PKA is a ubiquitous cellular kinase, also known as cAMP-dependent protein kinase, and it is well-established that plays an important role in regulating several functions of cell processes, including regulation of glycogen, sugar, and lipid metabolism[1]. H-89 2HCl, as a newly synthesized isoquinolinesulfonamide, is a ATP-competitive, potent inhibitor of protein kinase A (PKA)(IC50 = 48 nM), and has weak inhibition on several other kinases with IC50 of 80, 120, 135, 270, 2600 and 2800 nM for S6K1, MSK1, PKA, ROCKII, PKBα and MAPKAP-K1b, respectively[2]. H-89 2HCl has since been used extensively for evaluation of the role of PKA in the heart, osteoblasts, hepatocytes, smooth muscle cells, neuronal tissue, epithelial cells, etc[3]. PC12D cells pretreatment with H-89 2HCl led to a dose-dependent inhibition of the forskolin-induced neurite outgrowth and protein phosphorylation. In vivo experimental method, in skinned EDL fibres of the rat, H-89 2HCl with 1-2 μM significantly slowed the repriming rate in rat skinned fibres. Moreover, rat brain ventricles following an injection with the PKA antagonist, H-89 2HCl, the body temperature is increased further due to the inhibition of TRPV1 phosphorylation[4].
作用机制
H-89 2HCl inhibit the significant modulator role of the cAMP-PKA intracellular signaling pathway mainly acting as a selective and potent inhibitor of protein kinase A (PKA).
H89 2HCl significantly inhibited PKA activity in Sf9 cells and reduced cell tolerance to permethrin.
Int J Mol Sci. 2019 Sep 3;20(17):4300.
TF-1 cells
10 μM
1 h
Inhibition of PKA activity to study its effect on CREB-1 phosphorylation. Results showed that H-89 significantly inhibited forskolin-induced nuclear accumulation of pCREB-1, indicating that PKA plays a central role in the cAMP-PKA-pCREB network.
Biomed Pharmacother. 2011 Jul;65(4):293-7.
TF-1 cells
10 μM
15 h
Study the effect of PKA inhibition on CCR5 transcription. Results showed that H-89 significantly inhibited forskolin-induced increase in CCR5 mRNA levels, indicating that PKA acts in concert with CREB to modulate cAMP-mediated CCR5 transcription.
Inhibition of PKA activity, preventing PTX-induced hyperphosphorylation of Shank3 S1615
Elife. 2022 Apr 26;11:e74277.
COX-2 KO and control cells
10 µM
6 days
To study the role of H-89 in COX-2 KO and control cells, it was found that H-89 attenuated the suppressing effect of PGE2 on adipocyte differentiation.
Cells. 2022 Jun 2;11(11):1819.
bone marrow-derived macrophages (BMDMs)
20 μM
1 h
To investigate whether db-cAMP-induced macrophage polarization is dependent on PKA.The results showed that pre-treatment with H89 inhibited db-cAMP-induced Arg-1 and CD206 expression and IL-10 levels, suggesting that PKA is involved in db-cAMP-induced macrophage polarization
Cells. 2020 Jan 6;9(1):128.
H-89 2HCl 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Culex quinquefasciatus
HAmCqG8 and MAmCqG6 resistant mosquito larvae
Water treatment
6.25, 12.5, 25, 50 µM
Single treatment, lasting 24 hours
H89 2HCl significantly reduced the resistance of mosquito larvae to permethrin.
Int J Mol Sci. 2019 Sep 3;20(17):4300.
BALB/c mice
LPS-induced pleurisy model
Intrapleural injection
4 mg/kg
Single injection, lasting 30 hours
To investigate the effect of db-cAMP on macrophage polarization in vivo, results showed that db-cAMP decreased the number of LPS-induced M1 macrophages and increased the expression of engulfment-related molecules AnxA1 and CD36.
Cells. 2020 Jan 6;9(1):128.
Mice
Acute brain slices
Intraperitoneal or subcutaneous injections
5 mg/kg
Single injection, lasting 30 minutes
H-89 pretreatment prevented the increase in mEPSC frequency and dendritic spine density induced by Drd1 agonist
Elife. 2015 Nov 9;4:e10111
Mice
High-fat diet-induced MAFL model
Intraperitoneal injection
1 mg/kg
Once daily for 8 weeks
To evaluate the effect of H-89 on MAFL formation, the results showed that H-89 significantly improved hepatic steatosis and reduced ALT activity, and decreased the nuclear level of mature SREBP1c.
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