ABSTRACT
Objective@#Glucagon-like peptide-1(GLP-1) and gastrin synergistically promote the differentiation of insulin-producing cells which differentiated from rat bone marrow mesenchymal stem cells (BMSCs).@*Methods@#(1)Prepare IPCs model: pancreatic duodenal homeobox 1 (Pdx-1), neurogenin 3 (Ngn3) combined with V-type tendon fibrosarcoma oncogene homolog A (MafA) co-transfected BMSCs differentiation into IPCs; (2)IPCs were divided into 4 groups: Group A(uninduced group), group B(GLP-1 induction group), group C(gastrin induction group), and group D(GLP-1 combined with gastrin induction group). Cultured in high glucose medium for 7 days, the expression levels of insulin2, Pdx-1, GK, nestin, and glucagon mRNA were detected by RT-PCR. The insulin secretion of each group was detected by ELISA.@*Results@#After cultured for 7 days under high glucose conditions, the morphology of IPCs in each induction group changed significantly, gradually aggregated and formed scattered cell masses, and the combined induction group formed large cell masses. The staining of disulfide brown was reddish brown; The levels of insulin secretion increased gradually on the 0, 3rd, 5th, 7th, and 9th day after induction, and the increase was the most significant in the combined induction group (P<0.05). Compared with group A, the expression of insulin2 and GK in group B and D was significantly up-regulated, the expression of glucagon was down-regulated in group D, the expression of Pdx-1 was down-regulated in group C, and the expression of glucagon was up-regulated (P<0.05). Compared with group B, The expression of insulin2 was down-regulated in group C, and the expression level of glucagon was up-regulated. The expression levels of Pdx-1 and Insulin2 were significantly up-regulated in group D, and the expression level of glucagon was down-regulated (P<0.05). Compared with group C, the expression level of Pdx-1, insulin2 and GK was significantly up-regulated in group D, and the expression level of glucagon was down-regulated (P<0.05).@*Conclusion@#GLP-1 and gastrin synergistically promote the differentiation of IPCs into islet β cells by up-regulating GK and insulin2 and down-regulating glucagon.
ABSTRACT
Objective:Glucagon-like peptide-1(GLP-1) and gastrin synergistically promote the differentiation of insulin-producing cells which differentiated from rat bone marrow mesenchymal stem cells (BMSCs).Methods:(1)Prepare IPCs model: pancreatic duodenal homeobox 1 (Pdx-1), neurogenin 3 (Ngn3) combined with V-type tendon fibrosarcoma oncogene homolog A (MafA) co-transfected BMSCs differentiation into IPCs; (2)IPCs were divided into 4 groups: Group A(uninduced group), group B(GLP-1 induction group), group C(gastrin induction group), and group D(GLP-1 combined with gastrin induction group). Cultured in high glucose medium for 7 days, the expression levels of insulin2, Pdx-1, GK, nestin, and glucagon mRNA were detected by RT-PCR. The insulin secretion of each group was detected by ELISA.Results:After cultured for 7 days under high glucose conditions, the morphology of IPCs in each induction group changed significantly, gradually aggregated and formed scattered cell masses, and the combined induction group formed large cell masses. The staining of disulfide brown was reddish brown; The levels of insulin secretion increased gradually on the 0, 3rd, 5th, 7th, and 9th day after induction, and the increase was the most significant in the combined induction group ( P<0.05). Compared with group A, the expression of insulin2 and GK in group B and D was significantly up-regulated, the expression of glucagon was down-regulated in group D, the expression of Pdx-1 was down-regulated in group C, and the expression of glucagon was up-regulated ( P<0.05). Compared with group B, The expression of insulin2 was down-regulated in group C, and the expression level of glucagon was up-regulated. The expression levels of Pdx-1 and Insulin2 were significantly up-regulated in group D, and the expression level of glucagon was down-regulated ( P<0.05). Compared with group C, the expression level of Pdx-1, insulin2 and GK was significantly up-regulated in group D, and the expression level of glucagon was down-regulated ( P<0.05). Conclusion:GLP-1 and gastrin synergistically promote the differentiation of IPCs into islet β cells by up-regulating GK and insulin2 and down-regulating glucagon.
ABSTRACT
Objective To investigate the effects of pirfenidone on orbital fibroblasts (OFs) from patients with thyroid-associated ophthalmopathy (TAO) and its underlying mechanisms.Methods OFs from patients with TAO were isolated and cultured in DMEM.Cells were divided into four groups and treated with 0,250,500 and 1 000 μg/ml pirfenidone for 24,48 or 72 hours,respectively.Cell proliferation was detected by tetramethyl azo salt (MTT) assay,and cell viability was determined by trypan blue.Transforming growth factor (TGF) β1 mRNA level was determined by real-time fluorescence quantitative PCR (RT-qPCR).Type Ⅰ and type 11Ⅲ collagen secreted from cultured cells were measured by enzyme-linked immuno sorbent assay (ELISA).Results (1) The primary cultured OFs had typical fibroblast spindle-like morphology.(2) MTT assay showed that pirfenidone treatment significantly inhibited the proliferation of OFs in a dose-dependent manner (P＜0.05) with the proliferation rates of pirfenidone treated groups of-15.31％,-24.92％,-48.53％ from 250,500,1 000 μg/ml after 72 h,respectively,in which the inhibition effect of 1 000 μg/ml pirfenidone was significantly different from the other two treated groups (P＜0.05).There were no significant differences in the inhibitory effect of the same concentration group among different time points at 24 h,48 h and 72 h (P＞0.05).Trypan blue showed that the survival rate of OFs in different concentrations of pirfenidone from 0,250,500,1 000 μ-g/ml at 72 h were 78.37％,79.21％,78.24％ and 76.28％,respectively.There were no significant differences between each drug treated and the control group (P＞0.05).(3) RT-qPCR results showed that the mRNA expression levels of TGFβ1 at 250,500,1 000 μg/ml pirfenidone treated groups at 72 h were 0.760±0.010,0.440±0.006,and 0.290±0.002,respectively.Compared with the control group (0.950±0.014),the differences were statistically significant (all P＜0.05).Moreover,TGFβ1 mRNA expression level in 1 000 μg/ml pirfenidone treated group was significantly lower than those in the other two treated groups (all P＜0.05).The secretion of type Ⅰ collagen (0.633 ± 0.006,0.527 ± 0.003 and 0.402±0.008) and type 11Ⅲ collagen (0.511±0.003,0.439±0.007 and 0.223±0.006) in 250,500 and 1 000 μg/ml pirfenidone treated groups at 72 h were significantly lower than those in the control group (0.794±0.005,0.527±0.007,all P＜0.05).Type Ⅰ and type Ⅲ collagen secretion in 1 000 μg/ml pirfenidone treated group were significantly lower than those in the other two groups (P＜0.05).Conclusions Pirfenidone inhibits the cell proliferation,TGFβ1 expression and collagen secretion of OFs,which may contribute to the anti-fibrotic effect of pirfenidone.
ABSTRACT
Objective To explore the mechanism of bone marrow-derived endothelial progenitor cells (EPCs)in the treatment of peripheral neuropathy of diabetic rats. Methods Rats with diabetic peripheral neuropathy (DPN)were induced by streptozotocin(60 mg/kg). Male SD rats were divided into normal control group(NC group),DPN group, DPN+saline group(DPN+S group), and DPN+ EPCs group. Sciatic nerve motor nerve conduction velocity(MNCV)was measured. The expressions of NF-κB and myelin basic protein(MBP)in sciatic nerve were detected by immunohistochemistry. Results Compared with DPN group,the expression of NF-κB was reduced in the sciatic nerve of DPN+EPCs group,while the expression of NF-κB was increased in the sciatic nerve of DPN+ S group. There was no statistical difference in the expression of MBP between DPN and DPN+ EPCs groups. Compared with DPN+S group, the expression of MBP was higher in DPN+EPCs group. Conclusion Transplantation of EPCs inhibits the expression of NF-κB and increases the expression of MBP, which might be conducive to the repairs of nerve injury.
ABSTRACT
Objective To explore whether endothelial progenitor cells (EPCs) exerts therapeutic effects on rats with diabetic peripheral neuropathy (DPN).Methods Diabetes was induced by streptozotocin.Male SD rats were divided into normal control group(NC group), diabetic peripheral neuropathy group(DPN group), DPN+saline group(DPN+S group), and DPN+EPCs group.Sciatic nerve motor nerve conduction velocity(MNCV) was measured by a electromyography and trigger potentiometer instrument.Pathological changes were observed under optical microscope and electron microscope.Results Compared with normal control group [(52.91 ± 4.53) m/s], both DPN group and DPN+S group had slower sciatic nerve MNCV [(36.51 ± 6.30 and 25.37 ± 5.48) m/s, P＜0.01].Compared with DPN group, the MNCV of sciatic nerve in DPN + EPCs group had improved significantly [(36.51 ± 6.30 vs 46.20 ± 11.70) m/s, P ＜ 0.01].Sciatic nerve pathological changes, characterized by demyelination and axonal degeneration under light microscope and electron microscope were observed in DPN group, DPN+EPCs group, and DPN+S group.Compared with DPN+S group, the sciatic nerve pathological changes of DPN+EPCs group were alleviated.Conclusion After fed for 12 weeks, diabetic rats could progress to DPN rats.Transplantation of EPCs could improve MNCV, demyelination and axonal degeneration changes of sciatic nerver of the DPN rats.
ABSTRACT
Objective To investigate the expression of secretagogin(SCGN) and chromogranin A(CgA) in pancreatic neuroen‐docrine tumors (PNETs) .Methods Totally 54 cases of PNETs hospitalized in our hospital from October 2007 to October 2013 were enrolled in our study .Immunohistochemical detection was used to determine secretagogin and CgA .Results Secretagogin and CgA were highly expressed in all PNETs specimens .Compared with CgA ,secretagogin were highly expressed in the non‐functional PNETs ,tumor volume>30 cm3 ,the enveloped ,vascular invasion and lymph node metastasis (P0 .05) .Conclusion The expression of secretagogin and CgA in tumor combined with PNETs pathologi‐cal characteristics can predict the severity of PNETs .
ABSTRACT
Objective To explore the relationship of the levels of adhesion molecules and cytokines with vascular disease in patients with type 2 diabetes (T2DM).Methods Ninety-six T2DM patients were selected as case group and divided into non-vascular group (28 cases),micro-vascular group (33 cases) and major vascular group(35 cases) according to vascular disease.Meanwhile,30 healthy persons were selected as control group.Serum levels of serum soluble intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α) and yon willebrand factor (vWF) were determined by Enzyme-Linked Immuno Sorbent Assay (ELISA),and the expression of adhesion molecule and cytokines of patients with T2MD at 2 weeks after treatment.Results Compared with control group,the levels of ICAM-1,VCAM-1,TNF-α and IL-6 in all subgroup of T2DM patients were significantly elevated(P ＜ 0.05),and the levels showed an increasing trend with function deterioration (P ＜ 0.05).The levels of ICAM-1,VCAM-1,IL-6 and TNF-α in case group before treatment were (505.34 ± 56.42) μg/L,(570.85 ±59.54) μg/L,(94.51 ± 18.04) ng/L,(70.57 ± 18.34) ng/L respectively,significant different from those of after treatment ((390.53 ±45.23) μg/L,(482.93 ± 69.85) μg/L,(77.31 ± 15.49) ng/L,(50.45 ± 12.66) ng/L; t =16.281,8.712,7.697,8.445; P ＜ 0.05).There were significant positive correlation between ICAM-1,VCAM-1,IL-6,TNF-α and vWF in the case group(r =0.482,0.453,0.576,0.534 respectively,P ＜ 0.05).Conclusion Adhesion molecules and cytokines are take part in the process of the deterioration of T2DM and the occurrence of vascular disease.