To study the effect of Apigenin (AP) on the proliferation, cell cycle and apoptosis of mouse T cells in vitro.
The lymphocytes were prepared from lymph nodes and thymus of mice. The effect of AP on the proliferation of T in response to ConA at different concentrations (25, 50, 100, 150, 200 micromol/L) was detected by MTT. Cell cycle was measured by PI staining and FCM. The effect of Apigenin and Apigenin with DEX on T cell apoptosis was measured by Annexin V-FITC/PI double staining and FCM. The effect of different concentrations of AP cytotoxicity to T cells was measured by MTT.
25-200 micromol/L of AP didn't have cytotoxicity to T cells, but it had some inhibitory effect on T cells in response to ConA(P<0.01), arresting cell cycle at G0/G1 in a dose-dependent manner. Different concentrations of AP inhibited the apoptosis of T cells, especially those induced by DEX(P<0.01).
AP can inhibit the proliferation of mouse T cells in response to ConA, arrest cell cycle at G0/G1 and inhibit the apoptosis of T cells.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 04/2008; 24(4):337-40.
To study the changes of mitochondrial potential (delta psi m) and cardiolipin (CL) content during thymocyte apoptosis mediated by nitric oxide (NO).
SNAP was used as NO's donor to induce thymocyte apoptosis in mice and dexamethasone (DEX) was used as positive control drug. Three experiment groups were set, which were blank control group, SNAP group and DEX group. The ectropion of cell phosphatidylserine (PS) was detected by flow cytometry after stained with FITC-anti-annexin V mAb/PI. The changes of delta psi m and CL content during cell apoptosis were detected by DiOC6(3)/PE-anti-annexin V mAb and NAO/PE-anti-annexin V mAb, respectively.
6 hours after SNAP treatment, the thymocytes exhibited typical cell apoptotic characteristics and crenation appeared in most annexin V-positive cells. In DEX group, the delta psi m decreased and the rate of non-apoptotic cells was significantly higher than that in blank control group, but there was no significant difference between SNAP and control groups. 40%-50% of DiOC6(3)-negative cells in each group had the same size as normal cells. The percentage of the cells with reduced CL content in SNAP group was significantly higher than that in blank control group (P<0.01). Non-apoptotic cells with reduced CL content were not found. In blank control group and SNAP group, percentage of normal-sized cells with reduced CL was (48.32+/-3.96)% and (43.64+/-4.90)%, respectively.
The process of thymocyte apoptosis mediated by NO in mice includes successively PS ectropion, mitochondrial depolarization, CL oxidation and cell crenation. As compared with DEX group, the mitochondrial change mediated by NO is a later event in cell apoptosis.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 06/2006; 22(3):302-5.
To study the changes of mitochondrial mass and membrane potential (deltapsi(m)) during apoptosis of human promyelocytic leukemia cells (HL-60) induced by camptothecine(CPT).
HL-60 cells were stimulated with 4x10(-6) mol/L CPT. Apoptosis and necrosis of HL-60 cells were detected by annexin V-FITC/PI staining. Mitochondrial mass and membrane potential (deltapsi(m)) were measured by NAO and DiOC(6)(3) stanings, respectively.
After 12 hour treatment with 4x10(-6) mol/L CPT, the early apoptotic rate of HL-60 cells was significantly increased compared with the control group (12.75+/-4.61)% vs (2.88+/-2.49)%,(P<0.01), and the necrotic rate was significantly elevated too, (3.48+/-1.67)% and (0.71+/-1.10)%, (P<0.01). The result of PI staining showed that the apoptotic rate of HL-60 cells at late phase (12 h) in CPT group was (3.52+/-1.07)%, whereas that in control group was (0.46+/-1.06)%. At the same time, we observed that the cells in G(2)/M phase were arrested. The percentage of cells in G(2)/M phase in CPT group and the control was (13.45+/-1.91)% and (22.46+/-2.19)% (P<0.01) respectively. The percentage of cells with low mitochondrial mass in CPT and control groups was (25.74+/-2.09)% and (4.53+/-1.26)%, respectively (P<0.01). Mitochondrial membrane potential in CPT group and control group was (17.71+/-5.23)% and (1.64+/-2.00)%, respectively.
During CPT-induced apoptosis of HL-60 cells, mitochondrial mass and membrane potential drop significantly, but its depolarization heightens.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 04/2006; 22(2):144-7.