This study was undertaken to isolate and characterize the protease activity of human eye lens sample of mature and hyper mature cataract. Samples were collected just after surgery of the cataract lens and were stored at -20 degrees C. The total protein extract was isolated from 5 samples in each case (mature and hyper mature cataract) and clear supernatant obtained after centrifugation was used as an enzyme source. The optimum pH for the proteases of mature cataract was 7.5 while the proteases of hyper mature cataract were recorded for maximum activity at pH 5.5 and 7.5. The optimum temperature for both enzyme sources was 50 degrees C. Effect of different metal ions such as potassium, lead, silver, zinc and borate was studied. In each case protease activity was increased. Reducing agent e.g. beta mercaptoethanol also caused an increase in activity indicating the involvement of sulfhydryl groups. Protease activity was also located on agar plates.
Aged ; Amino Acids ; metabolism ; Cataract ; enzymology ; pathology ; Endopeptidases ; metabolism ; Enzyme Activation ; drug effects ; Humans ; Hydrogen-Ion Concentration ; Ions ; chemistry ; Metals ; chemistry ; pharmacology ; Middle Aged ; Substrate Specificity

OBJECTIVETo detect the expression of basic fibroblast growth factor (bFGF) in human ocular tissues, and to assess the effect of bFGF on the proliferation of human cataract lens epithelial cells (LECs) and its correlation with age.

METHODSEnucleated eyes were subjected to immunostaining for bFGF protein. Human cataract LECs were cultured in vitro, and treated with bFGF for 48 hr. Proliferation was estimated by the positive area ratio of proliferating cell nuclear antigen (PCNA) in immunohistochemistry.

RESULTSbFGF protein was found in various human ocular tissues. bFGF stimulated human cataract LEC proliferation, and there was an age-related decrease in responsiveness of human cataract LECs to bFGF (P < 0.05).

CONCLUSIONbFGF might play an important role in the proliferation of residual human cataract LECs after cataract surgery.

Adolescent ; Adult ; Age Factors ; Cataract ; metabolism ; pathology ; Child ; Child, Preschool ; Epithelial Cells ; chemistry ; pathology ; Fibroblast Growth Factor 2 ; biosynthesis ; Humans ; Immunohistochemistry ; Infant ; Infant, Newborn ; Lens, Crystalline ; chemistry ; pathology ; Middle Aged ; Proliferating Cell Nuclear Antigen ; analysis

We investigated whether amyloid beta (Abeta) aggregates have transforming growth factor beta- like cytokine activity and cause transdifferention of lens epithelial cells, leading to certain types of cataract. In order to mimic Abetaaggregates, Abeta- (1-40) was crosslinked to bovine serum albumin (BSA) with disuccinimidyl suberate according to a previously described procedure. When human lens epithelial B-3 (HLE B-3) cells were treated with the Abeta- (1-40) -BSA conjugates, we observed the translocation of Smad-3, as well as the induced mRNA levels of fibronectin (FN), collagen type I (Col I), smooth muscle actin (SMA) and matrix metalloproteinase-2 (MMP-2). In addition, we investigated the morphology of rat whole lens cultured for 5 days in the presence of Abeta- (1-40) -BSA, and the immunohistochemical localizations of Abeta- (1-40) /amyloid precursor protein (APP) in human clinical tissues beneath the anterior capsules. In rat whole lens cultures, treatment with Abeta- (1-40) -BSA produced a transformed morphology that had multiple layers of lens epithelial cells. To compare the anterior capsules in anterior subcapsular cataracts with those in nuclear cataracts, immunohistochemical studies of Abeta/APP in human clinical tissues revealed that the predominant immunostaining of Abeta occurs in the anterior epithelial plaques, which likely produces the abnormal extracellular matrix. Thus, these findings suggest that Abeta aggregates in vivo are possibly involved in the regulatory process by which lens epithelial cells may transdifferentiate into fibroblast-like cells, as well as help understand the mechanisms which lead to certain types of cataractogenesis.
Amyloid beta-Protein/*pharmacokinetics ; Animals ; Cataract/*metabolism/pathology ; Cell Differentiation ; Cell Line ; Epithelial Cells/*cytology/*metabolism ; Human ; Lens, Crystalline/*cytology ; Peptide Fragments/*pharmacokinetics ; Rats ; Serum Albumin, Bovine/*pharmacokinetics ; Support, Non-U.S. Gov't

OBJECTIVETo observe the the protective effection of polysaccharides-2b of mudan cortex of Paeonia suffruticosa andr (PSM2b) on diabetic cataract.

METHODThe animal model of diabetic cataract in rats was induced by streptozotocin (STZ) and freund's adjuvant complete (CFA). The initial opacity occurrence time in lens was investigated with cranny lamp, and opacity degree of lens was compared too. The activities of glutathione peroxidase (GSH-pX), superoxide dismutase (SOD), catalase (CAT), the content of malonaldehyde (MDA) in serum and lens were detected. At the same time, the activities of Na(+) -K(+) -ATPase, the content of macromolecular weight protein and infusibility protein in lens were detected too.

RESULTThe results examinated by cranny lamp showed that PSM2b could significantly postpone the occurrence and alleviate opacity degree of lens. Compared with model group, every treatment group of PSM2b could lower the level of MDA, high and middle dose groups could increase the levels of SOD, GSH-pX, CAT in serum and lens in evidence, and enhance the activity of Na(+) -K(+) -ATPase. These indexes present favorable positive correlation between dose and effect.

CONCLUSIONAll these results demonstrated that PSM2b had apparently protective effection on diabetic cataract in rats.

Animals ; Catalase ; blood ; metabolism ; Cataract ; etiology ; metabolism ; prevention & control ; Diabetes Mellitus, Experimental ; blood ; chemically induced ; complications ; Glutathione Peroxidase ; blood ; metabolism ; Lens, Crystalline ; drug effects ; metabolism ; pathology ; Male ; Malondialdehyde ; blood ; metabolism ; Paeonia ; chemistry ; Phytotherapy ; Plant Bark ; chemistry ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Streptozocin ; Superoxide Dismutase ; blood ; metabolism

Epalrestat is the unique aldose reductase inhibitor on the market, which was mainly used for the diabetic neuropathy. Lenses osmotic expansion could be induced by galactose to mimic the pathological process of diabetic cataract in vitro. In present study, we mainly investigated whether epalrestat possesses inhibitory effect on the lens osmotic expansion. The results indicated that epalrestat could not only markedly inhibit rat lens osmotic expansion in vitro, but also significantly reduced the high expression of the osmotic expansion-related genes such as AR and AQP1 in mRNA and protein levels. The findings may provide an important reference to epalrestat in the clinical application for the treatment of diabetic cataract.
Aldehyde Reductase ; antagonists & inhibitors ; biosynthesis ; genetics ; Animals ; Aquaporin 1 ; genetics ; metabolism ; Cataract ; etiology ; metabolism ; pathology ; Diabetes Mellitus, Experimental ; complications ; Enzyme Inhibitors ; pharmacology ; Galactose ; antagonists & inhibitors ; Lens, Crystalline ; drug effects ; metabolism ; pathology ; Male ; Osmosis ; drug effects ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rhodanine ; analogs & derivatives ; pharmacology ; Thiazolidines ; pharmacology

OBJECTIVETo evaluate the aldose reductase inhibitory (ARI) activity of different fractions of Hybanthus enneaspermus for potential use in diabetic cataract.

METHODSTotal phenol and flavonoid content of different fractions was determined. ARI activity of different fractions in rat lens was investigated in vitro.

RESULTSThe results showed significant level of phenolic and flavonoid content in ethyl acetate fraction [total phenol (212.15±0.79 mg/g), total flavonoid (39.11±2.27 mg/g)] and aqueous fraction [total phenol (140.62±0.57 mg/g), total flavonoid (26.07±1.49 mg/g)] as compared with the chloroform fraction [total phenol (68.56±0.51 mg/g), total flavonoid (13.41±0.82 mg/g)] and petrolium ether fraction [total phenol (36.68±0.43 mg/g), total flavonoid (11.55±1.06 mg/g)]. There was a significant difference in the ARI activity of each fraction, and it was found to be the highest in ethyl acetate fraction [IC50 (49.26±1.76 µg/mL)] followed by aqueous extract [IC50 (70.83±2.82 µg/mL)] and it was least in the petroleum ether fraction [IC50 (118.89±0.71 µg/mL)]. Chloroform fraction showed moderate activity [IC50 (98.52±1.80 µg/mL)].

CONCLUSIONSDifferent fractions showed significanct amount of ARI activity, where in ethyl acetate fraction it was found to be maximum which may be due to its high phenolic and flavonoid content. The extract after further evaluation may be used in the treatment of diabetic cataract.

Aldehyde Reductase ; antagonists & inhibitors ; Animals ; Cataract ; drug therapy ; prevention & control ; Diabetes Complications ; drug therapy ; prevention & control ; Diabetes Mellitus ; pathology ; Flavonoids ; analysis ; Lens, Crystalline ; enzymology ; Phenols ; analysis ; Plant Extracts ; pharmacology ; Rats ; Rats, Wistar ; Violaceae ; metabolism