Aim To investigate the expression of G protein-coupled receptor TGR5 and its effects on FN and TGF-β1 expression cultured under high glucose condition in rat glomerular mesangial cells , and then to explore the role of TGR5 in diabetic nephropathy. Methods INT-777 and TGR5 plasmid were used to activate TGR5 under high glucose(HG,30 mmol·L - 1 glucose ) condition, and anti-TGR5 small interfering RNA(TGR5 siRNA) was used to knock down TGR5. The protein expression of FN and TGF-β1 in rat me-sangial cells was detected by Western blot. Results TGR5 could be detected in rat glomerular mesangial cells. Both FN and TGF-β1 protein levels could be in-creased by high glucose compared with control group(P < 0. 05),and be inhibited by activiation of TGR5(P <0. 05). On the other hand,knockdown of TGR5 could increase FN and TGF-β1 protein to abnormal levels(P< 0. 01,P < 0. 05). Conclusion TGR5 suppresses HG-induced FN and TGF-β1 expression in rat glomer-ular mesangial cells,suggesting a protective role in the process of diabetic nephropathy.

A poly( GMA-EGDMA) coated SPME fiber was prepared using an in-situ polymerization by direct bonding to the surface of a polydopamine-modified stainless steel wire. Then the fiber was modified by sulfuric acid. A novel solid phase microextraction coating coupled to high performance liquid chromatography ( HPLC) method based on the as-prepared fiber was developed for the determination of four pharmaceuticals and personal care products ( PPCPs) in water samples. The influences of extraction parameters, including pH, extraction time, extraction temperature and salt addition were investigated. 3 mL water sample was extracted by the as-prepared fiber for 60 min at 30 ℃, and then desorbed with mobile phase for 30 min, respectively. Desorption solution was analyzed by HPLC-DAD ( diode array detection ) . The results indicated that the extraction yield of the fiber was good for four PPCPs. The linear correlation coefficients were>0. 997 with the linear range of 2-200 μg/L. The limits of detection (S/N=3) were 0. 5-5 μg/L with RSD (n=5) of 4. 1%-11. 9%. The recoveries of four PPCPs at spiked level of 20, 50, 100 μg/L were within the range of 70. 6%-105. 5%. The results showed that this method was easy, green, accurate and precise, and could be used to assay the four PPCPs in real water samples.

Macroautophagy (referred to as autophagy hereafter) is a major intracellular lysosomal degradation pathway that is responsible for the degradation of misfolded/damaged proteins and organelles. Previous studies showed that autophagy protects against acetaminophen (APAP)-induced injury (AILI) via selective removal of damaged mitochondria and APAP protein adducts. The lysosome is a critical organelle sitting at the end stage of autophagy for autophagic degradation via fusion with autophagosomes. In the present study, we showed that transcription factor EB (TFEB), a master transcription factor for lysosomal biogenesis, was impaired by APAP resulting in decreased lysosomal biogenesis in mouse livers. Genetic loss-of and gain-of function of hepatic TFEB exacerbated or protected against AILI, respectively. Mechanistically, overexpression of TFEB increased clearance of APAP protein adducts and mitochondria biogenesis as well as SQSTM1/p62-dependent non-canonical nuclear factor erythroid 2-related factor 2 (NRF2) activation to protect against AILI. We also performed an unbiased cell-based imaging high-throughput chemical screening on TFEB and identified a group of TFEB agonists. Among these agonists, salinomycin, an anticoccidial and antibacterial agent, activated TFEB and protected against AILI in mice. In conclusion, genetic and pharmacological activating TFEB may be a promising approach for protecting against AILI.