Journal: International Journal of Molecular Medicine

Article Title: Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

doi: 10.3892/ijmm.2023.5312

Figure Lengend Snippet: TSN alleviates ferroptosis of A/R-induced H9c2 cardiomyocytes via downregulation of VDAC1. (A) Cell Counting Kit-8 detection of viability in A/R-induced cells following TSN or Fer-1 pretreatment. (B) LDH, (C) MDA, (D) total iron, (E) GSH, GSSG, GSH/GSSG and (F) ROS were determined by quantitative kits in A/R-induced cells following TSN or Fer-1 treatment (magnification, ×200; scale bar, 50 μ m). (G) Expression of (H) ferroptosis-related proteins and VDAC1 were detected by western blot analysis in A/R-induced cells following TSN or Fer-1 pretreatment. Data are expressed as the mean ± SD (n=3). *** P<0.05. TSN, Tanshinone IIA; A/R, Anoxia/reoxygenation; VDAC1, Voltage-dependent anion channel 1; Fer-1, ferrostatin-1; LDH, lactate dehydrogenase; MDA, malondialdehyde; GSH, Glutathione; GSSG, Glutathione disulfide; ROS, reactive oxygen species; PTGS2, Prostaglandin endoperoxide synthase 2; GPX, Glutathione peroxidase 4.

Article Snippet: Next, CMV-ADV6-VDAC1 was transduced into 293 cells (ATCC, cat. no. CRL-1573) using RNAi-mate (Suzhou GenePharma Co., Ltd.) and a 2nd generation system under 95% O 2 and 5% CO 2 at 37°C for 6 h (quantity of adenovirus plasmid used for transfection is 10 μ g and the ratio used for adenovirus, packaging and envelope plasmids is 1:3:4).

Techniques: Cell Counting, Expressing, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

doi: 10.3892/ijmm.2023.5312

Figure Lengend Snippet: TSN binds to VDAC1. (A) Chemical structure of TSN. (B) Molecular structure of VDAC1. (C) 3D and (D) 2D diagram of the interaction between TSN and VDAC1. TSN, Tanshinone IIA; VDAC1, Voltage-dependent anion channel 1.

Article Snippet: Next, CMV-ADV6-VDAC1 was transduced into 293 cells (ATCC, cat. no. CRL-1573) using RNAi-mate (Suzhou GenePharma Co., Ltd.) and a 2nd generation system under 95% O 2 and 5% CO 2 at 37°C for 6 h (quantity of adenovirus plasmid used for transfection is 10 μ g and the ratio used for adenovirus, packaging and envelope plasmids is 1:3:4).

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

doi: 10.3892/ijmm.2023.5312

Figure Lengend Snippet: TSN inhibits ferroptosis of A/R-induced H9c2 cardiomyocytes by downregulating VDAC1. (A) Cell Counting Kit-8 detection of viability in A/R-induced cells after TSN, pAd/VDAC1 and pAd/NC pretreatment. (B) LDH, (C) MDA, (D) total iron, (E) GSH, GSSG, GSH/GSSG and (F) ROS were determined by quantitative kits in A/R-induced cells following TSN, pAd/VDAC1 and pAd/NC treatment (magnification, x200; scale bar, 50 μ m). (G) Expression of (H) ferroptosis-associated proteins and VDAC1 were detected by western blot analysis in A/R-induced cells after TSN, pAd/VDAC1 and pAd/NC pretreatment. Data are expressed as the mean ± SD (n=3). *** P<0.05. TSN, tanshinone IIA; A/R, anoxia/reoxygenation; VDAC1, voltage-dependent anion channel 1; NC, negative control; LDH, lactate dehydrogenase; MDA, malondialdehyde; GSH, Glutathione; GSSG, Glutathione disulfide; ROS, reactive oxygen species; PTGS2, Prostaglandin endoperoxide synthase 2; GPX4, Glutathione peroxidase 4.

Article Snippet: Next, CMV-ADV6-VDAC1 was transduced into 293 cells (ATCC, cat. no. CRL-1573) using RNAi-mate (Suzhou GenePharma Co., Ltd.) and a 2nd generation system under 95% O 2 and 5% CO 2 at 37°C for 6 h (quantity of adenovirus plasmid used for transfection is 10 μ g and the ratio used for adenovirus, packaging and envelope plasmids is 1:3:4).

Techniques: Cell Counting, Expressing, Western Blot, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

doi: 10.3892/ijmm.2023.5312

Figure Lengend Snippet: TSN improves mitochondrial function and integrity in H9c2 cardiomyocytes exposed to A/R by downregulating VDAC1. (A) Fluorescent probe BBcellProbe M61 indicating mPTP opening was detected by flow cytometry with the FL1-A: B525-FITC channel. (B) mPTP flow cytometry. (C) Flameng score and (D) transmission electron microscopy of H9c2 cells (magnification, ×8,000; scale bar, 2 μ m). Data are expressed as the mean ± SD (n=3). *** P<0.05. TSN, tanshinone IIA; A/R, Anoxia/reoxygenation; VDAC1, Voltage-dependent anion channel 1; mPTP, Mitochondrial permeability transition pore; NC, negative control.

Article Snippet: Next, CMV-ADV6-VDAC1 was transduced into 293 cells (ATCC, cat. no. CRL-1573) using RNAi-mate (Suzhou GenePharma Co., Ltd.) and a 2nd generation system under 95% O 2 and 5% CO 2 at 37°C for 6 h (quantity of adenovirus plasmid used for transfection is 10 μ g and the ratio used for adenovirus, packaging and envelope plasmids is 1:3:4).

Techniques: Flow Cytometry, Transmission Assay, Electron Microscopy, Permeability, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

doi: 10.3892/ijmm.2023.5312

Figure Lengend Snippet: TSN inhibits apoptosis of A/R-induced H9c2 cardiomyocytes by downregulating VDAC1. (A) Expression of (B) apoptosis-associated proteins was detected by western blot analysis in A/R-induced cells following TSN, pAd/VDAC1 and pAd/NC pretreatment. (C) Caspase-3 activity was measured using a Caspase-3 kit in A/R-induced cells after TSN, pAd/VDAC1 and pAd/NC treatment. (D) MMP and (E) apoptosis were detected by flow cytometry. (F) MMP levels detected by JC-1 in H9c2 cells indicated by the red/green fluorescence ratio. (G) Apoptotic rate measured by Annexin V-FITC/PI flow cytometry. Data are expressed as the mean ± SD (n=3). *** P<0.05. TSN, Tanshinone IIA; A/R, Anoxia/reoxygenation; VDAC1, Voltage-dependent anion channel 1; NC, negative control; MMP, mitochondrial membrane potential.

Article Snippet: Next, CMV-ADV6-VDAC1 was transduced into 293 cells (ATCC, cat. no. CRL-1573) using RNAi-mate (Suzhou GenePharma Co., Ltd.) and a 2nd generation system under 95% O 2 and 5% CO 2 at 37°C for 6 h (quantity of adenovirus plasmid used for transfection is 10 μ g and the ratio used for adenovirus, packaging and envelope plasmids is 1:3:4).

Techniques: Expressing, Western Blot, Activity Assay, Flow Cytometry, Fluorescence, Negative Control, Membrane

Journal: International Journal of Molecular Medicine

Article Title: Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

doi: 10.3892/ijmm.2023.5312

Figure Lengend Snippet: Potential mechanism of TSN in myocardial ischemia/reperfusion injury. TSN pretreatment upregulates the expression of VDAC1, thereby decreasing the accumulation of ROS and iron and abnormal lipid metabolism, maintaining mitochondrial function and protecting the myocardium against anoxia/reoxygenation-induced ferroptosis and apoptosis. TSN, tanshinone IIA; VDAC1, Voltage-dependent anion channel 1; ROS, reactive oxygen species; MDA, malondialdehyde; GSH, Glutathione; GSSG, Glutathione disulfide; LDH, lactate dehydrogenase; MMP, Mitochondrial membrane potential.

Article Snippet: Next, CMV-ADV6-VDAC1 was transduced into 293 cells (ATCC, cat. no. CRL-1573) using RNAi-mate (Suzhou GenePharma Co., Ltd.) and a 2nd generation system under 95% O 2 and 5% CO 2 at 37°C for 6 h (quantity of adenovirus plasmid used for transfection is 10 μ g and the ratio used for adenovirus, packaging and envelope plasmids is 1:3:4).

Techniques: Expressing, Membrane

Journal: iScience

Article Title: Glycosylated Delta-receptor-binding domain mucosal vaccine elicits broadly neutralizing antibodies with protection against SARS-CoV-2 challenge

doi: 10.1016/j.isci.2023.108033

Figure Lengend Snippet:

Article Snippet: Each of the above plasmids was respectively co-transfected with PS-PAX2 and pLenti-CMV-luciferase plasmids into 293T cells (ATCC) using the Polyetherimide (PEI) transfection method, followed by collection of pseudovirus-containing supernatants 72 h post-transfection.

Techniques: Recombinant, Software, Virus, Variant Assay, Adjuvant, Luciferase