Effects of Heparin on Transforming Growth Factor-β_1 and Extracellular Matrix Components in the Glomeruli of Diabetic Rats

The effects of heparin on the expression of transforming growth factor-β 1 (TGF-β 1) and two extracellular matrix components laminin (LN) and fibronectin (FN) in diabetic rat glomeruli were investigated. Twenty-six rats were randomly divided into control group (C, n=8), diabetic group (D, n=9), and diabetes+heparin group (DH, n=9). After 8-week therapy of heparin (200 U once daily by abdominal injection), TGF-β 1, LN and FN expression in glomeruli was detected by immunohistochemical method. The results showed that the expression levels of TGF-β 1, LN and FN were higher in group D than in group C. It was found that heparin could reduce 24-h urinary albumin excretion and inhibit overexpression of TGF-β 1, LN and FN in glomeruli of diabetic rats. It suggested that the inhibitory effect of heparin on diabetic glomerular sclerosis was at least partly related with the inhibition of TGF-β 1 expression.

AAV-mediated expression of p65shRNA and bone morphogenetic protein 4 synergistically enhances chondrocyte regeneration

BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.

Matrix metalloproteinases contribute to kidney fibrosis in chronic kidney diseases

Matrix metalloproteinases(MMPs) are members of the neutral proteinase family. They were previously thought to be anti-fibrotic because of their ability to degrade and remodel of extracellular matrix. However, recent studies have shown that MMPs are implicated in initiation and progression of kidney fibrosis through tubular cell epithelial–mesenchymal transition(EMT) as well as activation of resident fibroblasts, endothelial-mesenchymal transition(Endo MT) and pericyte-myofibroblast transdifferentiation. Interstitial macrophage infiltration has also been shown to correlate with the severity of kidney fibrosis in various chronic kidney diseases. MMPs secreted by macrophages, especially MMP-9, hasbeen shown by us to be profibrotic by induction of tubular cells EMT. EMT is mainly induced by transforming growth factor-β(TGF-β). However, MMP-9 was found by us and others to be up-regulated by TGF-β1 in kidney tubular epithelial cells and secreted by activated macrophages, resulting in EMT and ultimately kidney fibrosis. Therefore, MMP-9 may serve as a potential therapeutic target to prevent kidney fibrosis in chronic kidney disease. This review, by a particular focus on EMT, seeks to provide a comprehensive understanding of MMPs, especially MMP-9, in kidney fibrosis.

Transforming growth factor-α promotes mouse blastocyst outgrowth and secretion of matrix metalloproteinases

Objective To study the effect of transforming growth factor-α (TGF-α) on early stage of embryo implantation.Methods Mouse blastocysts were cultured in vitro in medium containing various concentrations of TGF-α. Blastocyst implantation capacity was evaluated by calculating the percentage of embryos with attachment or outgrowth. Matrix metalloproteinases (MMPs) secretion of blastocysts was observed using gelatin zymography. Results There was no significant difference in the percentage of attachment between control and TGF-α treated groups, but the percentage of outgrowth of TGF-α treated groups was significantly higher than that of the control group after 24h culturing. Gelatin zymography showed that blastocysts cultured in TGF-α treated groups started secreting MMPs earlier than those in the control group.Conclusion TGF-α is involved in regulating the mouse embryo implantation process by promoting blastocyst outgrowth and secreting matrix matalloproteinases.

Prognostic significance and relationship of SMAD3 phosphoisoforms and VEGFR-1 in gastric cancer:A clinicopathological study

BACKGROUND The TGF-β/SMAD3 and VEGFR-1 signaling pathways play important roles in gastric cancer metastasis.SMAD3 phosphorylation is a crucial prognostic marker in gastric cancer.AIM To determine the prognostic value and relationship of SMAD3 phospho-isoforms and VEGFR-1 in gastric cancer.METHODS This was a single-center observational study which enrolled 98 gastric cancer patients and 82 adjacent normal gastric tissues from patients aged 32-84 years(median age 65)between July 2006 and April 2007.Patients were followed up until death or the study ended(median follow-up duration of 28.5 mo).The samples were used to generate tissue microarrays(TMAs)for immunohistochemical(IHC)staining.The expressions of TGF-β1,pSMAD3C(S423/425),pSMAD3L(S204),and VEGFR-1 in gastric cancer(GC)tumor tissue and normal tissue were measured by IHC staining using TMAs obtained from 98 GC patients.Prognosis and survival information of the patients was recorded by Outdo Biotech from May 2007 to July 2015.The relationship between TGF-β1,pSMAD3C(S423/425),pSMAD3L(S204),and VEGFR-1 protein expression levels was analyzed using Pearson's correlation coefficient.The relationship between protein expression levels and clinicopathological parameters was analyzed using the Chi-squared test.A survival curve was generated using the Kaplan-Meier survival analysis.RESULTS TGFβ-1 and VEGFR-1 expression was significantly upregulated in gastric cancer tissue compared to adjacent noncancerous tissue.The positive expression of phosphorylated isoforms of Smad3 varied depending on the phosphorylation site[pSMAD3C(S423/425):51.0%and pSMAD3L(S204):31.6%].High expression of pSMAD-3L(S204)was significantly correlated with larger tumors(P=0.038)and later N stages(P=0.035).Additionally,high expression of VEGFR-1 was closely correlated with tumor size(P=0.015)and pathological grading(P=0.013).High expression of both pSMAD3L(S204)and VEGFR-1 was associated with unfavorable outcomes in terms of overall survival(OS).Multivariate analysis indicated that high expression of pSMAD3L(S204)and VEGFR-1 were independent risk factors for prognosis in GC patients.VEGFR-1 protein expression was correlated with TGF-β1(r=0.220,P=0.029),pSMAD3C(S423/425)(r=0.302,P=0.002),and pSMAD3L(S204)(r=0.201,P=0.047),respectively.Simultaneous overexpression of pSMAD3L(S204)and VEGFR-1 was associated with poor OS in gastric cancer patients.CONCLUSION Co-upregulation of pSMAD3L(S204)and VEGFR-1 can serve as a predictive marker for poor gastric cancer prognosis,and pSMAD3L(204)may be involved in enhanced gastric cancer metastasis in a VEGFR-1-dependent manner.

Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice

AIM:To investigate the role of human platelets in liver fibrosis.METHODS:Severe combined immunodeficiency(SCID)mice were administered CCl4and either phosphate-buffered saline(PBS group)or human platelet transfusions(hPLT group).Concentrations of hepatocyte growth factor(HGF),matrix metallopeptidases(MMP)-9,and transforming growth factor-β(TGF-β)in the liver tissue were compared between the PBS and the hPLT groups by enzyme-linked immunosorbent assay(ELISA)and Western blotting.The effects of a human platelet transfusion on liver fibrosis included the fibrotic area,hydroxyproline content,and-smooth muscle actin(α-SMA)expression,which were evaluated by picrosirius red staining,ELISA,and immunohistochemical staining using an anti-mouse-SMA antibody,respectively.Phosphorylations of mesenchymal-epithelial transition factor(Met)and SMAD3,downstream signals of HGF and TGF-β,were compared between the two groups by Western blotting and were quantified using densitometry.Hepatocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling.Furthermore,the accumulation of human platelets in the liver 2 h after platelet transfusion was compared between normal and fibrotic livers by immunohistochemical staining using an anti-human CD41 antibody.RESULTS:The fibrotic area and hydroxyproline content in the liver were both significantly lower in the hPLT group when compared to the PBS group(fibrotic area,1.7%±0.6%vs 2.5%±0.6%,P=0.03;hydroxyproline content,121±26 ng/g liver vs 156±47 ng/g liver,P=0.04).There was less α-smooth muscle actin staining in the hPLT group than in the PBS group(0.5%±0.1%vs 0.8%±0.3%,P=0.02).Hepatic expression levels of mouse HGF and MMP-9were significantly higher in the hPLT group than in the PBS group(HGF,109±13 ng/g liver vs 88±22 ng/g liver,P=0.03;MMP-9,113%±7%/GAPDH vs 92%±11%/GAPDH,P=0.04).In contrast,the concentration of mouse TGF-β in the liver tissue was significantly lower in the hPLT group than in the PBS group(22±5ng/g liver vs 39±6 ng/g liver,P=0.02).Phosphorylation of Met was more prevalent in the hPLT group than in the PBS group(37%±4%/GAPDH vs 20%±8%/GAPDH,P=0.03).Phosphorylation of SMAD3was weaker in the hPLT group than in the PBS group(60%±12%/GAPDH vs 84%±12%/GAPDH,P=0.1),although this difference was not significant.Furthermore,a lower rate of hepatocyte apoptosis was observed in the hPLT group than in the PBS group(5.9%±1.7%vs 2.9%±2.1%,P=0.02).Significant human platelet accumulation was observed in the fibrotic liver tissues,whereas few platelets accumulated in the normal liver.CONCLUSION:Human platelets inhibit liver fibrosis in SCID mice.Increased concentration of HGF in the liver suppresses hepatic stellate cell activation,induces MMPs,and inhibits hepatocyte apoptosis.

Danhong injection A modulator for Golgi structural stability after cerebral ischemiareperfusion injury

The cerebral ischemia-reperfusion model was established using the suture occlusion method, and rats were intraperitoneally given 8 mL/kg Danhong injection once a day prior to model establishment Rat brain tissues were harvested at 6, 24, 48, 72 hours after reperfusion. Immunohistochemical staining showed that transforming growth factor-J31 expression increased, while Golgi matrix protein GM130 expression decreased after cerebral ischemia-reperfusion. Danhong injection was shown to significantly up-regulate the expression of transforming growth factor-131 and GM130, and expres- sion levels peaked at 7 days after reperfusion. At 7 days after cerebral ischemia-reperfusion, Golgi morphology was damaged in untreated rats, while Golgi morphology breakage was not observed after intervention with Danhong injection. These experimental findings indicate that Danhong injec- tion can up-regulate the expression of transforming growth factor-131 and GM130, and maintain Golgi stability, thus playing a neuroprotective role in rats after cerebral ischemia-reperfusion.

Macrophage secretory products induce an inflammatory phenotype in hepatocytes

AIM:To investigate the influence of macrophages on hepatocyte phenotype and function.METHODS:Macrophages were differentiated from THP-1 monocytes via phorbol myristate acetate stimulation and the effects of monocyte or macrophageconditioned medium on HepG2 mRNA and protein expression determined.The in vivo relevance of these findings was confirmed using liver biopsies from 147 patients with hepatitis C virus(HCV)infection.RESULTS:Conditioned media from macrophages,but not monocytes,induced a transient morphological change in hepatocytes associated with upregulation of vimentin(7.8±2.5-fold,P=0.045)and transforming growth factor(TGF)-β1(2.6±0.2-fold,P<0.001)and downregulation of epithelial cadherin(1.7±0.02-fold,P=0.017)mRNA expression.Microarray analysis revealed significant upregulation of lipocalin-2(17-fold,P <0.001)and pathways associated with inflammation,and substantial downregulation of pathways related to hepatocyte function.In patients with chronic HCV,realtime polymerase chain reaction and immunohistochemistry confirmed an increase in lipocalin-2 mRNA(F0 1.0 ±0.3,F1 2.2±0.2,F2 3.0±9.3,F3/4 4.0±0.8,P= 0.003)and protein expression(F1 1.0±0.5,F2 1.3± 0.4,F3/4 3.6±0.4,P=0.014)with increasing liver injury.High performance liquid chromatography-tandem mass spectrometry analysis identified elevated levels of matrix metalloproteinase(MMP)-9 in macrophageconditioned medium,and a chemical inhibitor of MMP-9 attenuated the change in morphology and mRNA expression of TGF-β1(2.9±0.2 vs 1.04±0.1,P<0.001) in macrophage-conditioned media treated HepG2 cells.In patients with chronic HCV infection,hepatic mRNA expression of CD163(F0 1.0±0.2,F1/2 2.8±0.3,F3/4 5.3±1.0,P=0.001)and MMP-9(F0 1.0±0.4,F1/2 2.8±0.3,F3/4 4.1±0.8,P=0.011)was significantly associated with increasing stage of fibrosis.CONCLUSION:Secreted macrophage products alter the phenotype and function of hepatocytes,with increased expression of inflammatory mediators,suggesting that hepatocytes actively participate in liver injury.

All-trans retinoic acid regulates the expression of MMP-2 and TGF-β2 via RDH5 in retinal pigment epithelium cells

·AIM: To investigate the effect of all-trans retinoic acid(ATRA) on retinol dehydrogenase 5(RDH5), matrix metalloproteinase-2(MMP-2) and transforming growth factor-β2(TGF-β2) transcription levels, and the effect of RDH5 on MMP-2 and TGF-β2 in retinal pigment epithelium(RPE) cells.·METHODS: After adult RPE cell line-19(ARPE-19 cells) intervened with gradient concentrations of ATRA(0-20 μmol/L) for 24h, flow cytometry was used to detect the proliferation and apoptosis of cells in each group, and quantitative realtime polymerase chain reaction(q RT-PCR) was used to detect RDH5, MMP-2 and TGF-β2 m RNA expression. Then, after ARPE-19 cells transfected with three different si RNA targets for 48h, the RDH5 knockdown efficiency of each group and expression of MMP-2 and TGF-β2 m RNA within them was detected by q RT-PCR. ·RESULTS: Flow cytometry results showed that ATRA could inhibit the proliferation of RPE cells and promote the apoptosis of RPE cells, and the difference of apoptosis was statistically significant when the ATRA concentration exceeded 5 μmol/L and compared with the normal control group(P=0.027 and P=0.031, respectively). q RT-PCR results showed that ATRA could significantly inhibit the expression level of RDH5 m RNA(P<0.001) and promote the expression of MMP-2 and TGF-β2 m RNA(P=0.03 and P<0.001, respectively) in a dose-dependent manner, especially when treated with 5 μmol/L ATRA. The knockdown efficiency of RDH5 si RNA varies with different targets, among which RDH5 si RNA-435 had the highest knockdown efficiency, i.e., more than 50% lower than that of the negative control group(P=0.02). When RDH5 was knocked down for 48h, the results of q RT-PCR showed that the expressions of MMP-2 and TGF-β2 m RNA were significantly up-regulated(P<0.001).·CONCLUSION: ATRA inhibits the expression of RDH5 and promotes MMP-2 and TGF-β2, and further RDH5 knockdown significantly upregulates MMP-2 and TGF-β2. These findings suggest that RDH5 may be involved in an epithelial-mesenchymal transition of RPE cells mediated by ATRA.

Regulation Mechanism of TFP on TGF-β1/STAT3 Signaling Pathway in Immune-mediated Liver Injury in Mice

[Objectives]To study the effect of total flavonoids extracted from Polygonum perfoliatum L.(TFP)on immune-mediated liver injury induced by bacillus Calmette-Guerin plus lipopolysaccharide(BCG+LPS)in mice,and to explore its action mechanism.[Methods]60 Kunming mice were divided into normal group,model group,control group(bifendate)and TFP low,medium and high dose groups according to random number table method,with 10 mice in each group.On the first day of modeling,mice were injected with 0.2 mL of BCG solution(12.5 mg/mL)through the tail vein,and on the eleventh day,0.2 mL of LPS(37.5μg/mL)were injected into the tail vein to prepare a mouse model of immune-mediated liver injury;from the first day of modeling,the normal group and the model group were administered intragastrically with the corresponding volume of distilled water,and the bifendate group and the TFP high,medium,and low dose groups were administered intragastrically with the corresponding doses once a day for 11 d.After the last time administration,fasting but giving water for 16 h,took blood from eyes,then collected the liver tissue.The levels of alanine transaminase(ALT)and aspartate transaminase(AST)in serum were detected by biochemical method;transforming growth factor-β1(TGF-β1),intercellular adhesion molecule-1(ICAM-1),interleukin-6(IL-6)and interleukin-1β(IL-1β)expression levels in liver tissue were detected by enzyme-linked immunosorbent assay(ELISA);phosphorylated protein tyrosine kinase JAK-2(p-JAK2),phosphorylated signal transducer and activator of transcription 3(p-STAT3)protein expression levels were detected by Western Blot method;the degree of liver tissue lesions was detected by HE staining.[Results]Compared with the model group,the levels of ALT and AST in the serum of mice in each dose group of TFP(high dose 600 mg/kg,medium dose 400 mg/kg,and low dose 200 mg/kg)were reduced,and the activities of T-SOD and GSH-Px were increased;the content or expression ofβ1,ICAM-1,IL-6,IL-1βdecreased,and the expression of p-JAK2 and p-STAT3 protein decreased;pathological sections showed that the degree of inflammatory necrosis and the degree of lesions in the liver tissues of each dose group of TFP were reduced by varying degrees.[Conclusions]TFP has a protective effect on BCG+LPS-induced immune-mediated liver injury in mice.The mechanism may be related to regulating the phosphorylation level of JAK2 and inhibiting the inflammatory reaction,thereby regulating the TGF-β1/STAT3 signaling pathway and improving the immune-mediated liver injury.

AngⅡ、TGF-1β、MMP-1及T IMP-1与原发性高血压左心室肥厚的相关性研究

目的探讨血管紧张素Ⅱ(A ngⅡ)、转化生长因子1β(TGF-1β)、基质金属蛋白酶-1(MM P-1)及其抑制剂(T IM P-1)与原发性高血压(EH)左心室肥厚(LVH)的关系。方法对81例EH患者行超声心动仪检测,根据其结果计算左心室重量指数(LVM I),分为左心室肥厚(LVH)组和非肥厚(NLVH)组;检测其血中A ngⅡ、TGF-1β、MM P-1、T IM P-1水平,并与对照组进行对比分析。结果EH患者血中A ngⅡ、TGF-1β、T IM P-1水平均较对照组显著增高,而MM P-1水平显著降低;LVH组尤甚。LVM I与A ngⅡ、TGF-1β、T IM P-1呈显著正相关,与MM P-1呈显著负相关。结论MM P-1水平降低、T IM P-1水平升高是导致心肌胶原代谢紊乱,促进LVH形成的重要因素;TGF-1β不仅直接抑制MM P-1表达,而且介导A ngⅡ作用,共同促进LVH形成。

TGF-β1对甲状腺相关性眼病眼眶成纤维细胞外基质基因表达的影响

目的探讨转化生长因子-β1(transforming growth factor-β1,TGF-β1)对甲状腺相关性眼病(thyroid-associated ophthalmopathy,TAO)患者眼外肌来源成纤维细胞(orbital fi-broblasts,OF)细胞外基质(extracellular matrix,ECM)主要成分金属蛋白酶组织抑制剂-1(tissue inhibitors of metalloproteinases-1,TIMP-1)、胶原Ⅰ(collagen typeⅠ,COL-Ⅰ)、胶原Ⅲ(collagen typeⅢ,COL-Ⅲ)及纤维连接蛋白(fibronectin,FN)基因表达水平的影响。方法采用实时荧光定量逆转录聚合酶链反应(RT-PCR)观察10μg·L-1TGF-β1刺激OF后不同时间点(0h、3h、6h、12h、24h、48h)及不同浓度TGF-β1(1μg·L-1、5μg·L-1、10μg·L-1、20μg·L-1)刺激OF24h后TIMP-1、COL-I、COL-Ⅲ及FN mRNA表达的变化。结果 10μg·L-1TGF-β1刺激OF的时间效应:TIMP-1mRNA在3h和6h分别为对照组的1.50倍和2.46倍(P<0.01);COL-ⅠmRNA在24h和48h分别为对照组的5.49、3.69倍(P<0.01);COL-ⅢmRNA在24h和48h分别为对照组的2.14、1.63倍(P<0.01);FN mRNA在12h和24h后分别为对照组的2.45、1.53倍(P<0.01)。不同浓度TGF-β1刺激OF24h后剂量效应:与空白对照组相比,TIMP-1mRNA在5μg·L-1和10μg·L-1时分别为空白对照组的1.79、1.46倍(P<0.01);COL-ⅠmRNA在1μg·L-1和10μg·L-1分别空白对照组的1.94、3.29倍(P<0.01);COL-ⅢmRNA在5μg·L-1和10μg·L-1分别空白对照组的1.52、3.28倍(P<0.01);FN mRNA在1μg·L-1和10μg·L-1分别为空白对照组的1.30、2.45倍(P<0.01)。结论 TGF-β1以剂量和时间依赖的方式刺激OF表达TIMP-1、COL-Ⅰ、COL-Ⅲ及FN mRNA,TGF-β1可能在TAO眼外肌纤维化机制中发挥了重要的作用。

慢性阻塞性肺疾病患者应用异丙托溴铵联合乙酰半胱氨酸治疗前后血清中TGF-β和MMP-9水平的变化及其临床意义

目的:研究异丙托溴铵联合乙酰半胱氨酸(NAC)对慢性阻塞性肺疾病(COPD)稳定期患者血清中转化生长因子(TGF-β)和基质金属蛋白酶9(MMP-9)水平的影响,探讨其对COPD患者气道重塑的作用。方法:COPD稳定期患者80例,随机分为NAC组(27例)、异丙托溴铵组(27例)和NAC与异丙托溴铵联合用药组(26例),同时选取健康志愿者26人做为对照组。监测各组患者治疗前后症状和体征评分及肺功能指标[用力肺活量(FVC)、第1秒钟用力呼气容积(FEV1)、第1秒钟用力呼气容积占预计值的百分比(FEV1%)及第1秒钟用力呼气容积占用力肺活量的百分比(FEV1/FVC)],检测各组患者血清中TGF-β和MMP-9水平,FEV1值与血清中TGF-β和MMP-9水平的相关性采用Spearman相关分析。结果:与治疗前比较,治疗后各用药组患者症状和体征评分均明显降低(P<0.05);与NAC组和异丙托溴铵组比较,联合用药组患者治疗后症状和体征评分降低更明显(P<0.05)。与治疗前比较,治疗后各用药组患者肺功能指标明显改善(P<0.05);与NAC组和异丙托溴铵组比较,联合用药组患者肺功能指标改善更明显(P<0.05)。与对照组比较,治疗前后各用药组患者血清中TGF-β和MMP-9水平均明显升高(P<0.05);与治疗前比较,治疗后各用药组患者血清中TGF-β和MMP-9水平均明显降低(P<0.05);与NAC组和异丙托溴铵组比较,联合用药组患者血清中TGF-β和MMP-9水平降低更明显(P<0.05)。联合用药组患者治疗前后FEV1值与血清中TGF-β和MMP-9水平呈负相关关系(治疗前:r=-0.554,r=-0.477,均P<0.05;治疗后:r=-0.415,r=-0.654,均P<0.05)。结论:异丙托溴铵联合NAC治疗可明显改善COPD患者症状、体征及肺功能,降低血清中TGF-β和MMP-9水平,在一定程度上抑制气道重塑。

香青兰总黄酮对哮喘大鼠肺组织基质金属蛋白酶及其抑制剂的影响

目的观察香青兰总黄酮对哮喘大鼠肺组织基质金属蛋白酶9(MMP-9)、基质金属蛋白酶抑制剂1(TIMP-1)和转化生长因子β(1TGF-β1)的影响。方法采用卵白蛋白致敏和激发建立大鼠哮喘模型,给予香青兰总黄酮进行干预后,ELISA法检测大鼠肺组织MMP-9、TIMP-1和TGF-β1的水平。结果香青兰总黄酮180、360 mg.kg-1可降低哮喘大鼠肺组织中MMP-9、TIMP-1、TGF-β1含量及MMP-9/TIMP-1比值(P<0.05或P<0.01)。结论香青兰总黄酮可纠正哮喘大鼠MMP-9/TIMP-1失衡,降低TGF-β1水平,改善哮喘气道重塑。

纤溶酶原激活物抑制因子-1对肝星状细胞活化及细胞外基质合成的影响

目的探讨外源性纤溶酶原激活物抑制因子-1(PAI-1)对人肝星状细胞(LX-2)活化及细胞外基质合成的影响。方法采用四甲基偶氮唑盐法(MTT)检测培养液中加入PAI-1后LX-2细胞的增殖变化,确定PAI-1的最佳干预浓度。将LX-2培养液中加入PAI-1培养12h,ELISA法检测细胞上清液中转化生长因子(TGFβ1)、透明质酸(HA)的变化;免疫细胞化学法检测LX-2细胞内α-平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶抑制剂-1(TIMP-1)、尿激酶型纤溶酶原刺激物(PLAU)及PAI-1蛋白变化;逆转录聚合酶链反应(RT-PCR)检测LX-2细胞内TGFβ1 mRNA和PAI-1 mRNA变化,分析其相关关系。结果 PAI-1对LX-2细胞增殖刺激作用明显,PAI-1浓度为10ng/mL时刺激作用最为明显(F=11.697,P<0.01)。加入PAI-112h后(刺激组)较未加入PAI-1(对照组),细胞上清液中TGFβ1、HA表达显著增加(t=5.474,t′=5.785,P<0.01);LX-2细胞内ɑ-SMA、TIMP-1、PAI-1蛋白表达均显著增加(t=5.438、9.511、4.857,P<0.01),而MMP-2、PLAU蛋白表达差异无统计学意义(t=0.473、0.581,P>0.05);LX-2内TGFβ1 mRNA、PAI-1 mRNA增加显著(t=4.683,t′=4.135,P<0.01),TGFβ1与PAI-1 mRNA呈显著正相关(r=0.827,P<0.05)。结论 PAI-1可通过活化LX-2,刺激LX-2TGFβ1蛋白及mRNA的表达,促进细胞外基质生成,从而促进肝纤维化、肝硬化的形成和发展。

红景天提取物对肾小球系膜细胞株增殖及分泌细胞外基质的影响

目的：观察红景天提取物对高糖诱导的大鼠肾小球系膜细胞株（MCs）增殖及分泌纤维连接蛋白（FN）、层黏连蛋白（LN）和转化生长因子-β1（TGF-β1）的影响，探讨红景天治疗糖尿病肾病的作用机制。方法：体外培养大鼠肾小球系膜细胞株，用不同浓度的红景天提取物干预高糖培养的系膜细胞株，用4-甲基偶氮四唑蓝（MTT）法观察系膜细胞的增殖情况。ELISA法检测系膜细胞分泌FN、LN和TGF-131的水平。结果：在高糖（30mmol·L-1^）培养24h、48h时段，可观察到刺激系膜细胞刺激性的增殖，在48h时红景天高浓度组系膜细胞增殖水平为（0．819±0．043），与模型组（1．306±0．076）相比较，有显著的统计学意义（P〈0．05）。红景天提取物在1×1（3-8^-1×10-4mol·L-1^范围内对系膜细胞株具抑制作用，与模型组比较有统计学意义（P〈0．05），且随着浓度的升高，其抑制作用增强。正常培养的大鼠肾小球系膜细胞可分泌一定量的FN、LN和TGF-81，高糖刺激后以上各成分分泌增加。随着培养时间的延长，FN、LN和TGF-131的分泌增加，而红景天提取物对系膜细胞分泌物的抑制随其浓度的升高而显著增强。结论：高糖可以刺激系膜细胞增殖，红景天提取物能够抑制高糖诱导的系膜细胞株增殖及细胞外基质FN、LN和TGF-／31分泌的水平。

氨基胍对非酒精性脂肪性肝炎大鼠TGF-β1及MMP2、TIMP2表达的影响

目的:研究实验性非酒精性脂肪性肝炎(NASH)大鼠转化生长因子-β1(TGF-β1)、基质金属蛋白酶-2(MMP2)、基质金属蛋白酶抑制因子-2(TIMP2)表达情况及氨基胍(AG)的干预作用。方法:SD大鼠28只,随机分为3组,模型组(M组)、氨基胍组(A组)、正常组(N组)。检测各组大鼠肝湿重、血糖(GLU)、谷丙转氨酶(ALT)、三酰甘油(TG)、胆固醇(CHO)及透明质酸(HA)含量。免疫组化法判断各组大鼠肝脏病理学变化。竞争性ELISA测定血清糖基化终产物(AGEs)及TGF-β1含量。Western-blot检测肝组织TGF-β1及MMP2、TIMP2蛋白表达情况。结果:M组大鼠肝湿重和血清GLU、ALT、TG、CHO水平比N组升高。A组较N组轻度增高,与M组无明显差异。M组及A组肝脏炎症活动度及纤维化半定量计分较N组升高,而A组较M组降低。M组大鼠血清AGEs、HA、TGF-β1含量较N组升高。而A组较M组降低。肝组织TGF-β1蛋白表达与各组血清检测结果平行,而MMP2/TIMP2值则相反。结论:NASH大鼠血清及肝组织中TGF-β1上调,MMP2/TIMP2值下降,AG可抑制TGF-β1、上调MMP2/TIMP2值,减缓NASH大鼠肝损伤进程。

Sphingosine Kinase-1/sphingosine 1-phosphate pathway in diabetic nephropathy

Objective Diabetic nephropathy （DN） is the major cause of end-stage renal disease worldwide and its prevalence continues to increase.Currently,therapies for DN provide only partial renoprotection; hence new targets for therapeutic intervention need to be identified.In this review,we summarized the new target,sphingosine kinase-1/sphingosine 1-phosphate （SphK1/S1P） pathway,explored its potential therapeutic role in the prevention and treatment of DN.Data sources Most relevant articles were mainly identified by searching PubMed in English.Study selection Mainly original articles and critical review articles by major pioneer investigators in this field were selected to be reviewed.Results SphK1/S1P pathway can be activated by hyperglycemia,advanced glycation end products,and many proinflammatory cytokines,which leads to fibronectin,transforming growth factor-31 up-regulation and AP-1 activation.And then it could promote glomerular mesangial cells proliferation and extracellular matrix accumulation,mediating the initiation and progression of diabetic renal fibrosis.Conclusions SphK1/S1P pathway is closely correlated with the pathogenesis of DN.The results suggest that SphK1/ S1P pathway as a new target for clinically improving DN in future is of great prospect.

Anti-aging and anti-carcinogenic effects of 1α,25-dihyroxyvitamin D_(3) on skin

Photoaging and carcinogenesis are facilitated by oxidative stress,inflammation,angiogenesis,and extracellular matrix(ECM)remodeling.Oxidative effects include DNA damage,membrane oxidation,lipid peroxidation,and alterations in the expression of p53 and antioxidant enzymes.The inflammatory and angiogenesis mediators include interleukin-1,tumor necrosis factor-α,interleukin-8,transforming growth factor-β,and vascular endothelial growth factor.ECM remodeling includes alterations in the expression and organization of collagen,elastin,matrix metalloproteinases,and elastase.1α,25-dihydroxy-vitamin D_(3) has antioxidant,anti-inflammatory,and ECM regulatory properties,and can counteract the processes that facilitate photoaging and carcinogenesis.This review provides an overview of the beneficial effects of vitamin D supplementation at a molecular level,followed by a brief discussion regarding its use as a supplement.