CN104940205A - Application of pteroylglutamic acid in preventing and curing offspring diabetes caused by environmental endocrine disruptors (EED) - Google Patents

Abstract

The invention belongs to the field of medicine, relates to the application of vitamin medicine and health care, in particular to the preventive and health care effects of pteroyl glutamic acid on the abnormal glucose metabolism of offspring caused by environmental endocrine disruptors during pregnancy. The present invention uses an animal experimental model to conduct pteroylglutamate intervention experiments on pregnant and lactating female SD rats, and determines the effect of pteroylglutamate on the islet structure and function abnormalities caused by typical environmental endocrine disruptors. The prevention and protection effect of the high incidence of type 2 diabetes, the results of the present invention show that pteroyl glutamate supplementation during pregnancy has a preventive effect on the high incidence of glucose metabolism disorders in offspring caused by environmental endocrine disruptors. Pteroyl glutamic acid is harmless to the human body when used within a controlled dose range and can be used safely. This compound is added to health products or used as a food fortifier and taken orally, which can prevent abnormal glucose metabolism in offspring caused by environmental endocrine disruptors .

Description

Application of pteroylglutamic acid in prevention and treatment of offspring diabetes induced by environmental endocrine disruptors

technical field

The invention belongs to the field of medicine, relates to the application of vitamin medicine and health care, in particular to the preventive and health care effects of pteroyl glutamic acid on the abnormal glucose metabolism of offspring caused by environmental endocrine disruptors during pregnancy.

Background technique

Environmental endocrine disrupting chemicals (EDCs) refer to exogenous compounds that interfere with the human endocrine system, can affect the reproductive system, immune system, and even interfere with the normal development of tissues and organs, eventually causing health damage. EDCs are widely used in plastic products and polymer materials, which can pollute the food chain through different media such as soil, water and air, or penetrate into food from food packaging and drinking containers and ingest them into the body, making people in EDCs direct exposure. A large number of studies have confirmed that exposure to environmental pollutants during the embryonic period can affect the development of various organs and tissues and eventually lead to an increased risk of chronic diseases in adulthood, the most notable of which is the high incidence of type 1 diabetes caused by impaired islet structure and function . This type of health loss caused by exposure to environmental pollutants during critical developmental periods is difficult to recover into adulthood, and this type of chemical exposure is almost unavoidable in modern life. Therefore, we need to intervene early in the critical period of development to prevent or mitigate this kind of health loss in adulthood caused by early environmental pollution. However, there is still no clear protection method for the high incidence of type I diabetes caused by the damage of islets caused by environmental endocrine disruptors.

Pteroyl glutamic acid is an important vitamin, most of which are converted into 5-methyltetrahydrofolate after being ingested in the body, and participate in the transfer and transfer of one-carbon units in organisms. Processes such as chemicalization play a decisive role. Pteroylglutamic acid plays a vital role in maintaining health in the course of life, and has complex and important effects on genome stability, tumor susceptibility, and the occurrence and development of cardiovascular and metabolic diseases. As an essential micronutrient, it is very important for pregnant women. Supplementing pteroyl glutamate during pregnancy can prevent fetal neural tube defects, intrauterine growth retardation, premature delivery and low birth weight, etc., to protect its growth and health after birth. Epidemiological studies have confirmed that pteroyl glutamate insufficiency during pregnancy and 3 months after delivery can increase the offspring's metabolic syndrome such as impaired glucose tolerance, decreased insulin tolerance, and obesity at the age of 6-8. Glutamic acid-based multivitamin supplementation can significantly improve this phenomenon. However, there is still no clear report on whether pteroylglutamate intervention during pregnancy and how to intervene can protect offspring islet function changes caused by EDCs exposure.

Contents of the invention

The task of the present invention is to provide a method for reducing the risk of offspring type I diabetes caused by environmental endocrine disruptors.

The technical solution to realize the present invention is: use pteroyl glutamic acid for the preparation of drugs, health products or functional foods for preventing or treating offspring diabetes caused by environmental endocrine disruptors, that is, pteroyl glutamic acid can be used for preventing or treating environmental endocrine disruptors Endocrine disruptor-induced diabetes in offspring.

This patent uses animal experimental models to determine the preventive and protective effect of pteroyl glutamate on the high incidence of type I diabetes caused by the abnormal structure and function of islets caused by typical environmental endocrine disruptors.

Bisphenol A (Bisphenol A, BPA), chemical name: 2,2-bis(4-hydroxyphenyl) propane, chemical formula: C ₁₅ H ₁₆ O ₂ , basic structural formula: It is a typical environmental endocrine disruptor with estrogen-like effects widely manufactured and distributed around the world. Its monomers are often used in the production and processing of polymer materials such as polycarbonate and epoxy resin, and are widely used in the manufacture of plastic containers. Direct oral exposure to BPA in mice can disrupt the insulin pathway related to energy metabolism and insulin secretion, increasing the risk of diabetes. BPA is used in the present invention as representative of typical exposures of EDCs. Pteroylglutamic acid, also known as folic acid, chemical name: (2S)-2-[[4-[(2-amino-4-oxo-1H-6-pteridinyl)methylamino]benzyl]amino ] Glutaric acid, chemical formula: C ₁₉ H ₁₉ N ₇ O ₆ , basic structural formula: Exposure to phthalates (another typical EDCs) and hairspray use during critical developmental periods has been reported to be associated with neonatal hypospadias.

This patent uses animal experimental models to determine the preventive and protective effect of pteroyl glutamate on the high incidence of type I diabetes caused by the abnormal structure and function of islets caused by typical environmental endocrine disruptors. The experimental data are as follows:

Pregnancy was detected by vaginal wash smear method; female SD rats were intervened with pteroylglutamate and exposed to BPA during pregnancy and lactation, and were divided into the following three groups: control group, BPA at 40 μg/kg/day group (BPA-no folate), 300μg/kg/day pteroylglutamate+40μg/kg/day BPA pteroylglutamate intervention group (BPA-folate); the pteroylglutamate used in the experiment was purchased from the United States SIGMA company, purity 99%. The growth rate of the mother mice during pregnancy and the survival rate and body weight of the offspring were recorded. Relevant experiments were carried out in the weaning period of the offspring, that is, 21 days after birth, and in the juvenile period (9 weeks old) of the offspring rats. Determination of offspring body weight, pancreatic organ coefficient, fasting blood sugar, fasting insulin; intraperitoneal injection of 2g/kg body weight glucose for glucose tolerance test; intraperitoneal injection of 0.75IU/kg body weight insulin for insulin tolerance test; immunofluorescence double staining Morphological analysis of the pancreas was performed using the method; the state of the organelles of the islet β cells was observed by transmission electron microscopy; the glucose-stimulated insulin release experiment was performed on the isolated and purified islets in order to evaluate their insulin secretion function. In order to clarify its protective mechanism, SYBR relative quantitative method was used to analyze the expressions of imprinted genes Igf2, H19, Plagl1 and DNA methyl transfer-related genes Dnmt1, Dnmt3a, Dnmt3b in offspring pancreas; ), that is, BSP-PCR product purification combined with sequencing after plasmid cloning to detect the DNA methylation status of Igf2/H19 differentially methylated regions (differentially methylated regions, DMR).

The experiment of the present invention clarifies the preventive effect of pteroyl glutamic acid on the glucose metabolism disorder of progeny caused by environmental endocrine disruptors, and clarifies its mechanism.

The present invention reveals that pteroylglutamic acid can effectively improve the abnormal body weight of offspring at birth and 9 weeks of age caused by BPA, and significantly improve the pancreas organ coefficient, glucose tolerance, BCF, BCM, pancreatic structure, and β cells of offspring caused by BPA. Abnormalities in ultrastructural and high glucose-stimulated insulin secretion from isolated islets. In terms of its mechanism of action, the expressions of islet imprinted gene Igf2/H19 and DNA methyltransferase-related genes (Dnmts) in the offspring rats of the BPA group were significantly abnormal compared with the control group, and the pteroylglutamate intervention group was significantly higher than that of the BPA group. There is an obvious answer. At the same time, DNA methylation in the Igf2/H19DMR2 area of islets in the BPA group was abnormally increased, while that in the pteroylglutamate intervention group was relatively normal. The results of the present invention show that pteroylglutamic acid supplementation during pregnancy has a preventive effect on the high incidence of glucose metabolism disorders in offspring caused by environmental endocrine disruptors. Pteroylglutamic acid is harmless to the human body when used within the controlled dose range (the daily intake of pregnant women is less than 0.8mg), and can be used safely. The compound can be added to health products or used as a food fortifier and taken orally in the future to prevent abnormal glucose metabolism in offspring caused by environmental endocrine disruptors.

illustration

Figure 1 shows the glucose tolerance test. In the figure, the blood glucose response curve of offspring exposed to EDCs was abnormally elevated, and pteroylglutamate intervention can make the blood glucose response curve normal. The significance of the glucose tolerance test is that the glucose tolerance test is to measure the blood sugar at a fixed time after oral administration of glucose to evaluate the body's tolerance to glucose. If glucose intolerance occurs, the body is at a high risk of diabetes. Figure 1 shows that pteroyl glutamate intervention in the present invention can significantly reduce the abnormal glucose tolerance caused by typical EDCs exposure, and can protect the body from a high-risk state of diabetes.

Figure 2 is the assessment of islet structure and function. The results show that exposure to typical EDCs can cause islet structural abnormalities, manifested as decreased islet fraction and quality, and ultrastructural abnormalities, manifested as mitochondrial swelling and endoplasmic reticulum expansion. Intervention of pteroylglutamate can prevent the appearance of this islet structural abnormality, which is manifested by normal islet fraction and quality, complete mitochondrial structure, and normal endoplasmic reticulum arrangement. Pancreatic islets are the endocrine part of the pancreas. Abnormal islet function leads to insufficient insulin secretion and the main cause of diabetes. The pteroyl glutamate intervention in the present invention can significantly protect the abnormal structure of the pancreas and the ultrastructure of the islets caused by exposure to typical EDCs, thereby protecting the synthesis and secretion functions of the islets.

Figure 3 shows the protective mechanism-DNA methylation analysis. The results show that exposure to typical EDCs causes hypermethylation in the Igf2/H19DMR2 region, a key gene for islet development, and pteroylglutamic acid intervention can reduce this hypermethylation and make it normal to protect the development and function of islets. A large number of studies have shown that exposure to endocrine disruptors can affect the DNA methylation status of offspring, change gene expression, and affect their offspring phenotype and body state, making them susceptible to diseases. In the present invention, pteroylglutamic acid intervention can reduce and normalize the hypermethylation degree of the Igf2/H19DMR2 region caused by typical EDCs exposure, thereby protecting the development and function of islets. This is the internal mechanism of the preventive effect of pteroylglutamic acid on glucose metabolism disorders in offspring caused by environmental endocrine disruptors.

Detailed ways

The pteroylglutamic acid intervention experiment was carried out on pregnant and lactating female SD rats. The pteroylglutamic acid used in the experiment was purchased from SIGMA Company of the United States with a purity of 99%.

Pregnancy was detected by vaginal wash smear method; female SD rats were intervened with pteroylglutamate and exposed to BPA during pregnancy and lactation, and were divided into the following three groups: control group, BPA at 40 μg/kg/day group (BPA-no folate), 300μg/kg/day pteroylglutamate +40μg/kg/day BPA pteroylglutamate intervention group (BPA-folate); record the growth rate of the mother rats during pregnancy and the growth rate of the offspring after birth Survival and body weight. Relevant experiments were carried out in the weaning period of the offspring, that is, 21 days after birth, and in the juvenile period (9 weeks old) of the offspring rats. Determination of offspring body weight, pancreatic organ coefficient, fasting blood sugar, fasting insulin; intraperitoneal injection of 2g/kg bodyweight glucose for glucose tolerance test; intraperitoneal injection of 0.75IU/kg body weight insulin for insulin tolerance test; immunofluorescence double staining Morphological analysis of the pancreas was performed; transmission electron microscopy was used to observe the organelle state of the islet β cells; glucose-stimulated insulin release experiments were performed on the isolated and purified islets to evaluate their insulin secretion function. In order to clarify its protective mechanism, SYBR relative quantitative method was used to analyze the expressions of imprinted genes Igf2, H19, Plagl1 and DNA methyl transfer-related genes Dnmt1, Dnmt3a, Dnmt3b in offspring pancreas; ), that is, BSP-PCR product purification combined with sequencing after plasmid cloning to detect the DNA methylation status of Igf2/H19 differentially methylated regions (differently methylated regions, DMR). The invention discloses that pteroyl glutamic acid has a preventive effect on offspring glucose metabolism disorder caused by environmental endocrine disruptors.

Claims (2)

1. Application of pteroyl glutamic acid in the preparation of drugs for preventing or treating offspring diabetes caused by environmental endocrine disruptors. 2. Application of pteroyl glutamic acid in the preparation of health products or functional food for preventing or treating offspring diabetes caused by environmental endocrine disruptors.