Background and Objective: Sepsis is one of the most prevalent diseases that cause a chain reaction in the human system.Sepsis-related acute renal failure (ARF) exhibits a higher mortality risk.This research intends to study whether vanillin attenuates acute renal injury and enhances survival in a rat model of sepsis via regulating the NF-κB signaling pathway.Materials and Methods: A total of 24 Sprague-Dawley rats were split between 4 groups (sham, decease, vanillin 100 mg kgG 1 and vanillin 200 mg kgG 1 ).Animals were kept in the cages after CLP induction, then blood was withdrawn to assess the serum creatinine (sCr) and blood urea nitrogen (BUN) levels, autacoids were assessed in the kidney tissue homogenate, TUNEL assays, qRT-PCR and western blot assessments were performed and finally, histopathological analysis was carried out.Results: Levels of Scr, BUN and autacoids were higher in the disease group than in the sham group but reversed in vanillin-treated animals.In the disease group, the ratio of apoptosis and the mRNA expression of caspase-3 and Bax were higher but the mRNA expression of Bcl-2 was lower.In the histopathological study, the disease groupʼs histopathological score was higher than the sham groupʼs but significantly lowered in the vanillin-treated group than in the disease group. Conclusion:The findings in the research demonstrated that vanillin lowers nephron apoptosis, which attenuates sepsis-induced ARF through lowering the NF-κB p65 pathway and oxidative stress.
Extracellular ATP regulates cellular function in an autocrine or paracrine manner through activating purinergic signalling. Studies have shown that purinergic receptors were expressed in mammalian ovaries and they have been proposed as an intra-ovarian regulatory mechanism. P2X7 was expressed in porcine ovarian theca cells and murine and human ovarian surface epithelium and is involved in ATP-induced apoptotic cell death. However, the role of P2X7 in corpus luteum is still unclear. The aim of this study was to investigate the role of ATP signalling in murine luteal cells and the possible mechanism(s) involved. We found that P2X7 was highly expressed in murine small luteal cells. The agonists of P2X7, ATP and BzATP, inhibited the proliferation of luteal cells. P2X7 antagonist BBG reversed the inhibition induced by ATP and BzATP. Further studies showed that ATP and BzATP inhibited the expression of cell cycle regulators cyclinD2 and cyclinE2. ATP and BzATP also inhibited the p38–mitogen-activated protein kinase (MAPK) signalling pathway. These results reveal that P2X7 receptor activation is involved in corpus luteum formation and function.
Objective:To observe the changes of the autoantibodies and β-cell' function of patients with LADA treating by NPH or Sulfonylureas(SU).Methods:We treated 103 LADA patients with NPH or SU respectively for 8 weeks at the same condition(e.g.the similar life-style and same dosage of other antihyperglycemics).To observe and compare all patients' metabolic indexes of glucose and lipid and uric acid,and their GAD-Ab and ICA levels and glycemia curves and c-peptide curves after standard meal at two treatment stages(e.g.NPH and SU).Results:Both NPH and SU could ameliorate all patients' metabolic indexes of glucose and lipid and uric acid combining with similar life-style and same dosage of other antihyperglymics (P0.01).The patients' serum GAD-Ab titre and ICA positive at SU stage was higher than those at NPH stage(P0.05).And their c-peptide curve at SU stage was lower than that at NPH stage(P0.05),but there was no difference at their glycemia curves between two stage (P0.1).Conclusion:SU can control the hyperglycemia of patients with LADA,but may be harmful to the function of their β-cells.NPH is also effective to treat their hyperglycemia and maybe favorable to prevent their β-cells from immunological lesion.
Objective To investigate factors influencing islet isolation and purification in dogs and to explore how to obtain massive purified viable islets from canine pancreas.To provide experimental evidence for pancreatic islet transplantation in clinical practice.Methods Islets were isolated from pancreas of 22 dogs by modified automated techniques,followed by purification using continuous density gradients.The purity of islets was determined by dithizone (DTZ) staining,and the function of islets was evaluated by glucose stimulating insulin release test.Results The crude islets yield was ( 155 040 ±310) IEQ/pancreas while the yield was (74 200 ±185) IEQ/pancreas after purification.The average purity was (89.4 ±2.6) ％.The recovery rate was(47.8 ±1.3) ％.Islet viability was(93.0 ± 1.7)％.The modified techniques improved both the yield and the quality in islet isolation and purification.The islets were morphologically intact after purification.Glucose stimulating insulin release test showed that the difference of the secreted insulin concentration had statistical significance ( P ＜0.001 ) between low concentration glucose group and high concentration glucose group.Conclusion It is of great value for clinical research to obtain enough islets through islet isolation and purification in dogs.
Key words:
Islet transplantation; Isolation; Purification; Islet
Objective To investigate the experimental condition of separate purification and directed differentiation potency of pancreatic stem cells of adult rats in vitro. Methods The pancreas of adult rats were digested by collagenase V in situ perfusion. Filtration was performed by 100 mesh filter. Ficoll 400 density gradient centrifugation was used to separate pancreatic stem cells and stem cells were for cultivation. Epidermal growth factor (EGF), fibroblast growth factor (bFGF) was added to medium for in vitro cultivation.Immunofluorescence staining method was used to detect the expression of CK19, Pdx-1, nestin, insulin and glucagon, and the positive cell rate was calculated. The differentiated cell was evaluated by dithizone (DTZ)staining; insulin excretion function was determined by optical enzyme labeled ELISA staining. Results The pancreatic stem cell obtained from the study could be cultivated in vitro for 8 generations. The expression of CK19, Pdx-1 and nestin of the tem cells were all positive, and the rate of positive cells were (88.6 ± 6.2)% ,(84.6 ±8.6)% and (81.3 ±7.5)%. The differentiated cell was in brownish red color by DTZ dye. After high concentration sugar stimulation, the expression of insulin secretion was increased in the supernatant.Conclusions This method can harvest highly purified, and large amount of pancreatic duct stem cell.Artificial induction may result directed differentiate for islet-like clusters and have insulin secrete function.
Key words:
Pancreas; Stem cells; Cell culture techniques; Cell differentiation; Identify
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To study the expression of hypoxia inducible factor 1α(HIF-1α) of iron-overloaded in irradiated mice and its effect on erythropoiesis.Twenty mice were randomly divided into 4 groups: Ctrl (control group), IR (irradiation group), IO (irradiation + iron overload group), and RAPA (rapamycin treatment group). The iron overload model was verified. The CFU-E (colony forming unit-erythroid) and BFU-E(burst colony forming unit-erythroid) were cultured; flow cytometry was used to detect the ratios of early stage (Ter119+CD71-) to late stage (Ter119+CD71+) of primitive erythroblasts; RT-PCR was used to detect the mRNA expression of HIF-1α and its related signal molecules in bone marrow cells.The expression of HIF-1α in IR and IO group was significantly higher than that in Ctrl group, and that in IO group was significantly higher than IR group (P<0.05). The ratio of late stage primitive erythroblasts, the number of CFU-E and BFU-E in both IR and IO group were lower than those in Ctrl group, and those in IO group were significantly lower than those in IR group (P<0.05). Compared with Ctrl group, the expression of HIF-1α related signal pathway molecules in both IR and IO group was significantly decreased (P<0.05). Compared with IO group, the expression of HIF-1α and its related signal molecules in RAPA(mTOR inhibitor) group was decreased significantly (P<0.05), the number of BFU-E was increased significantly(P<0.05).Irradiation induces the increase of HIF-1α and the decrease of the ability of hematopoietic colony formation and the ratio of late stage primitive erythroblasts. Iron overload can aggravate the injury. mTOR inhibitor rapamycin can partially alleviate the injury, suggesting that iron overload can lead to injury of erythropoiesis through HIF-1α.
Objective To investigate the potential of pancreatic stem cells (PSCs) directed differentiation in vitro, and to evaluate the effects of differentiated PSCs allograft on the treatment of diabetes.Methods The PSCs of adult Wistar rats were separated and purified in vitro. The surface of PSCs was determined by immunofluorescence staining, and then it was stimulated by hepatocyte growth factor (HGF) and nicotinamide to induce directed differentiation. Dithizone dyeing was used to determine the islet-like cells after induction, and ELISA staining method was used to detect the insulin levels. Streptozotocin peritoneal injection was used to induce the diabetic rat mode. 40 rats were randomly allocated into pancreatic islet cells allograft group (experiment group) and placebo group. The serum insulin and glucose levels 1 d before transplantation and 1, 2, 3, 4 week after transplantation were measured. Results PSCs of adult Wistar rats were successfully obtained, and the expression of CK19, Pdx-1 and Nestin on cell surface was positive. Dithizone dyeing for directed differentiation cells showed brownish red color. The cells could express and secrete insulin after hyperglycaemia stimulation. The serum insulin and glucose levels 4 week after transplantation were (11.41 ±1.52) mU/L and (8.22 ± 2.7) mmol/L, which were (9.30 ± 1.56) mU/L and (12.23 ± 3.8) mmol/L in the placebo group, and difference was statistically significant (P＜0.05). Conclusions PSCs can be induced and directed differentiated in vitro into islet-like clusters with insulin secretion function. And its allograft has the potential for the treatment of diabetes.
Key words:
Pancreatic; Stem cells; Cell differentiation; Homograft; Transplantation
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