CN110115755A - The new medical use of glutathione - Google Patents

Abstract

The present invention provides application of the glutathione in the beta Cell of islet damage medicine caused by preparation treatment Long-term Fluctuation hyperglycemia, highly reactive form of oxygen metabolic memory.In addition, the present invention also provides a kind of pharmaceutical composition, a kind of pharmaceutical composition for preventing diabetes and a kind of pharmaceutical compositions for treating diabetes skin ulcer for treating the damage of beta Cell of islet caused by highly reactive form of oxygen metabolic memory or fluctuant hyperglycemia.The above composition contains glutathione or glutathione pharmaceutically acceptable salt derivative.These drugs can be fabricated to various dosage forms to cope with the disease under different type, different diseases.

Description

New medicinal uses of glutathione

technical field

The invention relates to the technical field of medicine, in particular to the application of glutathione in the preparation of medicines for the treatment of pancreatic beta cell damage caused by long-term fluctuating hyperglycemia and high active oxygen metabolism and memory.

Background technique

As of 2015, the number of diabetic patients in my country has reached 109.6 million, and about 1.3 million people have died of diabetes and its complications; in 2017, there were 451 million diabetic patients worldwide. If there is no intervention, the number of patients with diabetes in my country will rise to 154 million by 2040, and the number of diabetic patients in the world will increase to 693 million in 2045. Therefore, the prevention and treatment of diabetes is urgent. Diabetes is mainly divided into type I diabetes and type II diabetes, both of which show decreased or relatively decreased insulin secretion caused by functional defects of pancreatic beta cells. Therefore, protecting islet β cells from damage is of great significance for the prevention of diabetes. Studies have shown that a long-term high-sugar diet will cause physiological blood sugar fluctuations, high reactive oxygen species (ROS) metabolic memory, and then reduce the FOXO1 (forkhead protein O1) pathway, resulting in dedifferentiation of pancreatic β cells and impaired insulin secretion. Hyperglycemia-induced ROS is considered to be an important cause of β-cell dysfunction, and removing excess intracellular ROS or preventing excessive intracellular ROS generation is an effective means to protect β-cell function and an important way to prevent and treat diabetes.

Glutathione GSH is a small molecular polypeptide naturally synthesized in human cells. It is composed of glutamic acid, cysteine and glycine, and is widely distributed in various organs of the body. Its main physiological functions are scavenging free radicals, anti-oxidation and anti-aging. Studies have shown that GSH is an important free radical scavenger in the liver. When the intracellular GSH decreases, it will lead to a decrease in the GSH uptake by the mitochondria, thereby inducing or aggravating the damage of oxidative stress. However, the role of GSH in scavenging ROS is still unclear.

Contents of the invention

The purpose of the present invention is to fill up the shortcomings and deficiencies of the prior art, and provide a new medical application of glutathione.

The application of glutathione in the preparation of anti-FOXO1 pathway-reducing drugs, glutathione is:

The invention also provides the application of glutathione in the preparation of medicine for treating the damage of pancreatic beta cells caused by high active oxygen metabolism and memory, and the application of glutathione in the preparation of medicine for treating the damage of pancreatic beta cells caused by fluctuating hyperglycemia.

The scheme of the present invention provides a pharmaceutical composition for treating pancreatic beta cell damage caused by high active oxygen metabolism memory or fluctuating hyperglycemia, a pharmaceutical composition for preventing diabetes, and a pharmaceutical composition for treating diabetic skin ulcer. The above compositions contain glutathione or a pharmaceutically acceptable salt derivative of glutathione.

Preferably, these pharmaceutical compositions have the following formulation forms: injections, tablets, capsules, controlled-release formulations, sustained-release formulations, and nanoscale formulations. The present invention can be administered to patients in need of such treatment in the form of compositions through gastrointestinal tract administration, injection administration, cavity administration and the like. For oral administration, it can be made into conventional solid preparations such as tablets, powders, granules, capsules, etc., or into liquid preparations such as water, oil suspensions, or other liquid preparations such as syrup, etc.; for parenteral administration When used as medicine, it can be made into solution for injection, water or oily suspension, etc.

Long-term high-sugar diet will cause physiologically high reactive oxygen species (ROS) fluctuations, thereby reducing the FOXO1 pathway, resulting in dedifferentiation of pancreatic β cells and impaired insulin secretion. Timely and dynamic administration of glutathione can completely prevent the occurrence of this phenomenon. Therefore, for people with long-term high-sugar eating habits, timely supplementation of glutathione after meals can effectively prevent islet β-cell damage caused by high-sugar. In addition, although diabetic patients can effectively control blood sugar through exogenous insulin therapy, they have acquired metabolic memory of ROS response during the development of the disease. After eating, these patients are more prone to sugar-ROS pathological reaction, resulting in diabetic organ damage. Therefore, GSH can also be used as an effective drug to block diabetic organ damage, and can be used in combination with hypoglycemic drugs to prevent or treat diabetes.

The present invention will be described in further detail below in conjunction with the accompanying drawings.

Description of drawings

Figure 1 Oral glucose tolerance test and fasting plasma insulin detection after 38 days of experiment;

Figure 2 Detection of islet ROS in pancreatic tissue, the central light-colored plaque-like area is ROS;

Figure 3 is the expression of insulin and glucagon in islets;

Figure 4 shows the expression of FOXO1, TXNIP and MafA in islets;

Figure 5 shows the expression of Nurog3 and Pdx1 in pancreatic tissue and islets;

Figure 6 is the structural formula of glutathione.

Detailed ways

In order to better understand the essence of the present invention, the new application of GSH in the field of diabetes treatment and pharmacy is provided below in the form of examples. At the same time, it must be noted that these examples are only used to illustrate the present invention and are not intended to limit the protection scope of the present invention.

GSH prevents the production of high active oxygen metabolism memory ROS caused by high-sugar diet through the FOXO1 signaling pathway, thereby protecting pancreatic β cells and other body cells from damage, and can effectively prevent the occurrence of diabetes and treat organ damage caused by diabetes.

Based on this, glutathione or a pharmaceutically acceptable salt thereof can be used to prepare related drugs: a pharmaceutical composition for islet β-cell damage caused by high active oxygen metabolism memory or fluctuating hyperglycemia; a pharmaceutical composition for preventing diabetes; Pharmaceutical composition for treating diabetes.

These pharmaceutical compositions have the following preparation forms: injections, tablets, capsules, controlled or sustained release agents, and nanometer preparations.

The following provides experimental examples to illustrate the medicinal effects of glutathione.

Experimental example:

Experimental Materials

Experimental animals: In this project, adult female SD rats were selected, purchased from Shanghai Slack Co., Ltd., raised in the animal room of the Institute of Cardiac Regenerative Medicine, School of Basic Medicine, Wenzhou Medical University for two weeks, and used for formal experiments after adapting to the environment. The animal room was illuminated by natural day and night light, 12/12 hours of light/dark treatment, well ventilated, free to eat and drink, the ambient temperature was kept at 23±2°C, and the relative humidity was kept at 50-70%.

Main reagents and antibodies: Chloral hydrate was purchased from Sinopharm Chemical Reagent Co., Ltd., heparin sodium was purchased from Nanjing Audofoni Biotechnology Co., Ltd., dextrose was purchased from Shanghai Yuanye Biotechnology Co., Ltd., GSH was purchased from Shanghai Yuanye Biological Technology Co., Ltd. The list of antibodies used is as follows:

Table 1. List of antibodies used

Main instruments: blood glucose meter and blood glucose test strips were purchased from Roche Diagnostic Products (Shanghai) Co., Ltd., fluorescence microscope was purchased from Leica Company of Germany, and ultrapure water meter was purchased from Millipore Company of the United States.

Experimental process: The rats were divided into three groups, namely the sham operation group (Sham), the long-term high-sugar diet group (LOsG) and the long-term high-sugar diet group given glutathione at the same time (LOsG-TdGSH).

In the LOSG-TDGSH group, glucose solution was administered intragastrically and GSH solution was injected intraperitoneally every 6 hours;

In the LOSG group, glucose solution was intragastrically administered every 6 hours and normal saline was injected intraperitoneally;

In the Sham group, deionized water was intragastrically administered and normal saline was injected intraperitoneally every 6 hours.

The above test was continued for 38 days.

Experimental results:

Result 1: As shown in Figure 1, compared with the Sham group, after 38 days, the OGTT test of the LOSG group showed impaired glucose tolerance, while the OGTT of the LOSG-TDGSH group was normal. Fasting plasma insulin in LOSG group was significantly lower than that in Sham and LOSG-TDGSH groups, and there was no statistical difference between LOSG-TDGSH group and Sham group.

This suggests that insulin secretion is affected by a long-term high-sugar diet and that GSH can completely prevent this phenomenon.

Result 2: As shown in Figure 2, compared with the Sham group, after 38 days of the experiment, the ROS production in the islets of the LOSG group was significantly increased. There was no difference between the LOSG-TDGSH group and the Sham group.

This suggests that a long-term high-sugar diet can cause excess ROS production by pancreatic β-cells, and that GSH can prevent excess ROS production.

Result 3: Compared with the Sham group, the expression of pancreatic insulin in the LOSG group decreased significantly after 38 days of experimentation. There was no significant difference between the LOSG-TDGSH group and the Sham group.

This indicates that the expression of insulin in pancreatic tissue is affected by a long-term high-sugar diet, and GSH can completely prevent the low expression of insulin caused by high-sugar.

Results 4: The dedifferentiation of pancreatic β-cells is one of the mechanisms leading to the damage of β-cells in diabetes. Studies have shown that low expression of FoxO1 induces β-cell dedifferentiation and causes hyperglycemia under physiological stress. It can be seen from Figure 4A-C, Figure 4D, and Figure 4E that compared with the Sham group, LOsG significantly decreased the expression of P38FoxO1 mRNA and protein by immunohistochemical analysis, qRT-PCR and Western blot. And TdGSH can prevent LOsG from producing a decrease in FoxO1 expression. FoxO1 is known to repress the expression of TXNIP, which in turn downregulates MafA. MafA This is a transcription factor that drives insulin expression.

As shown in Figure 4F-I, compared with the Sham group, LOsG led to a significant increase in the expression of TXNIP in nuclear β cells; as shown in Figure 4J-M, the expression of MafA in the nucleus of β cells was significantly reduced. TdGSH again blocked LOsG-induced increase/decrease in TXNIP/MafA expression, respectively. These results suggest that LOsG downregulates FoxO1, releasing its inhibitory effect on TXNIP. Elevated expression of TXNIP inhibited MafA expression, thereby suppressing insulin expression in β cells of the LOsG group.

Outcome five: β-cells lacking FoxO1 lose their identity and transform into pancreatic endocrine progenitor cells, characterized by high expression of Neurog3 and lack of insulin and MafA expression. The inventors found that, as shown in Figure 5A-C and Figure 5H-J, after LOsG/ROS stress, the β-cell nucleus began to re-express Neurog3 and significantly migrated Pdx1 from the cytoplasm to the nucleus, that is, β-cell dysfunction was caused by hyperglycemia Induced oxidative stress caused by downregulation of FoxO1.

TdGSH completely blocks LOsG-induced β-cell dedifferentiation and prevents Neurog3 expression and relocalization of nuclear Pdx1 immunoreactivity. In addition, as shown in Figure 5F and Figure 5M, some acinar cells also showed high expression of nuclear Neurog3 and Pdx1 under LOsG/ROS stimulation. During pancreatic development, Neurog3 and Pdx1 are expressed in pancreatic progenitor cells and control cell differentiation. In this study, re-expression of Neurog3 and Pdx1 in β-cells and acinar cells indicated that LOsG/ROS stress drives pancreatic cells to dedifferentiate and revert to their common progenitor stage.

The above experimental example is only an explanation of the present invention, but not a limitation of the present invention. Those skilled in the art can make modifications without creative contributions as needed after reading this specification, but as long as they are within the scope of the claims of the present invention, they are all protected by the patent law.

Claims (9)

1. glutathione is preparing the application in anti-FOXO1 access reduction drug, the glutathione are as follows:

。

2. glutathione treats the application in beta Cell of islet damage medicine caused by highly reactive form of oxygen metabolic memory in preparation.

3. glutathione treats the application in beta Cell of islet damage medicine caused by fluctuant hyperglycemia in preparation.

4. a kind of FOXO1 access for the treatment of reduces the pharmaceutical composition of caused beta Cell of islet damage, which is characterized in that the composition Contain glutathione or glutathione pharmaceutically acceptable salt derivative.

5. a kind of FOXO1 access for the treatment of according to claim 4 reduces the pharmaceutical composition of caused beta Cell of islet damage, It is characterized by: the drug combination preparation form is injection, tablet, capsule, controlled release agent, sustained release agent or nanometer system One of agent.

6. a kind of pharmaceutical composition for preventing diabetes, which is characterized in that the composition contains glutathione or glutathione exists Pharmaceutically acceptable salt analog derivative.

7. a kind of pharmaceutical composition for preventing diabetes according to claim 6, it is characterised in that: the pharmaceutical composition Object dosage form is one of injection, tablet, capsule, controlled release agent, sustained release agent or nanometer formulation.

8. a kind of pharmaceutical composition for treating diabetes, which is characterized in that the composition contains glutathione or glutathione exists Pharmaceutically acceptable salt analog derivative.

9. a kind of pharmaceutical composition for treating diabetes according to claim 8, it is characterised in that: the pharmaceutical composition Object dosage form is one of injection, tablet, capsule, controlled release agent, sustained release agent or nanometer formulation.