Simultaneous induction of apoptosis and necroptosis by Tanshinone IIA in human hepatocellular carcinoma HepG2 cells.

Tanshinone IIA (Tan IIA), a constituent of the traditional medicinal plant Salvia miltiorrhiza BUNGE, has been reported to possess anticancer activity through induction of apoptosis in many cancer cells. Surprisingly, the present study finds that Tan IIA simultaneously causes apoptosis and necroptosis in human hepatocellular carcinoma HepG2 cells. We further find that apoptosis can be converted to necroptosis by pan-caspase inhibitor Z-VAD-fmk, and the two death modes can be blocked by necroptotic inhibitor necrostatin-1. The underlying mechanisms are revealed by analysis of the signaling molecules using western blotting. In control cells, FLICE inhibitory protein in short form (FLIPS) is expressed in relatively high levels and binds to caspase 8 in ripoptosome, which supposedly sustains cell survival. However, in Tan IIA-treated cells, FLIPS is down-regulated and may thus cause homodimer formation of cleaved caspase 8, cleavage of receptor-interacting serine/threonine-protein kinases 1, 3 (RIP1, RIP3), and mixed-lineage kinase domain-like (MLKL), in turn leads to cell apoptosis. In parallel, Tan IIA causes necroptosis by forming a suggested necrosomal complex composed of RIP1/RIP3. Regarding the inhibitors, z-VAD-fmk diminishes the cleaved caspase 8, RIP1, RIP3, and MLKL induced by Tan IIA, and reconstructs the ripoptosome complex, which marks cells moving from apoptosis to necroptosis. Nec-1 recovers the Tan IIA down-regulated FLIPS, consequently causes FLIPS to form heterodimer with caspase 8 and thus block apoptosis. Meanwhile, cleaved forms of RIP1 and RIP3 were observed preventing necroptosis. Intriguingly, the cytotoxicity of tumor necrosis factor-related apoptosis-inducing ligand to HepG2 cells is enhanced by Tan IIA in a pilot study, which may be attributed to low FLIPS levels induced by Tan IIA. In short, Tan IIA simultaneously induces both Nec-1 inhibition and FLIPS regulation-mediated apoptosis/necroptosis, which has not been previously documented. Moreover, the involvement of the cleavage type of MLKL in executing necroptosis warrants further investigation.

Effects of glycyrrhizae and glycyrrhizic acid on cellular immunocompetence in low-dose gamma-ray irradiated mice.

BACKGROUND: For both animals and human beings, it is important to prevent damage from ionizing radiation and to restore immunocompetence following irradiation. The present study was conducted to investigate the effects of glycyrrhizae (GL) and glycyrrhetinic acid (GA) on cellular immunocompetence in low dose gamma-ray-irradiated mice. METHODS: Six- to 8-week-old ICR strain' Crl:CD-1-ICR (BR) strain male mice, bred in the Institute of Cancer Research, U.S.A., were chosen and divided into four groups. Group A was the normal control. Group B, the experimental control, received 1 Gy of whole body gamma-ray irradiation. Groups C and D, the experimental groups, were treated with 500 mg/kg of GL (orally) and 5 mg/kg body weight of GA (i.p.), respectively, once a day, 5 days a week for 2 weeks after gamma-irradiation. The tested mice were killed, at 6 different intervals to measure their leukocyte and differential counts. Cellular immunocompetence was measured by the 3H-thymidine uptake in each group. RESULTS: One gray of gamma-ray irradiation had evident inhibition on the leukocyte and differential counts and the cellular immunity of mice. GL and GA could help to restore the decreased leukocyte counts and the cellular immunocompetence in low dose gamma-irradiated mice. CONCLUSION: GL and GA could help to restore decreased leukocyte counts and the cellular immunocompetence in low-dose gamma-ray-irradiated mice.

Therapeutic effects of moxibustion on experimental tumor.

This work investigated the therapeutic effects of the improved form of moxibustion (MT) on experimental tumor. Sarcoma 180 cells (1 x 10(7)) were transplanted into the subcutaneous tissue in the breast area of female ICR mice. Mice bearing a tumor were divided into one control and four experimental groups. The experimental groups were treated with MT for 1, 2, 3 and 4 times (abbreviated as MT1, MT2, MT3, MT4, respectively). This study showed that the experimental group treated with MT3 displayed the optimal therapeutic response. The longest mean survival time (87.8 days) within 120 days after treatment of MT3 significantly differed from the control group (60.2 days). In addition, uptake of 86Rb-radioactive tracer significantly decreased in tumors treated with MT3. The improved form of moxibustion used in this study is a reliable model of localized hyperthermia in tumor therapy.

Effects of gypenosides on cellular immunity of gamma-ray-irradiated mice.

OBJECTIVE: To investigate the effects of gypenosides (Gs) on cellular immunocompetence in the gamma-ray-irradiated mice. MATERIALS AND METHODS: Tested mice of ICR strain were treated continuously with Gs for 10 days before or after 4 Gy gamma-irradiation. Body weight and splenic weight of mice were measured. The blastogenic response of splenocytes to mitogens, such as PHA, Con A and LPS were also detected. The cellular immunocompetence was measured by 3H-thymidine incorporation in each tested mouse. RESULTS: Body weight as well as splenic weight decreased in gamma-ray-irradiated mice. The blastogenic responses of splenocytes to mitogens were inhibited after gamma-ray irradiation. To treat with Gs was effective to enhance the recovery of body weight, splenic weight and immunocompetence in gamma-ray-irradiated mice from radiation damage. CONCLUSIONS: Four Gy gamma-ray irradiation could decrease splenic weight and cellular immunocompetence of mice. Gs could help the recovery of the splenic weight and cellular immunocompetence in gamma-ray-irradiated mice.

Effects of glycyrrhizae and glycyrrhizic acid on cellular immunocompetence of gamma-ray-irradiated mice.

OBJECTIVE: To investigate the effects of glycyrrhizae (GL) and glycyrrhizic acid (GA) on cellular immunocompetence in the gamma-ray-irradiated mice. MATERIALS AND METHODS: Tested mice of ICR strain were treated with GL and GA for 10 days continuously after 4 Gy gamma-irradiation. The counts of total leukocytes, lymphocytes, neutrophils and monocytes in peripheral blood of mice were measured. The blastogenic responses of splenocytes to mitogens, such as PHA, PWM, Con A and LPS were also detected. The cellular immunocompetence was measured by 3H-thymidine incorporation in each tested mouse. RESULTS: The leukocytes and their differential counts decreased in gamma-irradiated mice. The blastogenic responses of splenocytes to mitogens were inhibited by gamma-ray irradiation. GL and GA were effective in enhancing the recovery of leukocyte count and the blastogenic responses of splenocytes to mitogens, such as PHA, PWM, Con A and LPS from the inhibition of 4 Gy gamma-irradiation. CONCLUSIONS: Four Gy gamma-ray irradiation can decrease leukocytes and their differential counts, and the blastogenic responses of splenocytes of mice. GL and GA can help the recovery of the cellular immunocompetence in the gamma-irradiated mice.

Effects of Glycyrrhizae and Glycyrrhizic acid on radiation injury in mice.

The effects of Glycyrrhizae (GL) and Glycyrrhizic acid (GA) on promoting the recovery of mice from radiation injury were investigated. Two hundred ICR strain male mice, 6-8 weeks old, were randomly divided into four groups. Group A was the normal control. Group B, the experimental control, was treated with 4 Gy gamma-ray irradiation. Groups C and D, the experimental groups, were treated with 500 mg/kg of GL (oral administration) and 5 mg/kg body weight of GA (i.p.), respectively, for ten days after gamma-ray irradiation. Eight mice in each group were sacrificed on days 5, 12, 26 and 33 post irradiation to measure the weights of spleen, thymus and testes, and the biosynthetic rates of DNA in these organs. The results revealed that 4 Gy gamma-ray irradiation evidently inhibited the weights of spleen, thymus and testes, and their DNA biosynthetic rates. GL and GA enhanced the recovery of these organs in mice from the injury of gamma-ray irradiation.

Protective effects of Gynostemma pentaphyllum in gamma-irradiated mice.

Radiation protective effects of Gynostemma Pentaphyllum (Gp) were investigated in gamma-irradiated mice. Animals were sacrificed on days 5, 15, 25 and 35 after gamma-irradiation. GOT, GPT, serum IgG and leukocyte counts were measured. Proliferation of splenocytes stimulated by mitogens, such as PHA, Con A, and LPS were detected and compared. The results showed that all parameters measured in this study were decreased and proliferation of splenocytes stimulated by mitogens were repressed in gamma-irradiated mice. Gp helped to recover the decreased leukocyte counts, GOT, GPT and IgG in serum and the proliferation of splenocytes stimulated by PHA, LPS and Con A in the gamma-ray irradiated mice.

Effects of Ganoderma lucidum and krestin on subset T-cell in spleen of gamma-irradiated mice.

Effects of Ganoderma lucidum (Gl) and Krestin (PSK) extracts on spleen, thymus and splenocytes in gamma-irradiated mice were investigated in this study. ICR strain male mice were divided into five groups. Group A was the normal control. Group B, the experimental control, was treated with Gl. Group C, the radiation treatment control, was treated with whole body exposure to 4 Gy gamma-irradiation (RT). Group D was treated with RT and Gl. Group E was treated with RT and PSK. The dosage of Gl was 400 mg/day/kg body weight and PSK was 500 mg/day/kg body weight. Our results indicated that the relative thymus weight in groups D and E were higher than group C on day 28 after gamma-irradiation. Group D was the highest in all the experimental groups. CD4 and CD8 splenocytes in group D were higher than group C on days 7 and 28. Gl was better than PSK in repairing the damage of subset T-cells in the spleen of gamma-irradiated mice.