TRAM-34 is a highly selective blocker of intermediate-conductance calcium-activated K+ channel (IKCa1) (Kd=20 nM). TRAM-34 inhibits the cloned and the native IKCa1 channel in human T lymphocytes with a Kd of 20-25 nM and is 200- to 1,500-fold selective over other ion channels. Combinations of TRAM-34 and cyclosporin A are more effective in suppressing lymphocyte mitogenesis than either compound alone[3]. TRAM-34, a specific inhibitor of K(Ca)3.1 channels increased or decreased cell proliferation depending on the concentration. At intermediate concentrations (3-10 microM) TRAM-34 increased cell proliferation, whereas at higher concentrations (20-100 microM) TRAM-34 decreased cell proliferation. TRAM-34, as well as inhibiting K(Ca)3.1, directly interacts with the oestrogen receptor and mimics the effects of 17beta-oestradiol on MCF-7 cell proliferation and gene modulation[4]. TRAM-34 can inhibit the proliferation and invasion of HL-60 cells, and can induce cell apoptosis and G0/G1 arrest. The time and concentration of TRAM-34 have effect on the malignant behavior of HL-60 cells[5].
TRAM-34 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Murine CLL B cells from Eμ-TCL1 mice
10 μM
Reduced cell viability
J Exp Clin Cancer Res. 2022 Feb 16;41(1):64.
Bone marrow-derived macrophages (BMDMs)
5 µM
3 hours
To investigate the effect of TRAM-34 on DSS-enhanced ATP or nigericin-induced NLRP3 inflammasome activation. Results showed that TRAM-34 significantly inhibited DSS-enhanced NLRP3 inflammasome activation and subsequent pyroptosis.
Cell Mol Immunol. 2022 Aug;19(8):925-943.
Primary human chronic lymphocytic leukemia (CLL) cells
10 μM
Decreased CLL cell proliferation
J Exp Clin Cancer Res. 2022 Feb 16;41(1):64.
Human lung mast cells (HLMC)
20 or 200 nM
3 hours
TRAM-34 markedly inhibited HLMC migration towards CXCL10, SCF, and ASM supernatants in a dose-dependent manner.
Thorax. 2006 Oct;61(10):880-5.
Human cardiac micro-vascular endothelial cells (HCMVECs)
1 μM
30 minutes
To investigate the effect of TRAM-34 on IKCa oxidation/tyrosine nitration in HCMVECs treated with high glucose and homocysteine. Results showed that TRAM-34 inhibited EDHF-induced vascular relaxation.
Redox Biol. 2018 Jun;16:215-225.
Acutely activated T cells
1 µM
72 hours
TRAM-34 completely suppressed proliferation of acutely activated T cells
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13942-7.
Quiescent T cells
1 µM
72 hours
TRAM-34 completely suppressed proliferation of quiescent T cells
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13942-7.
PAS T cells
1 µM
48-72 hours
TRAM-34 only partially suppressed proliferation of PAS cells
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13942-7.
Human T84 colonic epithelial cells
22 nM
3–6 minutes
TRAM-34 blocks IKCa1 currents in human T84 colonic epithelial cells with a Kd of 22 nM
Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6.
Human T lymphocytes
25 nM
3–6 minutes
TRAM-34 selectively blocks the IKCa1 channel with a Kd of 25 nM
Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6.
COS-7 cells
20 nM
3–6 minutes
TRAM-34 selectively blocks the IKCa1 channel with a Kd of 20 nM
Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6.
Murine renal fibroblasts (TFB)
100 nM
24 hours
Inhibited bFGF-induced renal fibroblast proliferation via G0/G1 arrest
Proc Natl Acad Sci U S A. 2009 Aug 25;106(34):14518-23.
Human microglial cells
1 μM
TRAM-34 significantly reduced the ATP-induced current potentiation but did not completely abolish it
J Neuroinflammation. 2021 Feb 15;18(1):44.
TRAM-34 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6J mice
DSS-induced colitis model
Intraperitoneal injection
40 mg/kg body weight
Twice daily until the endpoint
To investigate the effect of TRAM-34 on DSS-induced colitis. Results showed that TRAM-34 significantly alleviated DSS-induced colitis symptoms, including weight loss, increased disease activity index (DAI), and colon shortening. Additionally, TRAM-34 reduced inflammasome activation in the colon.
Cell Mol Immunol. 2022 Aug;19(8):925-943.
Eμ-TCL1 mice
Chronic lymphocytic leukemia (CLL) model
Intraperitoneal injections
10 nmol/g
Once a day for 2 weeks
TRAM-34 did not show any beneficial effect in vivo
J Exp Clin Cancer Res. 2022 Feb 16;41(1):64.
Mice
Db/db mice (Type 2 diabetes model)
In vitro administration (vascular ring assay)
1 μM
Single dose, 30 minutes
To investigate the effect of TRAM-34 on EDHF-induced vascular relaxation in small mesenteric arteries (SMAs) of db/db mice. Results showed that TRAM-34 inhibited EDHF-induced vascular relaxation.
Redox Biol. 2018 Jun;16:215-225.
Rat
Mesenteric artery
In vitro perfusion
1 μM
To investigate the role of TRPC3 channels in rat mesenteric artery. Pyr3 significantly reduced the ACh-induced endothelium-dependent vasodilation.
Cardiovasc Res. 2012 Sep 1;95(4):439-47.
CF-1BR mice
Acute toxicity test
Intravenous injection
0.5 mg/kg
Single dose, observed for 7 days
TRAM-34 is not acutely toxic at ~500–1,000 times the channel-blocking dose
Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6.
Rat
Isolated perfused heart model
Perfusate
1 μM
25–30 minutes
To evaluate the effect of TRAM-34 on SKA-31-induced increases in coronary flow. Results showed that TRAM-34 alone or in combination with apamin significantly inhibited the increases in coronary flow induced by SKA-31, bradykinin, and adenosine.
Cardiovasc Res. 2013 Feb 1;97(2):339-48
Mice and rats
Unilateral ureteral obstruction (UUO) model
Intraperitoneal injection
120 mg/kg
Once daily for 3 weeks
Attenuated progression of UUO-induced renal fibrosis, reduced collagen deposition and αSMA-positive cells
Proc Natl Acad Sci U S A. 2009 Aug 25;106(34):14518-23.
Apoe–/– mice
Atherosclerosis model
Subcutaneous injection
120 mg/kg/d
Once daily for 12 weeks
To study the effect of TRAM-34 on the development of atherosclerosis. Results showed that TRAM-34 significantly reduced atherosclerotic lesion area in aortas and carotid arteries, suppressed VSMC proliferation, macrophage and T lymphocyte infiltration, and reduced oxidative stress.
J Clin Invest. 2008 Sep;118(9):3025-37
Lewis rats
Adoptive transfer experimental autoimmune encephalomyelitis (AT-EAE)
Intraperitoneal injection
170 µg/kg
Three more injections at 4-hr intervals on day 0, three injections on day 1, and two injections on days 2–5
TRAM-34 alone had no effect on preventing AT-EAE, but enhanced the effect when combined with ShK-Dap22
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13942-7.
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