Examples of using this product（for reference only）

In Vitro：

Cell（mouse microvascular cerebral endothelial cells/bEnd.3；lycopene（0.25% BHT in THF））:

For the In Vitro experiment, we grew mouse microvascular cerebral endothelial cells (bEnd.3) in Dulbecco’s modified Eagle's medium (DMEM) supplemented with 10% FBS and 1% 100 U/mL penicillin and 100 U/mL streptomycin. Cultures were maintained under 5% CO2 at 37℃. We dissolved lycopene (Solarbio, China) in tetrahydrofuran (THF) containing 0.25% butylated hydroxytoluene (BHT) and stored it at -80℃ in 4 mM stock. On Day 5 of bEnd. 3 culture, the medium was replaced with pre-warmed PBS and then divided into a control group (solvent group), Ly294002 treatment group (10 mM; inhibitor group), and lycopene t Ly294002 group (lycopene t inhibitor group). The control group was treated with the same dose of FBS and double antibody (0.25% BHT in THF) to offset any possible interference. The lycopene t inhibitor group was pretreated with Ly294002 for 30 min and then treated with lycopene for 24 h.

Reference：

Xu XD, Teng Y, Zou JY, Ye Y, Song H, Wang ZY. Effects of lycopene on vascular remodeling through the LXR-PI3K-AKT signaling pathway in APP/PS1 mice. Biochem Biophys Res Commun. 2020 Jun 4;526(3):699-705. doi: 10.1016/j.bbrc.2020.02.063. Epub 2020 Apr 3. PMID: 32253029.

Cell（H9c2 cells，2.5~40 μM LP, 12h）:

Firstly, H9c2 cells were randomly divided into 8 groups at the first phase as follows: (i) control group (Control): H9c2 cells were incubated in normoxic conditions at 37℃ in a cell culture incubator for 13 hrs; (ii) hypoxia/re-oxygenation group (H/R): H9c2 cells were subjected to hypoxia for 12 hrs followed by reoxygenation for 1 hr; (iii) solvent group (Vehicle): H9c2 cells were pretreated with 0.1% DMSO for 12 hrs and underwent HR group procedures; (iv)–(viii) 2.5 μM LP pretreatment group (LP 2.5 μM), 5 μM LP pretreatment group (LP 5 μM), 10 μM LP pretreatment group (LP 10 μM), 20 μM LP pretreatment group (LP 20 μM), 40 μM LP pretreatment group(LP 40 μM): H9c2 cells were pretreated with LP at the dose of 2.5 μM, 5 μM, 10 μM, 20 μM and 40 μM for 12 hrs and underwent HR group procedures.

In the second phase (according to the result of first phase), 10 μM LP was selected to explore the mechanism further. Therefore, H9c2 cells were further divided into 4 groups as follows: (i) Control group; (ii) HR group; (iii)LP 10 μM group; (iv) LP 10 μM combined with ATR pretreatment group (LP 10 μM + ATR): H9c2 cells were pretreated with 10 μM LP and 20 μM ATR for 12 hrs and underwent HR group procedures.

Reference：

Li X, Jia P, Huang Z, Liu S, Miao J, Guo Y, Wu N, Jia D. Lycopene protects against myocardial ischemia-reperfusion injury by inhibiting mitochondrial permeability transition pore opening. Drug Des Devel Ther. 2019 Jul 11;13:2331-2342. doi: 10.2147/DDDT.S194753. PMID: 31371925; PMCID: PMC6635826.

In Vivo：

Mice（9-month-old APP/PS1 mice，4 mg/kg lycopene）:

We randomly assigned 12 APP/PS1 transgenic mice to two groups: APP/PS1+solvent (APP/PS1; AD group; n=6) and AD+4 mg/kg lycopene (LY; drug group; n = 6). Similarly, we randomly selected six WT C57/BL6J mice as the wild control group (WT group).

Reference：

Xu XD, Teng Y, Zou JY, Ye Y, Song H, Wang ZY. Effects of lycopene on vascular remodeling through the LXR-PI3K-AKT signaling pathway in APP/PS1 mice. Biochem Biophys Res Commun. 2020 Jun 4;526(3):699-705. doi: 10.1016/j.bbrc.2020.02.063. Epub 2020 Apr 3. PMID: 32253029.

Rat（10~40 mg/kg LP，1次/d，共5 d，I.P；experimental group: 40 mg/ kg LP in Corn Oil）:

In the first phase, 30 rats were divided into the following 5 groups with 6 rats per group: (i) ischemia-reperfusion group (IR): As described above; (ii) solvent group (Vehicle): The rats were pretreated with 2 mL medicinal corn oil by intraperitoneal injection once per day for 5 days, and underwent IR procedures; (iii–v) 10 mg/kg LP pretreatment group (LP 10 mg/kg), 20 mg/kg LP pretreatment group (LP 20 mg/kg), 40 mg/kg LP pretreatment group (LP 40 mg/kg): The rats were pretreated with 10, 20, 40 mg/kg LP by intraperitoneal injection once per day, for a total of 5 days, and underwent IR group procedures.

In the second phase (according to the results in the first phase), 40 mg/kg LP was selected as the most appropriate dose. A total of 80 rats were divided into the following 4 groups with 20 rats per group in the second phase: (i) IR group; (ii) Vehicle group; (iii) LP 40 mg/kg group; (iv) 40 mg/kg LP in combination with ATR pretreatment group (LP 40 mg/kg + ATR): 5 mg/kg ATR was injected intraperitoneally 30 mins prior to extraction of the heart, and the remainder of the procedure was as described for the LP 40 mg/kg group.

Reference：

Li X, Jia P, Huang Z, Liu S, Miao J, Guo Y, Wu N, Jia D. Lycopene protects against myocardial ischemia-reperfusion injury by inhibiting mitochondrial permeability transition pore opening. Drug Des Devel Ther. 2019 Jul 11;13:2331-2342. doi: 10.2147/DDDT.S194753. PMID: 31371925; PMCID: PMC6635826.

Mice（healthy male kunming mice (6 weeks old) weighing 23–28 g；5 mg/kg LYC（1 mg/mL的悬浮液），Gavage；溶剂：橄榄油）:

After an acclimatization period of a week, the mice were divided into four groups, ten in each. The mice were treated for 30 days as follows: Group 1 (CG) was orally administered with the vehicle (0.2 mL olive oil). Group 2 (AG) was orally administrated with AFB1 (≥ 99.8%) at 0.75 mg/kg. Group 3 (ALG) were treated with LYC (≥ 98%, Solarbio, Beijing, China) at 5 mg/kg 2 h prior to AFB1 administration, then given AFB1 at 0.75 mg/kg. Group 4(LG) was orally administrated with LYC at 5 mg/kg. AFB1 and LYC were administered by gavage in olive oil and made into suspension at the concentrations of 0.2 mg/mL and 1 mg/mL, respectively. The general health was monitored daily.

Reference：

Zhang J, Wang P, Xu F, Huang W, Ji Q, Han Y, Shao B, Li Y. Protective effects of lycopene against AFB1-induced erythrocyte dysfunction and oxidative stress in mice. Res Vet Sci. 2020 Apr;129:103-108. doi: 10.1016/j.rvsc.2020.01.015. Epub 2020 Jan 13. PMID: 31954314.