Inhibitory Effect of Rotundarpene on Parkinsonian Neurotoxin 1-Methyl-4-Phenylpyridinium-Induced Apoptotic Cell Death

BACKGROUND: The extract and hemiterpene glycosides of Ilex Rotunda Thunb exert antioxidant and anti-inflammatory effects. The effect of rotundarpene on apoptosis in neuronal cells caused by the 1-methyl-4-phenylpyridinium (MPP⁺) has not been reported previously. METHODS: Using differentiated PC12 cells and human neuroblastoma SH-SY5Y cells, we investigated the effect of rotundarpene on MPP⁺-caused apoptosis in relation to the cell death process. RESULTS: MPP⁺-induced cell death was identified using the MTT and neutral red uptake tests. Apoptosis was induced by eliciting decreases in the cytosolic levels of Bid and Bcl-2 proteins, increases in the cytosolic levels of Bax and p53, disruption of the mitochondrial transmembrane potential, and the release of cytochrome c and the activation of caspase-8, -9, and -3 in differentiated PC12 cells and SH-SY5Y cells. Treatment with rotundarpene reduced the MPP⁺-induced changes in the levels of apoptosis-regulated proteins, formation of reactive oxygen species, depletion and oxidation of glutathione, and cell death in both PC12 and SH-SY5Y cells. CONCLUSIONS: Rotundarpene may reduce MPP⁺-induced apoptosis in neuronal cells by suppressing the activation of the mitochondria-mediated pathway and the caspase-8 and Bid pathways. Rotundarpene appears to act by inhibiting the production of reactive oxygen species and by the depletion and oxidation of glutathione.

Inhibition of Zn(PMFPCl) 2 on HepG2 cells and its mechanism / 中国生化药物杂志

Objective To explore the inhibition of Zn(PMFPCl) 2 on HepG2 cells and its mechanism.Methods The HepG2 cells were divided into control group and experimental group of 10, 30 and 70 μmol/L.The cell proliferation was detected by MTT assay, cell apoptosis and cell cycle was analysed by flow cytometry, cellular morphological change was observed with inverted microscope and the expressions of apoptosis-regulated proteins of p53, p21, caspase-3, bax and bcl-2 in HepG2 cells were detected by Western blot.Results The inhibitory rates of experimental groups (10, 30, 70μmol/L) at 24, 48 and 72h were significantly higher than those of control group (P<0.05), and the highest one was 63.29% of 70 μmol/L Zn (PMFPCl)2at 72 h.The apoptosis rates of each experimental group at 48h was significantly higher than that of control group (P<0.05).The cells were induced a remarkable G1 arrest by Zn(PMFPCl) 2 which could inhibit proliferation.The number of adherent cells reduced and cells shrank, convex on cytomembrane surface appeared and the cells changed to round and were brighter.Western blot results showed that the protein levels of p53, p21, caspase-3 and bax increased and bcl-2 decreased with the Zn(PMFPCl)2concentration increasing (P<0.05).Conclusion Zn(PMFPCl)2 could inhibit the proliferation and promote apoptosis of HepG2 cells whose mechanisms are promotation of p53, p21, caspase-3 and bax expressions and inhibition of bcl-2 expression.

Effect of Ruanjian Xiaoying Decoction on the Relative Expression of Cytokines and Apoptosis-regulated Proteins Fas/Fasl in Experimental Autoimmune Thyroiditis Rats / 中国中医药信息杂志

Objective To evaluate the therapeutic effect of Xiaoying Ruanjian Decoction on experimental autoimmune thyroiditis (EAT) from the levels of autoantibodies, interleukin-1, tumor necrosis factor-? and apoptosis-regulated proteins Fas/Fasl of EAT rats, and investigate the mechanism from the relation between cytokines and apoptosis-regulated proteins. Methods Sixty SD female rats of SPF level were divided into normal group and model group. An animal model of EAT in rats was developed by injecting thyroglobulin mixed with Freund’s adjuvant immunization and drinking periodate water. The rats modeled were divided into Chinese medicine group, western medicine group and model group. The levels of autoantibodies, interleukin-1, tumor necrosis factor-? were detected with enzyme-linked immunosorbent assay. The level change of Fas/Fasl in the thyroid of rat was observed with immunohistochemical staining. Results Ruanjian Xiaoying Decoction significantly reduced the levels of autoantibody, interleukin-1, tumor necrosis factor-? and lowered the expression of Fas and Fasl in thyroid cells. Conclusion Ruanjian Xiaoying Decoction alleviates the levels of apoptosis-regulated proteins Fas/Fasl by decreasing the relative expression of antibodies and cytokines in serum levels, which plays a role in reducing apoptosis of thyroid cells.