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Our previous study showed that docosahexaenoic acid (DHA) plays an important role in decreasing lipid accumulation by inducing apoptosis of the adipocytes in grass carp. However, the mechanism involved remains unclear. DHA has been reported as the natural ligand of PPARγ. The present study aimed to assess whether PPARγ mediates the pro-apoptotic effects by DHA. Adipocytes of grass carp were cultured until 2 days post-confluence and were treated with DHA at various concentrations—0, 25, 50, 100, 200, and 400 μmol/L for 24 h and at 200 μmol/L for various time periods (0, 12, 24, and 48 h, respectively). Besides, the adipocytes were exposed to 200 μM DHA and PPARγ antagonist or inhibitor of certain key enzymes of apoptosis, following which the expression levels of key genes of the cell apoptotic and mitochondrial apoptotic pathways were detected. We found that DHA induced apoptosis of grass carp adipocytes in a time- and dose-dependent manner (P < 0.05). In addition, DHA treatment significantly increased the protein and gene expression levels of PPARγ (P < 0.05), but the PPARγ antagonist significantly abolished this effect and the DHA pro-apoptotic effect (P < 0.05). Moreover, treatment with caspase 9 inhibitor significantly attenuated the DHA-induced preadipocytes apoptosis effects, while treatment with caspase 8 inhibitor showed no influence. These observations suggest that the DHA-induced apoptosis in adipocytes might be mediated by PPARγ and via the intrinsic apoptotic pathway in grass carp. [ABSTRACT FROM AUTHOR]

Dibutyl phthalate (DBP) is a typical phthalate (PAEs). The environmental health risks of DBP have gradually attracted attention due to the common use in the production of plastics, cosmetics and skin care products. DBP was associated with diabetes, but its mechanism is not clear. In this study, an in vitro culture system of rat insulinoma (INS-1) cells was established to explore the effect of DBP on insulin synthesis and secretion and the potential mechanisms. INS-1 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and treated with 15, 30, 60 and 120 μmol/L of DBP and dimethyl sulfoxide (vehicle, < 0.1%) for 24 h. The contents of insulin in the intracellular fluid and the extracellular fluid of the cells were measured. The results showed that insulin synthesis and secretion in INS-1 cells were significantly decreased in 120 μmol/L DBP group. The apoptosis rate and mitochondrial membrane potential of INS-1 cells were measured by flow cytometry with annexin V-FITC conjugate and PI, and JC-1, respectively. The results showed that DBP caused an increase in the apoptosis rate and a significant decrease in the mitochondrial membrane potential in INS-1 cells in 60 μmol/L and 120 μmol/L DBP group. The results of western blot showed that the expression of Bax/Bcl-2, caspase-3, caspase-9 and Cyt-C were significantly increased. Meanwhile, the level of oxidative stress in INS-1 cells was detected by fluorescent probes DCFH-DA and western blot. With the increase of DBP exposure, the oxidative stress levels (MDA, GSH/GSSG) were increased; and the antioxidant index (SOD) levels were decreased. Our experimental results provide reliable evidence that DBP induced apoptosis and functional impairment in INS-1 cells through the mitochondrial apoptotic pathway and oxidative stress. Therefore, we hypothesized that interference with these two pathways could be considered in the development of preventive protection measures. [Display omitted] • DBP reduced the insulin synthesis and secretion of INS-1 cells. • DBP altered the expression of GLUT2 and PDX-1 associated with insulin synthesis and secretion in INS-1 cells. • DBP induced the mitochondrial apoptotic pathway and oxidative stress in INS-1 cells. [ABSTRACT FROM AUTHOR]

Betulinic acid (BA), an occurring pentacyclic triterpenoid, has various biological activities, such as anti-inflammation and antioxidation. Previous studies found that BA attenuated cyclophosphamide (CYP)-induced intestinal mucosal damage by inhibiting intestinal mucosal barrier dysfunctions and cell apoptosis. However, the effects and regulation mechanisms of BA on CYP-induced renal damage has not been reported in literature. Here, we found that BA pretreatment alleviated the elevation of serum urea level and inhibited the increase in serum neutrophil gelatinase-associated lipocalin level induced by CYP. Meanwhile, BA ameliorated renal tubular epithelial cell edema, and vacuolization of renal cortical tubular and renal glomerulus. Moreover, pretreatment with BA inhibited the mRNA expressions of pro-inflammatory cytokines interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α, and increased mRNA expressions of anti-inflammatory cytokines such as IL-10 and transforming growth factor-β by inactivation nuclear factor kappa-B. Simultaneously, BA decreased the accumulation of reactive oxygen species and malondialdehyde, and lowered the levels of superoxide dismutase and glutathione, while increased the activity of glutathione peroxidase in CYP-induced kidney damage mice. Besides, BA reduced the phosphorylation of extracellular signal-regulated kinases (ERK), inhibited the ratio of Bcl-2/Bax and cell apoptosis in CYP-triggered kidney damage. Furthermore, BA and/or PD98059 (an inhibitor of ERK) regulated mitigation of CYP-elicited renal injury and deactivation of the ERK pathway and mitochondrial apoptotic pathway, indicating that the protective effect of BA on CYP-induced renal damage may be associated with the down-regulation of ERK-mediated mitochondrial apoptotic pathway. Thus, BA could be a candidate agent against chemotherapy drug-induced nephrotoxicity by reducing inflammation and oxidative stress through suppression of ERK-mediated mitochondrial apoptotic pathway. [Display omitted] • BA alleviated CYP-induced renal damage by suppressing inflammation and oxidative stress. • BA reduced inflammation by inactivating NF-κB pathway. • BA reduced oxidative stress by inhibiting ERK-mediated mitochondrial apoptotic pathway. [ABSTRACT FROM AUTHOR]

Highlights: [•] Bmbuffy is located on the outer-membrane of mitochondria and ER. [•] Bmbuffy inhibits apoptosis in HCPT-induced BmN-SWU1 cells. [•] In the absence of transmembrane domain, Bmbuffy has no effects on apoptosis. [•] Bmbuffy interacts with Bmp53 in HCPT-induced apoptosis. [•] Bmp53 over-expression promoted apoptosis can be blocked by Bmbuffy. [ABSTRACT FROM AUTHOR]

Highlights: [•] SAR induced apoptosis via the caspase-dependent mitochondrial apoptosis pathway. [•] SAR activated UPR signaling pathways. [•] SAR provoked the generation of ROS. [•] The generation of ROS was necessary for ER stress mitochondrial apoptotic pathways. [•] Sarsasapogenin (SAR) retarded the cell cycle at S and G2 phase. [Copyright &y& Elsevier]

Abstract: Doxorubicin (Dox) has demonstrated potent activity in treating malignant lymphomas but its therapeutic efficacy is hampered by induction of cardiotoxicity. This side effect is related to the ability of the drug to generate reactive oxygen species in cells. Previously, we demonstrated that coupling Dox to penetratin (Pen), a cell penetrating peptide, represent a valuable strategy to overcome drug resistance in CHO cells. In the present study, we evaluated the consequences of the conjugation of Dox to Pen in term of apoptosis induction. When tested on CHO cells, Dox–Pen generated a typical apoptotic phenotype but at lower dose that needed for unconjugated Dox. Cell death induction was associated with chromatin condensation, caspase activation, Bax oligomerisation and release of cytochrome c. By using reactive oxygen species and c-jun NH2-terminal kinase (JNK) inhibitors, we prevented Dox- and Dox–Pen-induced CHO cell death. The chimeric soluble DR5 receptor that inhibits TRAIL induced cell death does not prevent Dox or Dox–Pen-induced cytotoxicity. These observations indicate that conjugation of Dox to cell penetrating peptide does not impair the ability of the drug to trigger cell death through activation of the intrinsic pathway involving c-Jun NH2-terminal kinase but could exhibit less toxic side effects and could warrant its use in clinic. [Copyright &y& Elsevier]

[Display omitted] • Six new and six known lignans were isolated from the fruits of Leonurus japonicus. • New compounds 5 and 6 were a pair of unusual epimeric alkaloidal lignans. • Compounds 1 , 2 , 7 , and 8 increased the survival rates of injured HL-7702 cells. • Compounds 1 and 7 regulated the mitochondrial apoptotic pathway in HL-7702 cells. Twelve tetrahydrofuran lignans (1 – 12), including six new compounds (1 – 6), were isolated from the 70% EtOH extract of the fruits of Leonurus japonicus. Spectroscopic analyses and ECD and OR calculations were used to determine their structures. Compounds 5 and 6 were unusual alkaloidal lignans with a butyrolactam unit. Based on the beneficial effects of the fruits of L. japonicus (Chongweizi in Chinese) on the liver in traditional Chinese medicine (TCM), the hepatocyte protective activities of the isolates were studied by MTT, Hoechst 33,342 staining, and western blotting. The MTT results revealed that compounds 1 , 2 , 7 , and 8 significantly increased the survival rates of HL-7702 cells injured by acetaminophen, with EC 50 values of 10.41 ± 0.90 μM, 19.86 ± 3.13 μM, 9.68 ± 1.93 μM, and 21.35 ± 3.58 μM, respectively. In the Hoechst 33,342 fluorescence staining, compounds 1 and 7 suppressed the apoptosis of the injured HL-7702 cells. Furthermore, the western blot analysis showed that compounds 1 and 7 increased the Bcl-2/Bax protein expression ratio and procaspase-3 protein expression, indicating that compounds 1 and 7 may exert hepatoprotective activity by regulating the mitochondrial apoptotic pathway. [ABSTRACT FROM AUTHOR]