Ningxin-Tongyu-Zishen formula alleviates the senescence of granulosa cells on D-galactose-induced premature ovarian insufficiency mice.

Ningxin-Tongyu-Zishen formula (NTZF) is a clinical experience formula for the treatment of premature ovarian insufficiency (POI) in traditional Chinese medicine (TCM), and the potential mechanism is unknown. For in vivo experiments, POI mouse models (C57BL/6 mice), were constructed by subcutaneous injection of D-galactose (D-gal, 200 mg/kg). After treatment of NTZF (10.14, 20.27, 40.54 g/kg;) or estradiol valerate (0.15 mg/kg), ovarian function, oxidative stress (OS) and protein expression of Sirt1/p53 were evaluated. For in vitro experiments, H2O2 (200 µM) was used to treat KGN to construct ovarian granulosa cells (OGCs) cell senescence model. Pretreatment with NTZF (1.06 mg/mL) or p53 inhibitor (Pifithrin-α, 1 µM) was performed before induction of senescence, and further evaluated the cell senescence, OS, mRNA and protein expression of Sirt1/p53. In vivo, NTZF improved ovarian function, alleviated OS and Sirt1/p53 signaling abnormalities in POI mice. In vitro experiments showed that NTZF reduced the level of OS and alleviated the senescence of H2O2-induced KGN. In addition, NTZF activated the protein expression of Sirt1, inhibited the mRNA transcription and protein expression of p53 and p21. Alleviating OGCs senescence and protecting ovarian function through Sirt1/p53 is one of the potential mechanisms of NTZF in the treatment of POI.

[Enhancing effect of Tween 80 on degradation of triphenyltin by Bacillus thuringiensis].

So far, the information regarding enhanced degradation and biodegradation mechanisms of TPhT, an endocrine disruptor, is severely limited. Whether dearylation during TPhT degradation occurs successively or synchronously is not revealed clearly. To deal with these problems, this study focused on the biodegradation of TPhT and its metabolites by Bacillus thuringiensis through the acceleration of Tween 80. The results showed that Tween 80 obviously increased the TPhT solubility. After degradation by cells in the presence of 80 mg L-1 Tween 80 for 2 d, the residual TPhT at 1 mg L-1 initially was decreased to 48.4%. During the biodegradation process, Tween 80 significantly reduced intracellular Na+, NH+4: and Mg2+ release, and increased extracellular Cl- , PO(3-)4 and K+ utilization. Metabolites analysis revealed that phenyltin biodegradation initially proceeded by cleaving the aromatic ring, not by splitting the covalent bonds between the benzene rings and tin atom. Ring-cleavage reactions in the benzenes of TPhT occurred individually and synchronously, producing diphenyltin, monophenyltin and tin accordingly.