A kind of active peptide with blood lipid-lowering function derived from Chinese wood beetle and its preparation method and application

Abstract

The invention discloses an active peptide with blood lipid-lowering function derived from the Chinese soil beetle, a preparation method and application thereof. The amino acid sequence of the active peptide of the present invention is LAPAPGTL. The active peptide provided by the invention has the function of lowering blood fat and can be developed into medicine or health product.

Description

technical field

[0001] The invention relates to the technical field of active peptides. More specifically, it relates to an active peptide with blood lipid-lowering effect and its preparation method and application. Background technique

[0002] Hyperlipidemia is a common cardiovascular disease at present. With the improvement of people's living standards, high-calorie and high-fat diets make the incidence of hyperlipidemia continue to increase. Hyperlipidemia has many complications, such as fatty liver, atherosclerosis, diabetes, etc. Therefore, the prevention and treatment of hyperlipidemia is one of the current research hotspots.

[0003] Ground beetle is a traditional Chinese medicine, which has the effect of dispelling blood stasis, renewing tendons and bones, and is used for bruises, tendon injuries and fractures, amenorrhea due to blood stasis, abdominal pain due to postpartum stasis, and abdominal mass due to addiction. Modern studies have concluded that the ground...

Examples

Embodiment 1

[0037] Embodiment 1 preparation of active peptide

[0038] (1) Treatment of ground beetle medicinal materials

[0039] Add 10 times the weight of deionized water or distilled water to the fresh body of the wood beetle, and then shear it for 15 minutes at a shear rate of 5000 rpm and at room temperature to prepare a wood beetle slurry.

[0040] (2) Protease hydrolysis

[0041] Heat the ground beetle slurry to 100°C to sterilize for 15 minutes, then let it cool to 37°C, use 0.1mol / L HCl to adjust the pH of the solution to 2.5, add 1.5% by weight of pepsin and set it in a constant temperature enzymolysis instrument for enzymolysis, the enzymolysis time 60min, and the enzymatic hydrolysis temperature is 37°C. Then use 2.0% sodium hydroxide solution to adjust the pH to 8.5, add 1.5% by weight of trypsin and place it in a constant temperature enzymolysis instrument for enzymolysis, the enzymolysis time is 180min, and the enzymolysis temperature is 37°C to obtain the bionic ground ...

Embodiment 2

[0052] Based on the amino acid sequences of the LL-8 and LM-9 active peptides identified in Example 1, experimental samples were obtained by solid-phase synthesis for verification of blood lipid-lowering activity. Solid-phase synthetic peptide company: Shanghai Gil Biotechnology Co., Ltd.

[0053] 80 SD rats, male, were adaptively fed for 7 days, and then divided into nine groups, namely blank group, model group, Xueshuantong capsule group, ground beetle medicinal material group, LL-8 high-dose group (0.5g / kg), LL-8 low-dose group (0.25g / kg), LM-9 high-dose group (0.5g / kg), LM-9 low-dose group (0.25g / kg), except the blank group, every mouse in other groups Quantitatively administer 50 g of high-fat feed, and at the same time remove the rats in the blank group, give each rat 4.0 mL of high-fat emulsion at 9:00 am every day, feeding method of high-fat emulsion: gavage orally, rats in the blank group continue to be given ordinary feed, continuous For 45 days, after giving high-f...

Embodiment 3

[0059] Based on the amino acid sequence of the LL-8 active peptide identified in Example 1, experimental samples were obtained by solid-phase synthesis for safety experimental verification. Solid-phase synthetic peptide company: Shanghai Gil Biotechnology Co., Ltd.

[0060] Healthy SD rats, 250-300g, 40, were adaptively fed for 7 days, and then set up as blank group, normal saline group, LL-8 group, and LL-8 group were given a large dose of 20.0g / 200g of drugs, administration Method: subcutaneous injection; the normal saline group was also given the same volume of normal saline by subcutaneous injection for 12 days. body weight, body temperature. At the same time, the weight of immune organs such as thymus and spleen of rats in each group was measured to measure the safety of LL-8.

[0061] Table 2 Short-term safety evaluation results of body temperature changes in rats in each group

[0062]

[0063]

[0064] Table 3 Organ weight results of rats in each group

[006...

technical field

[0001] The invention relates to the technical field of active peptides. More specifically, it relates to an active peptide with blood lipid-lowering effect and its preparation method and application. Background technique

[0002] Hyperlipidemia is a common cardiovascular disease at present. With the improvement of people's living standards, high-calorie and high-fat diets make the incidence of hyperlipidemia continue to increase. Hyperlipidemia has many complications, such as fatty liver, atherosclerosis, diabetes, etc. Therefore, the prevention and treatment of hyperlipidemia is one of the current research hotspots.

[0003] Ground beetle is a traditional Chinese medicine, which has the effect of dispelling blood stasis, renewing tendons and bones, and is used for bruises, tendon injuries and fractures, amenorrhea due to blood stasis, abdominal pain due to postpartum stasis, and abdominal mass due to addiction. Modern studies have concluded that the ground...

Examples

Embodiment 1

[0037] Embodiment 1 preparation of active peptide

[0038] (1) Treatment of ground beetle medicinal materials

[0039] Add 10 times the weight of deionized water or distilled water to the fresh body of the wood beetle, and then shear it for 15 minutes at a shear rate of 5000 rpm and at room temperature to prepare a wood beetle slurry.

[0040] (2) Protease hydrolysis

[0041] Heat the ground beetle slurry to 100°C to sterilize for 15 minutes, then let it cool to 37°C, use 0.1mol / L HCl to adjust the pH of the solution to 2.5, add 1.5% by weight of pepsin and set it in a constant temperature enzymolysis instrument for enzymolysis, the enzymolysis time 60min, and the enzymatic hydrolysis temperature is 37°C. Then use 2.0% sodium hydroxide solution to adjust the pH to 8.5, add 1.5% by weight of trypsin and place it in a constant temperature enzymolysis instrument for enzymolysis, the enzymolysis time is 180min, and the enzymolysis temperature is 37°C to obtain the bionic ground ...

Embodiment 2

[0052] Based on the amino acid sequences of the LL-8 and LM-9 active peptides identified in Example 1, experimental samples were obtained by solid-phase synthesis for verification of blood lipid-lowering activity. Solid-phase synthetic peptide company: Shanghai Gil Biotechnology Co., Ltd.

[0053] 80 SD rats, male, were adaptively fed for 7 days, and then divided into nine groups, namely blank group, model group, Xueshuantong capsule group, ground beetle medicinal material group, LL-8 high-dose group (0.5g / kg), LL-8 low-dose group (0.25g / kg), LM-9 high-dose group (0.5g / kg), LM-9 low-dose group (0.25g / kg), except the blank group, every mouse in other groups Quantitatively administer 50 g of high-fat feed, and at the same time remove the rats in the blank group, give each rat 4.0 mL of high-fat emulsion at 9:00 am every day, feeding method of high-fat emulsion: gavage orally, rats in the blank group continue to be given ordinary feed, continuous For 45 days, after giving high-f...

Embodiment 3

[0059] Based on the amino acid sequence of the LL-8 active peptide identified in Example 1, experimental samples were obtained by solid-phase synthesis for safety experimental verification. Solid-phase synthetic peptide company: Shanghai Gil Biotechnology Co., Ltd.

[0060] Healthy SD rats, 250-300g, 40, were adaptively fed for 7 days, and then set up as blank group, normal saline group, LL-8 group, and LL-8 group were given a large dose of 20.0g / 200g of drugs, administration Method: subcutaneous injection; the normal saline group was also given the same volume of normal saline by subcutaneous injection for 12 days. body weight, body temperature. At the same time, the weight of immune organs such as thymus and spleen of rats in each group was measured to measure the safety of LL-8.

[0061] Table 2 Short-term safety evaluation results of body temperature changes in rats in each group

[0062]

[0063]

[0064] Table 3 Organ weight results of rats in each group

[006...

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