CN112972681A - Application of MT-ND6 as new target in medicines for diagnosing and treating metabolic syndrome - Google Patents
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Abstract
The MT-ND6 is used as a new target point to be applied to medicines for diagnosing and treating metabolic syndrome, the expression of MT-ND6 gene in peripheral blood leukocytes of type 2 diabetics and livers of obese mice is remarkably reduced, the methylation level of MT-ND6 gene is remarkably increased, linear correlation analysis shows that the methylation level of MT-ND6 gene is negatively correlated with the expression of mRNA thereof and positively correlated with insulin resistance index HOMA-IR, glycosylated hemoglobin content and serum triglyceride level, the reduction of MT-ND6 can cause mitochondrial dysfunction, increase liver lipid droplet accumulation and reduce insulin sensitivity, and the MT-ND6 gene is closely correlated with the occurrence of metabolic syndrome; the invention provides a new target for clinical diagnosis and treatment of metabolic syndrome and complications thereof, and has important application value.
Description
Technical Field
The invention belongs to the fields of biology and medicine, and particularly relates to application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome.
Background
The metabolic syndrome is a pathological state which clinically comprises abnormal aggregation of various metabolic disorder symptoms such as obesity, diabetes, dyslipidemia and the like, and insulin resistance is considered as a pathological basis and a core link of the metabolic syndrome. With the development of social economy and the change of life style of people, metabolic syndrome and chronic non-infectious diseases caused by the metabolic syndrome seriously threaten the health and the life quality of Chinese people, and the elucidation of the pathogenesis and the search of more effective prevention targets are important problems to be solved urgently.
Mitochondria are key organelles in the cell that regulate glycolipid metabolism, and the process of maintaining dynamic balance and keeping normal functions of mitochondria, such as generation, degradation, fusion, division, distribution and the like of mitochondria under physiological conditions, is called mitochondria homeostasis. Mitochondria serve as energy factories for cells and as a source for the production of free radicals, and homeostatic imbalance thereof is a key factor for driving the formation of insulin resistance in mitochondria-rich tissues such as liver, skeletal muscle, and the like. Therefore, maintenance of mitochondrial homeostasis is essential for improving insulin sensitivity, preventing metabolic syndrome.
The MT-ND6 gene (mitogenic NADH-dehydrogenase 6) is encoded by mitochondrial DNA, is a subunit constituting the mitochondrial electron transport chain complex I, and plays an important role in maintaining the normal activity of the complex I and regulating the mitochondrial homeostasis. The mutation of MT-ND6 can cause the occurrence of mitochondrial genetic diseases such as Leber optic neuropathy and Leigh syndrome. However, no report is available on whether the MT-ND6 gene can regulate the occurrence of metabolic diseases such as obesity, diabetes, hyperlipidemia and the like.
Disclosure of Invention
The invention provides a novel diagnosis and treatment target for metabolic syndrome, aims to provide application of MT-ND6 as a novel target in medicines for diagnosing and treating the metabolic syndrome, and proposes that the expression of MT-ND6 gene in peripheral blood leukocytes of type 2 diabetics and livers of obese mice is remarkably reduced, and the methylation level of MT-ND6 gene is remarkably increased for the first time.
The linear correlation analysis shows that the methylation level of the MT-ND6 gene is linearly and negatively correlated with the expression of mRNA thereof, and is linearly and positively correlated with an insulin resistance index (HOMA-IR), glycosylated hemoglobin content and serum triglyceride level, and the methylation modification of the MT-ND6 is suggested to be possibly used as a biomarker for early clinical screening, diagnosis and prognosis of metabolic syndrome; the knock-down of MT-ND6 can cause mitochondrial dysfunction, increase the accumulation of liver lipid droplets and simultaneously reduce insulin sensitivity, the research results show that the MT-ND6 gene is closely related to the occurrence of metabolic syndrome, and the development of a medicament targeting the gene (or protein) is expected to become a new strategy for treating the metabolic syndrome; the invention provides a new target for clinical diagnosis and treatment of metabolic syndrome and has important application value.
In order to achieve the purpose, the invention is realized by the following technical scheme:
the application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome has the following mechanism: the methylation modification level of the MT-ND6 gene in peripheral blood leukocytes is in linear positive correlation with the HOMA-IR index and the content of glycosylated hemoglobin, and the methylation of the MT-ND6 gene can be used as a biomarker for clinical early screening, diagnosis and prognosis of insulin resistance and type 2 diabetes; and because insulin resistance is the pathological basis of the metabolic syndrome, the methylation of the MT-ND6 gene can also be used as a biomarker for the clinical early screening, diagnosis and prognosis of the metabolic syndrome.
The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome has the following mechanism: the methylation modification level of the MT-ND6 gene in peripheral blood leukocytes is in linear positive correlation with the serum triglyceride level, and the methylation of the MT-ND6 gene can be used as a biomarker for clinical early screening, diagnosis and prognosis of hyperlipidemia or hyperlipidaemia.
The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome has the following mechanism: the expression level of the MT-ND6 gene is reduced in type 2 diabetes mellitus patients and obese mice, and the methylation of the MT-ND6 gene is linearly and negatively correlated with the mRNA expression of the MT-ND6 gene, so that the expression level of the MT-ND6 can be used as a biomarker for early clinical screening, diagnosis and prognosis of hyperlipidemia, obesity, type 2 diabetes mellitus and metabolic syndrome.
The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome has the following mechanism: the knock-down of MT-ND6 can increase liver lipid droplet accumulation, and the MT-ND6 gene can be used as a novel therapeutic target for fatty liver and related liver diseases.
The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome has the following mechanism: the MT-ND6 gene can regulate and control the functions of mitochondria and maintain the steady state of mitochondria, and has good application prospect in preparing medicaments for improving diseases related to mitochondrial dysfunction.
The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome has the following mechanism: the knock-down of MT-ND6 can reduce insulin sensitivity, and shows that the MT-ND6 gene can be used as a novel therapeutic target for insulin resistance, type 2 diabetes and metabolic syndrome.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention discovers for the first time that the methylation modification and the expression level of the MT-ND6 gene in the peripheral blood leucocyte can be used as the biomarker for the clinical early screening, diagnosis and prognosis of the metabolic syndrome, and the pathogenesis process of the metabolic syndrome can be diagnosed only by a small amount of peripheral blood (less than 50 mu L) and conventional equipment (PCR), and the invention has the advantages of simple operation, safety and no wound.
The invention discovers that the MT-ND6 gene is a novel potential target spot for treating the metabolic syndrome for the first time, and the research and development of the medicine targeting the MT-ND6 gene (or protein) is expected to become a novel strategy for treating the metabolic syndrome, thereby having important development value and good application prospect.
The development of drugs targeting the gene (or protein) is expected to become a new strategy for treating metabolic syndrome.
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FIG. 1 shows that the methylation level of MT-ND6 gene is remarkably increased and the mRNA expression level is remarkably reduced in peripheral blood leukocytes of type 2 diabetic patients and the liver of obese mice; FIG. 1A shows the methylation level of MT-ND6 gene in peripheral blood leukocytes of type 2 diabetic patients; FIG. 1B shows the mRNA expression level of MT-ND6 gene in peripheral blood leukocytes of type 2 diabetic patients; FIG. 1C is the methylation level of the liver MT-ND6 gene of obese mice; FIG. 1D shows the expression level of mRNA of MT-ND6 gene in liver of obese mice, Con, normal human control; t2DM, diabetic group; chow, normal diet group, HFD, high fat diet group.
FIG. 2 is a graph showing that MT-ND6 methylation in human peripheral blood leukocytes is negatively correlated with MT-ND6 mRNA levels and positively correlated with serum triglyceride, HOMA-IR, and glycated hemoglobin levels; FIG. 2A is a linear correlation analysis of MT-ND6 gene methylation level with MT-ND6 mRNA expression; FIG. 2B is a linear correlation analysis of MT-ND6 gene methylation levels with serum triglyceride levels; FIG. 2C is a linear correlation analysis of MT-ND6 gene methylation levels with HOMA-IR; FIG. 2D is a linear correlation analysis of the methylation level of MT-ND6 gene with the content of glycated hemoglobin.
FIG. 3 is a photograph showing that knock-down of MT-ND6 gene in HepG2 cells causes mitochondrial dysfunction; FIG. 3A is a graph of mitochondrial oxygen consumption following knock-down of the MT-ND6 gene; FIG. 3B is a histogram of mitochondrial oxygen consumption after knock-down of MT-ND6 gene, NC, negative control group; ND6 esiRNA, MT-ND6 siRNA treated group.
FIG. 4 shows that knocking down MT-ND6 gene causes increased liver lipid droplet accumulation in mice; FIG. 4A is HE staining of mouse liver following knock-down of MT-ND6 gene; FIG. 4B shows the hepatic triglyceride content of mice after knock-down of MT-ND6 gene. NC, negative control group; ND6 siRNA, MT-ND6 siRNA treated group.
FIG. 5 shows that knocking down MT-ND6 gene causes decreased insulin sensitivity in mice; FIG. 5A is the oral glucose tolerance (OGTT) in mice following knock-down of the MT-ND6 gene; FIG. 5B is the protein expression level of mouse liver insulin signaling pathway following knock-down of MT-ND6 gene; NC, negative control group; ND6 siRNA, MT-ND6 siRNA treated group; NC + INS, negative control group injected with insulin; ND6 siRNA + INS, MT-ND6 siRNA injected insulin group.
Detailed Description
The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.
1. Experimental Material
Antibodies beta-actin, Insulin Receptor beta (IR), p-IR (Tyr 1150/1151), Akt, p-Akt (Ser 473), GSK3 alpha/beta, p-GSK3 alpha/beta (Ser 21/9) were purchased from Cell signalling; horseradish peroxidase-conjugated secondary antibodies against mouse and rabbit were purchased from Jackson ImmunoResearch Laboratories; invivofectamine 3.0 siRNA transfection reagent was purchased from Thermo-Fisher; DNA extraction kits were purchased from Qiagen corporation; TRIzol reagent was purchased from Invitrogen corporation; RT-PCR kits were purchased from Takara; DNA methylation kits were purchased from Zymo Research; western and IP cell lysates were purchased from Biyuntian Biotech limited; the triglyceride detection kit is purchased from Nanjing to build a bioengineering institute; erythrocyte lysates were purchased from Tiangen Biochemical technology, Inc.
2. Collection and processing of human blood samples
The fasting peripheral venous blood of the normal people of 45-65 years old and the type 2 diabetics is obtained from Guizhou space hospital, and the gene background is Chinese Han population. The blood sample is centrifuged for 10min at 4 ℃ and 3000g, and the supernatant is plasma and the sediment is blood cells. And (3) treating blood cells by using erythrocyte lysate to obtain white blood cells, extracting DNA by using a DNA extraction kit, and extracting RNA by using a TRIzol reagent.
3. Establishment of mouse obesity model and MT-ND6 siRNA treatment
(1) Establishment of mouse obesity model
Six week old male C57BL/6J mice were purchased from beijing vintonia, and after one week of acclimation the mice were randomly divided into two groups: normal diet group (Chow) and high fat diet group (HFD). The normal diet group was given a normal diet of 10% caloric ratio, and the high fat diet group was given a high fat diet of 60% caloric ratio. After 8 weeks of feeding, mice were sacrificed after overnight fasting, and serum and liver were frozen to-80 ℃ for use.
(2) Mouse MT-ND6 siRNA treatment
The mouse MT-ND6 siRNA sequence is 5 '-GGGUUUGUUGGUUGUUUAATT, 5' -UUAAACAACCAACAAACCCTT, and the in vivo stability is increased through 3 'end cholesterol modification and 2' end methoxyl modification. 5mg/kg siRNA was injected intravenously from the mouse tail using Invivofectamine 3.0 transfection reagent every 4 days, 8 days later the mice were fasted overnight and sacrificed.
4. Experimental methods
(1) Preparation of mouse liver homogenate
Shearing mouse liver with ophthalmic scissors in precooled PBS, cleaning residual blood and blood clot of tissue, centrifuging for 10min at 1000g, and discarding supernatant; adding 1mL of precooled homogenizing medium into each tube of the rest washed tissue sediment, and fully homogenizing by using a homogenizer; centrifuging at 4 deg.C for 10min at 1000g to obtain supernatant as tissue homogenate; protein quantification is carried out by using a BCA method, and leveling is carried out; triglyceride detection was performed using tissue homogenates.
(2) Liver morphology assay
Histomorphometric detection was performed using hematoxylin-eosin staining (HE staining). Placing a part of fresh liver in paraformaldehyde for fixing overnight; after ethanol dehydration, placing the liver tissue in a transparent agent xylene, and dipping in wax for embedding; slicing into slices (4-5 μm thick) in a slicer, ironing in hot water, sticking onto glass slide, and oven drying at 45 deg.C; staining the slices in hematoxylin water solution for several minutes, separating color in acid water and ammonia water for several seconds, washing with running water for 1 hr, adding distilled water, loading 70% and 90% ethanol, and dehydrating for 10 min; dyeing for 2-3min in ethanol-eosin dyeing solution; dehydrating the dyed section by 100% ethanol, performing xylene transparentization, dripping Canadian gum, covering with a cover glass, sealing, observing the dyeing result under a microscope, and taking a picture.
(3) Extraction and detection of RNA
Ultrasonically crushing peripheral blood leucocyte or small mouse liver in 1mL of TRIzol reagent, adding 200 mu L of chloroform, shaking and uniformly mixing, and standing at room temperature for 2-3 min; centrifuging at 12000g for 15min at 4 deg.C, slowly transferring supernatant, adding equal volume of precooled isopropanol, reversing, mixing, and standing at room temperature for 10 min; centrifuging at 4 deg.C for 15min at 12000g, discarding supernatant, washing precipitate with 75% precooled ethanol for 2 times; centrifuging at 4 deg.C for 10min at 12000g, dissolving precipitate with DEPC water, quantifying, and reverse transcribing 1 μ g RNA into cDNA with Takara RT-PCR kit; the expression level of MT-ND6 mRNA was detected by real-time PCR.
(4) DNA extraction and methylation detection
Peripheral blood leukocyte DNA or mouse liver DNA is extracted according to the DNA extraction kit instruction of Qiagen company; after the concentration of the extracted DNA was detected, 500ng of the DNA was sulfite-treated with an EZ DNA methylation kit from Zymo Research; the resulting DNA was tested for methylation levels of MT-ND6 using methylation specific PCR primers.
(5) Extraction and detection of proteins
1) Protein extraction
Resuspending a HepG2 cell sample or a cut mouse liver sample in precooled PBS, washing off redundant fat or blood, centrifuging for 10min at 1000g, and discarding the supernatant; adding 300 μ L of Western and IP cell lysate into the rest cleaned precipitate, performing ultrasonic disruption, and placing on ice for further lysis for 0.5 h; centrifuging at 4 deg.C for 10min at 13000 g to obtain supernatant as protein; protein quantification is carried out by using a BCA method, and leveling is carried out; adding 5X loading buffer and mercaptoethanol, boiling for 10min to denature protein, and storing at-80 deg.C.
2)Western blot
Running electrophoresis by using 10% acrylamide gel, wherein the loading amount of protein is 15 mu g; rotating the film for 100min by adopting a nitrocellulose film under the current of 300 mA; sealing with 5% skimmed milk at room temperature for 1 hr; diluting the primary antibody with 5% skimmed milk or 1% BSA at a ratio of 1:1000, and incubating overnight at 4 ℃; washing the membrane with 1 × TBST on a decolorizing shaker for 3 times, 10min each time, and washing primary antibody; diluting the secondary antibody with 5% skimmed milk or 1% BSA 1:5000, and incubating at room temperature for 1 h; washing the membrane with 1 × TBST on a decolorizing shaker for 4 times, 15min each time, and washing the secondary antibody; using horseradish peroxidase to catalyze a chemiluminescent substance to carry out chemiluminescence; the Western Blot X-ray film results were scanned using an EPSON Perfection V350 scanner and the scanned images were subjected to gray scale analysis using Quantity One software (Bio-Rad).
(6) Statistical analysis
Statistical analysis and statistical mapping of experimental data were performed using GraphPad Prism 5 software. All values are expressed as mean ± sem. Statistical analysis between groups student t-test was used and analysis of differences between groups was done-way ANOVA (LSD post-hoc comparison). P <0.05 was used as a criterion for statistically significant differences.
Example one
Methylation of MT-ND6 gene can be used as a biomarker for clinical early screening, diagnosis and prognosis of metabolic syndromes such as obesity and type 2 diabetes based on insulin resistance as pathology and complications thereof
As shown in FIGS. 1A and C, methylation levels of MT-ND6 gene were significantly increased in peripheral blood leukocytes of type 2 diabetic patients and in the liver of obese mice. As shown in FIGS. 2C and D, linear correlation analysis showed that the methylation level of MT-ND6 gene in human peripheral blood leukocytes was positively correlated with HOMA-IR and glycated hemoglobin content, suggesting that the methylation of MT-ND6 could be used as a biomarker for insulin resistance. Because insulin resistance is the pathological basis and the core link of various metabolic diseases such as obesity, type 2 diabetes mellitus and the like, the methylation of the MT-ND6 gene can be used as a biomarker for clinical early screening, diagnosis and prognosis of metabolic syndromes such as obesity, type 2 diabetes mellitus and the like and complications thereof.
Example two
Methylation of MT-ND6 gene can be used as biomarker for early clinical screening, diagnosis and prognosis of hyperlipidemia (or hyperlipidaemia)
The increase of triglyceride level in serum is one of the most important characteristics of hyperlipidemia (or hyperlipemia), as shown in fig. 2B, the methylation modification level of MT-ND6 gene in human peripheral blood leukocyte is linearly and positively correlated with the serum triglyceride level, suggesting that the methylation of MT-ND6 gene can be used as a biomarker for early screening, diagnosis and prognosis of hyperlipidemia (or hyperlipemia) in clinic.
EXAMPLE III
The expression level of MT-ND6 can be used as a biomarker for clinical early screening, diagnosis and prognosis of hyperlipidemia (or hyperlipidaemia), obesity, type 2 diabetes mellitus and metabolic syndrome and complications thereof
As shown in FIGS. 1B and D, the mRNA expression level of the MT-ND6 gene was decreased in type 2 diabetic patients and obese mice, while the methylation of the MT-ND6 gene in human peripheral blood leukocytes was linearly inversely correlated with the mRNA expression thereof as shown in FIG. 2A. Since the methylation level of MT-ND6 is in linear positive correlation with the content of serum triglyceride, HOMA-IR and glycosylated hemoglobin (FIG. 2B-D), the mRNA expression of MT-ND6 gene is in linear negative correlation with the content of serum triglyceride, HOMA-IR and glycosylated hemoglobin, which suggests that the expression level (mRNA and protein level) of MT-ND6 can be used as a biomarker for early clinical screening, diagnosis and prognosis of hyperlipidemia, obesity, type 2 diabetes, metabolic syndrome and complications thereof.
Example four
The MT-ND6 gene can regulate and control the functions of mitochondria and maintain the steady state of mitochondria, and has good application prospect in preparing medicaments for improving diseases related to mitochondrial dysfunction.
Mitochondrial homeostasis imbalance is a key contributor to the development of metabolic syndrome. The MT-ND6 gene is encoded by mitochondrial DNA, is a subunit constituting the mitochondrial electron transport chain complex I, and plays an important role in maintaining the normal activity of the complex I and regulating the mitochondrial homeostasis. As shown in FIGS. 3A and B, after the expression of MT-ND6 is knocked down by siRNA in human hepatocyte HepG2, the oxygen consumption capacity of mitochondria is remarkably reduced, including that the basal respiration, ATP generation, proton leakage, maximum respiration and reserve respiration of mitochondria are all remarkably reduced, which shows that the MT-ND6 gene plays an important role in maintaining the mitochondria steady state.
EXAMPLE five
MT-ND6 gene can be used as novel therapeutic target for fatty liver and related liver diseases
Excessive fat accumulation in the liver is an important pathological feature of fatty liver. As shown in FIG. 1D, the mRNA expression of MT-ND6 gene was significantly reduced in the liver of obese mice, suggesting that MT-ND6 may play an important role in obesity-related fatty liver. Further, knocking down the expression of MT-ND6 gene by siRNA in normal mice could significantly increase lipid droplet accumulation in liver (FIG. 4A), and simultaneously increase triglyceride content in mouse liver (FIG. 4B). The results show that the MT-ND6 gene can be used as a novel therapeutic target for fatty liver and related liver diseases (such as hepatic fibrosis, liver cirrhosis and the like), and the fatty liver and the related liver diseases are expected to be improved by exogenously over-expressing MT-ND 6.
EXAMPLE six
MT-ND6 gene can be used as a novel therapeutic target for insulin resistance, type 2 diabetes mellitus, metabolic syndrome and complications thereof
As shown in FIGS. 1B and D, the mRNA expression level of the MT-ND6 gene was decreased in type 2 diabetic patients and obese mice, suggesting that the MT-ND6 gene may play an important role in insulin resistance and metabolic syndrome. Further knockdown of expression of MT-ND6 in normal mice could significantly reduce insulin sensitivity, including impaired oral glucose tolerance (fig. 5A) and down-regulated insulin signaling pathway (fig. 5B). The results show that the MT-ND6 gene can be used as a novel treatment target for insulin resistance, type 2 diabetes mellitus, metabolic syndrome and complications thereof, and the metabolic syndrome and the complications thereof are expected to be improved by exogenously over-expressing MT-ND 6.
The invention can be used for the following applications:
the methylation level of the MT-ND6 gene in the human peripheral blood leukocyte is in linear positive correlation with the HOMA-IR and the content of the glycosylated hemoglobin, and the methylation of the MT-ND6 is suggested to be used as a biomarker of insulin resistance. Furthermore, the methylation of the gene has good application value in the clinical early screening, diagnosis and prognosis of metabolic diseases based on the insulin resistance as the pathological base.
The methylation level of the MT-ND6 gene in the human peripheral blood leukocytes is in linear positive correlation with the content of serum triglyceride, and further, the methylation of the gene has good application value in early screening, diagnosis and prognosis of hyperlipidemia (or hyperlipidaemia) clinic.
The mRNA expression of the MT-ND6 gene in the human peripheral blood leukocytes is linearly and negatively correlated with the methylation level of the MT-ND6 gene, and further, the expression level of the gene has good application value in early clinical screening, diagnosis and prognosis of metabolic diseases based on insulin resistance, such as hyperlipidemia (or hyperlipidaemia), obesity, type 2 diabetes and the like.
The MT-ND6 gene has the regulation and control functions of enhancing the oxygen consumption capacity of mitochondria and maintaining the mitochondria steady state. Furthermore, the gene has good application prospect in preparing medicines for improving diseases related to mitochondrial dysfunction.
The MT-ND6 gene has the function of regulating liver lipid accumulation, is a novel potential target spot for treating fatty liver and related liver diseases (such as hepatic fibrosis, liver cirrhosis and the like), and has good application prospect in preparing medicines for improving the fatty liver and the related liver diseases.
The MT-ND6 gene has a key effect of regulating insulin sensitivity, is a novel potential target for treating metabolic diseases based on insulin resistance, such as obesity, type 2 diabetes mellitus and the like, and is expected to become a new strategy for treating metabolic syndrome and complications thereof through research and development of a medicament targeting the MT-ND6 gene, and has important development value and good application prospect.
The embodiments given above are preferable examples for implementing the present invention, and the present invention is not limited to the above-described embodiments. Any non-essential addition and replacement made by the technical characteristics of the technical scheme of the invention by a person skilled in the art belong to the protection scope of the invention.
Claims (6)
- The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome is characterized in that the mechanism is as follows: the methylation modification level of the MT-ND6 gene in peripheral blood leukocytes is in linear positive correlation with the HOMA-IR index and the content of glycosylated hemoglobin, and the methylation of the MT-ND6 gene can be used as a biomarker for clinical early screening, diagnosis and prognosis of insulin resistance and type 2 diabetes; and because insulin resistance is the pathological basis of the metabolic syndrome, the methylation of the MT-ND6 gene can also be used as a biomarker for the clinical early screening, diagnosis and prognosis of the metabolic syndrome.
- The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome is characterized in that the mechanism is as follows: the methylation modification level of the MT-ND6 gene in peripheral blood leukocytes is in linear positive correlation with the serum triglyceride level, and the methylation of the MT-ND6 gene can be used as a biomarker for clinical early screening, diagnosis and prognosis of hyperlipidemia or hyperlipidaemia.
- The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome is characterized in that the mechanism is as follows: the expression level of the MT-ND6 gene is reduced in type 2 diabetes mellitus patients and obese mice, and the methylation of the MT-ND6 gene is linearly and negatively correlated with the mRNA expression of the MT-ND6 gene, so that the expression level of the MT-ND6 can be used as a biomarker for early clinical screening, diagnosis and prognosis of hyperlipidemia, obesity, type 2 diabetes mellitus and metabolic syndrome.
- The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome is characterized in that the mechanism is as follows: the knock-down of MT-ND6 can increase liver lipid droplet accumulation, and the MT-ND6 gene can be used as a novel therapeutic target for fatty liver and related liver diseases.
- The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome is characterized in that the MT-ND6 gene can regulate mitochondrial function and maintain mitochondrial homeostasis.
- The application of MT-ND6 as a new target point in medicines for diagnosing and treating metabolic syndrome is characterized in that the mechanism is as follows: the knock-down of MT-ND6 can reduce insulin sensitivity, and shows that the MT-ND6 gene can be used as a novel therapeutic target for insulin resistance, type 2 diabetes and metabolic syndrome.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011144818A1 (en) * | 2010-05-19 | 2011-11-24 | Mas-Metabolic Analytical Services Oy | Method for detection of predisposition to atherosclerosis, coronary heart disease and related conditions |
| WO2015086913A1 (en) * | 2013-12-12 | 2015-06-18 | Mas-Metabolic Analytical Services Oy | Methods for detection of the risk of obesity, the metabolic syndrome and diabetes |
| CN109661468A (en) * | 2016-05-06 | 2019-04-19 | 新加坡国立大学 | The mitochondria of recombinant nucleic acid delivers |
| CN110022876A (en) * | 2016-09-28 | 2019-07-16 | 阿布拉科斯生物科学有限公司 | The method for treating mitochondria and metabolic disorder |
| CN110869494A (en) * | 2017-06-12 | 2020-03-06 | 西奈卫生系统公司 | Allograft tolerance without systemic immune suppression |
| CN111961666A (en) * | 2020-08-25 | 2020-11-20 | 上海市第六人民医院 | Genome composition related to difficult metabolic diseases and application thereof |
-
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011144818A1 (en) * | 2010-05-19 | 2011-11-24 | Mas-Metabolic Analytical Services Oy | Method for detection of predisposition to atherosclerosis, coronary heart disease and related conditions |
| WO2015086913A1 (en) * | 2013-12-12 | 2015-06-18 | Mas-Metabolic Analytical Services Oy | Methods for detection of the risk of obesity, the metabolic syndrome and diabetes |
| CN109661468A (en) * | 2016-05-06 | 2019-04-19 | 新加坡国立大学 | The mitochondria of recombinant nucleic acid delivers |
| CN110022876A (en) * | 2016-09-28 | 2019-07-16 | 阿布拉科斯生物科学有限公司 | The method for treating mitochondria and metabolic disorder |
| CN110869494A (en) * | 2017-06-12 | 2020-03-06 | 西奈卫生系统公司 | Allograft tolerance without systemic immune suppression |
| CN111961666A (en) * | 2020-08-25 | 2020-11-20 | 上海市第六人民医院 | Genome composition related to difficult metabolic diseases and application thereof |
Non-Patent Citations (7)
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116459340A (en) * | 2023-03-13 | 2023-07-21 | 山东大学 | Application of olfactory receptor Olfr110 in metabolic homeostasis and metabolic diseases |
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