Intracellular GSH and ROS levels may be related to galactose-mediated human lens epithelial cell apoptosis: Role of recombinant hirudin variant III

Add time:07/29/2019    Source:sciencedirect.com

Oxidative processes in the lenses are the most commonly found damaging factor for the development of cataracts. Hirudin, a most potent inhibitor of thrombin as an antithrombic drug, also have potential use in cataracts. In order to investigate the mechanisms of hirudin against galactose-induced cataract at the cellular level. We used recombinant hirudin variant III (rHV3) to study the protective effect of hirudin on galactose-mediated human lens epithelial cells injury. The human lens epithelial cells (hLECs) were cultured in D/F12–10% FBS medium containing 125 mM d-galactose with or without rHV3. Cell viability was assessed by methylthiazol tetrazolium (MTT) assay and propidium iodide (PI) staining in situ. Cell apoptosis was elevated with comet assay (single cell gel electrophoresis, SCGE), AO/EB double staining and Annexin-V/PI double staining assay. Reactive oxygen species (ROS) were quantified with 2′,7′-dichlorofluorescein (DCF), and free glutathione (GSH) levels were measured with a commercial GSH quantification kit. Decreased viability and increased apoptosis of the hLECs were observed when incubated with 125 mM galactose. These hLECs also demonstrated the increased presence of ROS, whereas GSH was reduced. rHV3 blocked the induction of cell death, apoptosis and oxidative stress in hLECs. One mechanism may be through regulating intracellular ROS and GSH levels to inhibit apoptosis of the human lens epithelial cells.

We also recommend Trading Suppliers and Manufacturers of hirudin HV3 (cas 134289-40-0). Pls Click Website Link as below: cas 134289-40-0 suppliers

Prev:High cell density cultivation of recombinant Escherichia coli for hirudin variant 1 production
Next:Investigation on recombinant hirudin via oral route)