CNKI引文数据库《园艺学报》高被引频次论文排序

(截至2006年6月,排名前100位)序号被引文献题名被引文献作者被引文献来源被引频次1蔬菜硝酸盐累积的研究———Ⅰ.不同蔬菜硝酸盐沈明珠,翟宝杰,东惠茹,李俊国园艺学报/1982/04266和亚硝酸盐含量评价2温室土壤次生盐渍化的形成和治理途径研究童有为,陈淡飞园艺学报/1991/021323     

Effects of sterigmatocystin on interferon-γ secretion of human peripheral blood mononuclear cellsin vitro

AIM:To explore the putative effects of sterigmatocystin (ST) on human help T lymphocyte(Th1)function. METHODS:The effects of ST on interferon-γ(IFN-γ)secretion of human peripheral blood mononuclear cells(HPBMc) in vitro were determined with ELISA method. RESULTS:The effects of ST on IFN-γ secretion of HPBMc in vitro were closely dependent on ST concentrations. ST at relatively lower concentrations (0.03125-0.12500 mg/L) showed inhibiting effects on IFN-γ secretion. While, stimulating effects could be found when ST concentration was above 0.25mg/L. The highest level was seen in ST 1 mg/L group (P＜0.05)。At concentration ranging from 0.25 mg/L to 1 mg/L, a positive dose- effects correlation was found between ST concentration and IFN-γ secretion (r=0.492, P＜0.01). Time-effects analysis from 1 h to 64 h after ST treatment (1 mg/L)showed that the effects of ST on IFN-γ secretion varied as the changes of treatment times. An inhibiting effect on IFN-γ secretion of HPBMc 4 h and 8 h after ST treatment was found (8 h, P＜0.05). As the treatment time prolonged from 16 h, IFN-γ level gradually increased (32 h, P<0.05). There was a positive correlation between treatment time of ST and IFN-γ level of HPBMc in vitro from 16 h to 64 h after ST treatment (r=0.736, P＜0.01).CONCLUSION:The effects of ST on IFN-γ secretion of HPBMc in vitro closely depended on concentration and treatment time of ST. Generally, inhibiting effects were found at relatively lower ST concentration and shorter treatment period, while stimulation effects could be seen at relatively higher ST concentration and longer ST treatment time period.     

Influences of protein kinase C inhibitors on the expression of IL-2 and IFN-γ by human T-lymphocytes

AIM:To investigate the influences of protein kinase C(PKC) inhibitors on the expression of interleukin-2(IL-2) and interferon-γ(IFN-γ) byin vitro activated T-lymphocytes. METHODS:Double fluorescent staining together with flow cytometry was adopted to detect intracellular cytokines and to analyze the effects of H7 and gossypol on IL-2 and IFN-γ expression levels of T-lymphocytes stimulated with phorbol ester (PDB)+ionomycin(I) in the presence of monensin.RESULTS:The expression rates of IL-2 and IFN-γ of CD3⁺ T cells stimulated with PDB+I for 4 h were 16.64±2.04 and 25.81±3.53(x±s), respectively, which were significantly higher than that of control (1.06±0.22 and 3.12±0.77)(P＜0.05). Gossypol was able to inhibit the expression of IL-2 and IFN-γ significantly, with the expression rates of 2.08±0.12 and 9.01±1.90, respectively. At the presence of 50 μmol/L H7, the rates of IL-2⁺ and IFN-γ⁺ CD3⁺ T cells were 0.43±0.06 and 2.40±0.27, respectively. The effect of H7 was stronger than that of gossypol. CONCLUSION:PKC plays an important role in the expression of IL-2 and IFN-γ of CD3⁺T cells and its inhibitors H7 and gossypol exert significant inhibitory effect on the expression of these two cytokines. It is suggested that H7 and gossypol may have modulatory effect on T-cell-dependent specific immune responses by inhibiting PKC activity.     

“2012年园艺植物染色体倍性操作与遗传改良学术研讨会”预备通知

自2008年11月在中国农业科学院郑州果树研究所成功召开园艺植物染色体倍性操作与遗传改良研讨会以来,我国科研人员在该领域的研究取得了可喜的成绩。为进一步总结近几年来在该领域的研究进展,促进园艺植物倍性育种研究,同时为该领域的专家、学者和同仁们提供良好的交流平台。中国园艺学会定于2012年4月中旬在重庆召开园艺植物染色体倍性操作与遗传改良学术研讨会。欢迎从事该研究领域及相关研究工作的人员参加。     

Effect of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 and expression of related cytokines in penetrating keratoplasty in mice

AIM: To evaluate the effects of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 (Th17 cells) and its related cytokines in a mouse model of corneal allograft transplantation. METHODS: C57BL/6 mice were transplanted with corneal grafts from BALB/c mice and treated intraperitoneally with 1.0 μg 1α, 25-dihydroxyvitamin D3 or soybean oil every other day after operation. The transparency of the corneal grafts was evaluated for potential rejection signs by slit lamp biomicroscopy and histopathology. The expression levels of IL-17, RORγt and IFN-γ in the spleen were measured by real-time PCR. Moreover, the protein expression of RORγt and IL-17 in the peripheral blood was analyzed by Western blotting. IL-17 and IFN-γ in peripheral blood were measured by flow cytometry. RESULTS: 1α, 25-dihydroxyvitamin D3 significantly inhibited the rejection of the corneal allograft and reduced the numbers of inflammatory infiltrates in the corneal graft. In the spleen, 1α, 25-dihydroxyvitamin D3 treatment reduced the expression levels of IL-17, RORγt and IFN-γ. In the peripheral blood, 1α, 25-dihydroxyvitamin D3 treatment downregulated the expression levels of RORγt, IL-17 and IFN-γ. CONCLUSION: The effects of 1α, 25-dihydroxyvitamin D3 on suppressing corneal transplantation-induced allograft rejection in mice are closely associated with its modulation on IL-17 and related cytokine RORγt.     

Effects of interferon-γ, tumor necrosis factor-α and interleukin-1β on apoptosis of human airway smooth muscle cellsin vitro

AIM:To clarify if interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)can induce apoptosis of human airway smooth muscle cells (ASMCs) in vitro.METHODS:Human ASMCs were isolated and cultured in DMEM containing 10% fetal bovine serum. Passage 4-6 cell was used in the experiment. IFN-γ,TNF-α and IL-1β, were used separately or together in the treatment of human ASMCs. The effects of IFN-γ,TNF-α and IL-1β on the growth of the cells was detected by MTT method at the hour 0,24,48 and 72. Light microscopy and electron microscopy were used to examine the morphological change. DNA fragmentation was analyzed by agarose gel electrophoresis. SP immunohistological staing method was performed to detect the change of expressions of p 53, bcl- 2 and bax gene. The apoptosis cell percentage were detected by in situ end labeling technique (TUNEL)of fragmental DNA. RESULTS:(1)IFN-γ or IFN-γ together with TNF-α and IL-1β decreased the number of viable cells in a time dependent manner. (2) Light and electron microscopic examination showed cell shrinkage, membrane blebbing, nuclear contraction, chromatin condensation and nuclear fragmentation in human ASMCs. (3) Agarose gel electrophoresis showed a characteristic"ladder"of DNA bands representing integer multiples of the internucleosomal fragments (about 180-200 bp) in cytokine cotreated human ASMCs. (4)The expression of p 53 and bax gene in cytokine cotreated group was significantly higher than in control group, but the expression of bcl-2 gene was lower than in control group. (5)Stimultaneous treatment with IFN-γ(4×10⁵ U/L),TNF-α(4×10⁵ U/L)and /or IL-1β (10×10⁴ U/L) induced apoptosis of human ASMCs. Apoptotic index of human ASMCs in cytokine co-treated group was significantly higher than in control group (P＜0.01).CONCLUSION:Stimultaneous treatment with IFN-γ,TNF-α and /or IL-1β induced apoptosis of human ASMCs. These immune cytokines may play an important role in airway remodeling of asthma and of chronic obstructive pulmonary disease.     

Functional expression of B7H1 on pancreatic carcinoma panc-1 cells

AIM: To observe the effect of B7H1 expression in pancreatic carcinoma cells on the proliferation and activation of co-cultured T lymphocytes. METHODS: B7H1 expression in panc-1 cells before and after interferon-γ(IFN-γ) treatment or B7H1-siRNA transfection was evaluated by RT-PCR and flow cytometry. The influence of B7H1 expression on co-cultured PHA-activated T lymphocytes was determined by the methods of MTT and enzyme-linked immunosorbent assay (ELISA). RESULTS: B7H1 was highly expressed in panc-1 cells and up-regulated after IFN-γ stimulation. Such up-regulation led to the significant inhibition of T cell proliferation and secretion of cytokines such as IFN-γ and interleukin-2(IL-2). However, IL-10 production was enhanced. In contrast, knockdown of B7H1 expression in panc-1 cells by RNA interference resulted in increased T cell proliferation as well as IFN-γ and IL-2 production. Meanwhile, the IL-10 secretion decreased. CONCLUSION: B7H1-expressing panc-1 cells suppress T cell function by inhibiting T cell proliferation and production of Th1 cytokines. Suppression of B7H1 expression through siRNA restores T cell immune functions, indicating a potential strategy for immunotherapy against pancreatic cancer.     

Analysis of cytokines derived from murine yolk sac endothelial cells cultured in vitro

AIM:To purify murine yolk sac endothelial cells (mYS-EC) and investigate the cytokines mRNA expression in mYS-EC. METHODS:The murine yolk sacs were digested with 0.1% collagenase, resuspended in DMEM and counted after digestion and centrifugation. The yolk sac adherent cells were cultured in DMEM containing 15% FBS with 10% mBMEC-CM or 5μg/L VEGF, ECGF and bFGF. The phagocytose function and expression of vWF were evaluated via particle phagocytosis and immunohistochemistry method. Atlas cDNA expression array was used for analysis of cytokine expression in mYS-EC. RESULTS:Colonies consisting of pure yolk sac endothelial cells were obtained in liquid culture system containing 15% FBS and 10% mBMEC-CM or 5μg/L VEGF, ECGF and bFGF. For complete purification of the endothelial cells, subsequent passage was also necessary. Cellular cord formed during passage culture. The endothelial cells were round or oval sharp in morphology, positive in phagocytosis and factor VIII related antigen (von Willebrand's Factor, vWF). The mRNA expressions of cytokines, such as TGF-β2, TNF-α, IFN-γ, FL, BMP-4, MIP-1β, BMP-2A, FLT2, endothelin 2, thymosin β10, IL-6, IL-13, IL-9, SCYA5 and ACBP were detected in mYS-ECs. CONCLUSION:mYS-EC was purified and expanded in vitro. The mRNA expression of 15 kinds of cytokines was detected in mYS-ECs by Atlas arrays.     

Effects of PFOA exposure on inflammatory mediators IL-4, IFN-γ and glucocorticoid receptor in asthmatic mice

AIM: To investigate the effects of perfluorooctanoic acid (PFOA) exposure on the changes of asthmatic mouse airway inflammation, inflammatory mediators interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum, and glucocorticoid receptor (GR) expression in the lung tissue.METHODS: BALB/c mice (n=30) were randomly divided into 5 groups:normal control (C) group, asthma (A) group, asthma+low-dose PFOA (AP10) group, asthma+ mode-rate-dose PFOA (AP50) group and asthma+high-dose PFOA (AP100) group. Asthma model and PFOA exposure model of mice were established according to the grouping. The animals were sacrificed and their lungs were collected for HE staining, transmission electron microscopy, Western blot and immunohistochemical staining. ELISA was applied to detect the levels of IL-4 and IFN-γ in the serum.RESULTS: HE staining of the lungs showed that the asthmatic mice, compared with the normal control mice, had obvious mucus secretion around the airways and infiltration of inflammatory cells around airways and blood vessels, and the effects were much more marked in AP groups. Ultrastructural alteration of the lung tissues in the asthmatic mice were indicated by transmission electron microscopy. Compared with C group, the results of ELISA in A group and AP groups proved that IL-4 in the serum was increased and IFN-γ was decreased significantly (P<0.05). Compare with A group, IL-4 was significantly increased and IFN-γ was decreased in AP100 group (P<0.05), and no difference of those between AP10 group and AP50 group was found. The results of Western blot indicated that GR protein expression in the asthmatic mice were decreased compare with the normal mice (P<0.05), and no difference of that among A group and AP groups was observed. Immunohistochemical staining manifested that GR protein was mainly located in the cytoplasm of bronchial columnar epithelial cells, airway smooth muscle cells and vascular smooth muscle cells.CONCLUSION: Acute airway PFOA exposure in asthmatic mice dose-dependently exacebates lung inflammation by inducing Th2 type immune responses, promotes infiltration of inflammatory cells and mucus secretion around the airways and blood vessels, and destroys the ultrastructure of the lung tissues.     

Effect of adoptive transfer of CD4+ T cells with miR-7 knockdown on acute liver injury in mice

AIM:To study the effect of adoptive transfer of CD4⁺ T cells with microRNA-7 (miR-7) knockdown (KD) on mouse acute liver injury model and to investigate its significance. METHODS:CD4⁺ CD62L⁺ T cells were purified from the spleen of normal wild-type (WT) mice and miR-7KD mice by magnetic bead sorting, and were stained with CFSE. These 2×10⁶ CFSE-labeling cells were injected into normal mice via tail vein, and then the mouse acute liver injury model was induced by intraperitoneal injection of 30 mg/kg concanavalin A. After 72 h, the appearance, weight and weight index of the liver were investigated. The pathological change of the liver tissues was observed by HE staining. Real-time PCR was used to examine the mRNA expression of Bax and P53. The expression levels of CD62L, interleukin-4 (IL-4) and interferon-γ (IFN-γ) in the CD4⁺ T cells were analyzed by flow cytometry. RESULTS:We found that the liver tissue became lighter, and the weight (P<0.01) and weight index (P<0.05) were changed significantly in miR-7KD mice compared with control group. Moreover, HE staining showed that the liver cell damage was increased in the liver of miR-7KD mice. Meanwhile, the expression levels of Bax and P53 were significantly increased in miR-7KD group (P<0.05). The percentage of CD62L in CD4⁺ T cells was significantly decreased (P<0.01) in miR-7KD mice, with high expression of IFN-γ (P<0.05) and low expression of IL-4 (P<0.01) in CD4⁺T cells. CONCLUSION:These findings suggest that miR-7 knockdown significantly promotes the pathology of CD4⁺ T cell-mediated acute liver injury, which provides a preliminary experimental basis for further exploration on the mechanism of acute liver injury occurrence.     

Recombinant human endostatin regulates proliferation and related protein expression in human multiple myeloma cell line RPMI 8226

AIM: To investigate the effects of recombinant human endostatin (Endostar) on proliferation, cell cycle constitution and regulation of related protein expression in RPMI 8226 cells. METHODS: CCK-8 assay was used to explore the effect of Endostar on the proliferation of RPMI 8226 cells. The action of Endostar on apoptosis and cell cycle distribution were determined by flow cytometry. The expression of Bcl-2 and caspase-3 was evaluated by Western blotting analysis. The production of vascular cell adhesion molecule 1 (VCAM-1), interleukin 6 (IL-6) and vascular endothelial growth factor (VEGF) in response to Endostar was measured by real-time PCR and Western blotting. The influence of Endostar on the secretion of IL-6 and VEGF in the culture supernatants was detected by ELISA. RESULTS: Endostar inhibited the proliferation of RPMI 8226 cells with the highest inhibitory rate of 59.5±5.6% at dose of 250 mg/L, which was associated with slight up-regulation in the proportion of G1 phase. No significant modulation of apoptotic rate was observed with Endostar, as well as the expression of Bcl-2 or caspase-3. Endostar declined the expression of VCAM-1 and also attenuated the production and secretion of IL-6 and VEGF in RPMI 8226 cells. CONCLUSION: Endostar suppresses the proliferation of RPMI 8226 cells by enhancing the proportion of G1 phase and decreasing VCAM-1 expression and self-secretion of IL-6 and VEGF. No apoptosis of RPMI 8226 cells is induced by Endostar.     

Detection and enrichment of T lymphocytes based on cytokine secretion in human mixed lymphocyte reaction

AIM: Detection and enrichment of T lymphocytes after allogeneic PBMNC stimulation according to secreted cytokine were performed in order to explore a new approach for studying allogeneic reactive T lymphocytes. METHODS: The novel cytokine secretion assay (CKSA) was applied to detect T lymphocyte secreting IFN-γ, IL-4 and IL-10 at single cell level in human mixed lymphocyte reaction. IFN-γ secreting T cells were enriched by means of magnetic sorting system. RESULTS: Allogeneic PBMNC stimulation didn't alter the proportion of IL-4 and IL-10 secreting T lymphocytes (which were 0.12%±0.03% and 0.10%±0.03%, respectively), but increased proportion of IFN-γ secreting T lymphocytes (1.12%±0.13%). These IFN-γ- secreting T lymphocytes could be further enriched to 67.3%±10.5% . CONCLUSION: It is feasible to detect significantly increased IFN-γ-secreting T cells after allogeneic PBMNC stimulation based on the novel CKSA technique, and these cells could be efficiently enriched for further use.     

Suppressive effect of interferon γ on IL-13-induced fibrosis in fibroblasts

AIM：To investigate the suppressive effect of interferon γ (IFN-γ) on fibrosis induced by interleukin 13 (IL-13) in fibroblasts. METHODS：The fibroblasts were divided into IFN-γ (4×105U/L) group, IL-13 (100 μg/L) group, IFN-γ+IL-13 group and blank control group. At 24 h, 48 h and 72 h, the secreted collagen from fibroblasts was measured by hydroxyproline release assay. The mRNA expression of collagen type I α1 (Col1A1) in fibroblasts was examined by RT-PCR. The protein level of collagen type I synthesized in fibroblasts was analyzed by Western blotting. RESULTS：IFN-γ at 4×105U/ L significantly inhibited the proliferation of fibroblasts and down-regulated Col1A1 mRNA and cellular collagen. The mRNA expression of Col1A1 and the protein level of collagen type I in IFN-γ group were lower than those in blank control group at 48 h and 72 h. At 72 h, the mRNA expression of Col1A1 and the protein level of collagen type I in IL-13 group were substantially higher than those in blank control group, those in IFN-γ + IL-13 group were remarkable lower than those in blank control group, and those in IFN-γ group were also lower than those in blank control group. CONCLUSION：IFN-γ inhibits the fibrotic effect of IL-13 in fibroblasts.     

The expression of IFN-γ and IL-4 on T lymphocytes that infiltrate in nasal polyps

AIM:To investigate the expression of Th1-typed cytokine IFN-γ and Th2-typed cytokine IL-4 on T lymphocytes that infiltrate in nasal polyps for searching the pathogenesis of nasal polyps. METHODS:Nasal polyps tissue samples and peripheral blood were obtained from 21 patients. Normal human inferior turbinate mucosa and peripheral blood were obtained as well. Flow cytometry was adopted to detect the expression of IFN-γ and IL-4 of T lymphocytes. RESULTS: Th cytokines were rarely detected in inferior turbinate from normal human. Nasal polyps tissue consisted of abundant T lymphocytes. The expression of IL-4 and IFN-γ increased in peripheral blood from patients compared with normal human (P<0.05). The expression of IL-4 increased but the expression of IFN-γ decreased in nasal polyps compared with that of peripheral blood from the same patient (P＜0.05). CONCLUSION:There were generous of T lymphocytes infiltrating in nasal polyps. There was abnormal immune status in the local nasal mucosa from the patients, and the predomination of Th cytokine secretion changed compared with peripheral blood from the same patients, which resulted in the change of microenvironment of nasal mucosa and possibly close related to the formation of nasal polyps.     

Anti-Sonic hedgehog blocking antibody enhances killing effect of PBMCs on cervical carcinoma HeLa cells

AIM: To investigate the effect of anti-Sonic hedgehog(Shh) blocking antibody on the killing effect of peripheral blood mononuclear cells(PBMCs) on cervical carcinoma HeLa cells. METHODS: The expression levels of Shh and Shh signaling molecules in HeLa cells were detected by immunocytochemistry and RT-PCR. PBMCs from health peoples were isolated by the method of Ficoll density gradient centrifugation, and then co-cultured with HeLa cells in vitro. The expression of CD3, CD69 and CD71 was assayed by flew cytometry. The concentrations of IFN-γ, IL-10 and IL-4 in culture supernatants were detected by ELISA. The killing effect of PBMCs on HeLa cells was observed under microscope. RESULTS: Shh and Shh signaling molecules were expressed in HeLa cells. The level of Shh expression didn't change significantly in the 6th passage of HeLa cells. CD3⁺ cells were increased in the co-culture system. The expression of CD69 and CD 71, and the secretion of IFN-γ were increased, while the secretion of IL-10 was decreased in the co-culture system treated with anti-Shh blocking antibody. Anti-Shh blocking antibody has no effect on the secretion of IL-4. The killing effect of PBMCs on HeLa cells was strengthened by anti-Shh blocking antibody. CONCLUSION: Anti-Shh blocking antibody promotes the activation of PBMCs and enhances the killing effect of PBMCs on cervical carcinoma HeLa cells.     

Allogeneic antigen-pulsed murine immature CD8α+ dendritic cells prevent T-cells proliferation in vitro

AIM: To explore the optimal condition of predominant immature CD8α⁺ dendritic cells(predo-iDCs) preventing T-cell proliferation pulsed by allogeneic antigen. METHODS: Predo-iDCs, induced with GM-CSF +IL-4 +SCF +Flt3L from SPF healthy C57BL/6 murine bone marrow cells, were pulsed by different doses (0, 2.5, 5, 10 and 20 mg/L)of allogeneic murine splenocyte antigen. Syngeneic T-cells were co-incubated with Ag-pulsed DCs (DCs/T=1∶ 1, 2∶ 1, 4∶ 1) and the T-cell proliferation was measured by MTT. The secretion of cytokines (IFN-γ and IL-10) in the co-incubated supernatants was detected by ELISA. The effect of prevention of T-cell proliferation generated by murine predo-iDCs pulsed by allogeneic antigen was detected. The control derived from mature dendritic cells(mDCs)induced by GM-CSF +IL-4 +TNF-α. RESULTS: The effect of Ag-pulsed predo-iDCs for stimulating T-cell proliferation was the slightest in predo-iDCs/T 1∶ 1 group, compared with that in predo-iDCs/T 2∶ 1 and 4∶ 1 group (P<0.05). The secretion of IFN-γ in mixed lymphocyte reaction(MLR) was significantly lower than the one in mDCs control group, while the secretion of IL-10 was higher than that in control group when low dose of antigen (<2.5 mg/L) was added into MLR. CONCLUSION: Predominant iDCs pulsed by low dose of allogeneic antigen (2.5 mg/L) mixed 1∶ 1 with T-cells is the optimal condition for the prevention of T-cells proliferation.     

Effects of arsenic trioxide on T-bet/GATA3 signal pathway in MRL/lpr mice

AIM: To investigate the effect of arsenic trioxide (ATO) on T-bet/GATA3 signal pathway in MRL/lpr mice.METHODS: MRL/lpr mice and C57BL/6J mice at the age of 20 weeks were chosen and then divided in 2 different sub-groups, respectively. The mice in 2 sub-groups received ATO (0.4 mg·kg^-1·d^-1) and sodium chloride (NS, volume weight-determined) by intraperitoneal injection respectively for 2 months. Afterward, the spleens were isolated from the MRL/lpr and C57BL/6J mice under pathogen-free condition and the suspensions were prepared. The mRNA level of T-bet, GATA3, IFN-γ,IL-4 and the mRNA ratio of T-bet/GATA3 were detected by RT-qPCR. The protein expression of T-bet and GATA3 was determined by Western blot. The serum levels of IFN-γ and IL-4 were measured by ELISA.RESULTS: The mRNA and protein levels of T-bet, IFN-γ and the mRNA ratio of T-bet/GATA3 in NS group of MRL/lpr mice were higher than those in NS group of C57BL/6J mice (P<0.05). However, the GATA3 and IL-4 were lower in NS group of MRL/lpr mice in both mRNA and protein level (P<0.05). In MRL/lpr mice, the mRNA and protein levels of T-bet, IFN-γ and the mRNA ratio of T-bet/GATA3 were lower in ATO group compared with NS group (P<0.05), no difference was found in GATA3 and IL-4. No difference of the indexes mentioned above between ATO group and NS group in C57BL/6J mice was observed.CONCLUSION: ATO may affect the signaling pathway of T-bet/GATA3 to down-regulate the mRNA expression and the protein secretion of IFN-γ by decreasing the expression of T-bet in MRL/lpr mice.     

Activation of renal tubular epithelial cells by monocytes/macrophages and the relative mechanisms

AIM: To observe the direct effects of peripheral blood monocytes/macrophages (MO/MAC) on renal tubular epithelial cells (RTEC),and further probe into the possible mechanisms. METHODS: Conditioned medium(M-CM) of human peripheral blood MO/MAC was collected and added to HK-2,a human renal proximal tubular cell line.After incubation with M-CM for 24 hours,HK-2 cells were detected for DNA synthesis by [³H]-TdR incorporation,osteopontin (OPN) and α-smooth muscle actin (α-SMA) expression by Western blot,and fibronectin(FN) secretion by ELISA.Furthermore,anti-TGFβ₁ neutralizing antibody and interlukin-10(IL-10) were used separately to antagonize the effects of M-CM on HK-2 cells. RESULTS: ①DNA synthesis,α-SMA expression and FN secretion were all increased in HK-2 cells when incubated with M-CM.②When adding with anti-TGFβ₁ neutralizing antibody (5 mg/L) in the M-CM,the degree of upregulation of α-SMA and FN in HK-2 cells was much lower than that stimulated by M-CM alone.③M-CM added with IL-10 (20 μg/L) had a weaker ability to induce the increasing in α-SMA expression and FN excretion in HK-2 cells, compared with M-CM itself alone.M-CM from MO/MAC preincubated with IL-10 caused a lower upregulation of α-SMA expression in HK-2 cells than M-CM from non-preincubated MO/MAC. CONCLUSION: MO/MAC can directly induce proliferation,transdifferentiation and extracellular matrix secretion in RTEC.TGFβ₁ and proinflammatory cytokines secreted by MO/MAC might be involved in the aboveeffects.     

Effect of recombinant rat CC16 protein on LPS-induced expression of TNF-α, IL-6 and IL-8 in rat tracheal epithelial cells

AIM: To explore the effect of recombinamt rat CC16 protein (rCC16) on LPS-induced expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-8 in the rat tracheal epithelial (RTE) cells.METHODS: The RTE cells were incubated with rCC16 at concentrations of 0.5, 1.0 and 2.0 mg/L in serum-free media for 2 h prior to LPS (0.1 mg/L) treatment for further 24 h. The cells were harvested for assessing the mRNA levels of TNF-α, IL-6 and IL-8 by RT-qPCR. The cell culture supernatants were collected for analyzing the protein levels of TNF-α, IL-6 and IL-8 by ELISA. In addition, the nuclear translocation of nuclear factor-κB (NF-κB) p65 was tested by Western blot.RESULTS: rCC16 inhibited LPS-induced IL-6 and IL-8 expression at both mRNA and protein levels in the RTE cells in a concentration-dependent (0~2 mg/L) manner, as demonstrated by RT-qPCR and ELISA. However, no concentration-dependent manner between the dose of rCC16 and TNF-α expression was observed, and rCC16 inhibited LPS-induced TNF-α expression at lower concentration (0.5 mg/L). rCC16 concentration-dependently inhibited the effects of LPS on the level of nuclear translocation of NF-κB p65.CONCLUSION: rCC16 suppresses LPS-mediated TNF-α, IL-6 and IL-8 production through inactivation of NF-κB activity in RTE cells.[KEY WORDS] CC16 protein; Airway inflammation; LPS; Inflammatory mediators; Nuclear factor-κB