Effects of fibrobl ast growth factor 4 on the proliferation of fibroblast-like synoviocytes in rheumatoid arthritis / 中华风湿病学杂志

Objective To investigate the expression of fibroblast growth factor 4 (FGF4) in serum of active rheumatoid arthritis (RA) and its role in RA synoviocyte proliferation. Methods The serum level of FGF4 were detected by protein arrays in 20 patients with RA, and 20 age and gender matched healthy controls. FLSs were isolated from RA synovium,and were co-cultured with recombinant human FGF4 (rhFGF4). Cell proliferation was quantified by Cell Counting Kit-8 assay and cell cycle distribution was evaluated by flow-cytometry. The protein levels of cyclin D1, phospho-Akt (p-Akt) and phospho-p38 (p-p38) were measured by western blot. Results The serum expression of FGF4 in RA group was higher than that in control group (P=0.041). After being treated with different concentrations of rhFGF4 (12.5, 25, 50, and 100 ng/ml), RA-FLS showed significant increase in cell proliferation, with different rates of [(121 ±8)％], [(126 ±12)％], [(129 ± 12)％], a nd [(134 ±14)％] respectively, comparing with that of the controls [(100 ±0)％, (P12.5=0.049, P25=0.009, P50=0.004, P100=0.001).]. Among them, the percentage of G2/M+S phase cells were [(12.6±3.6)％], [(15.3±4.5)％], [(17.1±5.1)％], [(19.6±4.1)％] respectively, and except the lowest rhFGF4 concentration treatment group of 12.5 ng/ml, G2/M+S phase cells in other groups was significantly increased compared with the controls [(5.4±2.4)％] (P12.5=0.159, P25=0.042, P50=0.018, P100=0.005). And the protein expression of cyclin D1 was up-regulated after being treated with 50 ng/ml and 100 ng/ml rhFGF4 (P50=0.035, P100=0.027). FGF4 transiently increased the expression of p-Akt and p-p38 protein at the concentration of 50 ng/ml. Comparisons of data between groups were performed by independent sample Student's t-test. Statistical significant differences among groups were tested by one-way analysis of variance (ANOVA) or the Kruskal-Wallis test. The Dunnett's t-test was used for multiple comparisons. A P-value of <0.05 was considered statistically significant. Conclusion Our results suggest that FGF4 is highly expressed in the serum of active RA patients. FGF4 may promote the proliferation of RA-FLS via modulating PI3K/Akt and p38-MAPK signaling pathways, which subsequently contributs to synovial hyperplasia.

Induced maturation of hepatic progenitor cells in vitro

Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.

Influence of artificial liver support system on bone marrow stem cell differentiation factors in patients with chronic severe hepatitis B / 中华传染病杂志

Objective To study the influence of artifical liver support system (ALSS) on bone marrow stem cell (BMSC) differentiation factors in patients with chronic severe hepatitis B.Methods Fifty patients with chronic severe hepatitis B were divided into ALSS treatment group (n=25) and control group (n=25).The patients in control group received combined medical treatment and those in ALSS treatment group received ALSS treatment within 1 week of admission on the basis of combined medical treatment.The concentrations of hepatocyte growth factor(HGF),fibroblast growth factor-4 (FGF-4),epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) were detected before and 2 weeks after therapy.The data were analyzed by t test.Results The serum levels of HGF,FGF-4,EGF and bFGF of patients before ALSS treatment in treatment group were (689.10± 337.68) ng/L,(124.88±87.67) ng/L,(323.85±44.40) ng/L and (9.29± 1.38) ng/L,respectively; while the levels of those cytokines after ALSS treatment were (1081.50±356.66) ng/L,(110.76±79.71) ng/L,(347.80±71.73) ng/L and (9.57±1.15) ng/L,respectively,among which HGF level increased significantly after ALSS treatment (t =10.042,P＜0.01) and was higher than control group(t=6.670,P＜0.01).However,the levels of HGF,FGF-4,EGF and bFGF were not significantly different from those in the control group.And levels of all HGF,FGF-4,EGF and bFGF in control group were not statistically different before and after treatment.ConclusionALSS treatment can increase the serum HGF level but not FGF-4,EGF and bFGF,which may contribute to BMSC transdifferentiation that is involved in the hepatocyte repair and regeneration in chronic severe hepatitis B.

Evaluation of Spinal Fusion Using Bone Marrow Derived Mesenchymal Stem Cells with or without Fibroblast Growth Factor-4

OBJECTIVE: In this study, the authors assessed the ability of rat bone marrow derived mesenchymal stem cells (BMDMSCs), in the presence of a growth factor, fibroblast growth factor-4 (FGF-4) and hydroxyapatite, to act as a scaffold for posterolateral spinal fusion in a rat model. METHODS: Using a rat posterolateral spine fusion model, the experimental study comprised 3 groups. Group 1 was composed of 6 animals that were implanted with 0.08 gram hydroxyapatite only. Group 2 was composed of 6 animals that were implanted with 0.08 gram hydroxyapatite containing 1 x 10(6)/ 60 microliter rat of BMDMSCs. Group 3 was composed of 6 animals that were implanted with 0.08 gram hydroxyapatite containing 1 x 10(6)/ 60 microliter of rat BMDMSCs and FGF-4 1 microgram to induce the bony differentiation of the BMDMSCs. Rats were assessed using radiographs obtained at 4, 6, and 8 weeks postoperatively. After sacrifice, spines were explanted and assessed by manual palpation, high-resolution microcomputerized tomography, and histological analysis. RESULTS: Radiographic, high-resolution microcomputerized tomographic, and manual palpation revealed spinal fusion in five rats (83%) in Group 2 at 8 weeks. However, in Group 1, three (60%) rats developed fusion at L4-L5 by radiography and two (40%) by manual palpation in radiographic examination. In addition, in Group 3, bone fusion was observed in only 50% of rats by manual palpation and radiographic examination at this time. CONCLUSION: The present study demonstrates that 0.08 gram of hydroxyapatite with 1 x 10(6)/ 60 microliter rat of BMDMSCs induced bone fusion. FGF-4, added to differentiate primitive 1 x 10(6)/ 60 microliter rat of BMDMSCs did not induce fusion. Based on histologic data, FGF-4 appears to induce fibrotic change rather than differentiation to bone by 1 x 10(6)/ 60 microliter rat of BMDMSCs.