Exposure to a low concentration of mixed organochlorine pesticides impairs glucose metabolism and mitochondrial function in L6 myotubes and zebrafish.

More realistic effects on glucose metabolic dysfunction can be evaluated by applying organochlorine (OCP) mixtures than individual OCPs. We formulated an equal ratio mixture of five OCPs (chlordane, heptachlor, p,p'-dichlorodiphenyltrichloroethane, ß-hexachlorocyclohexane, and hexachlorobenzene) and treated L6 myotubes with this OCP mixture to investigate effects on glucose uptake and the underlying mechanism. Exposure to the OCP mixture reduced 2-NBDG staining, representing glucose uptake, and stimulated the excessive production of reactive oxygen species (ROS). Reduced 2-NBDG uptake and ROS overproduction were compensated by insulin treatment. The expression of proteins such as IRß, PI3K, and AKT was downregulated, indicating that ROS overproduction contributed to the inhibition of insulin-dependent glucose uptake. Reduction in mitochondria quantity and decreased expression levels of PGC-1α, PDH, and GLUT4 proteins were observed, suggesting that mitochondrial dysfunction played a causative role in the disruption of glucose uptake. The inhibition of glucose uptake and ROS overproduction caused by the OCP mixture were also found in zebrafish as an in vivo model. We demonstrated that exposure to the OCP mixture, even at the lowest concentration, perturbed glucose uptake, which was associated with mitochondrial dysfunction, suggesting that an OCP mixture could be a potential environmental factor in type 2 diabetes-related effects on skeletal muscles.

Tris(1,3-dichloro-2-propyl)phosphate (TDCIPP) disrupts zebrafish tail fin development.

The flame retardant, tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), is one of the most developmentally toxic organophosphate flame retardants (OPFRs). However, few mechanistic studies on phenotypic malformation caused by TDCIPP have been conducted. This study investigates the molecular mechanism underlying abnormal tail fin development consistently observed in zebrafish embryos exposed to TDCIPP. The results show that the defects in the tail fin (e.g., bent spine, defective caudal fin, and damaged tip) were associated with altered expression of transcription factors. The significant up-regulation of mmp9 and, among insulin-growth factor (IGF) families, igfbp-1a and igfbp1b was observed, whereas alterations in the expression of cdx4, igf1a, ifg1b, igf2b, and vegaa regulating tail development were dependent on time points. In accordance with changes in mRNA gene expression, TDCIPP impaired vessel formation and disorganized muscle in transgenic Tg(fli-GFP) zebrafish larvae. Furthermore, we found that the overexpression of mmp9 caused by TDCIPP was not linked to igfbp-1. Overall, these findings demonstrate that TDCIPP disrupts the progression of tail fin development, accompanied by defects in vessel and muscle formation in developing zebrafish embryos.

Selaginellin and biflavonoids as protein tyrosine phosphatase 1B inhibitors from Selaginella tamariscina and their glucose uptake stimulatory effects.

As part of an ongoing search for new antidiabetic agents from medicinal plants, the methanol extract of the aerial parts of Selaginella tamariscina was found to possess stimulatory effect on glucose uptake in 3T3-L1 adipocyte cells. Thus, bioassay-guided isolation of this active extract yielded two new compounds (1 and 2) along with five known biflavonoids (3-7). Their structures were elucidated by extensive analysis of spectroscopic and physicochemical data. The absolute configuration of compound 2 was determined by specific rotation and CD data analysis. All isolates exhibited potent inhibitory effects on PTP1B enzyme with IC50 values ranging from 4.5±0.1 to 13.2±0.8µM. Furthermore, the isolates (1-7) showed significant stimulatory effects on 2-NBDG uptake in 3T3-L1 adipocyte cells. Of these, compounds (1, 6, and 7) which exhibited mixed-competitive inhibition modes against PTP1B, showed potent stimulatory effects on 2-NBDG uptake. This result indicated the potential of these biflavonoids as lead molecules for development of antidiabetic agents and the beneficial use of S. tamariscina against hyperglycemia.

Insulin-mimetic selaginellins from Selaginella tamariscina with protein tyrosine phosphatase 1B (PTP1B) inhibitory activity.

As part of an ongoing search for new antidiabetic agents from medicinal plants, three new (2, 4, and 5) and two known selaginellin derivatives (1 and 3) were isolated from a methanol extract of Selaginella tamariscina. The structures of the new compounds were determined by spectroscopic data analysis. All isolates showed strong glucose uptake stimulatory effects in 3T3-L1 adipocyte cells at a concentration of 5 µM. Furthermore, these compounds were found to possess inhibitory effects on PTP1B enzyme activity with IC50 values ranging from 4.6 ± 0.1 to 21.6 ± 1.5 µM. Compound 2 showed the greatest potency, with an IC50 value of 4.6 ± 0.1 µM, when compared with the positive control (ursolic acid, IC50 = 3.5 ± 0.1 µM). Therefore, these selaginellin derivatives may have value as new lead compounds for the development of agents against type 2 diabetes.