Destruction of gastric cancer cells to mesothelial cells by apoptosis in the early peritoneal metastasis

This study examined the mechanism by which the gastric cancer cells lead to early peritoneal metastasis. HMrSV5 cells, a human peritoneal mesothelial cell line, were co-incubated with the supernatants of gastric cancer cells. Morphological changes of HMrSV5 cells were observed. The cell damage was quantitatively determined by MTT assay. The apoptosis of HMrSV5 cells was observed under transmission electron microscope. Acridine orange/ethidium bromide-stained condensed nuclei was detected by fluorescent microscopy and flow cytometry. The expressions of Bcl-2 and Bax was immunochemically evaluated. The results showed that conspicuous morphological changes of apoptosis were observed in HMrSV5 cells 24 h after treatment with the supernatants of gastric cancer cells. The supematants could induce apoptosis of HMrSV5 cells in a time-dependent manner. The supernatants could up-regulate the expression of Bax and suppress that of Bcl-2 in HMrSV5 cells. These findings demonstrated that gastric cancer cells can induce the apoptosis of HPMCs through supernatants in the early peritoneal metastasis, The abnormal expressions of Bcl-2 and Bax may contribute to the apoptosis. Anti-apoptosis drugs promise to be adjuvant chemotherapeutic agents in the treatment of peritoneal metastasis of gastric cancer.     

Inhibition effect of small interfering RNA of connective tissue growth factor on the expression of vascular endothelial growth factor and connective tissue growth factor in cultured human peritoneal mesothelial cells

Background The peritoneum response to peritoneal dialysis can lead to fibrosis. The transforming growth factor β1 （TGF-β1 ） plays a key role in regulating tissue repair and remodelling after injury. Connective tissue growth factor （CTGF）, a downstream mediator of TGF-β1 inducing fibrosis, has been implicated in peritoneal fibrosis. Vascular endothelial growth factor （VEGF） plays a key role in angiogenesis that can hasten peritoneal fibrosis. In this study, we investigated the effect of small interfering RNA （siRNA） of CTGF by pRETRO-SUPER （PRS） retrovirus vector on the expression of CTGF and VEGF in human peritoneal mesothelial cells. Methods Retrovirus producing CTGF siRNA were constructed from the inverted oligonucleotides and transferred into packaging cell line PT67 with lipofectamine, and the virus supernatant was used to infect human peritoneal mesothelial cell （HPMC）. The cells were divided into seven groups： low glucose DMEM, low glucose DMEM ＋ TGF-β1 5 ng/ml, low glucose DMEM ＋ TGF-β1 5 ng/ml ＋ PRS-CTGF-siRNA1-4 and low glucose DMEM ＋ TGF-β1 5 ng/ml ＋ PRS. The expression of CTGF and VEGF were measured by semiquantitative RT-PCR and Western blot. Results Low levels of CTGF and VEGF were detected in confluent HPMCs. Following stimulation with TGF-β1 , the levels of CTGF and VEGF were significantly upregulated （P〈0.01）. Introduction of PRS-CTGF-siRNA1-4 resulted in the significant reduction of CTGF mRNA and protein, and VEGF mRNA （P〈0.01）, especially in groups PRS-CTGF-siRNA, and PRS-CTGF-siRNA4. The introduction of PRS void vector did not have these effects （P〉0.05）. Conclusions The expression of CTGF siRNA mediated by PRS retrovirus vector can effectively reduce the level of CTGF and VEGF induced by TGF-β1 in cultured HPMCs. This study may provide potential therapeutic strategies to prevent the peritoneal fibrosis.     

Pentoxifylline ameliorates pulmonary damage caused by Streptococcus pneumoniae infection in mouse.

Streptococcus pneumoniae stimulated mouse peritoneal macrophagc to release tumor necrosis factor-α (TNFα) in vitro. When penicillin was added into the medium with bacteria, TNFα release was accelerated. Pentoxifylline (PTX), a phosphodiesterase inhibitor, significantly attenuated TNFα release caused either by Streptococcus pneumoniae or by its lysates. In this experiment, 150 Kunming mice were infected with Streptococcus peumoniae through inspiration. Dynamic changes of TNFα concentration in serum and bronchoalveolar lavage fluid were determined, and pulmonary pathological changes were also observed. It was found that PTX significantly attenuated TNFα activity in serum and bronchoalveolar lavage fluid, and inhibited white blood cell chemotaxis, emigration and infiltration. In conclusion, Streptococcus pneumoniae infection stimulates the release of TNFα which is probably the major mediater that causes tissue damage during Streptococcus pneumoniae infection. The mechanism is probably that Steptococc     

Over-expression of hypoxia-inducible factor 1 alpha increases angiogenesis of LNCaP cells

Objective:To evaluate the effect of HIF-1 α over-expression on angiogenesis in human prostate cancer cells. Methods:LNCaP cells(a human prostate cancer cell line) were transfected with the recombinant plasmid pcDNA3.1(-)-HIF-1α with Lipofectamine 2000 system. The positive clones were selected by G418 being further confirmed by Western blot and immunofluorescence. The expression levels of VEGF, iNOS and Ang- Ⅱ were determined. Results:The expression of HIF-1α in the LNCaP/HIF1α cells was significantly increased in transfected cells, which induced the up-regulation of VEGF, iNOS, whereas Ang- Ⅱ expression remained un- changed. Conclusion :Over-expression of HIF-1α can induce angiogenesis proteins and may improve the angiogenesis potency of prostate cancer.     

EFFECT OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR ACTIVATORS ON TUMOR NECROSIS FACTOR-αL EXPRESSION IN NEONATAL RAT CARDIAC MYOCYTES A

Objecfive To investigate the effect ofperoxisome proliferator-αctivated receptor-α (PPARα) and PPARγ activators on tumor necrosis factor-α (TNFα) expression in neonatal rat cardiac myocytes. Primary cultures of cardiac myocytes from 1- to 3-dayold Wistar rats were prepared, and myocytes were ex-posed to lipopolysaccharide (LPS) and varying concentrations of PPARα or PPARγ activator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγ expression in cultured cardiac myocytes. Transient transfection of TNFα promoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed. Performed Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFα mRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARα or PPARα mRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFα promoter activity was observed when myocytes was transiently transfected with whole length of TNFα promoter (-721/ 17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFα reporter construct in deletion of NF-κBbinding site (- 182/ 17). Conchusions PPARα and PPARγ activators may inhibit cardiac TNFα expression but not accompanied by change of PPARα or PPARγ mRNA expression. Therefore PPARα and PPARγ activators appear to play a role in anti-inflammation.The mechanism may partly be involved in suppression of the NF-κBpathway.     

EFFECT OF PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR ACTIVATORS ON TUMOR NECROSIS FACTOR-α EXPRESSION IN NEONATAL RAT CARDIAC MYOCYTES

Objective To investigate the effect ofperoxisome proliferator-activated receptor-α (PPARα) and PPARγ activators on tumor necrosis factor-α (TNFα) expression in neonatal rat cardiac myocytes.Methods Primary cultures of cardiac myocytes from 1- to 3-day-old Wistar rats were prepared, and myocytes were exposed to lipopolysaccharide (LPS) and varying concentrations of PPARα or PPARγ activator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγexpression in cultured cardiac myocytes. Transient transfection of TNFα promoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed.Results Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFα mRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARα or PPARγ mRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFαpromoter activity was observed when myocytes was transiently transfected with whole length of TNFα promoter (-721/ 17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFα reporter construct in deletion of NF-κB binding site (-182/ 17).Conclusions PPARα and PPARγ activators may inhibit cardiac TNFα expression but not accompanied by change of PPARα or PPARγmRNA expression. Therefore PPARα and PPARγ activators appear to play a role in anti-inflammation.The mechanism may partly be involved in suppression of the NF-κB pathway.     

Effects of IL-17 on expression of GRO-α and IL-8 in fibroblasts from nasal polyps

Recent studies indicated that interleukin（IL）-17, growth-related oncogene（GRO）-α and IL-8 play an important role in the pathogenesis of nasal polyps. However, the effects of the increased amount of IL-17 and the production of GRO-α and IL-8 in human nasal polyp fibroblasts are not completely understood. This study aimed to determine the effects of the increased IL-17 on the changes of GRO-α and IL-8 expression in human nasal polyp fibroblasts and further investigate the mechanism of neutrophil infiltration in nasal polyps. Nasal polyp fibroblasts were isolated from six cases of human nasal polyps, and the cells were stimulated with five different concentrations of IL-17. Real-time fluorescence quantitative polymerase chain reaction（RT-PCR） was used to detect the mRNA expression of GRO-α and IL-8. The mRNA of GRO-α and IL-8 was expressed in unstimulated controls and remarkably increased by stimulation with IL-17. Moreover, the levels of GRO-α and IL-8 produced by fibroblasts were increased gradually with the increases in IL-17 concentrations. The present study showed that nasal fibroblasts can produce GRO-α and IL-8, and their production is remarkably enhanced by IL-17 stimulation, thereby clarifying the mechanism of the IL-17 mediated neutrophil infiltration in nasal polyps. These findings might provide a rationale for using IL-17 inhibitors as a treatment for nasal inflammatory diseases such as nasal polyps.     

Induction of chondrogenesis of adipose-derived stem cells by novel recombinant TGF-β3 fusion protein

A new type of TGF-β3 fusion protein with targeted therapy function was constructed,and its feasibility and target specificity of inducing chondrogenesis were investigated by transfecting LAP-MMP-mTGF-β3 gene into adipose-derived stem cells (ADSCs).The recombinant pIRESEGFP-MMP was constructed by inserting the sense and antisense DNA of encoding the amino acid of the synthetic MMP enzyme cutting site into the eukaryotic expression vector pIRES-EGFP.LAP and mTGF-β3 fragments were obtained by using RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively,and the recombinant plasmid of pIRES-EGFPLAP-MMP-mTGF-β3 was constructed,which was transferred to ADSCs.The ADSCs were cultured and divided in three groups:experimental group (MMP group),negative control group (no MMP) and non-transfection group.The morphological changes were observed microscopically,and the expression of proteoglycan and type Ⅱ collagen (ColⅡ) was detected by using Alcian blue staining and immunohistochemistry staining at 7th,14th and 21st day after culture.The recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3 was correctly constructed by methods of enzyme cutting and sequencing analysis.The mTGF-β3 fusion protein was successfully expressed after transfection,and in the presence of the MMP,active protein mTGF-β3 was generated,which significantly promoted differentiation of ADSCs into chondrocytes and the expression of cartilage matrix.The novel fusion protein LAP-MMP-mTGF-β3 can targetedly induce differentiation of ADSCs into chondrocytes,which would open up prospects for target therapy of cartilage damage repair in future.     

Effect of retinoid X receptor alpha (RXRα) transfection on the proliferation and phenotype of rat hepatic stellate cells in vitro

Objective To study the effect of retinoid X receptor alpha (RXRα) transfection plus treatment with the RXRα ligand, 9-cis-RA, on the proliferation and phenotype of platelet-derived growth factor (PDGF)-activated hepatic stellate cells (HSCs). Methods PDGF activated rat hepatic stellate cells were transfected with eukaryotic expression vector pcDNA3.1- human RXRα, and confirmed by Western blot. Proliferation of transfected HSC was assayed by bromodeoxyuridine (BrdU) incorporation as well as MTT, and the phenotype (α-smooth muscle actin, desmin) was observed by immunocytochemistry with image analysis. Results Transfection of the RXRα gene and treatment with ligand 9-cis-RA of PDGF-activated HSCs extended the increased expression of RXRα protein for at least 168 hours. Cell proliferation and expressions of alpha-smooth muscle actin (α-SMA) and desmin were blocked, compared with groups of sham-transfected, PDGF-activated, no transfection, no ligand treatment, and irrelevant ligand treated HSCs. Conclusion Transfection with the RXRα gene followed by 9-cis-RA ligand treatment will inhibit the proliferation and reverse the phenotype of activated HSC.     

人腹膜间皮细胞的培养及特征

目的：建立人腹膜间皮细胞（HPMC）培养模型,检测HPMC细胞外基质蛋白及白细胞介素（IL－8）的表达。方法：采用胰蛋白酶－EDTA消化法,从人的腹膜组织中分离HPMC。采用形态学和链霉菌抗生物素蛋白－过氧化物酶连接法（即SP法）,对培养细胞进行鉴定。采用SP法检测HPMC纤维连接蛋白（FN）、胶原Ⅲ和胶原Ⅴ及转化生长因子（TGF）β1表达。采用逆转录多聚酶链反应（RT－PCR）检测HPMC IL－8 mNRA和FN mRNA的表达。结果：光镜示细胞融合后呈多边形,超微结构下可见丰富的微绒毛和内质网,免疫组化染色结果示HPMC可表达角蛋白,波形蛋白,胶原Ⅲ,FN,TGFβ1蛋白,Ⅷ因子、胶原Ⅴ表达阴性。人腹膜间皮细胞亦表达FN mRNA和IL－8 mRNA。结论：HPMC培养模型的建立,可为进一步研究腹膜纤维化的防治及IL－8在腹膜透析中的作用提供理论依据。     

丹参酮ⅡA磺酸钠注射液改善脂多糖-高糖腹膜透析液诱导的人腹膜间皮细胞损伤的实验研究

目的 评估丹参酮ⅡA磺酸钠注射液（STS）减轻脂多糖-高糖腹膜透析液（LPS-PDS）诱导的人腹膜间皮细胞（HPMCs）损伤以及抑制腹膜纤维化的作用。方法 HPMCs来自腹腔外科手术患者腹膜细胞的原代培养。以LPS-PDS诱导的HPMCs损伤为模型,通过观察STS对HPMCs增殖活性、转化生长因子-β（TGF-β1）分泌、基质金属蛋白酶-9（MMP-9）、组织基质金属蛋白酶抑制因子-1（TIMP1）等的影响,阐明STS对HPMCs的保护作用及抗腹膜纤维化的机理。结果 LPS-PDS可造成HPMCs损伤,表现为活性下降,STS可明显减轻LPS-PDS对HPMCs的毒性,改善其造成的细胞活性下降,减轻LPS-PDS诱导的TGF-β1、TIMP1 mRNA上调和MMP-9 mRNA表达下降。结论 STS可以减轻LPS-PDS诱导的HPMCs损伤,可能是通过调节TGF-β1、TIMP1、MMP-9等蛋白的表达,发挥保护腹膜细胞和拮抗腹膜纤维化的作用。      

人腹膜间皮细胞的培养及特征

目的 :建立人腹膜间皮细胞 (HPMC)培养模型 ,检测HPMC细胞外基质蛋白及白细胞介素 (IL - 8)的表达。方法 :采用胰蛋白酶 -EDTA消化法 ,从人的腹膜组织中分离HPMC。采用形态学和链霉菌抗生物素蛋白 过氧化物酶连接法 (即SP法 ) ,对培养细胞进行鉴定。采用SP法检测HPMC纤维连接蛋白 (FN)、胶原Ⅲ和胶原V及转化生长因子 (TGF) β1表达。采用逆转录多聚酶链反应 (RT -PCR)检测HPMCIL - 8mRNA和FNmRNA的表达。结果 :光镜示细胞融合后呈多边形 ,超微结构下可见丰富的微绒毛和内质网 ,免疫组化染色结果示HPMC可表达角蛋白 ,波形蛋白 ,胶原Ⅲ ,FN ,TGFβ1蛋白 ,Ⅷ因子、胶原V表达阴性。人腹膜间皮细胞亦表达FNmRNA和IL - 8mRNA。结论 :HPMC培养模型的建立 ,可为进一步研究腹膜纤维化的防治及IL - 8在腹膜透析中的作用提供理论依据。     

Galectin-1 在高糖腹透液诱导人腹膜间皮细胞转分化中的作用

目的：研究人腹膜间皮细胞(human peritoneal mesothelial cells,HPMCs) 中galectin-1 的表达和在高糖腹透液(peritoneal dialysate solution, PDS) 刺激下galectin-1 表达的变化及其与上皮细胞- 间充质细胞转分化(epithelial-to-mesenchymal transition,EMT) 的关系。方法：以HPMCs 为研究对象,实验分为：正常对照组(5.5 mmol/L 葡萄糖),含1.5% 葡萄糖的PDS(1.5% PDS) 组,2.5%PDS 组及4.25% PDS 组。培养48 h 后,采用实时定量PCR 和Western 印迹检测galectin-1,vimentin 和zo-1 mRNA 和蛋白的表达水平,分析galectin-1 与vimentin 和zo-1 表达的相关性。使用阳离子脂质体转染galectin-1 siRNA 至HPMCs 内,实验分为：正常对照组(5.5mmol/L 葡萄糖),高糖组(4.25% PDS) 和转染组(galectin-1 siRNA +4.25% PDS)。观察galectin siRNA 干预后vimentin 和zo-1 mRNA 和蛋白的表达水平。结果：不同浓度PDS 刺激HPMCs 48 h 后,与对照组比,galectin-1mRNA 表达上调,尤以4.25%PDS 组明显(P<0.05),其蛋白水平的表达均明显上调(P<0.05),且具有浓度依赖性;同时 vimentin mRNA 表达上调,以2.5%PDS 组和4.25%PDS 组更明显(P<0.05),其蛋白水平的表达均明显上调(P<0.05);而zo-1 mRNA 和蛋白水平的表达均明显下调(P<0.05)。不同浓度高糖PDS 刺激下galectin-1 mRNA 和蛋白表达水平与vimentin mRNA 和蛋白表达水平呈正相关(P<0.05);与zo-1 mRNA 和蛋白表达水平呈负相关(P<0.05)。Galectin-1 siRNA 干预后vimentin mRNA 和蛋白水平的表达较高糖组明显下调(P<0.05),而zo-1 mRNA 和蛋白水平的表达较高糖组明显上调(P<0.05)。结论：Galectin-1 与高糖PDS诱导的HPMCs EMT 相关,galectin-1 siRNA 能够抑制高糖PDS 诱导的HPMCs 转分化。 Galectin-1 可能参与腹膜纤维化的发生环节,成为防治腹膜纤维化的新靶点。     

咪达普利与福辛普利对糖尿病大鼠心肌间质重构的影响

目的:用咪达普利(达爽)及福辛普利(蒙诺)两种血管紧张素转换酶抑制剂(ACEI)对实验性糖尿病大鼠进行早期干预,观察两药对糖尿病大鼠心肌间质重构的影响,了解两药对糖尿病大鼠心脏的保护作用有无区别。方法:以链脲佐菌素对30只SD大鼠进行腹腔注射,建立实验性糖尿病大鼠模型3组,包括咪达普利干预组,福辛普利干预组,糖尿病对照组,另外取10只SD大鼠作为正常对照组。用免疫组化法检测Ⅰ和Ⅲ型胶原含量;免疫组化法检测转化生长因子β1(TGF-β1)、基质金属蛋白酶1(MMP-1)、基质金属蛋白酶组织抑制剂1(TIMP-1)蛋白含量的表达,逆转录-聚合酶链反应(RT-PCR)检测MMP-1和TIMP-1的mRNA表达。结果:糖尿病对照组心脏指数及心室指数显著大于正常对照组,反映间质重构的指标Ⅰ和Ⅲ型胶原、TGF-β1、TIMP-1蛋白及TIMP-1 mRNA的表达均增加,而MMP-1蛋白及mRNA的表达减少。咪达普利干预组和福辛普利干预组的上述各种异常指标均有所改善,但两组的改善程度无显著差异。结论:含羧基的咪达普利与含磷酰基的福辛普利作为ACEI类药物,均可通过对TGF-β1和基质金属蛋白酶的影响而改善实验性糖尿病大鼠心肌间质的重构,两者对心脏间质的保护作用无明显差异。     

高糖腹透液对腹膜间皮细胞分泌VEGF的影响及氟伐他汀的干预作用

目的:观察氟伐他汀(fluvastatin,Flu)对高糖腹透液(high-glucose peritoneal dialysate,HGPDS)诱导的人腹膜间皮细胞(human peritoneal mesothelial cells,HPMCs)分泌血管内皮生长因子(vascular endothelial growth factor,VEGF)的影响?方法:体外培养HPMCs,同步化48 h后,分组如下:正常对照组?HGPDS组?HGPDS+Flu组?单纯Flu组?倒置显微镜观察各组细胞形态,RT-PCR法检测各组VEGF的mRNA表达,Western blot检测各组VEGF蛋白的表达?结果:与正常对照组相比,在HGPDS作用下,48 h后细胞由铺路石样转变为长梭形,1 × 10-6 mol/L Flu可部分逆转HPMCs的形态改变?与正常对照组相比,在HGPDS作用下,人腹膜间皮细胞VEGF mRNA及蛋白表达明显增多(P < 0.05),并呈时间依赖性,VEGF mRNA和蛋白分别在HGPDS干预后12 h和48 h达高峰(P < 0.05)?Flu可明显抑制HGPDS诱导的VEGF的表达(P < 0.05),并呈浓度依赖性?结论:适当浓度的Flu可以抑制HGPDS刺激体外培养人腹膜间皮细胞VEGF表达增加?     

肝细胞生长因子对ADPKD囊肿衬里上皮细胞合成细胞外基质、基质金属蛋白酶及其抑制剂的作用

目的；观察重组人肝细胞生长因子（rhHGF)对常染色体显性遗传性多囊肾病（ADPKD)囊肿衬里上皮细胞合成细胞外项质（ECM），基质金属蛋白酶及其抑制剂的作用。方法：采用放免法测定细胞合成层粘连蛋白（LN）和Ⅲ型前胶原氨基末端肽（PⅢNP），采用ELISA法检测细胞合成Ⅳ型胶原(ColⅣ），采用RT－PCR方法测定细胞表达转化生长因子β1（TGFβ1），基质金属蛋白酶2（MMP－2），金属蛋白酶组织抑制剂1（TIMP－1），金属蛋白酶组织抑制剂2（TIMP－2）mRNA的情况，用蛋白质印迹方法检测细胞上清液中MMP－2蛋白表达，明胶酶谱法检测MMP－2活性。结果：rhHGF及rhHGF 抗TGFβ1抗体组ADPKD囊肿衬里上皮细胞合成LN，ColⅣ显著增多，但两组比较无显著差异。对照组，rhHGF组，rhHGF 抗HGF抗体组及rhHGF 抗TGFβ1抗体组各间合成PⅢNP值均无显著差异。对照组，rhHGF组，rhHGF＋抗HGF抗体组各组比较TGFβ1mRNA表达无显著差异。与对照比较，rhHGF显著上调细胞MMP－2 mRNA和细胞上清液中MMP－2蛋白表达及MMP－2活性，显著下调细胞TIMP－1，TIMP－2 mRNA表达。结论：HGF促进了ADPKD囊肿衬里上皮细胞合成LN和ColⅣ，增加MMP－2表达，抑制TIMP－1，TIMP－2的表达。这些改变可能参与了ADPKD囊肿的发生及发展。     

Troglitazone对人腹膜间皮细胞TGF-β1和纤维连接蛋白表达的影响

目的:研究过氧化物增生体激活受体γ(PPAR-γ)激动剂troglitazone(曲格列酮,TGZ)对高浓度葡萄糖刺激下体外培养的人腹膜间皮细胞(HPMCs)中TGF-β1和细胞外基质纤维连接蛋白(Fn)表达的影响.方法:采用胰蛋白酶消化法从人大网膜组织中分离间皮细胞,建立稳定的体外培养模型.根据细胞增殖与毒性实验选择troglitazone的最佳浓度15 μmol/L,干预高浓度葡萄糖(30 mmol/L D-葡萄糖)刺激下的人腹膜间皮细胞.采用逆转录多聚酶链式反应(RT-PCR)半定量检测HPMCs中PPAR-γ,TGF-β1以及Fn的mRNA表达;采用双抗夹心法酶联免疫吸附实验检测HPMCs培养液中TGF-β1蛋白质水平;Western印迹检测Fn蛋白质水平.结果:高糖(30 mmol/L D-葡萄糖)刺激可在mRNA和蛋白质水平明显上调HPMCs表达TGF-β1和Fn(P＜0.01),troglitazone干预高糖孵育的HPMCs,可使TGF-β1和FnmRNA和蛋白质的表达明显下调(P＜0.05).结论:Troglitazone能够明显抑制在高糖刺激下的HPMCsTGF-β1和Fn的表达,这可能为临床防治长期腹膜透析患者的腹膜纤维化提供一种较为有效的方法.     

丹参素对高糖刺激腹膜间皮细胞分泌纤维连接蛋白和I型胶原的影响

目的:探讨丹参素对高糖刺激培养的腹膜间皮细胞(HPMCs) 分泌纤维连接蛋白(FN)和I型胶原(Col-I)的影响.方法:以高糖刺激培养HPMCs,分别加用浓度为10,5,2.5,1.25,0.625 mg/L的丹参素进行干预,并选取浓度为10 mg/L的丹参素干预0,12,24,48,72 h;应用RT-PCR,EL...