阅读说明：本技术 氨基酸组合物在胎儿神经系统发育疾病药物中的应用 (Application of amino acid composition in medicine for treating fetal nervous system development diseases ) 是由 吕国荣 于 2021-09-01 设计创作，主要内容包括：本发明公开的是一种氨基酸组合物在胎儿神经系统发育疾病药物中的应用,包括氨基酸组合物的制药用途,氨基酸组合物为用于制备预防和治疗胎儿神经系统发育疾病的药物组合物,还公开披露了一种氨基酸组合物鉴定和统计处理方法。本发明研究超声引导羊膜腔穿刺获得的中枢神经系统孤立性软指标异常胎儿羊水的氨基酸代谢特征,得到用于制备预防和治疗胎儿神经系统发育疾病的药物组合物,解决胎儿单纯性脉络丛囊肿和单纯性侧脑室扩张的疾病,促进胎儿脑神经系统的发育和完善。(The invention discloses an application of an amino acid composition in a medicament for treating fetal nervous system development diseases, which comprises the pharmaceutical application of the amino acid composition, the amino acid composition is used for preparing the medicinal composition for preventing and treating the fetal nervous system development diseases, and an amino acid composition identification and statistical treatment method is also disclosed. The invention researches the amino acid metabolism characteristics of fetal amniotic fluid with abnormal central nervous system isolated soft index obtained by the puncture of amniotic cavity guided by ultrasound to obtain the pharmaceutical composition for preventing and treating the development diseases of the fetal nervous system, solves the problems of simple choroid plexus cyst and simple lateral ventricle expansion of the fetus, and promotes the development and the improvement of the fetal cerebral nervous system.)

1. The pharmaceutical use of an amino acid composition, characterized in that: the amino acid composition is used for preparing a pharmaceutical composition for preventing and treating fetal nervous system development diseases.

2. The pharmaceutical use of claim 1, wherein: the fetal development disease of the nervous system comprises: simple choroid plexus cysts and simple lateral ventricular dilatation.

3. The pharmaceutical use of claim 1, wherein: the amino acid composition comprises: amino acids and vitamins, the amino acids including aspartic acid, lysine, ornithine, citrulline, and beta-aminoisobutyric acid; the vitamins include B₃、B₅、B₆、B₉And B₁₂。

4. The pharmaceutical use of claim 2, wherein: the amino acid metabolism characteristics of the simple choroid plexus cyst and the simple lateral ventricle expansion are derived from the amino acid metabolism characteristics of amniotic fluid of maternal, fetal and placental tissues.

5. A method for identifying and statistically processing amino acid compositions, comprising: the identification and statistical processing method comprises the following specific steps:

the method comprises the following steps: determining a research object, and performing amniotic fluid puncture on an Isolated Choroid Plexus Cyst (ICPC) fetus, an isolated ventricular dilatation (IVM) fetus and a normal group fetus at 20-22 gestational weeks by using ultrasonic guidance;

step two: detecting amniotic fluid by using an AB Sciex Exion LC chromatograph system and an AB Sciex QTRAP 4500 mass spectrometer;

step three: and (3) compound identification and statistical treatment, summarizing the detection results in the step two to obtain 31 amino acids, performing statistical analysis on the obtained amino acid peak surfaces by adopting a Principal Component Analysis (PCA), a partial least squares discriminant analysis (PLS-DA) and univariate statistical analysis respectively, and performing pathway analysis finally.

6. The method for the identification and statistical treatment of amino acid compositions according to claim 5, characterized in that: and the third step comprises ICPC group identification and statistical analysis and IVM group identification and statistical analysis.

7. The method for the identification and statistical treatment of amino acid compositions according to claim 6, characterized in that: the ICPC group identification and statistical analysis specifically comprises the following steps:

(1) performing partial least squares discriminant analysis (PLS-DA) on the ICPC group and the normal group to obtain PLS-DA model scores of metabolites of the ICPPC group and the normal group;

(2) screening according to the criterion that the variable weight projection (VIP) value is greater than 1: proline, 5-hydroxylysine, isoleucine, beta-aminoisobutyric acid, lysine, ethanolamine, valine, glutamic acid, citrulline, aspartic acid;

(3) carrying out Mann-Whitney U rank sum test on the screened 10 amino acids;

(4) analyzing the altered metabolic pathway of the ICPC group compared with the normal group;

(5) and obtaining the identification and statistical conclusion of the ICPC group.

8. A method for the identification and statistical treatment of amino acid compositions according to claim 7, characterized in that: the content of the aspartic acid, the citrulline and the ethanolamine in the step (3) is reduced compared with that in a normal group; the content of valine, 5-hydroxylysine, proline and isoleucine is increased compared with that of the normal group; the contents of the beta-aminoisobutyric acid, the glutamic acid and the lysine are not obviously different.

9. The method for the identification and statistical treatment of amino acid compositions according to claim 6, characterized in that: the IVM group identification and statistical analysis specifically comprises:

(1) obtaining PLS-DA model scores of metabolites of the IVM group and the normal group by a partial least squares discriminant analysis method (PLS-DA) of the IVM group and the normal group;

(2) screening according to the criterion that the variable weight projection (VIP) value is greater than 1: proline, 5-hydroxylysine, lysine, aspartic acid, beta-aminoisobutyric acid, ornithine;

(3) carrying out Mann-Whitney U rank sum test on the 6 screened amino acids;

(3) analyzing the altered metabolic pathway of the IVM group compared with the normal group;

(4) and obtaining the identification and statistical conclusion of the IVM group.

10. The method for the identification and statistical treatment of amino acid compositions according to claim 9, characterized in that: the ornithine, lysine, beta-aminoisobutyric acid and aspartic acid contents in the step (2) are reduced compared with those in a normal group; the proline and 5-hydroxylysine content is increased compared with the normal group.

Technical Field

The invention relates to the technical field of medicines, in particular to application of an amino acid composition in medicines for treating fetal nervous system development diseases.

Background

Abnormal ultrasound soft index refers to ultrasound detectable morphological variation, the etiology of which is still unknown, usually transient and non-pathological, but suggesting an increased risk of potential fetal aneuploidy. The amniotic fluid is derived from maternal, fetal and placental tissues, is in direct contact with multiple organs of the fetus, and can reflect the health conditions of both the mother and fetus, including fetal structural abnormalities, chromosomal abnormalities, and maternal diseases.

The development of the central nervous system of a fetus is closely related to amino acids, wherein excitatory amino acids play various physiological and pathophysiological roles in the central nervous system; methionine cycle inhibitors prevented neural tube closure in mouse embryos, believed to be neural tube; closure depends on the activity of the methionine and folate cycles; l-serine synthesis disorders lead to congenital microcephaly. Meanwhile, the change of the intrinsic and extrinsic metabolites of the living system can be evaluated through metabonomics, and the investigation of physiological states, disease diagnosis, the discovery of markers and the identification of metabolic disorder pathways are carried out. Since in vivo metabolites are downstream products of gene expression and protein synthesis, they more directly reflect cellular activity at the functional level and at specific time points.

The scheme researches the amino acid metabolism characteristics of the fetal amniotic fluid with abnormal central nervous system isolated soft indexes obtained by the puncture of amniotic cavities under the guidance of ultrasound to obtain the pharmaceutical composition for preventing and treating the development diseases of the fetal nervous system, solves the diseases of simple choroid plexus cyst and simple lateral ventricle expansion of the fetus, and promotes the development and the perfection of the fetal cerebral nervous system.

Disclosure of Invention

The invention discloses an application of an amino acid composition in a medicament for treating a fetal nervous system development disease, which mainly aims to overcome the defects and shortcomings in the prior art, is used for preparing a medicament composition for preventing and treating the fetal nervous system development disease, solves the problems of simple choroid plexus cyst and simple lateral ventricle expansion of a fetus, and promotes the development and the perfection of a fetal cerebral nervous system.

The technical scheme adopted by the invention is as follows:

the pharmaceutical use of an amino acid composition for preparing a pharmaceutical composition for preventing and treating fetal neurologic development diseases.

Still further, the fetal neurologic development disorder comprises: simple choroid plexus cysts and simple lateral ventricular dilatation.

Still further, the amino acid composition comprises: amino acids and vitamins, the amino acids including aspartic acid, lysine, ornithine, citrulline, and beta-aminoisobutyric acid; the vitamins include B₃、B₅、B₆、B₉And B₁₂。

Still further, the metabolic characteristics of amino acids of simple choroid plexus cysts and simple lateral ventricular dilatation are derived from the metabolic characteristics of amino acids of amniotic fluid of maternal, fetal and placental tissues.

A method for identifying and statistically processing amino acid compositions, which comprises the following specific steps:

the method comprises the following steps: determining a research object, and performing amniotic fluid puncture on an Isolated Choroid Plexus Cyst (ICPC) fetus, an isolated ventricular dilatation (IVM) fetus and a normal group fetus at 20-22 gestational weeks by using ultrasonic guidance;

step two: detecting amniotic fluid by using an AB Sciex Exion LC chromatograph system and an AB Sciex QTRAP 4500 mass spectrometer;

step three: compound identification and statistical treatment, summarizing the detection results in the step two to obtain 31 amino acids, then respectively carrying out statistical analysis on the obtained amino acid peak surfaces by adopting a Principal Component Analysis (PCA), a partial least squares discriminant analysis (PLS-DA) and univariate statistical analysis,

and then finally performing pathway analysis.

Further, the third step includes ICPC group identification and statistical analysis and IVM group identification and statistical analysis.

Further, the ICPC panel identification and statistical analysis specifically includes:

(1) performing partial least squares discriminant analysis (PLS-DA) on the ICPC group and the normal group to obtain PLS-DA model scores of metabolites of the ICPPC group and the normal group;

(2) screening according to the criterion that the variable weight projection (VIP) value is greater than 1: proline, 5-hydroxylysine, isoleucine, beta-aminoisobutyric acid, lysine, ethanolamine, valine, glutamic acid, citrulline, aspartic acid;

(3) carrying out Mann-Whitney U rank sum test on the screened 10 amino acids;

(4) analyzing the altered metabolic pathway of the ICPC group compared with the normal group;

(5) and obtaining the identification and statistical conclusion of the ICPC group.

Further, in the step (3), the content of the aspartic acid, the citrulline and the ethanolamine is reduced compared with that in the normal group; the content of valine, 5-hydroxylysine, proline and isoleucine is increased compared with that of the normal group; the contents of the beta-aminoisobutyric acid, the glutamic acid and the lysine are not obviously different.

Further, the IVM group identification and statistical analysis specifically includes:

(1) obtaining PLS-DA model scores of metabolites of the IVM group and the normal group by a partial least squares discriminant analysis method (PLS-DA) of the IVM group and the normal group;

(2) screening according to the criterion that the variable weight projection (VIP) value is greater than 1: proline, 5-hydroxylysine, lysine, aspartic acid, beta-aminoisobutyric acid, ornithine;

(3) carrying out Mann-Whitney U rank sum test on the 6 screened amino acids;

(3) analyzing the altered metabolic pathway of the IVM group compared with the normal group;

(4) and obtaining the identification and statistical conclusion of the IVM group.

Further, in the step (2), the content of ornithine, lysine, beta-aminoisobutyric acid and aspartic acid is reduced compared with the normal group; the proline and 5-hydroxylysine content is increased compared with the normal group.

As can be seen from the above description of the present invention, compared with the prior art, the present invention has the following advantages:

1. the amino acid composition overcomes the defects of branched chain amino acid metabolic pathway abnormality, arginine synthetic pathway abnormality and pantothenic acid and coenzyme A synthetic pathway abnormality, and provides scientific basis for supplementing amino acid and vitamin for pregnant women.

2. The amino acid composition effectively avoids the change of a plurality of metabolic pathways in the fetus, prevents the metabolic disorder of the amino acid in the fetus, enables the energy metabolism in the fetus to be normal, and avoids the possibility of oxidative stress in the fetus.

3. The amino acid composition can prevent arginine biosynthesis pathway in fetus from changing, so that the development of nervous system of fetus is normal and smooth.

4. The amino acid composition can prevent and treat development diseases of the fetal nervous system, solve the problems of simple choroid plexus cyst and simple lateral ventricle expansion of a fetus, and promote the development and the improvement of the fetal cerebral nervous system.

Drawings

FIG. 1 is a PLS-DA model score plot of ICPC group metabolites of the present invention.

FIG. 2 is a PLS-DA model score plot of normal group metabolites compared to the ICPC group in accordance with the present invention.

FIG. 3 is a graph of the variable weight projection (VIP) scores for the amino acids of the ICPC set of the present invention.

FIG. 4 is a graph showing the statistical analysis of the aspartic acid content in the ICPC group of the present invention and the aspartic acid content in the normal group.

FIG. 5 is a graph showing the statistical analysis of citrulline content in the ICPC group of the present invention and the normal group.

FIG. 6 is a graph showing the statistical analysis of ethanolamine content in the ICPC group of the present invention and the normal group.

FIG. 7 is a graph showing the statistical analysis of the valine content in the ICPC group of the present invention and that in the normal group.

FIG. 8 is a graph showing the statistical analysis of the 5-hydroxylysine content of the ICPC group of the present invention and that of the normal group.

FIG. 9 is a graph showing the statistical analysis of proline content in the ICPC group of the present invention and the normal group.

FIG. 10 is a graph showing the statistical analysis of isoleucine content in the ICPC group of the present invention and the normal group.

FIG. 11 is a graph showing altered metabolic pathways in the ICPC panel of the present invention compared with the normal panel.

FIG. 12 is a PLS-DA model score plot of IVM group metabolites of the present invention.

FIG. 13 is a PLS-DA model score plot of normal group metabolites compared to IVM of the present invention.

FIG. 14 is a graph of the variable weight projection (VIP) scores for group IVM amino acids of the present invention.

FIG. 15 is a graph of altered metabolic pathways in the IVM group of the present invention compared to the normal group.

Detailed Description

Embodiments of the present invention will be further described with reference to the accompanying drawings.

As shown in fig. 1 to fig. 15, the amino acid composition is used for preparing a pharmaceutical composition for preventing and treating fetal development of nervous system diseases.

Still further, the fetal neurologic development disorder comprises: simple choroid plexus cysts and simple lateral ventricular dilatation.

Still further, the amino acid composition comprises: amino acids and vitamins, the amino acids including aspartic acid, lysine, ornithine, citrulline, and beta-aminoisobutyric acid; the vitamins include B₃、B₅、B₆、B₉And B₁₂。

Still further, the metabolic characteristics of amino acids of simple choroid plexus cysts and simple lateral ventricular dilatation are derived from the metabolic characteristics of amino acids of amniotic fluid of maternal, fetal and placental tissues.

A method for identifying and statistically processing amino acid compositions, which comprises the following specific steps:

the method comprises the following steps: determining a research object, and performing amniotic fluid puncture on an Isolated Choroid Plexus Cyst (ICPC) fetus, an isolated ventricular dilatation (IVM) fetus and a normal group fetus at 20-22 gestational weeks by using ultrasonic guidance;

step two: detecting amniotic fluid by using an AB Sciex Exion LC chromatograph system and an AB Sciex QTRAP 4500 mass spectrometer;

step three: compound identification and statistical treatment, summarizing the detection results in the step two to obtain 31 amino acids, then respectively carrying out statistical analysis on the obtained amino acid peak surfaces by adopting a Principal Component Analysis (PCA), a partial least squares discriminant analysis (PLS-DA) and univariate statistical analysis,

and then finally performing pathway analysis.

The 31 amino acids include: leucine, arginine, tyrosine, citrulline, 3-methyl-histidine, serine, 5-hydroxylysine, asparagine, alanine, cystathionine, gamma-aminobutyric acid, sarcosine, isoleucine, alpha-aminobutyric acid, hydroxyproline, proline, valine,

ornithine, methionine, histidine, ethanolamine, lysine, aspartic acid, glutamic acid, 1-methyl-histidine, tryptophan, phenylalanine, beta-aminoisobutyric acid, homocysteine,

threonine and glycine.

Further, the third step includes ICPC group identification and statistical analysis and IVM group identification and statistical analysis.

Further, the ICPC panel identification and statistical analysis specifically includes:

(1) performing partial least squares discriminant analysis (PLS-DA) on the ICPC group and the normal group to obtain PLS-DA model scores of metabolites of the ICPPC group and the normal group;

(2) screening according to the criterion that the variable weight projection (VIP) value is greater than 1: proline, 5-hydroxylysine, isoleucine, beta-aminoisobutyric acid, lysine, ethanolamine, valine, glutamic acid, citrulline, aspartic acid;

(3) carrying out Mann-Whitney U rank sum test on the screened 10 amino acids;

the content of the aspartic acid, the citrulline and the ethanolamine is reduced compared with that of a normal group; the content of valine, 5-hydroxylysine, proline and isoleucine is increased compared with that of the normal group; the contents of the beta-aminoisobutyric acid, the glutamic acid and the lysine are not obviously different.

(4) Analysis of altered metabolic pathways in the ICPC group compared to the normal group, where a total of 4 metabolic pathways were significantly altered, including a: valine, leucine and isoleucine biosynthetic pathways; b: the arginine degradation pathway; c: pantothenic acid and coenzyme a biosynthetic pathways; d:

valine, leucine and isoleucine degradation pathways;

(5) and obtaining the identification and statistical conclusion of the ICPC group, wherein the conclusion comprises the following steps: 1. an abnormality in the branched chain amino acid metabolic pathway; 2. abnormalities in the arginine synthetic pathway; 3. pantothenic acid and coenzyme A synthesis pathways are abnormal, aspartic acid is an important substance in the tricarboxylic acid cycle, and the risk of abnormal lipid and glucose metabolism exists; 4. provides scientific basis for the pregnant women to supplement amino acid and vitamin.

Further, the IVM group identification and statistical analysis specifically includes:

(1) obtaining PLS-DA model scores of metabolites of the IVM group and the normal group by a partial least squares discriminant analysis method (PLS-DA) of the IVM group and the normal group;

(2) screening according to the criterion that the variable weight projection (VIP) value is greater than 1: proline, 5-hydroxylysine, lysine, aspartic acid, beta-aminoisobutyric acid, ornithine;

the ornithine, lysine, beta-aminoisobutyric acid and aspartic acid contents are reduced compared with those of a normal group;

the proline and 5-hydroxylysine content is increased compared with the normal group.

(3) Carrying out Mann-Whitney U rank sum test on the 6 screened amino acids;

(3) analysis of altered metabolic pathways in the IVM group compared to the normal group, wherein a total of 3 metabolic pathways were significantly altered, including: firstly, arginine biosynthesis pathway; (ii) lysine degradation pathway; ③ metabolic pathway of arginine and proline;

(4) and obtaining the identification and statistical conclusion of the IVM group, wherein the conclusion comprises the following steps:

1. the IVM fetus has various metabolic pathways changed, amino acid metabolism in the fetus is disturbed, and finally energy metabolism in the fetus can be influenced, and the possibility of oxidative stress in the fetus is reflected.

2. The arginine biosynthetic pathway in IVM fetuses is obviously changed, and the pathways are closely related to the development of the nervous system of fetuses, thereby providing a new idea for exploring the generation mechanism of the arginine biosynthetic pathway.

3. Ornithine, lysine, aspartic acid and beta-aminoisobutyric acid depletion may be new targets for IVM therapy.

As can be seen from the above description of the present invention, compared with the prior art, the present invention has the following advantages:

1. the amino acid composition overcomes the defects of branched chain amino acid metabolic pathway abnormality, arginine synthetic pathway abnormality and pantothenic acid and coenzyme A synthetic pathway abnormality, and provides scientific basis for supplementing amino acid and vitamin for pregnant women.

2. The amino acid composition effectively avoids the change of a plurality of metabolic pathways in the fetus, prevents the metabolic disorder of the amino acid in the fetus, enables the energy metabolism in the fetus to be normal, and avoids the possibility of oxidative stress in the fetus.

3. The amino acid composition can prevent arginine biosynthesis pathway in fetus from changing, so that the development of nervous system of fetus is normal and smooth.

4. The amino acid composition can prevent and treat development diseases of the fetal nervous system, solve the problems of simple choroid plexus cyst and simple lateral ventricle expansion of a fetus, and promote the development and the improvement of the fetal cerebral nervous system.

The above description is only an embodiment of the present invention, but the design concept of the present invention is not limited thereto, and any insubstantial modifications of the present invention using this concept shall fall within the scope of infringing the present invention.