阅读说明：本技术 支链氨基酸在制备缓解术后胰岛素抵抗制剂中的应用 (Application of branched chain amino acid in preparation of preparation for relieving postoperative insulin resistance ) 是由 孙鹏 张瑾 孙海鹏 张云鹏 于 2021-08-31 设计创作，主要内容包括：本发明涉及支链氨基酸在制备缓解术后胰岛素抵抗制剂中的应用,所述支链氨基酸包括：亮氨酸、缬氨酸以及异亮氨酸。本发明的制剂能够有效缓解患者术后胰岛素抵抗,降低术后空腹高血糖,减少伤口感染和围手术期其他并发症,适用于外科手术患者的加速康复；本发明为术后胰岛素抵抗及术后高血糖的预防和治疗提供了新的方法和思路,对于寻求新的术前干预措施和积极有效的分子靶点,有较大的理论价值和现实意义。(The invention relates to application of branched-chain amino acids in preparing a preparation for relieving postoperative insulin resistance, wherein the branched-chain amino acids comprise: leucine, valine, and isoleucine. The preparation can effectively relieve postoperative insulin resistance of patients, reduce postoperative fasting hyperglycemia, reduce wound infection and other complications in perioperative period, and is suitable for accelerated rehabilitation of patients in surgical operations; the invention provides a new method and thought for preventing and treating postoperative insulin resistance and postoperative hyperglycemia, and has greater theoretical value and practical significance for searching for new preoperative intervention measures and active and effective molecular targets.)

1. Use of branched chain amino acids in the manufacture of a formulation for alleviating post-operative insulin resistance, wherein the branched chain amino acids comprise: leucine, valine, and isoleucine.

2. The use of claim 1, wherein the branched chain amino acids are present in the formulation at a total concentration of 20mg/mL to 30 mg/mL.

3. The use according to claim 2, wherein the total concentration of branched chain amino acids in the formulation is 25 mg/mL.

4. Use according to claim 3, wherein the mass ratio of the three branched amino acids is selected from leucine: valine: isoleucine 1:1:1, leucine: valine: isoleucine 2:1:1 or leucine: valine: isoleucine is 4:1: 1.

5. The use according to claim 4, wherein the mass ratio of the three branched-chain amino acids is leucine: valine: isoleucine is 1:1: 1.

6. The use of claim 5, wherein the three branched-chain amino acids are each present in the formulation at a concentration of 8mg/mL to 9 mg/mL.

7. The use of claim 1, wherein the pre-operative administration of the formulation reduces post-operative hepatic gluconeogenesis levels, thereby alleviating post-operative insulin resistance.

Technical Field

The invention relates to the technical field of surgical clinical nutrition, in particular to application of branched chain amino acid in preparation of a preparation for relieving postoperative insulin resistance.

Background

Post-operative insulin resistance refers to the phenomenon of post-operative elevation of fasting glucose stress caused by decreased glucose uptake capacity of the body as a result of insulin action or decreased inhibition of hepatic glucose output by insulin following surgery.

A series of stress reactions caused by surgical wounds have influence on postoperative metabolism, organ functions and rehabilitation process of patients, and particularly the hyperglycemia phenomenon caused by postoperative insulin resistance is an important factor related to postoperative complications and rehabilitation speed. Post-operative stress hyperglycemia, particularly blood glucose levels in excess of 180mg/dL, has a clear correlation with perioperative adverse clinical outcomes including compromised immune function, surgical site infection, delayed wound healing, and prolonged hospital stays. Preoperative dietary deprivation, operative procedures, postoperative pain and reduced mobility, etc., all cause post-operative insulin resistance in the body.

The recovery acceleration measures such as a minimally invasive technique, good pain relief and preoperative carbohydrate oral administration can reduce the stress response of a patient to an operation to a certain extent, relieve the abnormal carbohydrate metabolism caused by postoperative insulin resistance and accelerate postoperative recovery of the patient, but can not completely prevent postoperative stress hyperglycemia. Therefore, the continuous exploration and improvement of perioperative accelerated rehabilitation intervention measures to further reduce post-operative stress insulin resistance is also an urgent clinical problem to be solved.

In recent years, studies on branched-chain amino acids and metabolic diseases have been hot. Branched chain amino acids refer to amino acids containing a branched side chain on the alpha carbon, including leucine, isoleucine and valine, which belong to amino acids that cannot be synthesized in the human body and must be taken in from an external source, are nutrient substances required for maintaining the growth of an animal body, and are also important nutrient metabolism signal molecules. In recent years, clinical researches show that the metabolic disorder of the branched-chain amino acid is closely related to the occurrence and development of diabetes, and the serum branched-chain amino acid level can even predict the occurrence of the diabetes and the prognosis of treatment means. Basic studies have reported that highly branched amino acids have two distinct effects of promoting or inhibiting sugar metabolism for reasons that are not clear.

Branched chain amino acids are associated with chronic insulin resistance, but studies of branched chain amino acids with post-operative insulin resistance (acute insulin resistance) have rarely been reported. There is currently no literature reporting measures to intervene in post-operative insulin resistance through preoperative branched chain amino acid intake.

Disclosure of Invention

Postoperative insulin resistance caused by perioperative stress, mainly manifested as elevated postoperative fasting glucose, is an independent risk factor for patient prognosis, and can increase the incidence of postoperative complications and patient mortality. The invention aims to overcome the defects in the prior art and provides application of branched chain amino acid in preparing a preparation for relieving postoperative insulin resistance.

In order to achieve the purpose, the invention adopts the technical scheme that:

there is provided the use of branched chain amino acids in the manufacture of a formulation for alleviating post-operative insulin resistance, the branched chain amino acids comprising: leucine, valine, and isoleucine.

Preferably, the total concentration of the branched chain amino acids in the formulation is 20mg/mL to 30 mg/mL.

More preferably, the total concentration of branched chain amino acids in the formulation is 25 mg/mL.

Preferably, the mass ratio of the three branched amino acids is selected from leucine: valine: isoleucine 1:1:1, leucine: valine: isoleucine 2:1:1 or leucine: valine: isoleucine is 4:1: 1.

More preferably, the mass ratio of the three branched amino acids is leucine: valine: isoleucine is 1:1: 1.

More preferably, the concentration of each of the three branched-chain amino acids in the formulation is 8mg/mL to 9 mg/mL.

Preferably, the pre-operative administration of the formulation reduces post-operative hepatic gluconeogenesis levels, thereby alleviating post-operative insulin resistance.

By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:

the preparation can effectively relieve postoperative insulin resistance of patients, reduce postoperative fasting hyperglycemia, reduce wound infection and other complications in perioperative period, and is suitable for accelerated rehabilitation of patients in surgical operations; the invention provides a new method and thought for preventing and treating postoperative insulin resistance and postoperative hyperglycemia, and has greater theoretical value and practical significance for searching for new preoperative intervention measures and active and effective molecular targets.

Drawings

FIG. 1 is a graph showing the results of blood glucose levels before and after the operation of rats (the time for sacrifice of rats is 4 hours after the start of partial excision of small intestine);

FIG. 2 is a graph showing the results of insulin levels before and after the operation of rats (the time for sacrifice of rats was 4 hours after the start of partial excision of small intestine);

FIG. 3 is a graph showing the results of the insulin signaling pathway (PI3K-AKT-mTOR-p70S6K) WB after rat surgery, in which (a) shows the results of liver tissue WB and (b) shows the results of muscle tissue WB;

FIG. 4 is a graph showing the results of relative mRNA expression levels after rat surgery;

FIG. 5 is a graph showing the results of blood glucose levels before and after the rat operation (the time for sacrifice is 0.5h after the start of the small intestine partial resection);

FIG. 6 is a graph showing the results of insulin levels before and after the operation of rats (the time for sacrifice of rats is 0.5h after the start of the small intestine partial resection);

wherein DDH2O represents pure water; BCAA represents a branched chain amino acid; ns indicates no significant difference; p < 0.05; p < 0.01.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.

The invention is further described with reference to the following drawings and specific examples, which are not intended to be limiting.

Example 1

The embodiment provides a branched amino acid preparation for relieving postoperative insulin resistance, which is prepared by the following steps: dissolving 100mg of each of leucine, valine and isoleucine branched chain amino acid powder in 12mL of water to obtain the branched chain amino acid preparation.

Example 2

1) Establishment of insulin resistance model after rat small intestine partial excision

With reference to the method of professor STROMMER of the college of medical sciences stropharaska, sweden: male SD rats of about 200g were selected, fasted for 17 hours before surgery, anesthetized by isoflurane gas induction, and then anesthetized by intraperitoneal injection using 350 μ L/100g of 10% chloral hydrate. Blood was collected from the heart at 300. mu.L after anesthesia and blood glucose was measured immediately. The pipe is put to rat right side carotid (reserving the blood sampling), it is fixed with rat supine position after the anesthesia, the neck unhairing, the disinfection, cut skin from the neck centre, isolate right side carotid, distal end ligature and press from both sides with the artery and carry the end of nearing the heart, cut a osculum on the arterial wall, insert diameter 0.9mm silicone tube (intraductal pouring into 25U/mL heparin normal saline solution anticoagulation in advance) toward the end of nearing the heart direction, the fixed intubate of ligature loosens the artery and presss from both sides, then seal the intubate export and prevent that blood from flowing. After each blood sampling, the anticoagulant effect is achieved by using a normal saline solution containing heparin, so that the intubation blockage is prevented. A5 cm incision was made at the midline of the abdomen, 2-3 mesenteric vessels were disconnected at a distance of 5 cm from the ligament of flexor, and the small intestine, 5 cm long, was removed. Small intestinal anastomosis was performed with 6/0 suture and then the abdomen was closed with 4/0 suture. The whole operation lasts about 40 minutes. The rats after surgery were placed on a 37 ℃ incubation blanket. Blood samples were collected at 300 microliters each before surgery and 30, 60, 90 and 120 minutes post anesthesia. Blood samples were centrifuged at 3000 rpm for 20 minutes at 4 ℃ and plasma was collected and stored at-80 ℃. Rats were sacrificed 4h after the start of surgery, skeletal muscle, liver tissue, adipose tissue were extracted in liquid nitrogen and stored at-80 ℃.

2) Effect of gastric gavage intervention with branched chain amino acids on post-operative insulin resistance in rats

20 male SD rats (200 g) at 6 weeks of age were divided into 2 groups according to the gavage composition, and 10 rats each group were gavage (2 hours before surgery and 3 mL). The rats were divided into a pure water gavage group and a branched chain amino acid gavage group (i.e., the formulation described in example 1), and pre-operative body weights of the rats were recorded. Rat small intestine partial excision was performed. Glucose oxidase method measures blood glucose value, and ELISA method measures plasma insulin value.

3) Results of the experiment

Blood glucose levels before and after rat surgery are shown in fig. 1, and insulin levels before and after rat surgery are shown in fig. 2; as shown in FIG. 3, there was no significant difference in the insulin pathways of the liver tissue and muscle tissue after the operation between the pure water gavage group and the branched chain amino acid gavage group; as shown in fig. 4, transcription of the key enzyme G6PC1 for hepatic gluconeogenesis was significantly reduced after the gastric lavage group with branched chain amino acids;

the time for killing the rats was changed to 0.5h after the start of the small intestine resection, the blood glucose levels before and after the rat surgery are shown in fig. 5, and the insulin levels before and after the rat surgery are shown in fig. 6;

the above results show that: the rat is subjected to intragastric administration of branched chain amino acid 2 hours before operation, so that the hepatic gluconeogenesis can be inhibited, and the postoperative hyperglycemia can be reduced, thereby relieving the postoperative insulin resistance.

In conclusion, the invention establishes an acute insulin resistance animal model of moderate wound in the abdomen by performing small intestine partial excision on rats, increases branched chain amino acid intake by gavage, explores feasibility of influencing postoperative insulin resistance by changing branched chain amino acid intake and establishes exact correlation between branched chain amino acid and postoperative insulin resistance, thereby providing experimental basis for developing new nutrients for relieving postoperative insulin resistance of patients.

While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.