Exploration of Multidimensional Regulation of Dahuang Ling Xian Capsules on Bile Duct Cells at Inflammatory State Based on miRNA Chip Technology / 广州中医药大学学报

Objective To investigate the effect of Dahuang Ling Xian Capsules(DLXC) on miRNAs in bile duct cells at inflammatory state. Methods The cultured well-grown bile duct cells at logarithmic phase were chosen for the experiment. Lipopolysaccharide (LPS , 5 μg/mL) was used to induce the model of bile duct cells at inflammatory state, and then DLXC (1 mg/mL) was added. After co-culturing for 72 h, the total RNA of bile duct cells was collected for miRNA chip detection and bioinformatics analysis. Results DLXC showed regulatory effect on the expression of 30 miRNAs in bile duct cells at inflammatory state, through which to affect biological processes involving with bile duct cell signal transduction, organic compound reaction and hypoxia stress, cell components such as cytoplasm , endoplasmic reticulum and membrane rafts , molecular biological functions of signal transduction activity , DNA binding in transcription regulation region and voltage-gated potassium channel activity , and multiple pathways of hypoxia-inducible factor-1(HIF-1) signaling pathway , uanosine 3 ' , 5 '-cyclic monophosphate-protein kinase C (cGMP-PKG) signaling pathway , tumor necrosis factor alpha (TNF-α) signaling pathway, chemokine signaling pathway, apoptosis signaling pathway, phosphatidylinositol-3-kinase (PI3K)-Akt signaling pathway and Toll-like signaling pathway. Conclusion DLXC exert the multidimensional regulation of bile duct cells at inflammatory state with miRNAs as targets.

Ca2 -activated K Currents of Pancreatic Duct Cells in Guinea-pig

There are numerous studies on transepithelial transports in duct cells including Cl and/or HCO3. However, studies on transepithelial K transport of normal duct cells in exocrine glands are scarce. In the present study, we examined the characteristics of K currents in single duct cells isolated from guinea pig pancreas, using a whole-cell patch clamp technique. Both Cl and K conductance were found with KCl rich pipette solutions. When the bath solution was changed to low Cl, reversal potentials shifted to the negative side, 75 4 mV, suggesting that this current is dominantly selective to K. We then characterized this outward rectifying K current and examined its Ca2 dependency. The K currents were activated by intracellular Ca2. 100 nM or 500 nM Ca2 in pipette significantly (P< 0.05) increased outward currents (currents were normalized, 76.8 7.9 pA, n=4 or 107.9 35.5 pA, n=6) at 100 mV membrane potential, compared to those with 0 nM Ca2 in pipette (27.8 3.7 pA, n=6). We next examined whether this K current, recorded with 100 nM Ca2 in pipette, was inhibited by various inhibitors, including Ba2, TEA and iberiotoxin. The currents were inhibited by 40.4 % (n=3), 87.0 % (n=5) and 82.5 % (n=9) by 1 mM Ba2, 5 mM TEA and 100 nM iberiotoxin, respectively. Particularly, an almost complete inhibition of the current by 100 nM iberiotoxin further confirmed that this current was activated by intracellular Ca2. The K current may play a role in secretory process, since recycling of K is critical for the initiation and sustaining of Cl or HCO3 secretion in these cells.

Stem Cell In Adult Pancreas: Its Activation and Induction of Beta Cell Differentiation / 대한해부학회지

Stem cells in adult pancreas and their specific marker are poorly characterized. We hypothesized that pancreatic stem cells could evolve from the duct system in response to neogenic stimulation. Following partial pancreatectomy (Px), we found extensive formation of ductules consisting of nestin-positive epithelial cells with higher replicating ability in the neogenic foci after Px. The neogenic ductules were isolated for the culture of nestin-positive duct cells. These nestinpositive duct cells were numerous and displayed extensive self-replication in the duct cell explants, thus depicted as nestin-positive duct stem (NPDS) cells. Endocrine cells, mostly insulin cells were present in the explants at day 2 as single cells or as small clusters adjacent to the NPDS cells, and formed islet-like masses at day 3 of culture, implying islet cell differentiation from NPDS cells. We found transient up-regulation of PDX-1 expression by RT-PCR at day 3 after Px in pancreatic tissue. We investigated the effect of clusterin overexpression on differentiation of insulin beta cells from duct cells We found that the number of insulin producing cells increased 11.5 fold when clusterin was overexpressed. Insulin expression, both insulin mRNA and peptide levels, was increased in clusterin cDNA transfected cells. In conclusion, we suggest that NPDS cells could be generated from adult pancreas by neogenic motivations and they may differentiate into insulin-secreting-cells, and clusterin could stimulate not only differentiation of precursor cells in the pancreatic duct, but also proliferation of predifferentiated beta cells. Those differentiated beta cells are functional cells secreting insulin in response to glucose stimulation.

Protective effect of peroxisome proliferator-activated receptor ? on TNF?-induced injury of inner medullary collecting duct cells / 第二军医大学学报

Objective:To observe the protective effect of peroxisome proliferator-activated receptor ?(PPAR?) on TNF?-induced injury of the inner medullary collecting duct(IMCD) cells.Methods: Cultured IMCD cells were randomly divided into blank control,TNF? stimulation,PPAR? transfection(pPPAR?-control) and PPAR? transfection + TNF? stimulation groups.Stimulation was induced with 50 ng/ml TNF? for 24 h and mouse wild-type PPAR? plasmid was used to transfect IMCD cells.Cell supernatant MCP-1 or TGF?_1 was detected by ELISA method.Results: IMCD cells transfected with wild-type PPAR? plasmid had high expression of PPAR? mRNA and protein.The contents of MCP-1 and TGF?_1 in the supernatant were similar in blank control group and pPPAR?-control group.Compared with blank control group,TNF? stimulation group had decreased contents of MCP-1 and TGF?_1in the supernatant(P