Mutant cell lines resistant to azetidine carboxylic acid: Quantitative and qualitative differences in pyrroline-5-carboxylate synthase activity

Mutant Chinese hamster lung fibroblasts were selected that are resistant to the proline analog L-azetidine-2-carboxylic acid. Resistance in the two mutant cell lines is associated with two distinct alterations in pyrroline-5-carboxylate synthase, the enzyme that catalyzes the proline biosynthetic step leading from glutamic acid to pyrroline-5-carboxylate. In one mutant cell line, pyrroline-5-carboxylate synthase specific activity is increased 30-fold over the level in control cells. In the other mutant line, pyrroline-5-carboxylate synthase activity is not increased, but the enzyme has become insensitive to inhibition by ornithine and proline.     

Downregulation of TRPC6 expression is a critical molecular event during FK506 treatment for overactive bladder

PurposeIt has been suggested that FK506 could improve some symptoms of OAB in both clinical settings and animal models; however, its mechanism of action is not well-understood. Here, we investigated the effect of FK506 on TRPC6 in bladder smooth muscle, and explored the possible involvement of TRPC6 in OAB.MethodsFK506 was injected intraperitoneally into rats in which OAB was induced via BOO, and urodynamic indices were recorded. Rats and human bladder smooth muscle tissues with or without OAB were examined for TRPC6 expression by western blot, RT-PCR and IF staining. Cultured BSMCs were treated with PDGF, TRPC6 siRNAs and FK506. Then the TRPC6 expression and cellular proliferation were examined, and the Ca²⁺ influx and contractility of BSMCs were examined by time-lapse Ca²⁺ imaging and collagen gel contraction. Finally, IF and Co-IP were performed to test the effects of FK506 on NFAT translocation to the nucleus and the interaction of TRPC6 with FKBP12, respectively.ResultsFK506 improved urodynamic indices of OAB rats, and TRPC6 was expressed in rats and human bladder tissues. TRPC6 elevation in OAB rats was inhibited by FK506, and this inhibition coincided with improvements in urodynamic indices. PDGF enhanced TRPC6 expression, cellular proliferation, Ca²⁺ influx and contractility of BSMCs, and these effects were inhibited by TRPC6 siRNAs and FK506. FK506 inhibited NFAT translocation to the nucleus and disrupted the interaction of TRPC6 with FKBP12.ConclusionsOur results collectively indicate that FK506 may be used to treat OAB, and that TRPC6 may serve as an attractive target for therapeutic intervention in OAB.     

双氯芬酸钠栓和索利那新片治疗TURP术后OAB的临床疗效观察

目的:探讨双氯芬酸钠栓和索利那新治疗TURP术后OAB的临床疗效及预防性用药的必要性。方法:采取随机区组单盲的临床试验设计,将入选患者随机分为预防用药组和观察用药组。预防性用药组分别别使用双氯芬酸钠栓、索利那新片单药处理及两种药物联合处理,观察用药组在出现症状后按照预防性用药组的相同方法给药。结果:观察用药组中,双氯芬酸钠栓治疗的完全缓解率为50%,索利那新为81.8%,联合用药为90.5%,显著高于双氯芬酸钠栓(P0.05),各组不良反应的发生情况比较无显著性差异(P0.05)。预防用药组中,索利那新、双氯芬酸钠栓及联合用药分别可使TURP术后OAB的发生率降低45.0%、50.9%、53.8%.,三组之间比较无统计学差异(P0.05),且三组不良反应的发生情况比较无显著性差异(P0.05)。结论:双氯芬酸钠栓和索利那新治疗TURP术后OAB均安全有效,联合用药效果更佳,且预防性用药可以显著降低TURP术后OAB症状的发生率。