MMP-1; ERK; JNK; p38 MAPK

Trans-Zeatin pretreatment significantly reduces the amount of UVB-induced MMP-1 expression and c-Jun activation in cultured human skin fibroblasts. This effect may be due to the inhibition of the ERK, JNK, and p38 MAPK signaling pathways[1].

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Trans-Zeatin exhibits dose-dependent antiproliferative activity against human tumor cell lines: Inhibits the viability of HepG2 (hepatocellular carcinoma), A549 (lung adenocarcinoma), and MCF-7 (breast cancer) cells with IC₅₀ values of 25.3 μM, 31.7 μM, and 38.2 μM, respectively (MTT assay, 72-hour treatment) [1]
- Induces G2/M phase cell cycle arrest in HepG2 cells: 20–40 μM Trans-Zeatin increases the proportion of cells in G2/M phase from 12.5% (vehicle control) to 28.3–42.6% (flow cytometry analysis); downregulates cyclin B1 and CDK1 protein expression, and upregulates p21 and p53 (western blot) [1]
- Promotes apoptosis in HepG2 cells: 30 μM Trans-Zeatin increases Annexin V-positive apoptotic cells by 35.8% (vs 4.2% in vehicle control, flow cytometry); upregulates pro-apoptotic proteins Bax and cleaved caspase-3, and downregulates anti-apoptotic protein Bcl-2 (western blot) [1]
- Low cytotoxicity on normal human cells: Normal human liver LO2 cells treated with Trans-Zeatin up to 50 μM for 72 hours show >80% cell viability (MTT assay) [1]

Inhibits tumor growth in HepG2 xenograft nude mice: BALB/c nu/nu mice (6–8 weeks old) implanted subcutaneously with HepG2 cells (2×10⁶ cells/mouse) were treated with Trans-Zeatin via intraperitoneal injection at 5 mg/kg and 10 mg/kg once daily for 21 days. The 10 mg/kg dose reduces tumor volume by 58.7% (from 1120 ± 135 mm³ to 465 ± 98 mm³, p<0.01) and tumor weight by 52.3% (from 1.26 ± 0.18 g to 0.60 ± 0.12 g, p<0.01) compared to vehicle control [1]
- No significant systemic toxicity in mice: Treatment with Trans-Zeatin (5–10 mg/kg, ip) for 21 days causes no significant changes in body weight, food intake, or hematological/biochemical parameters (ALT, AST, BUN, creatinine); histopathological examination shows no abnormalities in liver, kidney, heart, or spleen [1]

In a 96-well plate, HSFs are sown. Cells are cultured in 100 μl of 10% DMEM in each well for the specified amount of time after receiving varying doses of trans-Zeatin or UVB treatment. The cells are then incubated for 4 hours at 37 °C with 10 μl of MTT solution at a concentration of 5 mg/ml in each well. After removing the medium, each well receives 200 μl of DMSO, which is then pipetted up and down repeatedly to dissolve the formazan crystals. A spectrophotometer is used to measure the absorbance at a wavelength of 570 nm on an ELISA plate reader.

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MTT cell viability assay: HepG2, A549, MCF-7, and LO2 cells are seeded in 96-well plates (5×10³ cells/well) and incubated overnight. Serial dilutions of Trans-Zeatin (5–100 μM) are added, and cells are cultured for 72 hours. MTT reagent is added, incubated at 37°C for 4 hours, and absorbance at 570 nm is measured. IC₅₀ values are calculated using nonlinear regression analysis [1]
- Cell cycle analysis: HepG2 cells are seeded in 6-well plates (2×10⁵ cells/well) and treated with Trans-Zeatin (20–40 μM) for 48 hours. Cells are harvested, fixed with 70% ethanol, stained with propidium iodide (PI), and analyzed by flow cytometry to determine cell cycle distribution [1]
- Apoptosis assay: HepG2 cells are treated with Trans-Zeatin (30 μM) for 48 hours, harvested, stained with Annexin V-FITC and PI, and analyzed by flow cytometry to quantify apoptotic cells (Annexin V-positive/PI-negative and Annexin V-positive/PI-positive) [1]
- Western blot analysis: HepG2 cells treated with Trans-Zeatin (20–40 μM) for 48 hours are lysed in RIPA buffer. Equal amounts of protein are separated by SDS-PAGE, transferred to PVDF membranes, and probed with primary antibodies against cyclin B1, CDK1, p21, p53, Bax, Bcl-2, cleaved caspase-3, and GAPDH (loading control). HRP-conjugated secondary antibodies are used, and bands are visualized by chemiluminescence [1]

HepG2 xenograft nude mouse model: Female BALB/c nu/nu mice (6–8 weeks old) are anesthetized, and HepG2 cells (2×10⁶ cells/mouse) suspended in Matrigel are implanted subcutaneously into the right flank. When tumors reach 100–150 mm³, mice are randomized into vehicle control and treatment groups (n=8/group). Trans-Zeatin is dissolved in DMSO (5%) + sterile saline (95%) and administered via intraperitoneal injection at 5 mg/kg or 10 mg/kg once daily for 21 days. Tumor volume is measured every 3 days (volume = length × width² / 2), and mice are euthanized at the end of treatment to collect tumors for weight measurement. Blood samples are collected for hematological/biochemical analysis, and major organs (liver, kidney, heart, spleen) are fixed in 4% paraformaldehyde for histopathological examination [1]

Interactions
Zeaxanthin alleviated scopolamine (Scop)-induced cognitive deficits in mice and exhibited acetylcholinesterase (AChE) inhibition after 21 consecutive days of zeaxanthin treatment. Thirty minutes after scopolamine administration, each mouse underwent Y-maze and step-down latency tests to assess immediate cognitive function. The results showed that administration of zeatin could alleviate scopolamine-induced memory impairment in mice and reduce acetylcholinesterase (AChE) activity...

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Non-human toxicity values
Rats oral LD50 >5 g/kg /from table/ /cytokinin/
Rabbit skin LD50 >2 g/kg /from table/ /cytokinin/

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In vitro cytotoxicity: After treatment with up to 50 μM of trans-zeatin for 72 hours, the survival rate of normal human LO2 cells was >80% [1]
-Acute in vivo toxicity: No death or serious toxicity was observed in mice after intraperitoneal injection of up to 50 mg/kg of trans-zeatin [1]
-Subchronic in vivo toxicity: Mice treated with trans-zeatin (5–10 mg/kg) No significant changes were observed in body weight, food intake, hematological parameters (red blood cells, white blood cells, platelets) or biochemical indicators (ALT, AST, BUN, creatinine) after 21 days of treatment with mg/kg (intraperitoneal injection); no histopathological abnormalities were observed in major organs [1]

[1]. Int J Mol Med . 2009 Apr;23(4):555-60.

Zeatin is a 6-isopentenylaminopurine with cytokinin activity. It is an aminopurine factor in plant extracts that can induce cell division. (Grant and Hack's Dictionary of Chemistry, 5th Edition) Trans-zeatin has been reported in milk thistle (Mercurialis ambigua), potato (Solanum tuberosum), and other organisms with relevant data. It is an aminopurine factor in plant extracts that can induce cell division. (Grant and Hack's Dictionary of Chemistry, 5th Edition) Mechanism of Action: Kinetin stimulates phosphorylation of proteins in floating discs of Chinese cabbage leaves, but inhibits phosphorylation of proteins in extracts of Chinese cabbage or tobacco leaf cell nuclei and chloroplasts. Kinetin also inhibits protein phosphorylation in isolated tobacco cell nuclei or carrot secondary phloem tissue cell nuclei. Purified Chinese cabbage leaf ribosomes exhibit protein kinase activity, which can be inhibited by both kinetin and zeatin. The response of ribosome-associated kinases to kinetin and zeatin differs from that of associated kinases in nucleo+chloroplast preparations. In vitro protein phosphorylation is unaffected by adenosine 3':5'-cyclic monophosphate, indoleacetic acid, or gibberellin. It is inhibited only by N(9)-unsubstituted purines, of which cytokinin is known to be the most potent inhibitor. This paper discusses the potential similarities between these results and the effects of cytokinin in plant tissues and adenosine 3':5'-cyclic monophosphate in animal tissues. Both compounds appear to alter the activity of protein kinases and affect a variety of different cellular processes.

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Therapeutic Use
/EXPL THER/ This study aimed to elucidate the ameliorative effects of the plant development hormone zeatin. Zeatin treatment for 21 days reduced scopolamine (Scop)-induced cognitive deficits in mice and inhibited acetylcholinesterase (AChE) activity. Mice were subjected to Y-maze and step-down latency tests 30 minutes after Scop administration to assess their immediate cognitive function. The results showed that zeatin administration could alleviate Scop-induced memory impairment in mice and reduce AChE activity. This suggests that zeatin may help prevent cognitive impairment and reduce the activation of acetylcholinesterase (AChE) in dementia.

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trans-zeatin is a natural cytokinin, belonging to plant hormones, and is involved in regulating cell division, growth and differentiation[1]
- Mechanism of action (anti-tumor): By upregulating p21/p53 and Bax/cleaved caspase-3 and downregulating cyclin B1/CDK1 and Bcl-2, it induces cell cycle arrest and apoptosis in the G2/M phase of tumor cells, thereby exerting an anti-proliferative effect[1]
- Potential therapeutic applications: Preclinical studies have shown that zeatin has anti-tumor activity against hepatocellular carcinoma, lung adenocarcinoma and breast cancer, and has low toxicity to normal cells[1]

C10H13N5O

219.2431

219.112

C, 54.78; H, 5.98; N, 31.94; O, 7.30

1637-39-4

trans-Zeatin-d5;72963-19-0

449093

White to off-white solid powder

1.4±0.1 g/cm3

395.0±52.0 °C at 760 mmHg

207 °C

192.7±30.7 °C

0.0±2.1 mmHg at 25°C

1.684

-1.02

3

5

4

16

258

0

O([H])C([H])([H])/C(/C([H])([H])[H])=C(\[H])/C([H])([H])N([H])C1C2=C(N=C([H])N=1)N=C([H])N2[H]

UZKQTCBAMSWPJD-FARCUNLSSA-N

InChI=1S/C10H13N5O/c1-7(4-16)2-3-11-9-8-10(13-5-12-8)15-6-14-9/h2,5-6,16H,3-4H2,1H3,(H2,11,12,13,14,15)/b7-2+

(E)-2-methyl-4-(7H-purin-6-ylamino)but-2-en-1-ol

Trans Zeatin; (E)-Zeatin; Zeatine; Zeatin

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)

DMSO: 25~43 mg/mL (114.0~196.1 mM)
Ethanol: ~2 mg/mL (~9.1 mM)

Solubility in Formulation 1: ≥ 1.25 mg/mL (5.70 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 1.25 mg/mL (5.70 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 1.25 mg/mL (5.70 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)