ERK; Caspase-3; Caspase-12; Human Endogenous Metabolite

Tauroursodeoxycholate (TUDCA) reduces the viability and migration of vascular smooth muscle cells (VSMCs) by inhibiting the phosphorylation of ERK and by inducing mitogen-activated protein kinase phosphatase-1 (MKP-1) via PKCα. By blocking ERK through Ca2+-dependent PKC translocation, tauroursodeoxycholate prevents the VSMCs from proliferating and migrating. Platelet-derived growth factor (PDGF) and MMP-9 expression brought on by vascular injury are both prevented by tauroursodeoxycholate. Tauroursodeoxycholate (200 μM) reduced VSMC viability, which suggests that Tauroursodeoxycholate's ability to inhibit cell proliferation depended on MKP-1 expression[1]. MKP-1 expression can be reduced by using specific si-RNA to knock it down.

Immunohistochemistry is used to examine the impact of tauroursodeoxycholate (TUDCA) on the proliferation and apoptosis of VSMCs in vivo. Tauroursodeoxycholate (10, 50, and 100 mg/kg) increases the caspase 3 activity of injured tissues in a dose-dependent manner, suggesting that it causes VSMCs in the neointima to undergo apoptosis. At one week after injury, additional testing and comparison of the phosphorylation level of ERK and MMP-9 expression is done using the injured tissues in comparison to normal controls. Both ERK phosphorylation and MMP-9 expression were upregulated in the tissues after balloon injury. Tauroursodeoxycholate (10, 50, and 100 mg/kg) inhibits the expression of MMP-9 and ERK in a dose-dependent manner[1]. Tauroursodeoxycholate (TUDCA) is a hydrophilic bile acid. By lowering ER stress and apoptosis, tauroursodeoxycholate, a cytoprotective agent, enhances liver function and can prevent hepatocellular carcinoma. In Ang II-induced ApoE-/- mice, tauroursodeoxycholate significantly decreases expression of apoptosis molecules like caspase-3, caspase-12, C/EBP homologous protein, c-Jun N-terminal kinase (JNK), activating transcription factor 4 (ATF4), X-box binding protein (XBP), and eukaryotic initiation factor 2α (eIF2; p<0.05). In ApoE-/- mice, tauroursodeoxycholate prevents the development of abdominal aortic aneurysms (AAAs) brought on by Ang II. Tauroursodeoxycholate is administered to ApoE-/- mice (ER stress inhibitor group) at a dose of 0.5 g/kg/day. Total cholesterol levels (663.6±88.7 mg/dL vs 655.7±65.4 mg/dL; p>0 .05) and systolic blood pressure (141.3±5.6 mmHg vs 145.98.9 mmHg; p>0.05) were comparable between the AAA model group and the Tauroursodeoxycholate group. Additionally, when compared to the AAA model group, the Tauroursodeoxycholate group's maximum aortic diameter was significantly lower (0.95±0.03 mm vs 1.79±0.04 mm; p<0.05). The AAA lesion areas in the Tauroursodeoxycholate group are also smaller than those in the AAA model group (0.37±0.03 mm2 vs 1.51±0.06 mm2; p<0.05)[2].

Ez-Cytox is used to assess the viability and growth of cells. Smooth Muscle Cell Growth Medium 2 (SMCGM2) is used to seed and cultivate VSMCs (5×103 cells) on 96-well plates. After serum starvation, Tauroursodeoxycholate (0, 50, 100, and 200 μM) is added to the hVSMCs, with or without 1,2-bis(o-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA, 10 μM) and 7-hydroxystaurosporine (H7, 10 μM) and cultured for 24 h. HVSMCs are seeded onto 96-well plates and cultured to determine the impact of tauroursodeoxycholate on the PDGF-stimulated hVSMC proliferation. Tauroursodeoxycholate (0, 50, 100, and 200 μM) is added to the hVSMCs after serum starvation, with or without PDGF-BB (50 ng/mL), and the cells are then cultured. The optical density at 450 nm is used to assess cell viability after the addition of 10 μL of Ez-Cytox into each well[1].

Rats: Ketamine (70 mg/kg) and Xylazine (7 mg/kg) are combined to anesthetize Sprague-Dawley rats. For two weeks, tauroursodeoxycholate is given orally once daily at various concentrations (e.g., vehicle, 10, 50, and 100 mg/kg). The carotid arteries are preserved by perfusion with 4% formaldehyde, followed by paraffin embedding and H&E staining of sections (8 μm)[1].
Mice: Thirty C57BL/6 male ApoE-/- mice are divided into three groups at random, each with ten mice, and they are eight weeks old. (i) ApoE-/- mice are implanted with mini-osmotic pumps to release Ang II (1000 ng/kg/min) over the course of 28 days (AAA model group); (ii) AAA model mice are treated with Tauroursodeoxycholate daily for 4 weeks at a dosage of 0.5 g/kg/day in drinking water (Tauroursodeoxycholate group). Following a 28-day Ang II infusion, mice are sacrificed[2].

[1]. Tauroursodeoxycholate (TUDCA) inhibits neointimal hyperplasia by suppression of ERK via PKCα-mediated MKP-1 induction. Cardiovasc Res. 2011 Nov 1;92(2):307-16.

[2]. Tauroursodeoxycholic Acid Attenuates Angiotensin II Induced Abdominal Aortic Aneurysm Formation in Apolipoprotein E-deficient Mice by Inhibiting Endoplasmic Reticulum Stress. Eur J Vasc Endovasc Surg. 2017 Mar;53(3):337-345.

C26H44NNAO6S

521.69

C, 59.86; H, 8.50; N, 2.68; Na, 4.41; O, 18.40; S, 6.15

35807-85-3

Tauroursodeoxycholate;14605-22-2;Tauroursodeoxycholate-d4 sodium;2410279-95-5;Tauroursodeoxycholate dihydrate;117609-50-4

Solid powder

C[C@H](CCC(=O)NCCS(=O)(=O)[O-])[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2[C@H](C[C@H]4[C@@]3(CC[C@H](C4)O)C)O)C.[Na+]

IYPNVUSIMGAJFC-JUWYWQLMSA-M

InChI=1S/C26H45NO6S.Na/c1-16(4-7-23(30)27-12-13-34(31,32)33)19-5-6-20-24-21(9-11-26(19,20)3)25(2)10-8-18(28)14-17(25)15-22(24)29;/h16-22,24,28-29H,4-15H2,1-3H3,(H,27,30)(H,31,32,33);/q;+1/p-1/t16-,17+,18-,19-,20+,21+,22+,24+,25+,26-;/m1./s1

sodium;2-[[(4R)-4-[(3R,5S,7S,8R,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]ethanesulfonate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)