Dissertation > Excellent graduate degree dissertation topics show

Fluvastatin Decreases the Expression of Fibronectin by Inhibiting the Signal Passageway of Rho-kinase in HK-2 Cells

Author: ShuJie
Tutor: LiuJia
School: Nanjing Medical University
Course: Internal Medicine
Keywords: Fluvastatin HK-2 cells Rho kinase Diabetic nephropathy Fibronectin
CLC: R587.2
Type: Master's thesis
Year: 2010
Downloads: 70
Quote: 0
Read: Download Dissertation

Abstract

Diabetic nephropathy (diabetic nephropathy, DN) is a microvascular complication of diabetes, the most serious one, it has become ESRD (end stage renal disease, ESRD) and diabetes, the leading cause of major death cause of disability. Previous studies of diabetic nephropathy more emphasis on the glomeruli, recent research found that while the presence of early diabetic nephropathy glomerular and tubular dysfunction, renal tubular lesions in the occurrence and development of diabetic nephropathy also plays an important role. Recent studies have found that statins inhibit the mevalonate metabolites - compounds such as isoprene farnesyl pyrophosphate ester (FPP) and bovine Long ox (base) pyrophosphate (GGPP) synthesis, so dependent on FPP, GGPP small GTP modified proteins do not located in the cell membrane, thereby inhibiting intracellular signal transduction plays an anti-proliferative and other non-lipid inflammatory renal protection. Rho kinase is a small G protein RhoA downstream most important and most unique signaling molecule in the role of a variety of factors is activated in diabetic nephropathy development play a role. To further observation statins on diabetic nephropathy protective mechanisms, the present study, cultured human proximal tubular epithelial cells (HK-2 cells) were observed fluvastatin on high glucose-induced HK-2 cells extracellular matrix impact of Rho kinase signaling pathway and its inherent relationship to explore fluvastatin possible kidney protection mechanisms. [Objective] 1. Study of high glucose on human renal tubular epithelial cell fibronectin expression. (2) investigate fluvastatin inhibited high glucose-induced human renal tubular epithelial cell fibronectin expression mechanism. 3 To explore Rho kinase signaling pathway in high glucose-induced human renal tubular epithelial cells fibronectin expression in rats. 4 To explore Rho kinase agonists LPA, isoprene compounds - mevalonate, GGPP, FPP and Rho kinase signaling pathway and fluvastatin linked. [Methods] 1. HK-2 cells in culture and subculture: HK-2 cells were supplemented with 10% fetal bovine serum and blue / streptomycin (100U/ml) in DMEM medium, 37 ℃ incubator (5% CO2) culture. HK-2 cells were grown to the logarithmic growth phase, the culture fluid was aspirated, trypsin digestion at 37 ° C incubator for about 2min, containing 10% FBS in DMEM to terminate digestion, cells were collected by centrifugation, supernatant, containing 10% fetal bovine serum in DMEM resuspended in 5 * 104 / cm2 vaccination. 2. HK-2 cells grouped and Intervention: When the cells were grown to logarithmic phase serum-free culture medium sync 24 hours, the cells were synchronized in the G0 phase. The cell groups: ① 0h group; 6h group; 12 groups; 24 group ② normal glucose group; mannitol group; high glucose group; high glucose fluvastatin low-dose group; high glucose fluvastatin dose group; high glucose fluvastatin statin high-dose group ③ normal glucose group; high glucose group; high glucose fluvastatin 12h group; high glucose fluvastatin LPA group; high glucose fluvastatin 20h group ④ high glucose group; high glucose fluvastatin dose group; High sugar fluvastatin dose group mevalonate group; high glucose fluvastatin dose group GGPP group; high glucose fluvastatin dose group FPP group. 3.Western Blot Detection HK-2 cells p-MYPT1 (Rho kinase activation function symbol), FN expression [Results] 1 high glucose on HK-2 cells p-MYPT1, FN expression (Western Blot) with high 0h glucose control group, high glucose stimulated HK-2 cells 6h, 12h, 24h can cause p-MYPT1 significant activation (p lt; 0.05), stimulation after 12h p-MYPT1 activity reached its peak (p lt; 0.01) . High glucose-induced FN expression was time-dependent, with high glucose stimulation time, FN expression gradually increased (p lt; 0.05 or p lt; 0.01). 2 fluvastatin on HK-2 cells p-MYPT1, FN expression (Western Blot) cultured HK-2 cells 12h, and normal glucose control group compared to high glucose group p-MYPT1 and FN expression was significantly increased ( P lt; 0.01), fluvastatin (10-7, 10-6 and 10-5 mol / L) and treated with p-MYPT1 expression of FN were higher glucose decreased in a concentration-dependent manner (P lt; 0.05) 3. LPA on the p-MYPT1, FN expression (Western Blot) in the observation of LPA on the p-MYPT1, FN expression experiments, cultured HK-2 cells were divided into the following five groups: normal glucose group ; the high glucose group; high glucose 12h fluvastatin group; high glucose fluvastatin LPA group; high glucose 20h fluvastatin group. High sugar fluvastatin LPA group p-MYPT1, FN expression were higher than fluvastatin 12h group and fluvastatin 20h group (P lt; 0.05) 4. Fluvastatin and mevalonate, FPP, GGPP for p -MYPT1, FN expression (Western Blot) and high-sugar group, fluvastatin dose group and FPP group p-MYPT1, FN expression decreased (P lt; 0.05), and fluvastatin dose group phase ratio, mevalonate group and GGPP group p-MYPT1, FN expression was significantly higher (P lt; 0.05). [Conclusion] a high glucose can induce the expression of Rho kinase and FN. 2 fluvastatin can inhibit Rho kinase induced by high glucose and FN expression in a concentration-dependent manner. 3 fluvastatin may inhibit Rho kinase phosphorylation of extracellular matrix and inhibit the expression of fibronectin. 4. Rho kinase signaling pathway may be diabetic nephropathy renal interstitial fibrosis in the initiating signals.

Related Dissertations

CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Islet disease > Diabetic coma and other complications
© 2012 www.DissertationTopic.Net Mobile