CN103394073A - Application of perilipin in preparation of medicine - Google Patents
Application of perilipin in preparation of medicine Download PDFInfo
- Publication number
- CN103394073A CN103394073A CN2013103003083A CN201310300308A CN103394073A CN 103394073 A CN103394073 A CN 103394073A CN 2013103003083 A CN2013103003083 A CN 2013103003083A CN 201310300308 A CN201310300308 A CN 201310300308A CN 103394073 A CN103394073 A CN 103394073A
- Authority
- CN
- China
- Prior art keywords
- perilipin
- autophagy
- fat
- abt737
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the field of preparation of medicines and particularly relates to an application of perilipin in preparation of a medicine for inhibiting fat cell autophagy and treating lipid metabolism disorder. According to the study, berberine and Perilipin with inhibiting action on the fat cell autophagy are used for detection and a result shows that the autophagy promotes degradation of Perilipin; an autophagy agonist ABT737 is adopted for observing the degradation of the Perilipin, and after autophagy is activated by ABT737, the expression of the Perilipin is obviously reduced; and after treatment of the berberine, compared with the individual ABT737 group, a common treatment group of the berberine and the ABT737 has the advantage that the expression of the Perilipin is increased. in conclusion, the Perilipin has an inhibiting action on the fat cell autophagy, the reduction of the content of the Perilipin promotes lipolysis and causes disorder of energy metabolism of the whole organism; therefore, a new research direction is provided for treating the lipid metabolism disorder.
Description
Technical field
The invention belongs to medical preparation field, particularly perilipin suppresses the application in adipose cell autophagy medicine and treatment lipid metabolic disorder medicine in preparation.
Background technology
The structure that the cell lactone drips forms by triglyceride and a small amount of cholesteryl ester the core that neutral fat drips, and outside is covered with monolayer phospholipid, a large amount of protein localizations is arranged simultaneously in the monolayer phospholipid surface that fat drips, and is called fat and drips surface protein.For adipose cell, it is topmost part in its born of the same parents that fat drips, and from the typical adipose cell that is separated to the white adipose tissue, contains a large fat and drips, more than diameter can reach 100 μ m, occupy most of space in cell, be called single chamber fat and drip (unilocular lipid droplets); From the adipose cell that separates brown adipose tissue, contain the much little fat that is dispersed in and drip, be called multi-cavity fat and drip (multilocular lipid droplets).Increasing research shows, it is active, the highly dynamic organelle of a metabolism that the adipose cell lactone drips, be the core point of regulation and control body lipid balance and the energy balance, and fat drip surface protein played important function in formation, fusion and metabolism that fat drips is regulated.At fat, drip in surface protein, most importantly fat drips structural protein PAT family protein, comprising Perilipin, and ADRP, TIP47, S3-12 etc.
Perilipin (Perilipin) is that fat drips one of core member of protein family, is to be positioned the albumen that fat drips the high phosphatization on surface.When the body storage power is deficient in order to nutrient substance, utilize, unnecessary energy is stored in during fat drips with the form of triglyceride in the fat stores process.When the body requirement energy, the triglyceride of storing is hydrolyzed by the activation of steatolysis approach, and lipid stores and the coordination of utilization is to be subjected to fat to drip the regulation and control of surface protein perilipin family.
Recent study is found, autophagy is playing an important role aspect the lipid metabolism of liver and fatty tissue, autophagy is one and engulfs self cytoplasm protein or organelle and make its coated vesicle that enters, and merge and form the autophagy lysosome with lysosome, the degrade process of its content that wraps up, the metabolism that realizes by this cell itself need and the renewal of some organelle.So the autophagy phenomenon provides new thinking for the treatment of obesity.Under the normal growth condition, nutrient substance is abundant, autophagocytosis remains on a foundation level, be mainly used in house keeper's purpose, it for cell provide a kind of routine-the garbage-cleaning service, as the degraded of long-lived protein and damaged cell device, such continuous renewal is particularly important to the cell of static termination differentiation; Under the condition of hunger, autophagy is further induced, and for cells survival provides extra inside nutrition, autophagy can be by energy stress or other stress-induceds widely, great majority are the synthetic re-using aminoacid for important albumen, or provide catabolic substrate for the generation of ATP.
Perilipin expresses with the research of the relation of human obesity less, and due to the Body Mass Index span difference of object of study, sample size is less, the equal factors in the institute fatty tissue position of getting, and acquired results is not quite identical.And the relevant report of the correlation research of Perilipin and autophagy did not occur especially.
Summary of the invention
The purpose of this invention is to provide perilipin and suppress the application in adipose cell autophagy medicine in preparation.
Another object of the present invention is to provide the application of perilipin in preparation treatment lipid metabolic disorder medicine.
Autophagy (autophagy) is the distinctive biosis of eukaryotic cell, and cell, by the autophagosome parcel self impaired or aging organelle and the macromole that form duplicature, then is combined with lysosome, realizes the degraded to content.
Under base state, Perilipin is coated on the cell lactone and drips surface, forms one molecule barrier and stops that lipase contact fat drips and suppresses steatolysis, makes cell remain lower steatolysis state; Yet when hormonal stimulation, Perilipin is by the PKA phosphorylation, and this barrier is modified and certain variation is occurred, thereby promotes the HSL transposition and activate fatty tissue triglyceride enzyme (ATGL), decomposes triacylglycerol.Thereby Perilipin is the last one molecule gating device that starts the steatolysis reaction.Simultaneously, adipose cell is in the situation that Catecholamine matter continues to stimulate, and the cell lactone drips form to be reinvented, and large fat drips minimizing, and the quantity that little fat drips sharply increases, and this phenomenon is also referred to as cracked that fat drips.The cracked fat that can increase that drips due to fat drips surface area, so be considered to have all the time, promotes fat-splitting effect.Fat drips reinvents is the complex process that the various structures such as cytoskeleton participates in, and whether the steatolysis of Catecholamine matter mediation and fat exist inevitable contact between dripping and reinventing, and this is known little about it now.Only one piece of report discovery, in six phosphorylation sites of known Perilipin, what the phosphorylation in serine 492 sites participation fat dripped reinvents, and this site mutation can stop forskolin and IBMX to stimulate the fat that causes to drip cracked.In fruit bat, the Perilipin mutant can make the body lactone drip from multi-cavity fat to drip and change large single chamber fat into and drip, and causes the storage of lipid and mobilize unbalance.Think at present take Perilipin as main PAT family protein may fat drip synthetic fast, it is stable that newborn fat drips, fat drips the adjusting of maturation process, and fat drops in intracellular transportation and plays a role.
Our hypothesis autophagy under base state stimulates steatolysis by the degraded that promotes Perilipin, so adopt, the inhibited berberine of adipose cell autophagy and Perilipin are detected, our result is presented in the situation that autophagy suppressed by berberine, the increasing expression of Perilipin, this shows that autophagy promotes the degraded of Perilipin, that is to say that lysosomal pathway plays an important role in the degradation process of adipose cell Perilipin.
We have further adopted autophagy agonist ABT737 to observe the degraded of Perilipin, we find on the 3T3-L1 mature fat cell, after ABT737 activates autophagy, the expression of Perilipin obviously reduces, the release showed increased of glycerol, and namely the steatolysis of base state increases, and after berberine is processed, we find to compare with independent ABT737 processed group, the Perilipin increasing expression of berberine and ABT737 co-treatment group, and the release of glycerol reduces.
research shows: berberine has inhibitory action to the adipose cell autophagy, and the Perilipin of berberine group expresses significantly rising, illustrate that berberine is by the Perilipin effect, Perilipin has inhibitory action to the adipose cell autophagy, Perilipin plays barrier action between neutral fat and lipase, protection fat exempts from the attack of lipase, adiposis patient and rodent, the minimizing of Perilipin content promotes steatolysis to increase, and then cause the disorder of whole human body energy metabolism, this provides a kind of new method for preventing obesity and lipid metabolic disorder treatment.
The present invention has following beneficial effect:
1, show by experiment: Perilipin has inhibitory action to the adipose cell autophagy; Perilipin plays barrier action between neutral fat and lipase; protection fat exempts from the attack of lipase; adiposis patient and rodent; the minimizing of Perilipin content promotes steatolysis to increase; and then causing the disorder of whole human body energy metabolism, this provides a kind of new method for preventing obesity and relevant metabolism syndrome thereof.2, for clinically more reasonably by Perilipin for suppressing adipose cell autophagy medicine and the lipid metabolic disorder medicinal application provides experimental data and theoretical foundation.
The accompanying drawing explanation
Fig. 1 is after berberine is processed rear 3T3-L1 adipose cell, Perilipin protein expression figure.
Fig. 2 is Perilipin protein expression comparison diagram.
Fig. 3 is that glycerol discharges comparison diagram.
The specific embodiment
Below in conjunction with embodiment, the invention will be further described:
The mice 3T3-L1 fibroblast (front adipose cell) of using in experiment is purchased from U.S. ATCC (American Type Culture Collection).Dexamethasone (Dexamethasone, DEX), 3-isobutyl-1-methylxanthine (3-isobutyl-1-methylxanthine, IBMX), dimethyl sulfoxide (Dimethyl sulfoxide, DMSO) are purchased from U.S. Sigma company; (HumR, INS) purchased from Lilly Co., Eli. for the recombinant human insulin; Bovine serum albumin (Bovine serum albumin, BSA) is purchased from German Roche company; Hyclone (fetal calf serum, FCS) is purchased from Austrian PAA company; The 11965DMEM culture medium is purchased from U.S. Gibco BRL company; ABT737 is purchased from Selleck Chemicals LLC; The triglyceride determination test kit is purchased from Shanghai Kehua Bio-engineering Co., Ltd, and 21063DMEM Never Guess(is without phenol red) culture fluid is purchased from Gibco BRL company, and Perilipin antibody is purchased from Cell Signaling Technology.
The culture fluid of derivant A: volume fraction 10%FCS-DMEM, also contain IBMX0.5mmol/L, dexamethasone 1 μ mol/L, insulin 1.67 μ mol/L.
The culture fluid of derivant B: volume fraction 10%FCS-DMEM, also contain insulin 1.67 μ mol/L.
Embodiment
1. cell culture and induce differentiation
1), before 3T3-L1 the cultivation of adipose cell strain with go down to posterity
The 3T3-L1 fibroblast is incubated in the sugared 10%FCS-DMEM solution of normal height, is placed in 37 ℃, 5%CO
2Incubator.Treat that under microscope, observation of cell is adherent, be fusiformis, bright, change culture fluid and go down to posterity when cell to 90% merges.
By culture fluid from sucking-off culture bottle, add 0.25% pancreatin 4mL to digest, under microscope, visible cell is shrunk to circle by irregular polygon or fusiformis, this process approximately needs 2min to add normal culture fluid to stop the reaction of pancreatin, with pipet, repeatedly blow and beat cell residual on bottle wall, make cell detachment culture bottle wall, suck and be equipped with in the centrifuge tube of culture fluid, the centrifugal 3min of 1000rpm, sedimentation cell.After centrifugal, supernatant discarded, then add normal culture fluid, with pipet piping and druming, cell is scatter.According to inoculum density, getting the celliferous culture fluid of part is inoculated in 12 orifice plates or 6 orifice plates, in 5%CO
2Incubator in 37 ℃ of cultivations.
2), before 3T3-L1 the adipose cell strain induce differentiation
After passage, change every other day normal culture fluid (volume fraction 90%DMEM+10% hyclone), continue to cultivate 48h.After cell fusion, add induced liquid A to start to induce (the 0th day), cultivate 48h.
The 2nd day, remove induced liquid A, change with induced liquid B, then cultivate 48h.
The 4th day, remove induced liquid B, all to change with the sugared 10%FCS-DMEM culture fluid of normal height later, this moment, visible a few cell form was rounded, and there have a small amount of fat to ooze to be existing.
Change every other day culture fluid one time, the 8th day visible 90% above cell is rounded and occur that a large amount of fat drips later, namely represent 3T3-L1 before adipose cell induced successfully, become ripe adipose cell.
The adipose cell of inducing is hatched to 12h with 0.2%BSA-DMEM, can carry out subsequent experimental.
2, the impact of Perilipin on the 3T3-L1 mature fat cell
in step (1), be induced to differentiate in the front adipose cell of 3T3-L1 of mature fat cell, berberine (the 0 μ mol/L that adds variable concentrations, 0.2 μ mol/L, 1 μ mol/L, 5 μ mol/L) effect 12h, protein immunoblot detects (Western Blot), specifically as shown in Figure 1: autophagy marker protein LC3II/I reduces along with increasing of berberine concentration, illustrate that berberine has inhibitory action to cell autophagy, Perilipin increases along with increasing of berberine concentration, be dose dependent, wherein the Perilipin of 5 μ mol/L groups significantly raises, illustrate that 3T3-L1 adipose cell autophagy and Perilipin have dependency, autophagy promotes the degraded of Perilipin, that is to say that lysosomal pathway plays an important role in the degradation process of adipose cell Perilipin.
3, utilize autophagy agonist ABT737, observing Perilipin affects the 3T3-L1 mature fat cell
In step (1), be induced to differentiate in the front adipose cell of 3T3-L1 of mature fat cell, add respectively berberine (BBR), ABT737, after ABT737+BBR effect 12h, Western Blot result such as Fig. 2 show: (1) is compared with matched group (Control group), and the Perilipin of BBR group expresses and raises; ABT737 group Perilipin expresses significantly and reduces, and does not almost detect; (2) with the ABT737 group, compare, ABT737+BBR group Perilipin expresses significantly and raises.
In step (1), be induced to differentiate into mature fat cell 3T3-L1, with 0.2%BSA, process cell 12h, then change into without phenol red culture fluid, with BBR, ABT737, BBR+ABT737, process 12h.Get cell conditioned medium liquid, with the Triglyceride Reagent box, measure glycerol content in supernatant, result such as Fig. 3 show: (1) is compared with the Control group, and BBR group glycerol burst size significantly reduces; ABT737 group glycerol burst size significantly raises; (2) with the ABT737 group, compare, ABT737+BBR group glycerol burst size obviously reduces.(in Fig. 3, * represents P<0.05; * represents P<0.01; * * represents P<0.001).
Testing result explanation: on the 3T3-L1 mature fat cell, after ABT737 activates autophagy, the expression of Perilipin obviously reduces, the release showed increased of glycerol, the steatolysis that is base state increases, and after berberine is processed, we find to compare with independent ABT737 processed group, the Perilipin increasing expression of berberine and ABT737 co-treatment group, the release of glycerol reduces, and this explanation berberine suppresses increasing expression and the fat-splitting minimizing that autophagy is accompanied by Perilipin.
in sum, berberine has inhibitory action to the adipose cell autophagy, and the Perilipin of berberine group expresses significantly rising, illustrate that berberine is by the Perilipin effect, Perilipin has inhibitory action to the adipose cell autophagy, Perilipin plays barrier action between neutral fat and lipase, protection fat exempts from the attack of lipase, adiposis patient and rodent, the minimizing of Perilipin content promotes steatolysis to increase, and then cause the disorder of whole human body energy metabolism, this provides a kind of new method for preventing obesity and relevant metabolism syndrome thereof.
Claims (2)
1. perilipin suppresses the application in adipose cell autophagy medicine in preparation.
2. the application of perilipin in preparation treatment lipid metabolic disorder medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013103003083A CN103394073A (en) | 2013-07-17 | 2013-07-17 | Application of perilipin in preparation of medicine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013103003083A CN103394073A (en) | 2013-07-17 | 2013-07-17 | Application of perilipin in preparation of medicine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103394073A true CN103394073A (en) | 2013-11-20 |
Family
ID=49557926
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2013103003083A Pending CN103394073A (en) | 2013-07-17 | 2013-07-17 | Application of perilipin in preparation of medicine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103394073A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111057136A (en) * | 2019-12-30 | 2020-04-24 | 中国科学院生物物理研究所 | Fat body and application thereof in evaluating interaction between protein to be detected and fat droplets |
-
2013
- 2013-07-17 CN CN2013103003083A patent/CN103394073A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 张利红等: "一种调控脂解的重要蛋白-围脂滴蛋白（perilipin)", 《中国生物化学与分子生物学报》 * |
| 王李洁: "自噬在肥胖中的研究进展", 《中国病理生理杂志》 * |
| 邓其兰等: "围脂滴蛋白（perilipin)在纸质代谢中的功能", 《饲料工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111057136A (en) * | 2019-12-30 | 2020-04-24 | 中国科学院生物物理研究所 | Fat body and application thereof in evaluating interaction between protein to be detected and fat droplets |
| CN111057136B (en) * | 2019-12-30 | 2021-09-28 | 中国科学院生物物理研究所 | Fat body and application thereof in evaluating interaction between protein to be detected and fat droplets |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Aberrant hepatic expression of PPARγ2 stimulates hepatic lipogenesis in a mouse model of obesity, insulin resistance, dyslipidemia, and hepatic steatosis | |
| Christian et al. | Autophagy: Emerging roles in lipid homeostasis and metabolic control | |
| Li et al. | MicroRNA-328 as a regulator of cardiac hypertrophy | |
| Jaworski et al. | Regulation of triglyceride metabolism. IV. Hormonal regulation of lipolysis in adipose tissue | |
| Sun et al. | Aldehyde dehydrogenase 2 ameliorates doxorubicin-induced myocardial dysfunction through detoxification of 4-HNE and suppression of autophagy | |
| Watt et al. | Triglyceride metabolism in exercising muscle | |
| Meng et al. | Regulatory T cells prevent plaque disruption in apolipoprotein E-knockout mice | |
| Sun et al. | High free fatty acids level related with cardiac dysfunction in obese rats | |
| Fang et al. | Ceramide and sphingosine 1-phosphate in adipose dysfunction | |
| Poulsen et al. | Identification of inflammatory and proresolving lipid mediators in bone marrow and their lipidomic profiles with ovariectomy and omega‐3 intake | |
| Liu et al. | Oleanolic acid diminishes liquid fructose‐induced fatty liver in rats: role of modulation of hepatic sterol regulatory element‐binding protein‐1c‐mediated expression of genes responsible for de novo fatty acid synthesis | |
| Tian et al. | Electroacupuncture at ST‐36 protects interstitial cells of Cajal via sustaining heme oxygenase‐1 positive M2 macrophages in the stomach of diabetic mice | |
| Ishihata et al. | Vascular-and hepato-protective effects of passion fruit seed extract containing piceatannol in chronic high-fat diet-fed rats | |
| Wu et al. | Red yeast rice prevents atherosclerosis through regulating inflammatory signaling pathways | |
| Lu et al. | Pyridostigmine protects against cardiomyopathy associated with adipose tissue browning and improvement of vagal activity in high-fat diet rats | |
| Xie et al. | Zeaxanthin ameliorates obesity by activating the β3-adrenergic receptor to stimulate inguinal fat thermogenesis and modulating the gut microbiota | |
| Park et al. | Enantiomers of higenamine inhibit LPS-induced iNOS in a macrophage cell line and improve the survival of mice with experimental endotoxemia | |
| Wuchert et al. | Tumor necrosis factor-α, interleukin-1β and nitric oxide induce calcium transients in distinct populations of cells cultured from the rat area postrema | |
| US20240041813A1 (en) | Very-long-chain polyunsaturated fatty acids, elovanoid hydroxylated derivatives, and methods of use | |
| CN103394073A (en) | Application of perilipin in preparation of medicine | |
| Müller | Take-over: multiple mechanisms of inter-adipocyte communication | |
| Kajimoto et al. | Liver-Specific silencing of Lipin1 reduces fat mass as well as hepatic triglyceride biosynthesis in mice | |
| US20220339134A1 (en) | Very-long-chain polyunsaturated fatty acids, elovanoid hydroxylated derivatives, and methods of use | |
| El-Sayyad et al. | Effects of diabetes and/or hypercholesterolemia on skin development of rat fetuses | |
| Ogasawara et al. | Oligonol-induced degradation of perilipin 1 is regulated through lysosomal degradation machinery |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20131120 |