17β-estradiol inhibits TGF-β-induced collagen gel contraction mediated by human Tenon fibroblasts via Smads and MAPK signaling pathways

AIM:To investigate the impact of 17β-estradiol on the collagen gels contraction(CGC)and inflammation induced by transforming growth factor(TGF)-βin human Tenon fibroblasts(HTFs).METHODS:HTFs were three-dimensionally cultivated in type I collagen-generated gels with or without TGF-β(5 ng/mL),17β-estradiol(12.5 to 100μmol/L),or progesterone(12.5 to 100μmol/L).Then,the collagen gel diameter was determined to assess the contraction,and the development of stress fibers was analyzed using immunofluorescence staining.Immunoblot and gelatin zymography assays were used to analyze matrix metalloproteinases(MMPs)and tissue inhibitors of metalloproteinases(TIMPs)being released into culture supernatants.Enzyme-linked immunosorbent assay(ELISA)and reverse transcription-quantitative polymerase chain reaction(RT-PCR)were used to detect interleukin(IL)-6,monocyte chemoattractant proteins(MCP)-1,and vascular endothelial growth factor(VEGF)in HTFs at the translational and transcriptional levels.The phosphorylation levels of Sma-and Mad-related proteins(Smads),mitogen-activated protein kinases(MAPKs),and protein kinase B(AKT)were measured by immunoblotting.Statistical analysis was performed using either the Tukey-Kramer test or Student’s unpaired t-test to compare the various treatments.RESULTS:The CGC caused by TGF-βin HTFs was significantly inhibited by 17β-estradiol(25 to 100μmol/L),and a statistically significant difference was observed when comparing the normal control group with 17β-estradiol concentrations exceeding 25μmol/L(P<0.05).The suppressive impact of 17β-estradiol became evident 24h after administration and peaked at 72h(P<0.05),whereas progesterone had no impact.Moreover,17β-estradiol attenuated the formation of stress fibers,and the production of MMP-3 and MMP-1 in HTFs stimulated by TGF-β.The expression of MCP-1,IL-6,and VEGF mRNA and protein in HTFs were suppressed by 100μmol/L 17β-estradiol(P<0.01).Additionally,the phosphorylation of Smad2 Smad3,p38,and extracellular signal-regulated kinase(ERK)were downregulated(P<0.01).CONCLUSION:17β-estradiol significantly inhibits the CGC and inflammation caused by TGF-βin HTFs.This inhibition is likely related to the suppression of stress fibers,inhibition of MMPs,and attenuation of Smads and MAPK(ERK and p38)signaling.17β-estradiol may have potential clinical benefits in preventing scar development and inflammation in the conjunctiva.

Distribution and localization of fibroblast growth factor-8 in rat brain and nerve cells during neural stem/progenitor cell differentiation

The present study explored the distribution and localization of fibroblast growth factor-8 and its potential receptor, fibroblast growth factor receptor-3, in adult rat brain in vivo and in nerve cells during differentiation of neural stem/progenitor cells in vitro. Immunohistochemistry was used to examine the distribution of fibroblast growth factor-8 in adult rat brain in vivo. Localization of fibroblast growth factor-8 and fibroblast growth factor receptor-3 in cells during neural stem/progenitor cell differentiation in vitro was detected by immunofluorescence. Flow cytometry and immunofluorescence were used to evaluate the effect of an anti-fibroblast growth factor-8 antibody on neural stem/progenitor cell differentiation and expansion in vitro. Results from this study confirmed that fibroblast growth factor-8 was mainly distributed in adult midbrain, namely the substantia nigra, compact part, dorsal tier, substantia nigra and reticular part, but was not detected in the forebrain comprising the caudate putamen and striatum. Unusual results were obtained in retrosplenial locations of adult rat brain. We found that fibroblast growth factor-8 and fibroblast growth factor receptor-3 were distributed on the cell membrane and in the cytoplasm of nerve cells using immunohistochemistry and immunofluorescence analyses. We considered that the distribution of fibroblast growth factor-8 and fibroblast growth factor receptor-3 in neural cells corresponded to the characteristics of fibroblast growth factor-8, a secretory factor. Addition of an anti-fibroblast growth factor-8 antibody to cultures significantly affected the rate of expansion and differentiation of neural stem/progenitor cells. In contrast, addition of recombinant fibroblast growth factor-8 to differentiation medium promoted neural stem/progenitor cell differentiation and increased the final yields of dopaminergic neurons and total neurons. Our study may help delineate the important roles of fibroblast growth factor-8 in brain activities and neural stem/progenitor cell differentiation.

The hypoxia-inducible factor-1α activates ectopic production of fibroblast growth factor 23 in tumor-induced osteomalacia

Tumor-induced osteomalacia （TIO） is a rare paraneoplastic syndrome in which ectopic production of fibroblast growth factor 23 （FGF23） by non-malignant mesenchymal tumors causes phosphate wasting and bone fractures. Recent studies have implicated the hypoxia-inducible factor-la （HIF-la） in other phosphate wasting disorders caused by elevated FGF23, including X-linked hypophosphatemic rickets and autosomal dominant hypophosphatemia. Here we provide evidence that HIF-la mediates aberrant FGF23 in TIO by transcriptionally activating its promoter. Immunohistochemical studies in phosphaturic mesenchymal tumors resected from patients with documented TIO showed that HIF-la and FGF23 were co-localized in spindle- shaped cells adjacent to blood vessels. Cultured tumor tissue produced high levels of intact FGF23 and demonstrated increased expression of HIF-la protein. Transfection of MC3T3-E1 and Saos-2 cells with a HIF-la expression construct induced the activity of a FGF23 reporter construct. Prior treatment of tumor organ cultures with HIF-la inhibitors decreased HIF-la and FGF23 protein accumulation and inhibited HIF-la-induced luciferase reporter activity in transfected cells. Chromatin immunoprecipitation assays confirmed binding to a HIF-la consensus sequence within the proximal FGF23 promoter, which was eliminated by treatment with a HIF-la inhibitor. These results show for the first time that HIF-la is a direct transcriptional activator of FGF23 and suggest that upregulation of HIF-la activity in TIO contributes to the aberrant FGF23 production in these patients.

Local inhibition of matrix metalloproteinases reduced M2 macrophage activity and impeded recovery in spinal cord transected rats after treatment with fibroblast growth factor-1 and nerve grafts

Alternatively activated macrophages （M2 macrophages） promote central nervous system regeneration. Our previous study demonstrated that treatment with peripheral nerve grafts and fibroblast growth factor-1 recruited more M2 macrophages and improved partial functional recovery in spinal cord transected rats. The migration of macrophages is matrix metalloproteinase （MMP） dependent. We used a general inhibitor of MMPs to influence macrophage migration, and we examined the migration of macrophage populations and changes in spinal function. Rat spinal cords were completely transected at Ts, and 5 mm of spinal cord was removed （group T）. In group R, spinal cord-transected rats received treatment with fibroblast grow th factor- 1 and peripheral nerve grafts. In group RG, rats received the same treatment as group R with the addition of 200 μM GM6001 （an MMP inhibitor） to the fibrin mix. We found that MMP-9, but not MMP- 2, was upregulated in the graft area of rats in group R. Local application of the MMP inhibitor resulted in a reduction in the ratio of arginase-1 （M2 macrophage subset）/inducible nitric oxide synthase-postive cells. When the MMP inhibitor was applied at 8 weeks postoperation, the partial functional recovery observed in group R was lost. This effect was accompanied by a decrease in brain-derived neurotrophic factor levels in the nerve graft. These results suggested that the arginase-1 positive population in spinal cord transected rats is a migratory cell population rather than the phenotypic conversion of early iNOS^＋ cells and that the migration of the arginase-1^＋ population could be regulated locally. Simultaneous application of MMP in- hibitors or promotion of MMP activity for spinal cord injury needs to be considered if the coadministered treatment involves M2 recruitment.

Significance of serum fibroblast growth factor-23 and miR-208b in pathogenesis of atrial fibrillation and their relationship with prognosis

BACKGROUND The incidence and prevalence of atrial fibrillation are increasing each year,and this condition is one of the most common clinical arrhythmias.AIM To investigate the levels and significance of serum fibroblast growth factor 23(FGF-23)and miR-208 b in patients with atrial fibrillation and their relationship with prognosis.METHODS From May 2018 to October 2019,240 patients with atrial fibrillation were selected as an observation group,including 134 with paroxysmal atrial fibrillation and 106 with persistent atrial fibrillation;150 patients with healthy sinus rhythm were selected as a control group.The serum levels of FGF-23 and miR-208 b in the two groups were measured.In the observation group,cardiac parameters were determined by echocardiography.RESULTS The serum levels of FGF-23 and miR-208 b in the observation group were 210.20±89.60 ng/mL and 5.30±1.22 ng/mL,which were significantly higher than the corresponding values in the control group(P<0.05).In the observation group,the serum levels of FGF-23 and miR-208 b in patients with persistent atrial fibrillation were 234.22±70.05 ng/mL and 5.83±1.00 ng/mL,which were significantly higher than the corresponding values in patients with paroxysmal atrial fibrillation(P<0.05).The left atrial dimension(LAD)of patients with persistent atrial fibrillation was 38.81±5.11 mm,which was significantly higher than that of patients with paroxysmal atrial fibrillation(P>0.05).The serum levels of FGF-23and miR-208 b were positively correlated with the LAD(r=0.411 and 0.382,P<0.05).In the observation group,the serum levels of FGF-23 and miR-208 b in patients with a major cardiovascular event(MACE)were 243.30±72.29 ng/mL and 6.12±1.12 ng/mL,which were significantly higher than the corresponding values in patients without a MACE(P<0.05).CONCLUSION The serum levels of FGF-23 and miR-208 b are increased in patients with atrial fibrillation and are related to the type of disease,cardiac parameters,and prognosis.

Cross-talk between microRNA-let7c and transforming growth factor-β2 during epithelial-to-mesenchymal transition of retinal pigment epithelial cells

AIM: To explore the roles of microRNA-let7 c(miR-let7 c) and transforming growth factor-β2(TGF-β2) and cellular signaling during epithelial-to-mesenchymal transition(EMT) of retinal pigment epithelial cells. METHODS: Retinal pigment epithelial(ARPE-19) cells were cultured with no serum for 12 h, and then with recombinant human TGF-β2 for different lengths of time. ARPE-19 cells were transfected with 1×106 TU/mL miR-let7 c mimcs(miR-let7 cM), miR-let7 c mimcs negative control(miR-let7cMNC) and miR-let7 c inhibitor(miR-let7 cI) using the transfection reagent. The expression of keratin-18, vimentin, N-cadherin, IKB alpha, p65 were detected by Western blot, quantitative polymerase chain reaction and immunofluorescence. RESULTS: The expression of miR-let7c was dramatically reduced and the nuclear factor-kappa B(NF-κB) signaling pathway was activated after induction by TGF-β2(P<0.05). In turn, overexpressed miR-let7 c significantly inhibited TGF-β2-induced EMT(P<0.05). However, miR-let7 c was unable to inhibit TGF-β2-induced EMT when the NF-κB signaling pathway was inhibited by BAY11-7082(P<0.01). CONCLUSION: The miR-let7 c regulates TGF-β2-induced EMT through the NF-κB signaling pathway in ARPE-19 cells.

Expression of fibroblast growth factor-2 and fibroblast growth factor receptor-1 protein in the hippocampus in rats exhibiting chronic stress-induced depression

There is evidence that the expression of members of the fibroblast growth factor （FGF） protein family is altered in post-mortem brains of humans suffering from major depressive disorder. The present study examined whether the expression of fibroblast growth factor-2 （FGF2） and fibroblast growth factor receptor-1 （FGFR1） protein is altered following chronic stress in an animal model. Rats were exposed to 35 days of chronic unpredictable mild stress, and then tested using open-field and sucrose consumption tests. Compared with the control group, rats in the chronic stress group exhibited obvious depressive-like behaviors, including anhedonia, anxiety and decreased mobility. The results of western blot analysis and immunohistochemical analysis revealed a downregulation of the expression of FGF2 and FGFR1 in the hippocampus of rats, particularly in the CA1, CA3 and dentate gyrus. This decreased expression is in accord with the results of post-mortem studies in humans with major depressive disorder. These findings suggest that FGF2 and FGFR1 proteins participate in the pathophysiology of depressive-like behavior, and may play an important role in the mechanism of chronic stress-induced depression.

TRANSFORMING GROWTH FACTOR-β AND FIBROBLAST GROWTH FACTOR INDUCE LENS EPITHELIAL EXPLANT METAPLASIA:IMPLICATIONS FOR THE FORMATION OF SUBCAPSULAR OPACIFICATION

Objective. This study was to investigate the effects of transforming growth factor-β(TGFβ) and fi- broblast growth factor (FGF) in the subcapsular opacification formation of the lens. Methods. Lens epithelial explants from 10-day-old rats were cultured with TGFβ1 or TGFβ2 in the presence of FGF for 5 days, then were examined by light and electron microscopy, and by immunolocal- ization of smooth muscle(α-sm) actin and type I collagen. Results. In TGFβ/FGF-treated explants,extensive proliferation occured, with formation of spindle and star-shaped cells. These cells showed ultrastructure and biochemical features of fibroblast or myofibroblast. Prominent Golgi apparatus and rough endoplaic reticulum were observed in some cells. Intracellular micro- filaments with cytoplasmic dense babies and membrane associated dense bodies, features of smooth muscle cells, were also observed. Some cells showed reactivity to -sin actin antibody. TGFβ/FGF-treated ex- plants were strongly stained with type I collagen antibody. Condusion. In the presence of FGF, TGFβ1 and TGFβ2 induced lens epithelial cell (LEC ) proliferation and transformation into fibroblast or myofibroblast-like cells, with producing of abundant collagen matrix in the explants. The changes are similar to the metaplasia that occurrs in subcapsular opacification of the lens. The findings suggest that TGFβ and FGF plays a role in the pathogenesis of subcapsular opacification of the lens.

RNA interference affects tumorigenicity and expression of insulin-like growth factor-1,insulin-like growth factor-1 receptor,and basic fibroblast growth factor-2 in rat C6 glioma cells

BACKGROUND： Human gliomas are more likely to express basic fibroblast growth factor-2 （FGF-2） insulin-like growth factor-1（IGF-1）, and IGF-1 receptor （IGF-1R） than normal brain tissue. These factors activate signal transduction systems of Ras/MAPK and PI3K/Akl, which promote glioma growth. OBJECTIVE： To utilize RNA interference （RNAi） technique to down-regulate FGF-2, IGF-1, and IGF-1R gene expression, and to investigate the effects of these genes on rat C6 glioma cells, as well as the feasibility of RNAi for treating glioma. DESIGN, TIME AND SETTING： This neurooncological, randomized, controlled, in vivo and in vitro experiment, which used RNAi methodology, was performed at the Laboratory of Molecular Biology, Institute of Biochemistry, Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS： Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences. Small interfering RNA （siRNA） was synthesized by Shanghai GenePharma. Anti-IGF-1, anti-IGF-1R, anti-FGF-2, anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology, USA. Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center, Chinese Academy of Sciences. METHODS： C6 glioma cells were transfected with siRNA, which was chemically synthesized in vitro to correspond to endogenous FGF-2, IGF-1, and IGF-1R genes. The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR, and protein expression was determined by Western blot analysis. C6 glioma cell proliferation was observed using a growth curve C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry. C6 glioma cell growth regression was observed by transwell migration assay. In addition, nude mouse subcutaneous tumor models were used in this study. For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA, two blank control groups, with six mice each, were set up： A （2.5 μg siRNA was injected one week after C6 cells were inoculated, Le., when tumor volume reached 8 mm × 8 mm） and B （siRNA was injected at the same time with C6 cells were inoculated. To study the effects of FGF-2 siRNA, the groups consisted of a blank control group, negative control group, 2.6 μg siRNA group, 4 μg siRNA group, and 5.3 μg siRNA group, with six mice each. MAIN OUTCOME MEASURES： mRNA and protein inhibition ratio of FGF-2, IGF-1, and IGF-1 R; C6 glioma cell proliferation, apoptosis, and cycle growth arrest; C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS： All siRNA constructs proved to be effective. After 48 hours, transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio 〉 80% and an IGF-1 gene inhibition ratio of approximately 70%. Protein expression levels for FGF-2, IGF-1, and IGF-1R decreased in a dose-dependent manner following siRNA transfection, with an inhibition rate 〉 85%, 60%, and 50%, respectively. C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA. The apoptosis rate of C6 glioma cells induced by FGF-2, IGF-1, and IGF-1R siRNA was 39.96%, 15.07% and 22.47%, respectively （P 〈 0.01）. Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration. At 30 days after intratumoral injection of 2.6, 4, and 5.3 tJg FGF-2 siRNA, tumor growth regression rate of FGF-2 siRNA was 56%, 67%, and 86%, respectively. The tumor growth regression rate was 71.88% and 45.71%, respectively, when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation. When IGF-1 or IGF-1 R siRNA was intratumorally injected during C6 glioma cell transplantation, the tumor growth regression rate was 78.13% and 74.29%, respectively. CONCLUSION： siRNA transfection downregulated gene expression of FGF-2, IGF-1, and IGF-1R In addition, siRNA treatment markedly suppressed glioma cell proliferation, growth, and migration, and concomitantly reduced subcutaneous tumorigenicity.

Growth factor-and cytokine-driven pathways governing liver stemness and differentiation

Liver is unique in its capacity to regenerate in response to injury or tissue loss. Hepatocytes and other liver cells are able to proliferate and repopulate the liver. However, when this response is impaired, the contribution of hepatic progenitors becomes very relevant. Here, we present an update of recent studies on growth factors and cytokine-driven intracellular pathways that govern liver stem/pro-genitor cell expansion and differentiation, and the rel-evance of these signals in liver development, regeneration and carcinogenesis. Tyrosine kinase receptor signaling, in particular, c-Met, epidermal growth factor receptors or fibroblast growth factor receptors, contribute to prolifera-tion, survival and differentiation of liver stem/progenitor cells. Different evidence suggests a dual role for the trans-forming growth factor (TGF)-β signaling pathway in liver stemness and differentiation. On the one hand, TGF-βmediates progression of differentiation from a progenitor stage, but on the other hand, it contributes to the expan-sion of liver stem cells. Hedgehog family ligands are nec-essary to promote hepatoblast proliferation but need to be shut off to permit subsequent hepatoblast differentiation. In the same line, the Wnt family and β-catenin/T-cell fac-tor pathway is clearly involved in the maintenance of liver stemness phenotype, and its repression is necessary for liver differentiation during development. Collectively, data indicate that liver stem/progenitor cells follow their own rules and regulations. The same signals that are essential for their activation, expansion and differentiation are good candidates to contribute, under adequate conditions, to the paradigm of transformation from a pro-regenerative to a pro-tumorigenic role. From a clinical perspective, this is a fundamental issue for liver stem/progenitor cell-based therapies.

Tetrandrine inhibits activation of rat hepatic stellate cells in vitro via transforming growth factor-β signaling

AIM: To investigate the effect of various concentrations of tetrandrine on activation of quiescent rat hepatic stellate cells (HSCs) and transforming growth factor-β (TGF-β) signaling in vitro.METHODS: HSCs were isolated from rats by in situperfusion of liver and 18% Nycodenz gradient centrifugation, and primarily cultured on uncoated plastic plates for 24 hwith DMEM containing 20% fetal bovine serum (FBS/DMEM) before the culture medium was substituted with 2% FBS/DMEM for another 24 h. Then, the HSCs were cultured in 2% FBS/DMEM with tetrandrine (0.25, 0.5, 1,2 mg/L, respectively). Cell morphological features were observed under an inverted microscope, smooth muscleα-actin (α-SMA) was detected by immunocytochemistry and image analysis system, laminin (LN) and type Ⅲprocollagen (PCⅢ) in supernatants were determined byradioimmunoassay. TGF-β1 mRNA, Smad 7 mRNA and Smad 7 protein were analyzed with RT-PCR and Western blotting, respectively.RESULTS: Tetrandrine at the concentrations of 0.25-2 mg/L prevented morphological transformation of HSC from the quiescent state to the activated one, while α-SMA, LN and PCⅢ expressions were inhibited. As estimated by gray values, the expression of α-SMA in tetrandrine groups (0.25, 0.5, 1, 2 mg/L) was reduced from 21.3% to 42.2%(control: 0.67, tetrandrine groups: 0.82, 0.85, 0.96, or 0.96, respectively, which were statistically different from the control, P＜0.01), and the difference was more significant in tetrandrine at 1 and 2 mg/L. The content of LN in supernatants was significantly decreased in tetrandrine groups to 58.5%, 69.1%, 65.8% or 60.0% that of the control respectively, and that of PCⅢ to 84.6%, 81.5%,75.7% or 80.7% respectively (P＜0.05 vs control), with no significant difference among tetrandrine groups. RTPCR showed that TGF-β1 mRNA expression was reduced by tetrandrine treatments from 56.56% to 87.90% in comparison with the control, while Smad 7 mRNA was increased 1.4-4.8 times. The TGF-β1 mRNA and Smad 7 mRNA expression was in a significant negative correlation (r= -0.755, P＜0.01), and both were significantly correlated with α-SMA protein expression (r = -0.938, P＜0.01;r = 0.938, P＜0.01, respectively). The up-regulation of Smad 7 protein by tetrandrine (1 mg/L)was confirmed by Western blotting as well.CONCLUSION: Tetrandrine has a direct inhibiting effect on the activation of rat HSCs in culture. It up-regulates the expression of Smad 7 which in turn blocks TGF-β1 expression and signaling.

Semaphorin 7a participants in pterygium by regulating vascular endothelial growth factor

AIM: To investigate the relationship between semaphorin 7a expression and cell proliferation and migration in pterygium fibroblasts. METHODS: Twenty-six patients with surgically diagnosed pterygium were enrolled, including 15 cases of primary pterygium and 11 cases of recurrent pterygium. In addition, 12 cases of normal conjunctival tissue were collected. The expression of semaphorin 7a in normal conjunctival tissue, primary pterygium and recurrent pterygium was detected by real-time polymerase chain reaction. Recurrent pterygium fibroblasts were isolated and cultured, and the expression of semaphorin 7a was silenced by small interfering RNA(siRNA) interference technique. Furthermore, the effects of si-semaphorin 7a interference on the mRNA and protein levels of β1-integrin, vascular endothelial growth factor A(VEGFA) and vascular endothelial growth factor receptor(VEGFR), and on fibroblast proliferation were analyzed. Transwell assay was used to detect the effect of semaphorin 7a interference on fibroblast migration. RESULTS: Semaphorin 7a was highly expressed in the primary pterygium and recurrent pterygium samples than that of the normal conjunctival tissue. Compared with the primary pterygium, the expression of semaphoring 7a in the recurrent pterygium samples was significantly increased(P<0.05). The mRNA and protein expression levels of β1-integrin, VEGFA and VEGFR were decreased after si-semaphorin 7a transfection, and as well as the cell proliferation and migration. CONCLUSION: Semaphorin 7a might play important roles in the pathogenesis of pterygium by affecting the expression of β1-integrin, VEGFA and VEGFR.

心房颤动病人血清SLC7A11、FGF23水平检测及临床意义

目的:检测心房颤动(AF)病人与窦性心律者血清溶质载体家族7成员11(SLC7A11)、血清成纤维细胞生长因子23(FGF23)的水平,分析二者与AF之间的相关性及临床意义。方法:选取住院的AF病人118例作为观察组,根据相关指南分为阵发性AF组67例和非阵发性AF组51例。对照组选取窦性心律健康者96名。选择酶联吸附免疫实验法(ELISA)测出血清中SLC7A11、FGF23浓度;比较3组病人的临床资料及血清学指标,利用Pearson相关性分析血清SLC7A11、FGF23水平与超声心动图中左房内径(LAD)、左室舒张内径(LVD)、左心室射血分数(LVEF)和左心室缩短分数(FS)相关性。采用多元logsitic回归分析AF病人AF发生持续相关因素。结果:与对照组相比,血清SLC7A11在阵发性AF组和非阵发性AF组均下降(P<0.01),且非阵发性AF组中SLC7A11低于阵发性AF组(P<0.01),血清FGF23在阵发性AF组和非阵发性AF组均升高(P<0.01),且非阵发性AF组中FGF23高于阵发性AF组(P<0.01);Pearson相关性分析显示,LAD与血清SLC7A11呈负相关关系(r=-0.534,P<0.01),与血清FGF23呈正相关关系(r=0.532,P<0.01)。多元logsitic回归分析结果显示,SLC7A11是AF独立的保护因素(OR=0.231,P<0.01),而FGF23是独立危险因素(OR=1.097,P<0.01)。结论:SLC7A11、FGF23可能与AF的发病、进展有关。

成纤维细胞生长因子-7在巨噬细胞感染结核分枝杆菌中的免疫机制初步研究

巨噬细胞分泌的纤维细胞生长因子-7(fibroblast growth factor-7,FGF-7)具有一定的细胞修复作用及抗炎症作用。通过PCR技术、Western-blot及ELISA实验研究分析FGF-7在巨噬细胞感染结核分枝杆菌(Mycobacterium tuberculosis,M.avium)后的分子免疫机制。研究发现fgf-7基因在结核病患者外周血单个核细胞中表达增强,并且U973巨噬细胞在感染M.avium后,其fgf-7基因与FGF-7蛋白亦表达增强,同时U973巨噬细胞上清中的细胞因子TNF-α与IFN-γ分泌量显著增加。实验结果表明巨噬细胞受M.avium感染后,M.avium可增强巨噬细胞fgf-7基因及其蛋白质的表达,并促进细胞因子TNF-α与IFN-γ的分泌;提示FGF-7可能与TNF-α、IFN-γ等共同引起炎症反应从而参与对M.avium的抑制或杀伤作用,并修复损伤的巨噬细胞。

碳酸司维拉姆联合血液透析对尿毒症后肾性骨病患者的影响

目的:观察并探讨尿毒症后肾性骨病患者临床治疗过程中血液透析与碳酸司维拉姆联合应用效果及对成纤维细胞生长因子23(FGF-23)、骨形态发生蛋白7(BMP-7)的影响。方法:于2022年1月—2023年1月广州市第一人民医院南沙医院接收72例尿毒症后肾性骨病患者,运用随机数字表法给进行分组,对照组(n=36)、联合组(n=36)。其中对照组进行血液透析治疗,联合组则在血液透析基础上加用碳酸司维拉姆。比较两组临床治疗效果、骨碱性磷酸酶(BAP)、FGF-23、BMP-7、甲状旁腺激素(PTH)、C反应蛋白(CRP)、β_(2)微球蛋白(β_(2)-MG)、骨钙素(OCN)、不良反应。结果:联合组临床总有效率为91.67%,高于对照组的72.22%,差异有统计学意义(P<0.05)。治疗前,两组BAP、FGF-23、BMP-7、PTH、CRP、β_(2)-MG、OCN比较,差异均无统计学意义(P>0.05);治疗后,联合组BMP-7高于对照组,BAP、FGF-23、PTH、CRP、β_(2)-MG、OCN均低于对照组,差异均有统计学意义(P<0.05);联合组胃胀气、腹泻、反胃、干呕等不良反应发生率为11.11%与对照组的16.67%比较,差异无统计学意义(P>0.05)。结论:尿毒症后肾性骨病患者在血液透析期间联合碳酸司维拉姆治疗具有确切效果,可调节骨代谢,改善肾功能,减轻微炎症反应,安全性高,患者耐受。

促愈消肿汤对运动致肱骨远端骨折患者术后骨生长及功能恢复的影响

目的:探究促愈消肿汤对运动致肱骨远端骨折患者术后血清成纤维细胞生长因子2(FGF-2)、骨形态发生蛋白7(BMP-7)水平及骨愈合状态的影响。方法:选取2021年3月至2022年9月河南省中医院收治的运动致肱骨远端骨折患者86例作为研究对象,采用随机数字法分为对照组和观察组,每组43例。对照组术后采用常规康复治疗;观察组在对照组基础上服用促愈消肿汤治疗。对比治疗后2组临床疗效及术后恢复情况,术后1 d与术后7 d时2组肘关节功能、关节活动度及血清FGF-2、BMP-7水平差异。结果:观察组临床疗效为93.02%,显著高于对照组的76.74%(P<0.05);治疗后观察组疼痛消除时间、术处消肿时间及骨折愈合时间显著短于对照组(均P<0.05);术后7 d时,观察组Mayo肘关节功能量表,关节活动度(ROM)评分以及血清FGF-2、BMP-7水平显著升高,且高于对照组(均P<0.05);术后7 d时,观察组视觉模拟评分法(VAS)得分较术后1 d时显著降低,且低于对照组(均P<0.05)。结论:运动致肱骨远端骨折患者术后服用促愈消肿汤能有效促进血清FGF-2、BMP-7水平表达上调,改善肘关节活动功能,缓解术后疼痛,加速骨折愈合。

扶肾降浊方含药血清对肾小管间质损害大鼠成纤维细胞抗纤维化因子HGF和BMP-7的影响

目的:探讨扶肾降浊方对系膜增生性肾小球肾炎(Ms PGN)大鼠肾小管间质损害的疗效及机制。方法:采用扶肾降浊方水溶液灌胃Wistar大鼠常规制备含药血清,在Ms PGN动物模型的基础上,延长造模时间至20周,使其自然发展为肾小管间质损害模型,体外培养造模12、16和20周末大鼠间质成纤维细胞,采用real-time PCR和Western blotting法检测扶肾降浊方含药血清对病理状态间质成纤维细胞中抗纤维化因子肝细胞生长因子(HGF)、骨形态发生蛋白-7(BMP-7)mRNA及蛋白表达的影响。结果:病理状态间质成纤维细胞中抗纤维化因子HGF、BMP-7 mRNA和蛋白表达下调,扶肾降浊方含药血清随着给药周期的延长可部分逆转间质损害造成的上述mRNA和蛋白表达异常。结论:扶肾降浊方含药血清对Ms PGN大鼠间质成纤维细胞的保护作用可能与调节抗纤维化因子HGF、BMP-7 mRNA和蛋白表达有关。

3I、7b型腺病毒感染对人胚肺成纤维细胞转化生长因子β_1 mRNA及其蛋白表达的影响

目的探讨3I、7b型腺病毒型感染对体外培养的人胚肺成纤维细胞转化生长因子β1(TGF-β1)mRNA及其蛋白表达的影响。方法以3I、7b型腺病毒分别攻击体外培养的人胚肺成纤维细胞,另设正常细胞组。采用酶联免疫吸附试验及原位杂交法检测各组细胞TGF-β1蛋白及TGF-β1mRNA表达。结果3I、7b型病毒感染组较正常细胞组TGF-β1蛋白及mRNA表达均明显增强(Pa<0.01),而3I、7b型病毒感染组间比较无显著性差异(Pa>0.05)。结论肺成纤维细胞和TGF-β1可能参与腺病毒肺炎的发病过程。

重组hFGF-7腺病毒对角质形成细胞的生物学效应

目的:研究重组腺病毒导入人成纤维细胞生长因子7(hFGF-7)对人皮肤角质形成细胞(HaCat)的生物学效应。方法:通过倍比稀释和感染实验,测定重组腺病毒rAd-hFGF-7的滴度;流式细胞术检测重组腺病毒感染HaCat细胞的感染效率;MTT法检测rAd-hFGF-7对HaCat细胞增殖的影响;重组腺病毒对HaCat细胞周期的影响通过流式细胞术进行检测。结果:高滴度重组腺病毒可高效感染HaCat细胞,其促细胞增殖作用随着重组腺病毒MO I值的增加而增强,当MO I值为50时,感染细胞进入S期和G2期的细胞比率显著增加。结论:重组腺病毒rAd-hFGF-7可促进HaCat细胞的增殖,改变HaCat细胞周期。

rhKGF、CBLB502和WR2721对放射性口腔黏膜炎的防治效果观察

目的比较r h KG F、CB L B 5 0 2、W R 2 7 2 1对放射性口腔黏膜炎(R O M)的防治作用。方法健康雄性C57BL/6J小鼠随机分为正常对照组、照射对照组、rh KGF组、CBLB502组及WR2721组。其中24只小鼠观察17Gy头颈部照射小鼠的30d存活率及体重变化,另外20只小鼠利用1%甲苯胺蓝染色法观察照射小鼠舌组织溃疡情况,HE染色观察舌组织病理学改变,Ki-67免疫组化检测黏膜角化上皮细胞增殖情况。结果与照射对照组相比,rh KGF和WR2721给药组小鼠30d存活率显著提高,体重恢复较快,角化上皮细胞增殖显著,未发生明显的口腔黏膜炎;CBLB502组小鼠的存活率、体重变化及组织病理学指标与照射对照组比较差异无统计学意义(P>0.05)。结论 rh KGF和WR2721对放射性口腔黏膜炎有较好的防治作用,而CBLB502并不能减轻放射性口腔黏膜炎的发生。