CN109289039A - Purposes of the transcription factor YY1 in preparation treatment diabetes targeted drug - Google Patents
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Abstract
The invention discloses purposes of the transcription factor YY1 in preparation treatment diabetes targeted drug, further disclose purposes of the transcription factor YY1 in the drug of preparation regulation beta Cell of islet development；The development for regulating and controlling beta Cell of islet by transcription factor YY1 is used to prepare treatment diabetes targeted drug；The development of regulation beta Cell of islet is realized by the expression of 5 α of chondriogen cytochrome C oxidase subunit, the isocitric dehydrogenase subunit α and cromoci of regulation core coding, and then islet β cell insulin is influenced, new way is provided for treatment diabetes especially diabetes B.
Description
Technical field
The present invention relates to the function of gene and applied technical fields, and in particular to a kind of transcription factor YY1 is treated in preparation
Purposes in diabetes targeted drug, further to transcription factor YY1 in the drug of preparation regulation beta Cell of islet development
Purposes.
Background technique
In recent years, with the raising of the development of social economy and living standards of the people, the illness rate of diabetes is also year by year
Rise, wherein diabetes B (T2DM) has accounted for 95% or more.Diabetes B has become a kind of human health of seriously affecting
Global public health problem.Chinese population data show Chinese Adult diabetes prevalence up to 11.6%, diabetes
Early period, illness rate was more up to 50.1%, and diabetes B is a kind of complex disease, and definite pathogenesis is still unclear, at present
Research shows that diabetes B is result caused by heredity, environment etc. are multifactor.
The insulin of islet β cell is the hormone that whole body uniquely can be hypoglycemic, maintain the normal quantity of β cell and
Function is to maintain the necessary condition of body glucostasis.The link of diabetes B occurrence and development most critical is exactly beta Cell of islet function
Energy obstacle, cannot secrete the insulin for meeting body needs.Therefore beta Cell of islet, which becomes, finds pathogenesis of diabetes mellitus and controls
Treat the key of target spot.
Transcription factor plays an important role in terms of the development of regulation beta Cell of islet and function in pancreas.In recent years, Pdx-
The transcription factor that some pairs of β cells such as 1, Ngn3, Pax4, Nkx2.2, Nkx6.1, MafA and NueroD play a decisive role is successive
It is found.2013, by the way that the full exon group of the insulinoma distributed is sequenced, discovery turned in south Shanghai Ruijin Hospital Cao Ya etc.
There is high frequency body cell T372R gene mutation in record factor YY1, and T372R gene mutation changes the expression of YY1 target gene, this
Research has prompted transcription factor YY1 that may play an important role in beta Cell of islet.
YY1 is zinc finger class transcription factor GLI-Kr ü ppel family member, and combs a member in protein family (PcG) more.
YY1 gene very high homology between different genera, the DNA binding structural domain of YY1 can form zinc fingers die body, and identification is specific
DNA sequence dna.YY1 is a kind of multifunctional protein, and the table of target gene is regulated and controled and interacting from different albumen confactors
It reaches, participates in the normal physiology courses such as regulating cell development, differentiation, duplication and proliferation.YY1, which has, inhibits transcription and activated transcription
Dual function, just because of this, YY1 is referred to as the yin-yang factor.
A large amount of related YY1 existing at present are swollen in lymthoma, leukaemia, prostate cancer, breast cancer, cervical carcinoma, colon cancer etc.
Expression and the research of regulatory mechanism in tumor tissue, the results showed that YY1 overexpression in most tumours, and it is swollen with it
The formation of tumor, progress and prognosis are related.In metabolic organs such as liver, muscle, fat, YY1 has been found to participate in regulation mitochondria
Function and insulin/insulin-like growth factor signal path, these are same for the existence of beta Cell of islet and the secretion of insulin
Sample is of great significance.However up to the present, there are no the relevant researchs of effect of the transcription factor YY1 in beta Cell of islet.
Summary of the invention
The purpose of the present invention is to provide the drugs that a kind of transcription factor YY1 is used to prepare regulation beta Cell of islet development.
The purpose of the present invention is achieved through the following technical solutions:
In a first aspect, purposes of the transcription factor YY1 in preparation treatment diabetes targeted drug.
Preferably, transcription factor YY1 is realized by the development of regulation beta Cell of islet.
Second aspect, purposes of the transcription factor YY1 in the drug of preparation regulation beta Cell of islet development.
Preferably, transcription factor YY1 is realized by the mitochondrial gene expression of regulation core coding.
It is furthermore preferred that the chondriogen of the core coding is 5 α of cytochrome C oxidase subunit, isocitric acid dehydrogenation
Enzyme subunit α and cromoci.
Preferably, in the drug, using transcription factor YY1 as active constituent, further include pharmaceutically acceptable auxiliary material or
Complementary ingredient；It is furthermore preferred that the mass percentage content of the transcription factor YY1 be >=98%, application dose be 1~
5000mg/kg。
Preferably, the pharmaceutical dosage form is tablet, pulvis, granule, injection, capsule, oral solution or sustained release agent.
Compared with prior art, the beneficial effects of the present invention are:
It is a discovery of the invention that transcription factor YY1 can be used for preparing treatment diabetes by the development of regulation beta Cell of islet
Targeted drug；It is sub- single by 5 α of chondriogen cytochrome C oxidase subunit of regulation core coding, isocitric dehydrogenase
The development of regulation beta Cell of islet is realized in the expression of position α and cromoci, and then influences islet β cell insulin, to control
It treats diabetes especially diabetes B and provides new way.
Detailed description of the invention
Upon reading the detailed description of non-limiting embodiments with reference to the following drawings, other feature of the invention,
Objects and advantages will become more apparent upon:
Fig. 1 is the forming types and identification schematic diagram of YY1 β KO transgenic mice；Wherein, (A) YY1flox mouse and Mip-
The forming types and mating strategy ideograph of cre mouse (or Rip-cre mouse)；(B) genotype identification of transgenic mice, M
Representation DNA marker (DL2000, Takara), the mouse that number is 1-7,1 represents YY1flox/flox mouse, and 2 represent
YY1-/- mouse, 3 represent YY1flox/- mouse, and 4 represent Mip-Cre- mouse, and 5 represent Mip-Cre+ mouse, and 6 represent Rip-
Cre+ mouse, 7 represent Rip-Cre- mouse；(C) the mRNA expression (n=3) of Mip-YY1 β KO mouse islets YY1；(D)
The mRNA expression (n=3) of Rip-YY1 β KO mouse islets YY1；(E) protein expression of Mip-YY1 β KO mouse islets YY1
Horizontal (n=2), compared with the control group p < 0.01；
Fig. 2 is that YY1 β KO mouse hyperglycemia and impaired glucose tolerance schematic diagram one occurs；Wherein, (A) Rip-YY1 β KO
The random blood glucose level (n=20) of mouse and control mice；(B) random blood glucose level of Mip-YY1 β KO mouse and control mice
(n=13:14)；(C) 6 hours blood glucose levels (n=20) of empty stomach of Rip-YY1 β KO mouse and control mice；(D)Mip-YY1β
6 hours blood glucose levels (n=11) of empty stomach of KO mouse and control mice；
Fig. 3 is that YY1 β KO mouse hyperglycemia and impaired glucose tolerance schematic diagram two occurs；Wherein, (E) IPGTT shows 6
The glucose tolerance curve (n=7) of week old Rip-YY1 β KO mouse and control mice；It (F) is the area under the curve (AUC) of (E)；
(G) IPGTT shows that the glucose tolerance curve (n=10) (H) of 8 week old Mip-YY1 β KO mouse and control mice is (G)
AUC；(I) IPGTT shows the glucose tolerance curve (n=7) of 16 week old Mip-YY1 β KO mouse and control mice；(J) it is (I)
AUC；(K) ITT shows the insulin sensitivity implementations (n=8:11) of 16 week old Mip-YY1 β KO mouse and control mice；(L)
For the AUC of (K)；(M) survivorship curve (n=18) of Rip-YY1 β KO mouse and control mice；
Fig. 4 is that impaired glucose tolerance phenotype schematic diagram occurs in Mip-YY1 β KO mouse under HFD is fed；Wherein, (A) IPGTT is aobvious
Show the glucose tolerance curve (n=5) of Mip-YY1 β KO mouse and control mice when giving HFD and feeding 16 weeks；(B) it is (A)
Area under the curve；(C) it is quick to be shown in the insulin of Mip-YY1 β KO mouse and control mice when giving HFD nursing 16 weeks by ITT
Perceptual situation (n=5)；It (D) is the area under the curve of (C)；
Fig. 5 is the insulin secretion schematic diagram that specific knockdown YY1 reduces glucose stimulation in mature beta Cell of islet；Its
In, (A) shows the insulin level in serum (n=11) of 6 week old Rip-YY1 β KO mouse and the stimulation of control mice glucose,
It (B) is the area under the curve of (A)；(C) in the serum for showing 16 week old Mip-YY1 β KO mouse and the stimulation of control mice glucose
Insulin level (n=7:10), (D) are the area under the curve of (C)；(E) GSIS detects Rip-YY1 β KO mouse and control mice
The insulin secreting ability of primary pancreas islet extracorporeal glucose stimulation；(F) GSIS detects Mip-YY1 β KO mouse and control mice is former
For the insulin secreting ability of pancreas islet extracorporeal glucose stimulation；
Fig. 6 is that specific knockdown YY1 reduces insulin content schematic diagram in beta Cell of islet；Wherein, (A) 16 week old Mip-
The immunohistochemical staining of insulin, scale bar represent 100 μm in the pancreatic tissue of YY1 β KO mouse and control mice slice,
(C) (n=18) insulin content is surveyed in (B) (n=4) and pancreas islet in the pancreas of 16 week old Mip-YY1 β KO mouse and control mice
It is fixed；It (D) is Mip-YY1 β KO mouse and the typical Electronic Speculum schematic diagram of control mice beta Cell of islet；It (E) is each of statistics Electronic Speculum
Percentage (the n of different types of insulin vesica (mature vesica, prematurity vesica, empty pocket bubble, rodlike vesica) number in section
=3)；
Fig. 7 is YY1 β KO mouse islets morphological analysis schematic diagram；Wherein, (A) 16 week old Mip-YY1 β KO mouse and right
According to mice pancreatic histotomy immunofluorescence dyeing, green indicates insulin, and red indicates Ki67, and blue indicates Dapi；(B) it unites
Count in (A) percentage shared by Ki67 positive cell, insulin positive cells >=3000/mouse in the cell of insulin positive
(n=3)；(C) the pancreas islet primary cell AnnexinV/PI bivariate stream that 16 week old Mip-YY1 β KO mouse and control mice are broken up
Formula cell instrument scatter plot；(D) ratio (n=3) (E) pancreatic tissue shared by apoptosis early stage cell (FITC+/PI-) in C is counted to cut
Piece TUNEL dyeing counts percentage shared by TUNEL positive cell, insulin positive cells > 8000 in insulin positive cells
A/mouse (n=3)；(F) 6 week old Rip-YY1 β KO mouse and control mice pancreatic tissue are sliced immunofluorescence dyeing, green table
Show insulin, red indicates glucagon, and blue indicates Dapi；(G) glucagon positive cell and pancreas islet in F figure are counted
The ratio of plain positive cell, insulin positive cells >=3000/mouse (n=3)；(H) 16 week old Mip-YY1 β KO are counted
The cell of glucagon positive cell and insulin positive in mouse and control mice pancreatic tissue slice immunofluorescence dyeing
Ratio, insulin positive cells >=3000/mouse (n=3)；(I) pancreas of 6 week old Rip-YY1 β KO mouse and control mice
Island β cell quality (n=3)；(J) the beta Cell of islet quality (n=4) of 16 week old Mip-YY1 β KO mouse and control mice；(K)
It counts every pancreatic tissue of 16 week old Mip-YY1 β KO mouse and control mice and is sliced upper pancreas islet number (n=4)；
Fig. 8 is to knock out YY1 in beta Cell of islet to can inhibit the transcription schematic diagram of insulin；Wherein, (A) Rip-YY1 β KO is small
The mRNA expression (n=4) of Ins1, Ins2 in the pancreas islet of mouse, (B) Mip-YY1 β KO mouse and corresponding control mice；
Fig. 9 is to knock out YY1 in beta Cell of islet to reduce cellular ATP content schematic diagram；Wherein, (A) Mip-YY1 β KO mouse
With ATP content (n=4) in the pancreas islet of control mice；(B) by the siRNA of YY1 (si-YY1) or control (si-NC) transfect into
Enter in MIN6 cell, the content (n=3) of ATP is detected after 48 hours；
Figure 10 is to knock out YY1 in beta Cell of islet to make cell mitochondrial that dysfunction schematic diagram occur；Wherein, (A) streaming
Cell instrument detects TMRE positive cell (n=3) in the pancreas islet primary cell of Mip-YY1 β KO mouse and control mice；(B)
Seahorse detects Mip-YY1 β KO mouse and the pancreas islet primary cell of control mice aoxidizes respiration capability curve (n=5:7)；
(C) the pancreas islet primary cell of Seahorse detection Mip-YY1 β KO mouse and control mice average oxygen consumption in high sugared low sugar
It measures (n=5:7)；
Figure 11 is the mitochondrial gene transcription schematic diagram of YY1 regulation core coding；Wherein, (A) ChIP analysis is by YY1's
SiRNA (si-YY1) or control (si-NC) transfection enter MIN6 cell in, after 48 hours transcriptional level detection Cox5a,
IDH3a and CytC promoter combines (n=3)；(B) the protein bound DNA of detected through gel electrophoresis YY1 (n=3)；(C) by YY1's
It is overexpressed plasmid or control plasmid transfection enters in MIN6 cell, the luciferase reporting of cromoci is detected after 48 hours
Gene activity (n=3)；(D) siRNA of YY1 (si-YY1) or control (si-NC) transfection are entered in MIN6 cell, 48 is small
When after detect cromoci luciferase reporter gene it is active (n=3)；(E) pancreas islet mitochondrial protein immunoblotting (n=
3)。
Specific embodiment
The following describes the present invention in detail with reference to examples.Following embodiment will be helpful to those skilled in the art
The present invention is further understood, but the invention is not limited in any way.It should be pointed out that those skilled in the art
For, without departing from the inventive concept of the premise, it can also make certain adjustments and improvements.These belong to guarantor of the invention
Protect range.
Embodiment 1: beta Cell of islet specific knockdown transcription factor YY1 mouse experiment
Experimental animal and raw material explanation:
SPF grades of YY1flox mouse buyings are in U.S. Jackson Lab, and line numbers are Jax No.014649, by Nanjing
Model animal research institute conservation.
SPF grades of Mip-Cre mouse are constructed by Ruijin Hospital, Shanghai Jiao Tong University School of Medicine doctor Cao Yanan.
SPF grades of Rip-Cre mouse are granted by French Universit é ClaudeBernardLyon professor Zhang Changxian.
SPF grades of C57BL/6J females and male mice, common mouse feed buying are limited in Shanghai Si Laike experimental animal
Responsible company；60kcal%Fat high lipid food (D12492) is purchased in Research Diets company of the U.S..
Beta Cell of islet specific knockdown transcription factor YY1 mouse knocks out referring to attached drawing 1, the tissue specificity of gene target
(Fig. 1-A) mates Rip-cre and Mip-cre mouse with YY1flox mouse respectively, and obtaining genotype after two generations is
YY1lox/lox Cre+Mouse, as YY1 β KO mouse, choosing brood genotype is YY1lox/lox Cre-Mouse be compare it is small
Mouse, genotype identification result is as shown if figure 1-b；Separating mouse pancreas islet extracts pancreas islet RNA and albumen, in transcription and translation respectively
The knockout efficiency of level identification YY1.It can be seen that Mip-YY1 β KO mouse knocks out efficiency in 40% or so (Fig. 1-C), and Rip-
YY1 β KO mouse knocks out efficiency and reaches 70% or so (Fig. 1-D), and the protein expression of YY1 also obviously lowers (Fig. 1-E), and two groups
Transgenic mice constructs success.
For beta Cell of islet specific knockdown transcription factor YY1 mouse glycometabolism phenotype referring to attached drawing 2 and 3, Mip-YY1 β KO is small
There is different degrees of hyperglycemia (Fig. 2-A, B, C, D) and impaired glucose tolerance (Fig. 3-in mouse and Rip-YY1 β KO mouse
E, F, G, H, I, J).Wherein, Rip-YY1 β KO mouse due to blood glucose it is excessively high, mouse survival rate is significantly affected (Fig. 3-M).Machine
There is the phenomenon that hyperglycemia and impaired glucose tolerance in body, on the one hand may be the missing or secreting function of beta Cell of islet quality
It is impaired, cause insulin level in serum deficiency to cause, on the other hand may be since the target organ of periphery insulin action is sent out
Insulin resistance has been given birth to, or has been caused by both sides reason collective effect.Mip-YY1 β KO mouse islets element tolerance test
(ITT) show compared with control mice, Mip-YY1 β KO mouse islets element sensibility do not occur and substantially change (Fig. 3-K and
L), illustrating that hyperglycemia and the phenotype of impaired glucose tolerance occurs in Mip-YY1 β KO mouse is drawn by peripheral insulin resistance
It rises, and being likely to pancreas islet form or functional defect causes hypoinsulinism to cause.
The beta Cell of islet specific knockdown YY1 mouse glycometabolism phenotype of High-fat diet referring to fig. 4, is fed giving HFD
When supporting the 16th week, the impaired glucose tolerance phenomenon of Mip-YY1 β KO mouse is obvious (Fig. 4-A, Fig. 4-B)；Insulin tolerance test
The result shows that Mip-YY1 β KO mouse at High-fat diet 16 weeks its periphery insulin sensitivity compared with control mice
Property there is no significant difference (Fig. 4-C, Fig. 4-D), the results showed that HFD nursing under Mip-YY1 β KO mouse impaired glucose tolerance
It still remains, main cause is not peripheral insulin resistance.
The insulin secretion of beta Cell of islet specific knockdown transcription factor YY1 mouse glucose stimulation is reduced referring to attached drawing
5, the insulin secretion test (IRT) of glucose stimulation finds 15 minutes and 30 points of YY1 β KO mouse after glucose stimulation
Clock, insulin level in serum are substantially less than control group, and wherein Rip-YY1 β KO mouse is almost without insulin secretion peak
Occur (Fig. 5-A, B, C, D)；Mouse islets are separated, have carried out the pancreas of glucose stimulation in primary pancreas islet level in vitro
Element secretion in island tests (GSIS) to assess the insulin secretion function of YY1 β KO mouse β cell, the results showed that in the pancreas with pancreas islet
After the correction of island cellulose content, under the conditions of low sugar (2.8mM) and height sugared (16.7mM), the insulin water of YY1 β KO mouse islets secretion
It is flat to be significantly lower than control group (Fig. 5-E and F).
Beta Cell of islet specific knockdown transcription factor YY1 mouse islets cellulose content is reduced referring to 6,16 week old Mip- of attached drawing
Insulin content obviously reduces (Fig. 6-C) than control group in YY1 β KO mouse primary pancreas islet, the pancreas of Mip-YY1 β KO mice pancreatic
Island cellulose content also decreases (Fig. 6-B) than control group.Mice pancreatic histotomy immunohistochemical staining the results show that
Insulin coloring in YY1 β KO mice pancreatic slice obviously shoals (Fig. 6-A)；Islet tissue is taken to carry out transmission electron microscope observing, it will
Insulin vesica carries out statistic of classification according to vesica form, and Mip-YY1 β KO mouse β koilocytosis steeps compared with the control group for discovery
The percentage that shape insulin vesica accounts for total vesica number dramatically increases, and the ratio of mature insulin vesica is declined (Fig. 6-E)；
Illustrate that insulin content in beta Cell of islet, insulin can be significantly reduced in specific knockdown transcription factor YY1 in beta Cell of islet
Vesica maturation is affected.
YY1 β KO mouse islets morphological analysis is referring to attached drawing 7, and compared with control mice, the Mip-YY1 β KO of 16 week old is small
The percentage of insulin positive cells shared by Ki67 positive cell is (Fig. 7-A and B) in a slight decrease in mouse pancreas slice.Apoptosis is early
A series of variations can occur for phase, cell membrane surface.Phosphatidylserine (PS) can be exposed to outside after birth out of cell membrane, and
Annexin V has the compatibility of height to PS, and the AnnexinV of FITC label can be used as detection and be exposed to cell membrane surface PS
Responsive probe.Apoptosis later period, cell membrane damage, and DNA can be contaminated by PI generates red fluorescence, normal cell and in withering
Die early stage cell will not dye PI and generate red fluorescent；The bivariate flow cytometer showed that flow cytometry analysis generates dissipates
(Fig. 7-C) does cross door on point diagram, and left lower quadrant shows the living cells of FITC-/PI-；Right upper quadrant is the non-live of FITC+/PI+
Cell, i.e. non-viable non-apoptotic cell；And right lower quadrant is the apoptotic cell of FITC+/PI-.Fig. 7-D display, the Mip-YY1 β KO of 16 week old
The cell proportion that mouse islets are in apoptosis early stage is relatively higher than control group.
Pancreas is sliced immunofluorescence dyeing TUNEL experiment detection β Apoptosis, shows the Mip-YY1 β KO mouse of 16 week old
Permillage shared by TUNEL positive cell is significantly higher than control group (Fig. 7-E) in insulin positive cells；Pancreas is sliced and is carried out
Insulin (Insulin) and glucagon (Glucagon) dyeing (Fig. 7-F), find in Mip-YY1 β KO mouse islets
The ratio of Glucagon positive cell and Insulin positive cell is risen (Fig. 7-H), in Rip-YY1 β KO mouse islets
In, this ratio has significant difference (7-G) compared with the control group.In addition, the beta Cell of islet quality to two groups of mouse carries out
Statistics finds to have dropped half (Fig. 7-I) in the Mip-YY1 β KO mouse islets β cell quality of 16 week old than control group, and 6
The β cell quality of the Rip-YY1 β KO mouse islets of week old has dropped about 2/3rds (Fig. 7-J) compared with the control group, between two groups
There is apparent difference.The experimental results showed that, the specific knockdown YY1 in beta Cell of islet, Apoptosis increases, β cell ratio above
Example is reduced, and causes the Mass lost of beta Cell of islet, and pancreatic beta cell mass and pancreatic insulin content are closely related, pancreas
The reduction of insulin content may be caused by being reduced due to pancreatic beta cell mass.
Embodiment 2: transcription factor YY1 regulates and controls the verifying of beta Cell of islet by influencing mitochondrial
Referring to attached drawing 8, in Rip-YY1 β KO and Mip-YY1 β KO mouse islets, two transcripts of insulin gene
The mRNA level in-site of Ins1 and Ins2 is substantially reduced (Fig. 8-A and B) compared to control group.
The ATP content that beta Cell of islet specific knockdown YY1 reduces pancreas islet is small referring to the Mip-YY1 β KO of 9,16 week old of attached drawing
Compared with the control group, the content of ATP is greatly reduced the content of ATP in pancreas islet in mouse pancreas islet, illustrates after YY1 is knocked out, pancreas islet β
Cellular ATP content decline, cell be in " starvation " state, this may be the reason of leading to the reduction of β cell insulin secretion,
After YY1 gene 48h is fallen in the interference of MIN6 cell-specific, the ATP level of si-YY1 group is significantly reduced compared to si-NC group.
Beta Cell of islet specific knockdown YY1 reduces pancreas islet mitochondria referring to attached drawing 10: separating mouse pancreas islet, insulating box are incubated
Acquisition primary cell is broken up after educating, and is stimulated pancreas islet primary cell using the glucose of 2.8mM and 16.7mM respectively, is carried out simultaneously
TMRE dyeing, uses the TMRE tinctorial strength and counting of flow cytomery individual cells after 30 minutes.TMRE is a kind of band
There is the dyestuff of the fluorescent red-orange of positive charge, the selective mitochondrial inner membrane for being positioned at living cells can be used for mitochondrial membrane electricity
The detection of potential difference.Mitochondria activity is stronger, and proton-pump is stronger, and film two sides potential difference is just more obvious, and TMRE coloring is more
Obviously.The results show that under the stimulation of 16.7mM glucose there is more the fluorescence intensity cell peak of TMRE in control group mice pancreas islet
It significantly moving to right, illustrates after high sugar stimulation, the mitochondria activity in beta Cell of islet increases, and in Mip-YY1 β KO group mouse
In pancreas islet, the phenomenon that TMRE positive cell peak moves to right (Figure 10-A) is not observed.
Using Seahorse XF24 mitochondrial oxidation respiratory function analysis system, it is small to detect 16 week old Mip-YY1 β KO groups
The mitochondria consumption rate of mouse and control group mice pancreas islet.After XF24 test board aquation, balance, two groups of mouse islets are tested respectively
3mM glucose (basic condition), 20mM glucose (high sugar stimulation) and successively be added oligomycin (oligomycin),
Cell consumption rate (OCR) (Figure 10-B) when FCCP, antimycin A (antimycin A) and rotenone (Rotenone).Oligomycin
It is ATP synthetase inhibitors, OCR declines after this medicine is added, and the oxygen demand of reduction represents body for synthesizing the oxygen consumption of ATP
Amount, shows the ATP yield of cell indirectly.Uncoupler FCCP can make proton flow back, largely as proton carrier
Oxygen consumption, OCR rises at this time, but cannot generate ATP after the reflux of these protons, but water is generated in conjunction with oxygen, and FCCP is added
The increase of oxygen consumption afterwards represents the maximum oxygen-consuming capacity of mitochondria, shows maximum breathing ability indirectly.Being eventually adding is antimycin
A and rotenone, both this is respiratory chain inhibitor, can prevent completely mitochondria oxygen consumption, the results showed that, with control group mice pancreas islet
Compare, the Baseline oxygen consumption of Mip-YY1 β KO group mouse islets, oxygen demand (figure, 10-C) after high sugar stimulation, ATP generate ability and
Maximum breathing ability decreases, and this further illustrates cell Mitochondria functions after beta Cell of islet specific knockdown YY1
Weaken.
Transcription factor YY1 regulates and controls the mitochondrial gene expression of core coding referring to attached drawing 11, interferes in MIN6 cell-specific
After falling YY1 gene 48h, Cox5a, IDH3a and CytC promoter region gene that si-YY1 group combines compared with si-NC group obviously subtract
Few (Figure 11 A, B).Cromoci participates in the electron transmission of mitochondria, finds the transcriptional level of cromoci obviously by YY1's
Regulation, when MIN6 cell-specific is overexpressed YY1, the transcriptional activity of cromoci is remarkably reinforced (Figure 11-C), and
After YY1 gene is fallen in the interference of MIN6 cell-specific, the transcriptional activity of cromoci is substantially reduced (Figure 11-D).Separating mouse pancreas
Island carries out immunoblot experiment as shown in Figure 11-E, represents in Mip-YY1 β KO group mouse islets Mitochondria electron transport chain complex
Property protein expression level reduce.
The above is presently preferred embodiments of the present invention, but the present invention should not be limited to disclosed in the embodiment
Content.So all do not depart from the lower equivalent or modification completed of spirit disclosed in this invention, the model that the present invention protects is both fallen within
It encloses.
Claims (8)
1. purposes of the transcription factor YY1 in preparation treatment diabetes targeted drug.
2. purposes as described in claim 1, which is characterized in that transcription factor YY1 is real by the development of regulation beta Cell of islet
It is existing.
3. purposes of the transcription factor YY1 in the drug of preparation regulation beta Cell of islet development.
4. purposes as claimed in claim 3, which is characterized in that the chondriogen that transcription factor YY1 passes through regulation core coding
Expression is realized.
5. purposes as claimed in claim 4, which is characterized in that the chondriogen of the core coding is cromoci oxidation
5 α of enzyme subunit, isocitric dehydrogenase subunit α and cromoci.
6. purposes as claimed in claim 3, which is characterized in that the drug further includes using transcription factor YY1 as active constituent
Pharmaceutically acceptable auxiliary material or complementary ingredient.
7. purposes as claimed in claim 6, which is characterized in that the mass percentage content of the medicament transfer record factor YY1
It is >=98%, application dose is 1~5000mg/kg.
8. the purposes as described in claim 3 or 7, which is characterized in that the pharmaceutical dosage form is tablet, pulvis, granule, needle
Agent, capsule, oral solution or sustained release agent.
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| CN201811473083.0A CN109289039A (en) | 2018-12-04 | 2018-12-04 | Purposes of the transcription factor YY1 in preparation treatment diabetes targeted drug |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070141072A1 (en) * | 2002-12-20 | 2007-06-21 | Ernst-Moritz-Arndt-Universitat Greifswald | Use of multifunctional transcription factor yin-yang-1 and variants thereof for treating illnesses, especially type 1 diabetes |
| CN105031619A (en) * | 2015-08-13 | 2015-11-11 | 上海交通大学医学院附属瑞金医院 | Application of excreted factor GREM2 to 2-type diabetic therapy drug preparation |
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2018
- 2018-12-04 CN CN201811473083.0A patent/CN109289039A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070141072A1 (en) * | 2002-12-20 | 2007-06-21 | Ernst-Moritz-Arndt-Universitat Greifswald | Use of multifunctional transcription factor yin-yang-1 and variants thereof for treating illnesses, especially type 1 diabetes |
| CN105031619A (en) * | 2015-08-13 | 2015-11-11 | 上海交通大学医学院附属瑞金医院 | Application of excreted factor GREM2 to 2-type diabetic therapy drug preparation |
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| Title |
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| DALONG SONG等: "YY1 deficiency in β-cells leads tomitochondrial dysfunction and diabetes in mice", 《METABOLISM CLINICAL AND EXPERIMENTAL》 * |
| FRANCISCO VERDEGUER等: "Decreased Genetic Dosage of Hepatic Yin Yang 1 Causes Diabetic-Like Symptoms", 《MOLECULAR ENDOCRINOLOGY》 * |
| SHARON M. BLÄTTLER等: "Yin Yang 1 deficiency in skeletal muscle protects against rapamycin-induced diabetic-like symptoms through activation of insulin/IGF signaling", 《CELL METAB》 * |
| 张伟等: "转录因子YY1的作用机制", 《生命的化学》 * |
| 王淑秋等: "《病理生理学》", 31 October 2010, 世界图书出版社西安公司 * |
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