Kisspeptin-10 - GPR54 Ligand - MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEKisspeptin-10Cat. No.: HY-P0254CAS No.: 374675-21-5分式: CHNO分量: 1302.44Sequence: Tyr-Asn-Trp-Asn-Ser-Phe-Gly-Leu-Arg-Phe-NH2Sequence Shortening: YNWNSFGLRF-NH2作靶点: Others作通路: Others储存式: Please store the product under the recommen

2、ded conditions inthe COA.BIOLOGICAL ACTIVITY物活性 Kisspeptin-10种有效的内 性 GPR54 激动剂，作管收缩剂和管成抑制剂。IC50 & Target GPR54 1Angiogenesis 1体外研究 Kisspeptin-10 (KP-10) significantly increases adhesion of THP-1 cells to HUVECs by 10-fold at 10 M.Pretreatment with the GPR54 antagonist, P234 (20 M), significantly inh

3、ibits Kisspeptin-10 (10 M)-inducedadhesion of THP-1 cells to HUVECs. These results indicate that Kisspeptin-10 increases the adhesion ofTHP-1 cells to HUVECs by GPR54. Kisspeptin-10 markedly enhances mRNA expression for TNF-, IL-6,MCP-1, ICAM-1, VCAM-1, and E-selectin in HUVECs. Kisspeptin-10 signif

4、icantly increases the proteinexpression of ICAM-1 and VCAM-1 in HUVECs in parallel with the increases in mRNA levels 1.体内研究The effects of Kisspeptin-10 (KP-10) and the GPR54 antagonist P234 are evaluated on the development ofatherosclerotic lesions in 2 different strains of ApoE-/- mice (C57/B6 and

5、BALB/c). In ApoE-/- mice (C57/B6),the (entire) surface and cross-sectional area of the root (plaque size) of aortic atherosclerotic lesions withpentraxin-3-positive area, monocyte/macrophage infiltration, and VSMC content as well as plasma totalcholesterol concentration are significantly increased a

6、t 17 weeks of age compared with 13 weeks of age. By17 weeks of age, plasma Kisspeptin-10 concentration is significantly elevated in mice infused with a highdose of Kisspeptin-10 (12.5 g/kg per hour) compared with the vehicle control. A high dose of Kisspeptin-10(12.5 g/kg per hour) significantly enh

7、ances the aortic atherosclerotic lesion area and atheromatous plaque1/3 Master of Small Molecules 您边的抑制剂师www.MedChemEsize, with significant increases in pentraxin-3-positive area and monocyte/macrophage infiltration and asignificant decrease in VSMC content. There are no significant differences in b

8、ody weight, food intake,systolic and diastolic BP, and plasma concentrations of either total cholesterol or glucose among the 3groups of ApoE-/- mice at 17 weeks of age 1. The injection of Kisspeptin-10 can slow down microvascularcutaneous blood flow in mice. The changes in cardiac metabolites in ra

9、ts treated with Kisspeptin-10 isinvestigated using a metabonomics approarch based on GC/TOF-MS to the rats. The identification ofmetabolic pathways and biomarkers may contribute to understanding the mechanism by which Kisspeptin-10treatment alters cardiac functions. Mitochondria in which energy meta

10、bolism occurs is observed throughtransmission electron microscope (TEM), and the perturbations in energy metabolism can be inferred fromchanges in mitochondrial structure. There is a clear separation between the control (N) and Kisspeptin-10 (K)groups in the plot regarding serum samples 2.PROTOCOLCe

11、ll Assay 1 HUVECs, HUVEC-derived EA.hy926 cells, or HASMCs at passage 2 to 8 are seeded into 96-well plates(1104 cells/100 L/well) and incubated for 24 hours in DMEM or SmGM-2 containing 10% or 5% FBS,respectively. Cells are then incubated for a further 48 hours with the indicated concentrations of

12、Kisspeptin-10 in fresh media. Ten microliters of WST-8 solution are then added to each well. After 1 hour of incubation,the quantity of formazan product is determined by reading absorbance at 450 nm using a Sunrise Remote R-micro plate reader 1.MCE has not independently confirmed the accuracy of the

13、se methods. They are for reference only.Animal Mice 1Administration 12 A total of 66 of male spontaneously hyperlipidemic ApoE-/- mice in 2 strains, C57/B6 (KOR/StmSlc-Apoeshlmice) and BALB/c (KOR/StmSlc-Apoeshl mice), are used. Mice are fed a high-cholesterol diet containing16.5% fat, 1.25% cholest

14、erol, and 0.5% sodium cholate, starting at 13 weeks of age. Mice at 13 weeks ofage are infused for 4 weeks with Kisspeptin-10 and/or P234, a GPR54 antagonist, using osmotic minipumps.The time course and dose of Kisspeptin-10 and P234 infusion are decided. Experiment 1 is performed toevaluate the dos

15、e-dependent effects of Kisspeptin-10 on atherogenesis in 17 ApoE-/- mice (C57BL/6). At 13weeks of age, 3 mice are euthanized as preinfusion controls. The remaining 14 are divided into 3 groups of5, 4, and 5 and then infused with saline (vehicle) or Kisspeptin-10 at doses of 5 and 12.5 g/kg per hour,

16、respectively. Experiment 2 is performed to evaluate the suppressive effects of P234 on Kisspeptin-10-induced atherogenesis in 38 ApoE-/- mice (BALB/c). At 13 weeks of age, 7 mice are euthanized as a controlbefore infusion. The remaining 31 are divided into 3 groups of 10, 10, and 11 and are then inf

17、used withsaline, Kisspeptin-10 (12.5 g/kg per hour) or Kisspeptin-10 (12.5 g/kg per hour)+P234 (50 g/kg per hour),respectively. Experiment 3 is performed to evaluate the preventive effects of P234 on endogenousKisspeptin-10-induced atherogenesis (the natural course of atherogenesis) in 10 ApoE-/- mi

18、ce (BALB/c).From 13 to 17 weeks of age, 5 and 5 mice are infused with saline and P234 (50 g/kg per hour), respectively1.Rats 2Twenty eight-week-old male Sprague-Dawley rats weighing 20020 g are used. They are maintained at constant temperature (232C) and humidity (4515%), with controlled lighting (l

19、ight 12 h-dark 12 h), and arefed at a standard diet, with free access to tap water. They are adapted to the environment for two weeks to2/3 Master of Small Molecules 您边的抑制剂师www.MedChemEovercome the stress of transportation before the animal experiments began. Then, all of the rats are dividedinto tw

20、o groups randomly. Each rat in Kisspeptin-10 group receives subcutaneous injection with 200 LKisspeptin-10 (40 nmol/200L) every day, each rat of the control group receives subcutaneous injection with200 L saline every day, both of the two groups are treated for 7 days continuously. Rats are anesthet

21、izedwith urethane (1.5 g/kg, i.p.). Under general anesthesia, blood samples are collected and rats areeuthanized. All efforts are made to minimize the discomfort and stress of animals. Then all the samples areanalyzed. Seven days later, the 20 rats are decapitated after exposure to anesthesia. Cardiac tissue andserum samples are collected and stored at -80C until being used for total RNA and protein extraction as wellas metabonomics analysis. The rest of the heart tissue is fixed with 4% paraformaldehyde and