The Study Of Aldose Reductase Inhibitors And Activity Of Extracts From Natural Products

Natural products refer to plants, animals, minerals and other naturally occurringpharmacologically active natural products. Application of natural products has a longhistory in China, and compared to synthetic drugs, natural products are moreaccessible and less costly. A long time ago it was used as a natural product to treatdiabetes,and has a broad application base. Aldose reductase enzyme is a sign ofdiabetes complications,which is active in high glucose conditions, opening polyolpathway, catalyzed conversion of glucose into sorbitol, sorbitol accumulation in cellscause a variety of complications. So the aldose reductase is a natural target for drugscreening, hoping to get some of the aldose reductase inhibitors.By detecting aldose reductase NADPH coenzyme critical to measure theabsorbance change aldose reductase inhibition, screened on aldose reductaseinhibitory effect of the drug are prominent lens and mesangial cell culture, lens andglomerulars are more typical of diabetic complications affecting the site, throughphysiological and biochemical studies of these two parts to better reflect the naturalproducts of aldose reductase inhibitory activity. In this experiment, a plurality ofangles from natural drugs inhibit the complications of diabetes drug screening fordiabetes and its complications look for new drugs, especially natural products ofinhibitors which provide a method and basis.The experiment is divided into four parts:1. Aldose reductase extracted from fresh cow lens, in phosphate buffer manuallytriturated gradient salting out, centrifugation, dialysis, to obtain a crude enzyme, bymeasuring the activity and protein concentration measuring specific activity of theenzyme for the enzyme2.93U/mg, suitable for further experiments.2. Select the80kinds of natural products, extraction with water of activeingredients, from screening for aldose reductase inhibitors, get rhodiola, brucea, olive,silkworm, radix and other effects based on the value of screening than the IC50gooddrugs, respectively, with lens and HBZY-1cells in high glucose which were co-cultured, measuring soluble protein and a variety of antioxidant index,comprehensive evaluation of drugs on aldose reductase caused by diabetes mellitusdisease treatment.3. Drug co-cultured in high glucose lens observation of drug active role for thebody intuitively by paraffin. These drugs for the lens in high glucose can recover to acertain extent after injury.4. The high glucose and oxidative damage can lead to advanced glycation endproducts (AGEs), and AGEs will directly lead to various complications of diabetes,and the experiments are also discussed that aldose reductase inhibition with lowerIC50value drugs of rhodiola, brucea, olive can also inhibite advanced glycation endproducts, consistent with aldose reductase inhibitors.