse serum cholinesterase [J]. Journal of the First Military Medical University, 1983(03):101-105. DOI:CNKI:SUN:DYJD.0.1983-03-035".

[0005] Horse serum cholinesterase is also an enzyme, and the steps of the catalytic reaction also roughly include several processes of substrate binding, catalytic reaction and product release. Increasing the activity of horse serum cholinesterase will help improve the efficiency of enzyme catalytic reaction, but the existing research in the prior art mostly focuses on inhibiting the activity of cholinesterase, such as CN112940075B, CN113956320B, and CN103420942B, and there is no method of increasing the activity of cholinesterase. Summary of the invention

[0006] In order to activate cholinesterase and improve the activity of cholinesterase, the present invention provides an equine serum cholinesterase activator and application thereof. The activator can significantly improve the activity of cholinesterase, especially improve the activity of acetylcholinesterase.

[0007] The present invention provides an equine serum cholinesterase activator, the formula of which is as follows: 5-10 g of nano silicon dioxide, 0.05-0.1 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L;

[0008] Wherein, the particle size of nano-silicon dioxide is 20 to 60 nm.

[0009] Since the essence of serum cholinesterase participating in catalytic reaction is that it has special substrate binding pocket region and active site, the present invention improves the catalytic activity of serum cholinesterase by improving the activity of active site. In the present invention, nano silicon dioxide has adsorption aggregation effect on cholinesterase, and cholinesterase gathers with nano silicon dioxide as crystal nucleus, namely has fixation effect on free cholinesterase, and our research shows that the adsorption amount of nano silicon dioxide to cholinesterase can reach more than 41.56mg / 100mg. In addition, after nano silicon dioxide adsorbs cholinesterase, the nucleophilic activity of the serine hydroxyl active site of cholinesterase is enhanced, so that it is easy to combine choline compound substrates such as acetylcholine.

[0010] In order to enhance the effect of nano silicon dioxide, the present invention also uses sodium hexametaphosphate, which has surface activity, both hydrophilic and hydrophobic groups, and is an excellent dispersant for dispersing nano silicon dioxide. On the other hand, sodium hexametaphosphate can also activate the activity of cholinesterase and accelerate the catalytic reaction.

[0011] The pH of the equine serum cholinesterase activator is adjusted to 7.0-7.4 with sodium hydroxide or hydrochloric acid; pH 7.0-7.4 is the optimal pH for cholinesterase.

[0012] Preferably, the formula of the equine serum cholinesterase activator is as follows: 8 g of nano-silicon dioxide, 0.07 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L;

[0013] Alternatively, the formula of the equine serum cholinesterase activator is as follows: 7 g of nano-silicon dioxide, 0.07 g of sodium hexametaphosphate, and an inorganic liquid adjusted to 1 L.

[0014] The enzyme activity of these two formulas tested was the highest, reaching over 701U / mL.

[0015] Preferably, the inorganic liquid is double distilled water or a buffer.

[0016] The buffer solution formula is as follows: NaH2PO4·2H2O (sodium dihydrogen phosphate dihydrate) 0.24 g, Na2HPO4·12H2O (sodium dihydrogen phosphate dodecahydrate) 3.63 g, NaCl (sodium chloride) 8 g, KCl (potassium chloride) 0.2 g, double distilled water to 1 L;

[0017] The buffer contains many ions, which can dissolve and protect reagents. When nano-silica and sodium hexametaphosphate increase the activity of serum cholinesterase, the buffer can stabilize the structure and activity of serum cholinesterase.

[0018] Preferably, the equine serum cholinesterase activator is prepared according to the following method:

[0019] Prepare various raw materials according to the formula amount of equine serum cholinesterase activator;

[0020] First, disperse the nano-silicon dioxide in the inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.0-7.4. Sodium hexametaphosphate is easily soluble, so it is added after the nano-silicon dioxide to ensure the dispersion of the nano-silicon dioxide first.

[0021] The present invention also provides an application of an equine serum cholinesterase activator, wherein the equine serum cholinesterase is cholinesterase, and the application is used to increase the activity of equine serum cholinesterase. The substrate of the cholinesterase is acetylcholine. After the equine serum cholinesterase activator is mixed with cholinesterase, it is allowed to stand for 5 minutes to increase the enzyme activity.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] In order to activate cholinesterase and improve the activity of cholinesterase, the present invention provides an activator, which can significantly improve the activity of cholinesterase, especially improve the activity of acetylcholinesterase.

[0024] In the present invention, nano-silicon dioxide has an adsorption and aggregation effect on cholinesterase. Our research shows that the adsorption amount of nano-silicon dioxide on cholinesterase can reach 41.56mg / 100mg. In addition, after nano-silicon dioxide adsorbs cholinesterase, it enhances the nucleophilic activity of the serine hydroxyl active site of cholinesterase, making it easy to bind to choline compound substrates such as acetylcholine.

[0025] In order to enhance the effect of nano-silicon dioxide in activating the activity of cholinesterase, the present invention also uses sodium hexametaphosphate. In addition, sodium hexametaphosphate has surface activity, has both hydrophilic and hydrophobic groups, and is an excellent dispersant for dispersing nano-silicon dioxide. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is the technical roadmap of the present invention. DETAILED DESCRIPTION

[0027] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments and drawings.

[0028] In the description of the present invention, unless otherwise specified, all reagents used are commercially available and all methods used are conventional techniques in the art.

[0029] Horse serum cholinesterase was purchased from Shanghai Yansheng Industrial Co., Ltd.

[0030] The activity of horse serum cholinesterase was detected using a cholinesterase (CHE) colorimetric test kit, brand: Elabscience.

[0031] The enzyme activity test method used in the test of the present invention is as follows: horse serum choline ester is prepared into a 0.5 mg / mL dispersion liquid with sterile deionization to obtain a standard product, which is allowed to stand at room temperature for 5 minutes, and the enzyme activity of the standard product is tested using a cholinesterase (CHE) colorimetric test kit after the sample is mixed. Then, sterile water is replaced with an equal amount of activator to prepare a 0.5 mg / mL dispersion liquid to obtain a test sample, which is allowed to stand at room temperature for 5 minutes, and the enzyme activity is tested using a cholinesterase (CHE) colorimetric test kit after the sample is mixed to obtain a test enzyme activity. Three parallel tests are performed in each group, and the average value is taken.

[0032] The acetylcholine content was tested using an acetylcholine (ACH) enzyme-linked immunosorbent assay kit, brand Elabscience.

[0033] The acetylcholine hydrolysis efficiency method used in the test of the present invention is as follows: horse serum choline ester is prepared into a 0.5 mg / mL dispersion liquid with sterile deionization to obtain a standard product, which is allowed to stand at room temperature for 5 minutes, and then mixed with acetylcholine, which is recorded as a standard product control group. Then, an equal amount of activator is used to replace sterile water, and a 0.5 mg / mL dispersion liquid is prepared to obtain a test sample, which is allowed to stand at room temperature for 5 minutes, and then mixed with acetylcholine, which is recorded as a test sample group. Three parallel tests are performed in each group, and the average value is taken.

[0034] The inventive concept of the present invention is as follows:

[0035] The present invention provides an equine serum cholinesterase activator, which has the following formula: 5-10 g of nano silicon dioxide, 0.05-0.1 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of neutral or alkaline. Preferably, the pH of the equine serum cholinesterase activator is 7.0-7.4.

[0036] Specific embodiments and effects are shown as follows.

[0037] 1. Effect of different nano-silicon dioxide dosage on cholinesterase activity

[0038] Example 1

[0039] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 5 g of nano silicon dioxide, 0.05 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH value of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0040] The equine serum cholinesterase activator is prepared according to the following method:

[0041] Prepare the raw materials according to the recipe quantity;

[0042] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0043] Example 2

[0044] A cholinesterase activator for equine serum, the formula of which is as follows: 6g of nano silicon dioxide, 0.05g of sodium hexametaphosphate, and an inorganic liquid fixed to 1L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20nm.

[0045] The equine serum cholinesterase activator is prepared according to the following method:

[0046] Prepare the raw materials according to the recipe quantity;

[0047] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0048] Example 3

[0049] A cholinesterase activator for equine serum has the following formula: 7 g of nano silicon dioxide, 0.05 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0050] The equine serum cholinesterase activator is prepared according to the following method:

[0051] Prepare the raw materials according to the recipe quantity;

[0052] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0053] Example 4

[0054] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 8g of nano silicon dioxide, 0.05g of sodium hexametaphosphate, and an inorganic liquid fixed to 1L, with a pH value of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20nm.

[0055] The equine serum cholinesterase activator is prepared according to the following method:

[0056] Prepare the raw materials according to the recipe quantity;

[0057] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0058] Example 5

[0059] A serum cholinesterase activator for equine animals has the following formula: 9 g of nano silicon dioxide, 0.05 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0060] The equine serum cholinesterase activator is prepared according to the following method:

[0061] Prepare the raw materials according to the recipe quantity;

[0062] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0063] Example 6

[0064] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 10 g of nano silicon dioxide, 0.05 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH value of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0065] The equine serum cholinesterase activator is prepared according to the following method:

[0066] Prepare the raw materials according to the recipe quantity;

[0067] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0068] The difference between the activator formulas of Examples 1 to 6 is the different dosage of nano-silicon dioxide, and horse serum choline ester is used as the test object. The relevant dosage and enzyme activity test results are shown in Table 1. The results show that compared with the enzyme activity of the standard product, after using the activator, the test enzyme activity is increased by 25% to 33%.

[0069] Table 1 shows the results of different nano-silica dosages and enzyme activity tests

[0070]

[0071] 2. Effects of different nano-silica particle sizes on cholinesterase activity

[0072] Example 7

[0073] A cholinesterase activator for equine serum, the formula of which is as follows: 5g of nano-silicon dioxide, 0.05g of sodium hexametaphosphate, and an inorganic liquid fixed to 1L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano-silicon dioxide is 40nm.

[0074] The equine serum cholinesterase activator is prepared according to the following method:

[0075] Prepare the raw materials according to the recipe quantity;

[0076] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0077] Example 8

[0078] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 5 g of nano silicon dioxide, 0.05 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH value of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 60 nm.

[0079] The equine serum cholinesterase activator is prepared according to the following method:

[0080] Prepare the raw materials according to the recipe quantity;

[0081] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0082] The difference between the activator formulas of Example 1, Example 7 and Example 8 is that the particle size of nano-silicon dioxide is different. Horse serum choline ester is used as the test object, and the enzyme activity test results are shown in Table 2. The results show that compared with the enzyme activity of the standard product, after using the activator, the test enzyme activity at the particle size of 20nm, 40nm, and 60nm is increased by 26%, 26%, and 25%, respectively.

[0083] Table 2 shows the test results of different nano-silica particle sizes and enzyme activity

[0084]

[0085] 3. Effects of different dosages of sodium hexametaphosphate on cholinesterase activity

[0086] Example 9

[0087] A cholinesterase activator for equine serum, the formula of which is as follows: 5g of nano-silicon dioxide, 0.06g of sodium hexametaphosphate, and an inorganic liquid fixed to 1L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano-silicon dioxide is 20nm.

[0088] The equine serum cholinesterase activator is prepared according to the following method:

[0089] Prepare the raw materials according to the recipe quantity;

[0090] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0091] Example 10

[0092] A cholinesterase activator for equine serum has the following formula: 5 g of nano silicon dioxide, 0.07 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0093] The equine serum cholinesterase activator is prepared according to the following method:

[0094] Prepare the raw materials according to the recipe quantity;

[0095] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0096] Embodiment 11

[0097] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 5 g of nano silicon dioxide, 0.08 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH value of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0098] The equine serum cholinesterase activator is prepared according to the following method:

[0099] Prepare the raw materials according to the recipe quantity;

[0100] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0101] Example 12

[0102] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 5 g of nano silicon dioxide, 0.09 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0103] The equine serum cholinesterase activator is prepared according to the following method:

[0104] Prepare the raw materials according to the recipe quantity;

[0105] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0106] Example 13

[0107] A cholinesterase activator for serum of equine animals, the formula of which is as follows: 5 g of nano silicon dioxide, 0.1 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.4, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0108] The equine serum cholinesterase activator is prepared according to the following method:

[0109] Prepare the raw materials according to the recipe quantity;

[0110] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0111] The difference between the activator formulas of Examples 1 and 9 to 13 is the different dosage of sodium hexametaphosphate, and the test results of enzyme activity are shown in Table 3 using horse serum cholinesterase as the test object. The results show that compared with the enzyme activity of the standard product, after using the activator, the test enzyme activity increased by 26% to 31%.

[0112] Table 3 shows the results of different sodium hexametaphosphate dosages and enzyme activity tests

[0113]

[0114] 4. Effects of different inorganic liquids on serum cholinesterase activity in equine animals

[0115] Embodiment 14

[0116] A equine serum cholinesterase activator has the following formula: 5 g of nano silicon dioxide, 0.07 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.4, wherein the inorganic liquid is a buffer solution.

[0117] The buffer solution formula is as follows: NaH2PO4·2H2O 0.24 g, Na2HPO4·12H2O 3.63 g, NaCl 8 g, potassium chloride 0.2 g, and double distilled water to 1 L.

[0118] The particle size of nano-silica is 20nm.

[0119] The equine serum cholinesterase activator is prepared according to the following method:

[0120] Prepare the raw materials according to the recipe quantity;

[0121] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.4.

[0122] Horse serum cholinesterase was used as the test object, and the results of the enzyme activity test are shown in Table 4. The results showed that compared with the enzyme activity of the standard product, the test enzyme activity was improved after using the activator.

[0123] Table 4 shows the test results of inorganic liquid and enzyme activity

[0124]

[0125] 5. Effect of different pH on cholinesterase activity

[0126] Embodiment 15

[0127] A serum cholinesterase activator for equine animals has the following formula: 5 g of nano silicon dioxide, 0.05 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, with a pH of 7.0, wherein the inorganic liquid is double distilled water. The particle size of the nano silicon dioxide is 20 nm.

[0128] The equine serum cholinesterase activator is prepared according to the following method:

[0129] Prepare the raw materials according to the recipe quantity;

[0130] First, disperse the nano-silicon dioxide in an inorganic liquid, then add sodium hexametaphosphate to disperse it, and adjust the pH to 7.0.

[0131] Horse serum cholinesterase was used as the test object, and the results of the enzyme activity test are shown in Table 5. The results showed that compared with the enzyme activity of the standard product, the enzyme activity under weak alkaline conditions was higher after using the activator.

[0132] Table 5 is the pH and enzyme activity test results

[0133]

[0134] 6. Controlled Trials

[0135] Comparison 1

[0136] A control activator has the following formula: 5 g of nano-silicon dioxide, inorganic liquid fixed to 1 L, pH 7.4, the inorganic liquid is double distilled water, and the particle size of the nano-silicon dioxide is 20 nm.

[0137] The control activator was prepared according to the following method:

[0138] Prepare the raw materials according to the recipe quantity;

[0139] Disperse nano-silicon dioxide in an inorganic liquid and adjust the pH to 7.4.

[0140] Comparison 2

[0141] A control activator has the following formula: 0.07 g of sodium hexametaphosphate, an inorganic liquid fixed to 1 L, a pH of 7.4, and the inorganic liquid is double distilled water.

[0142] The control activator was prepared according to the following method:

[0143] Prepare the raw materials according to the recipe quantity;

[0144] Sodium hexametaphosphate was added to the inorganic liquid to dissolve it and the pH was adjusted to 7.4.

[0145] Comparison 3

[0146] A control activator is an inorganic liquid with a pH of 7.4, and the inorganic liquid is double distilled water.

[0147] The results in Table 6 show that although the activators of Control 1 and Control 2 respectively use nano-silicon dioxide and sodium hexametaphosphate, the effects of improving the enzyme activity are not as good as those of Example 1. Therefore, the activation effect is better when the activator of nano-silicon dioxide and sodium hexametaphosphate is used.

[0148] Table 6 shows the activator formula and enzyme activity test results

[0149]

[0150]

[0151] VII. Adsorption test of nano-silica on horse serum cholinesterase

[0152] The adsorption conditions of horse serum cholinesterase and nano-silicon dioxide used in the present invention are: the mass ratio of enzyme / nano-silicon dioxide is 1:1, adsorption at room temperature for 1h, and adsorption pH is 7.4. The adsorption amount test results are shown in Table 7, and the results show that the adsorption amount of horse serum cholinesterase by nano-silicon dioxide is 41.56mg / 100mg to 44.87mg / 100mg.

[0153] Table 7 is the adsorption test results

[0154] Nano-silica particle size (nm) Adsorption capacity (mg / 100mg) 20 44.87 40 41.56 60 41.90

[0155] 8. Acetylcholine hydrolysis efficiency test

[0156] The test sample was mixed with acetylcholine, reacted for a period of time, and the acetylcholine residue was tested using an acetylcholine enzyme-linked immunosorbent assay kit to calculate the acetylcholine hydrolysis efficiency, that is, to count the amount of acetylcholine that can be catalyzed and decomposed by one enzyme molecule within one second. The results are shown in Table 8. The results in Table 8 show that the acetylcholine hydrolysis efficiency of Examples 1 to 15 is higher than that of the standard control group.

[0157] Table 8 is the test results of acetylcholine hydrolysis efficiency

[0158]

[0159]

[0160] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes a preferred embodiment. Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they know the basic creative concept. Therefore, the attached claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0161] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. An application of an equine serum cholinesterase activator, characterized in that: The equine serum cholinesterase is cholinesterase, and the application is used to increase the activity of equine serum cholinesterase; The formula of the equine serum cholinesterase activator is as follows: 5-10 g of nano-silicon dioxide, 0.05-0.1 g of sodium hexametaphosphate, and an inorganic liquid fixed to 1 L, wherein the particle size of the nano-silicon dioxide is 20-60 nm, and the pH of the equine serum cholinesterase activator is neutral or alkaline; The inorganic liquid is double distilled water or buffer; The buffer solution formula is as follows: 0.24 g sodium dihydrogen phosphate dihydrate, 3.63 g disodium hydrogen phosphate dodecahydrate, 8 g sodium chloride, 0.2 g potassium chloride, and double distilled water to 1 L; The equine serum cholinesterase activator is mixed with cholinesterase and then allowed to stand for 5 minutes to increase the enzyme activity.

2. The use of the equine serum cholinesterase activator according to claim 1, characterized in that: The pH is 7.0-7.

4.

3. The use of the equine serum cholinesterase activator according to claim 2, characterized in that: 8g of nano-silicon dioxide, 0.07g of sodium hexametaphosphate, and inorganic liquid are adjusted to 1L.

4. The use of the equine serum cholinesterase activator according to claim 1, characterized in that: 7g of nano-silicon dioxide, 0.07g of sodium hexametaphosphate, and inorganic liquid are adjusted to 1L.

5. The use of the equine serum cholinesterase activator according to claim 1, characterized in that: The adsorption capacity of nano-silicon dioxide on serum cholinesterase of equine animals is ≥41.56mg / 100mg.

6. The use of the equine serum cholinesterase activator according to claim 1, characterized in that: The equine serum cholinesterase activator is prepared according to the following method: Prepare the raw materials according to the recipe quantity; Firstly, nano silicon dioxide is dispersed in an inorganic liquid, and then sodium hexametaphosphate is added to disperse it, and the pH is adjusted to 7.0-7.

4.

7. The use of the equine serum cholinesterase activator according to claim 1, characterized in that: The substrate of the cholinesterase is acetylcholine.

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