Phorbol 12-Myristate 13-Acetate；佛波酯
| MDL | MFCD00036736 |
| EC | EINECS 605-413-5 |
| InChIKey | PHEDXBVPIONUQT-RGYGYFBISA-N |
| InChI | InChI=1S/C36H56O8/c1-7-8-9-10-11-12-13-14-15-16-17-18-29(39)43-32-24(3)35(42)27(30-33(5,6)36(30,32)44-25(4)38)20-26(22-37)21-34(41)28(35)19-23(2)31(34)40/h19-20,24,27-28,30,32,37,41-42H,7-18,21-22H2,1-6H3/t24-,27+,28-,30-,32-,34-,35-,36-/m1/s1 |
| PubChem CID | 27924 |
| 别名 | PMA，phorbol ester，Phorbol myristate acetate，12-豆蔻酸-13-乙酸佛波醇，12-O-Tetradecanoylphorbol13-acetate |
| 英文名称 | Phorbol 12-Myristate 13-Acetate |
| CAS | 16561-29-8 |
| 分子式 | C36H56O8 |
| 分子量 | 616.83 |
| 纯度 | HPLC≥98% |
| 单位 | 瓶 |
| 生物活性 | Phorbol 12-myristate 13-acetate (PMA) 是一种佛波酯，是常用的 PKC 活化剂。[1-4] |
| In Vitro | 为了检查PKC在p38MAPK磷酸化中的作用，用PKC活化剂PMA（100nM）刺激细胞，其模拟PKC的天然活化剂DAG与PKC的C1区的结合。在与αT3-1细胞中GnRH观察到的类似的两种细胞类型中观察到PMA的p38MAPK磷酸化，即缓慢的持续激活（分别在30分钟时为3.2倍和3.6倍）。由GnRH和PMA激活的PKC在p38MAPK磷酸化中起不同作用的矛盾发现可以通过PKC的差异定位来解释。 αT3-1细胞中p38MAPK磷酸化的基础，GnRH-和PMA-刺激是通过电压门控Ca2 +通道和Ca2 +动员的Ca2 +流入介导的，而在分化的LβT2促性腺细胞中，它仅通过Ca2 +动员介导[2]。 |
| In Vivo | PMA是PKC激动剂，其逆转由5-羟基癸酸（5-HD）诱导的损伤。因此，mitoKATP的激活通过PKC途径保护SOD和MDA中的线粒体功能[3]。 |
| SMILES | CCCCCCCCCCCCCC(O[C@H]([C@H]1C)[C@]2(OC(C)=O)[C@@]([C@@](C=C(CO)C[C@]34O)([H])[C@@]1(O)[C@]4([H])C=C(C)C3=O)([H])C2(C)C)=O |
| 靶点 | PKC |
| 动物实验 | 大鼠[3]所有实验均用雄性Wistar大鼠（体重250-280g）进行。将125只Wistar大鼠随机分成7组。 （1）假手术组（n = 21）给予侧脑室注射0.9％生理盐水; （2）IR组大鼠（n = 21）在大脑中动脉闭塞（MCAO）前30 min给予侧脑室注射0.9％生理盐水; （3）在MCAO前30分钟，给予Carbenoxolone（CBX）组（n = 21）大鼠侧脑室注射CBX（5μg/ mL×10μL）; （4）在MCAO之前30分钟，给二氮嗪（DZX）组（n = 21）的大鼠腹侧注射DZX（2mM×30μL）; （5）对5-HD组（n = 21）的大鼠进行5-HD（100mM×10μL）的侧脑室注射，10分钟后，在MCAO前15分钟注射DZX; （6）给DZX + Ro组大鼠（n = 15）注射DZX侧脑室，10分钟后，在MCAO前15分钟注射Ro-31-8425（400μg/ kg）; （7）注射5-HD和DZX后，给5-HD + PMA组（n = 15）的大鼠腹膜内注射PMA（200μg/ kg）。 |
| 细胞实验 | αT3-1和LβT-2细胞在补充有10％胎牛血清（FCS）和L-谷氨酰胺2mM，青霉素和链霉素（100单位/ mL）的DMEM中在37℃加湿培养箱中培养5％CO2培养。 C。在同一培养基中血清饥饿为0.1％FCS，持续16小时。然后如所示，将GnRH和PMA添加一段时间。通常，αT3-1细胞通过ExGen 500或jetPRIME瞬时转染，而LβT2细胞仅通过jetPRIME转染试剂转染。对于显性阴性（DN）PKC的实验，用1.5μgp38α-GFP和3μg对照载体，pCDNA3或3μgDN-PKC构建体转染αT3-1细胞（在6cm平板中）。对于LβT2细胞，用4μgp38α-GFP以及9μg对照载体，pCDNA3或9μgDN-PKC构建体进行转染（在10cm平板中）。转染后约30小时，将细胞血清饥饿（0.1％FCS）16小时，然后用GnRH或PMA刺激，用冰冷的PBS洗涤两次，用裂解缓冲液处理，然后进行一次冻融循环。收获细胞;离心（15,000×g，15分钟，4℃）后，取上清液进行免疫沉淀实验[2]。 |
| 数据来源文献 | [1]. Xu F, et al. Protein kinase C-mediated Ca2+ entry in HEK 293 cells transiently expressing human TRPV4. Br J Pharmacol. 2003 Sep;140(2):413-21.
[2]. Mugami S, et al. Differential roles of PKC isoforms (PKCs) and Ca2+ in GnRH and phorbol 12-myristate 13-acetate (PMA) stimulation of p38MAPK phosphorylation in immortalized gonadotrope cells. Mol Cell Endocrinol. 2017 Jan 5;439:141-154. [3]. Hou S, et al. Mechanism of Mitochondrial Connexin43’s Protection of the Neurovascular Unit under Acute Cerebral Ischemia-Reperfusion Injury. Int J Mol Sci. 2016 May 5;17(5). pii: E679. [4]. Zhang T, et al. MPTP-Induced Dopamine Depletion in Basolateral Amygdala via Decrease of D2R Activation Suppresses GABAA Receptors Expression and LTD Induction Leading to Anxiety-Like Behaviors. Front Mol Neurosci. 2017 Aug 7;10:247. |
| 规格 | 1mg 5mg |
Phorbol 12-myristate 13-acetate是一种佛波酯，是常用的 PKC 活化剂。
使用本产品的应用案例（仅供参考）
In Vitro
Cell（HUEVC, NIH-3T3 and RAW-264.7 cells）
As for intracellular ROS scavenging ability, HUEVC, NIH-3T3 and RAW-264.7 cells (4 × 105 cells) were seeded in a 24-well plate,then PMA was added into medium for up-regulating intracellular ROS level. Subsequently, different concentrations of EGCG/Zn Ps were added into medium. After incubating for 4 h, intracellular ROS level was detected by DCFH-DA.
来源文献：Chen Z, Duan J, Diao Y, Chen Y, Liang X, Li H, Miao Y, Gao Q, Gui L, Wang X, Yang J, Li Y. ROS-responsive capsules engineered from EGCG-Zinc networks improve therapeutic angiogenesis in mouse limb ischemia. Bioact Mater. 2020 Aug 7;6(1):1-11. doi: 10.1016/j.bioactmat.2020.07.013. PMID: 32817909; PMCID: PMC7415630.
Cell（the peripheral blood lymphocytes, 50 ng/ml PMA, 5h）
Blood from the M6-immunized (three doses) mice or rhesus monkeys was collected at and 28 days post-infection. Red blood cells were removed, and the peripheral blood lymphocytes were washed and suspended in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin. Then, the cells were cultured for 5 h in the presence of phorbol 12-myristate 13-acetate (PMA) (50 ng/ml;Jinpan),ionomycin calcium salt (1 μg/ml; Jinpan) and GolgiStop (0.67 μl/ml)
来源文献：Xu X, Feng X, Wang L, Yi T, Zheng L, Jiang G, Fan S, Liao Y, Feng M, Zhang Y, Li D, Li Q. A HSV1 mutant leads to an attenuated phenotype and induces immunity with a protective effect. PLoS Pathog. 2020 Aug 10;16(8):e1008703. doi: 10.1371/journal.ppat.1008703. PMID: 32776994; PMCID: PMC7440667.
Cell（Tumor-associated macrophages (TAMs) induction，200 ng/mL PMA，24h）
THP-1 cells (1.5×105 cells/ml) were treated with 200 ng/ml phorbol myristate acetate (PMA, Jinpan Science & Technology Co., Ltd, Beijing, China.) for 24 h and polarized into macrophages. To induce TAMs, THP-1 macrophages were cultured with CM of GC cells for a further 24h prior to harvesting.
来源文献：Sun L , Li J , Yan W , et al. H19 contributes to aerobic glycolysis, proliferation and immune escape of gastric cancer cells via the miR-519d-3p/LDHA axis. 2020.
Cell（THP-1 AR silencing and differentiation，50 ng/mL PMA，24h）
THP-1 cells were infected for 24 h with lentiviruses harboring (AR scramble or AR shRNA) shRNAs that target human AR and cloned into the U6-shRNA-EF1α- EGFP-Puro vector. The U6-shRNA-EF1α- EGFP-Puro vector encoding a scrambled sequence not matching any mammalian sequence was used as control. Positively infected cells were sorted by flow cytometry 72 h after infection. Infected THP-1 monocytes (THP-1sc and THP-1ARsi) were induced to differentiate into macrophages (MФsc and MФARsi) by exposing to 50 ng/mL phorbol myristate acetate (PMA) (Jinpan, Beijing, China) for 24 h, then incubated for an additional 24 h in the absence of PMA in a conditioning medium (CM). The collected CM was used to treat HASMCs for investigating the role of AR in HASMC calcification.
来源文献：Pang H, Xiao L, Lu Z, Chen H, Shang Z, Jiang N, Wang X, Wei F, Jiang A, Chen Y, Niu Y. Targeting androgen receptor in macrophages inhibits phosphate-induced vascular smooth muscle cell calcification by decreasing IL-6 expression. Vascul Pharmacol. 2020 Jul;130:106681. doi: 10.1016/j.vph.2020.106681. Epub 2020 May 5. PMID: 32387336.
Cell（HepG2 cells，10 nmol/L PMA，1h，37 °C）
To monitor endogenous ClO−, HepG2 cells were treated with LPS(1 μg/mL) for 12 h and then coincubated with PMA (10 nmol/L) and the peptide@Ag/Au NCs solution at 37 °C for 1 h, followed by washing three times before imaging. In the control assay, HepG2 cells were treated with LPS (1 μg/mL) for 12 h and then incubated with PMA (10 nmol/L) for 1 h. The cells were subsequently cultured in medium containing uric acid (250 nmol/L) and DMSO (0.5%) for 15 min and then treated with the peptide@Ag/Au NCs solution at 37 °C for 1 h. The cells were washed three times with PBS to remove the unbound peptide@Ag/Au NCs and were then observed under a Nikon A1R MP multiphoton microscope with a 60× oil-immersion objective lens. The images of peptide@Ag/Au NCs were captured under excitation at409 nm.
来源文献：Jia M, Mi W, Guo S, Yang QZ, Jin Y, Shao N. Peptide-capped functionalized Ag/Au bimetal nanoclusters with enhanced red fluorescence for lysosome-targeted imaging of hypochlorite in living cells. Talanta. 2020 Aug 15;216:120926. doi: 10.1016/j.talanta.2020.120926. Epub 2020 Mar 14. PMID: 32456892.