Vascular endothelial growth factor A, secreted in response to transforming growth factor-β1 under hypoxic conditions, induces autocrine effects on migration of prostate cancer cells

Hypoxia and transforming growth factor-β1 （TGF-β1） increase vascular endothelial growth factor A （VEGFA） expression in a number of malignancies. This effect of hypoxia and TGF-β1 might be responsible for tumor progression and metastasis of advanced prostate cancer. In the present study, TGF-β1 was shown to induce VEGFA165 secretion from both normal cell lines （HPV7 and RWPE1） and prostate cancer cell lines （DU 145 and PC3）. Conversely, hypoxia-stimulated VEGFA165 secretion was observed only in prostate cancer cell lines. Hypoxia induced TGF-β1 expression in PC3 prostate cancer cells, and the TGF-β1 type I receptor （ALK5） kinase inhibitor partially blocked hypoxia-mediated VEGFA16s secretion. This effect of hypoxia provides a novel mechanism to increase VEGFA expression in prostate cancer cells. Although autocrine signaling of VEGFA has been implicated in prostate cancer progression and metastasis, the associated mechanism is poorly characterized. VEGFA activity is mediated via VEGF receptor （VEGFR） 1 （Fit-l） and 2 （FIk-I/KDR）. Whereas VEGFR-1 mRNA was detected in normal prostate epithelial cells, VEGFR-2 mRNA and VEGFR protein were expressed only in PC3 cells. VEGFA165 treatment induced phosphorylation of extracellular signal-regulated kinase 1/2 （ERKI/2） in PC3 cells but not in HPV7 cells, suggesting that the autocrine function of VEGFA may be uniquely associated with prostate cancer. Activation of VEGFR-2 by VEGFA165 was shown to enhance migration of PC3 cells. A similar effect was also observed with endogenous VEGFA induced by TGF-β1 and hypoxia. These findings illustrate that an autocrine loop of VEGFA via VEGFR-2 is critical for the tumorigenic effects of TGF-β1 and hypoxia on metastatic prostate cancers.

The Effect of Simvastatin on mRNA Expression of Transforming Growth Factor-β1,Bone Morphogenetic Protein-2 and Vascular Endothelial Growth Factor in Tooth Extraction Socket

Aim To determine the effect of local simvastatin application on the mRNA expression level of transforming growth factor-β1 （TGF-β1）, bone morphogenetic protein-2 （BMP-2） and vascular endothelial growth factor （VEGF） in the tooth sockets of rat. Methodology Forty-eight male Wistar rats were randomly divided into experimental and control groups （n=24）. Polylactic acid/polyglycolic acid copolymer carriers, with or without simvastatin, were implanted into extraction sockets of right mandibular incisors. The expression of TGF-β1, BMP-2 and VEGF mRNA was determined by in situ hybridization in the tooth extraction socket at five days, one week, two weeks and four weeks after implantation. Results The fusiform stroma cells in the tooth extraction socket began to express TGF-β1, BMP-2 and VEGF mRNA in both experimental and control groups from one week after tooth extraction until the end of experiment. The expression of TGF-131 and BMP-2 mRNA in the experimental group was significantly up-regulated after one, two and four weeks, and expression of VEGF mRNA was significantly increased after one and two weeks compared with that in the control group. Conclusion The findings indicate that local administration of simvastatin can influence alveolar bone remodeling by regulating the expression of a school of growth factors which are crucial to osteogenesis in the tooth extraction socket.

Genetic expression of Col-2A and Col-10A as a function of administration of IGF-1 &TGF-<i>β</i>with and without anterior mandibular repositioning appliance on the growth of mandibular condylar cartilage in young rabbit

New Zealand (NZ) young rabbits with the administration of insulin-like growth factor (IGF-1) and transforming growth factor-β (TGF-β) with and without mandibular anterior repositioning appliances are explored for the growth of the mandibular condylar cartilage (MCC). 32 growing NZ and rabbits were divided into 4 groups: the group with saline injection in TMJ, the group which received growth factor injection in TMJ, the group which received anterior positioning appliance and the group which received growth factors injection as well as mandibular repositioning appliance. Gene expression was studied by real-time RT-PCR and cartilage growth by histomorphometry. Administration of growth factors along with mandibular repositioning appliances has induced 1) 1.70-fold expression of Col-2Agene (p value < 0.0005) and 2) 1.47-fold expression of Col-10Agene (p value < 0.0005). In contrast, administration of only mandibular repositioning appliances induced 1) 1.28-fold expression of Col-2Agene (p value < 0.0005) and 2) merely 0.62-fold expression of Col-10Agene (p value < 0.0005), while administration of growth factors only induced 1) mere 0.56-fold expression of Col-2Agene (p value 10A gene (p value growth factors along with mandibular repositioning appliances causes an increase in genetic expressions which have been corroborated by histomorphometry and validated by statistical analysis, during an accelerated growth of mandibular condylar cartilage. Administration of growth factors in the TMJ could provide a synergistic role along with mandibular repositioning appliances for treatment of mandibular retrognathism as well as disorders on the MCC.

TGF-β1 alters microRNA profile in human gastric cancer cells

Objective： MicroRNAs （miRNAs） are important regulators that play a key role in tumorigenesis and rumor progression. Transforming growth factor-β1 （TGF-β1） is involved in invasion and metastasis in many tumors. In this study, we investigated the microRNAs （miRNA） profiles altered by TGF-β1 in gastric cancer （GC） cells. Methods： We detected the expression profiles of miRNA by miRNA microarray and quantitative real- time polymerase chain reaction. Migration and invasion, wound-healing assay, prediction of miRNA targets, Western blot and qRT-PCR analysis were carried out to determine the role of one selected miRNA, namely miR-193b, in affecting the biological behaviors of GC BGC823 cells. Results： Among 847 human miRNAs in the microarray, three miRNAs （miR-27a, miR-29b-1 and miR-194） were up-regulated and three （miR-574-3p, miR-193b and miR-130b） were down-regulated in BGC823 cells treated with TGF-β1 compared with control, miR-193b suppressed the invasion and metastasis of GC cells in vivo and in vitro, and down-regulated urokinase-type plasminogen activator （uPA） protein in GC cells. Conclusions： TGF-β1 altered miRNA expression profile in BGC823 cells. Among the altered miRNAs, TGF-β1 induced the down-regulation of miR-193b, which inhibited cell invasion and metastasis in vivo and in vitro, and down-regulated uPA protein in GC cells.

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.

新疆汉族、维吾尔族原发性高血压患者血管损害与血清中TGF-β1水平相关性研究

目的研究新疆汉族、维吾尔族原发性高血压患者血管损害与血清转化生长因子(TGF-β1)水平的关系。方法选取2014年6月-2016年3月新疆医科大学第一附属医院高血压科住院的原发性高血压患者152例,其中汉族80例,维吾尔族72例;采用酶联免疫吸附测定法(ELISA)检测患者血清中TGF-β1水平,应用四肢多普勒超声测定脉搏波传导速度(PWV)、踝臂指数(ABI),分析不同民族患者血清中颈动脉内膜中膜厚度(IMT)与各指标之间的相关性。结果维吾尔族患者IMT较汉族患者明显增厚,差异有统计学意义(P<0.05);维吾尔族患者血清中TGF-β1水平明显高于汉族患者,差异有统计学意义(P<0.05);相关性分析显示两民族总体IMT与年龄呈正相关(r=0.39 P=0.00),ABI独立于高血压为IMT增厚的独立危险因素(P=0.02);汉族患者IMT与年龄呈正相关(r=0.41 P=0.00),ABI独立于高血压为IMT增厚的独立危险因素(P=0.02);维吾尔族患者与各观察指标无明显相关性,IMT与血清中TGF-β1水平无明显相关性。结论维吾尔族人群中高血压所致的血管损害重于汉族人群,但与血清中TGF-β1水平无明显相关性。

Influence of ginsenoside Rg1, a panaxatriol saponin from Panax notoginseng, on renal fibrosis in rats with unilateral ureteral obstruction

Total saponins of Panax notoginseng （PNS） have been shown to ameliorate renal interstitial fibrosis. Ginsenoside Rg 1, a panaxatriol saponin, is one of the major active molecules from PNS. The present study was undertaken to investigate the effect of ginsenoside Rgl on renal fibrosis in rats with unilateral ureteral obstruction （UUO）. The rats were randomly divided into 3 groups： sham-operation （n=15）, UUO （n=15） and UUO with ginsenoside Rgl treatment （n=15, 50 mg per kg body weight, intraperitoneally （i.p.） injected）. The rats were sacrificed on Days 7 and 14 after the surgery. Histological examination demonstrated that ginsenoside Rgl significantly inhibited interstitial fibrosis including tubular injury as well as collagen deposition, u-smooth muscle actin （α-SMA） and E-cadherin are two markers of tubular epithelial-myofibroblast transition （TEMT）. Interestingly, ginsenoside Rgl notably decreased α-SMA expression and simultaneously enhanced E-cadherin expression. The messenger RNA （mRNA） of transforming growth factor-β1 （TGF-β1）, a key mediator to regulate TEMT, in the obstructed kidney increased dramatically, but was found to decrease significantly after administration of ginsenoside Rg 1. Further study showed that ginsenoside Rgl considerably decreased the levels of both active TGF-β1 and phosphorylated Smad2 （pSmad2）. Moreover, ginsenoside Rgl substantially suppressed the expression of thrombospondin-1 （TSP-1）, a cytokine which can promote the transcription of TGF-β1 mRNA and the activation of latent TGF-β1. These results suggest that ginsenoside Rgl inhibits renal interstitial fibrosis in rats with UUO. The mechanism might be partly related to the blocking of TEMT via suppressing the expression of TSP-1.

Unveiling the therapeutic potential:KBU2046 halts triple-negative breast cancer cell migration by constricting TGF-β1 activation in vitro

Background:Triple-negative breast cancer(TNBC)is a heterogeneous,recurring cancer characterized by a high rate of metastasis,poor prognosis,and lack of efficient therapies.KBU2046,a small molecule inhibitor,can inhibit cell motility in malignant tumors,including breast cancer.However,the specific targets and the corresponding mechanism of its function remain unclear.Methods:In this study,we employed(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium)(MTS)assay and transwell assay to investigate the impact of KBU2046 on the proliferation and migration of TNBC cells in vitro.RNA-Seq was used to explore the targets of KBU2046 that inhibit the motility of TNBC.Finally,confirmed the predicted important signaling pathways through RT-qPCR and western blotting.Results:In this study,we found that KBU2046 functioned as a novel transforming growth factor-β(TGF-β1)inhibitor,effectively suppressing tumor cell motility in vitro.Mechanistically,it directly down-regulated leucine-rich repeat-containing 8 family,member E(LRRC8E),latent TGFβ-binding protein 3(LTBP3),dynein light chain 1(DNAL1),and MAF family of bZIP transcription factors(MAFF)genes,along with reduced protein expression of the integrin family.Additionally,KBU2046 decreased phosphorylation levels of Raf and ERK.This deactivation of the ERK signaling pathway impeded cancer invasion and metastasis.Conclusions:In summary,these findings advocate for the utilization of TGF-β1 as a diagnostic and prognostic biomarker and as a therapeutic target in TNBC.Furthermore,our data underscore the potential of KBU2046 as a novel therapeutic strategy for combating cancer metastasis.

Inflammatory Mechanism of Total Flavonoids of Chrysanthemum and Medicated Serum on Castrated Dry Eye Animal and Cell Models

Objective To observe the effects of total flavonoids of chrysanthemum and medicated serum on the expression of related proteins in the lacrimal tissue and dry-eye cell models of male rabbits with dry eye caused by castration.Methods(1)150 male Japanese rabbits were randomly divided into five groups,with 30 rabbits in each group:normal control group(group A),sham group(group B),model group(group C),androgen control group(group D)and total flavonoids of chrysanthemum treatment group(group E).The androgen deficiency dry-eye model was established by bilateral castration in groups C,D and E.Normal saline was administered to groups A,B and C by gavage;androgen(testosterone propionate)was injected into muscle in group D;and group E was given total flavonoids of chrysanthemum by gavage.All white rabbits were tested the Schirmer I test(SIT)and tear break-up time(BUT).After euthanasia,tear gland tissue was harvested so that we could observe pathological changes in the expression of related inflammatory factors in the lacrimal gland tissue.The expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)was detected in the lacrimal gland tissue by immunohistochemistry.Reverse transcription PCR was used to quantitatively detect expression of TGF-β1 mRNA.(2)Male Wistar rat lacrimal epithelial cells were used to establish a model of eye stem cell apoptosis caused by androgen levels.The blank control group was set up without androgen culture,the control group with androgen culture,and the total flavonoids of chrysanthemum group without androgen.The MTT method was used to determine the optimal intervention dosage of drug-containing plasma.Western blot and QPCR were used to detect the expression of AR mRNA,NF-κB phosphorylated protein and TGF-β1 in lacrimal epithelial cells,and the androgen-like effect of total flavonoids of chrysanthemum was observed.Results(1)Immunohistochemistry showed that groups A,B,D and E had significantly lower expression of IL-1βand TNF-αthan group C(P<0.05);among these,group E had slightly higher expression than group D(P>0.05).RT-PCR results showed that the relative expression of TGF-β1 mRNA in groups A,B,D and E was significantly higher than in group C(P<0.05),and the relative expression of TGF-β1 mRNA in groups D and E was higher than that in groups A and B(P<0.05).(2)Using the MTT method,the final concentration of interfering cells was calculated to be 13.2%.The expression of AR protein,NF-κB and TGF-β1 in the chrysanthemum flavonoid plasma intervention and testosterone propionate intervention groups was enhanced,and there were significant differences relative to the blank group(P<0.01).The expression level of NF-κB in the total flavonoid containing plasma intervention group was lower than that in the testosterone propionate intervention group(P<0.01).Conclusions The total flavonoids of chrysanthemum can inhibit IL-1βand TNF-αexpression in the lacrimal gland tissue of castrated male rabbits with dry eye to increase synthesis of TGF-β1 mRNA and TGF-β1,thereby inhibiting the inflammatory response.The medicated plasma with total flavonoids of chrysanthemum promotes expression of AR mRNA,upregulating expression of NF-κB,further promoting upregulation of TGF-β1 protein expression in lacrimal epithelial cells,inhibiting inflammation by regulating related proteins,and ultimately alleviating the symptoms of dry eye.

Dual therapy of rosiglitazone/pioglitazone with glimepiride on diabetic nephropathy in experimentally induced type 2 diabetes rats

Diabetic nephropathy is a major cause of end-stage renal disease （ESRD） in the general population. It is estimated that diabetic nephropathy will eventually develop in about 40% of all patients with diabetes; therefore, prevention is critical for delaying the development and progression of diabetic kidney disease. Despite extensive efforts, medical advances are still not successful enough to prevent the progression of the disease. In the present study, we focused on the comparison of combination therapies and whether they offered additional renoprotection. Type 2 diabetes mellitus was induced by intraperitoneally administering streptozotocin （90 mg/kg） in neonatal rats and then these rats were treated with rosiglitazone （1.0 mg/kg） in combination with glimepiride （0.5 mg/kg） or with pioglitazone （2.5 mg/kg） in combination with glimepiride （0.5 mg/kg）. Diabetic nephropathy markers were evaluated by biochemical and ELISA kits and renal structural changes were examined by light microscopy and transmission electron microscopy. Results show that the combination of pioglitazone with glimepiride is more effective in amelioration of diabetic nephropathy than rosiglitazone with glimepiride drug therapy due to glycemic control, suppressing albumin excretion rate, total protein excretion rate and augmented TNF-α signaling during the development of streptozotocin induced type 2 diabetic nephropathy.

Experimental research of pulmonary injury on irradiation combined with pemetrexed

Objective: The aim of our study was to examine whether irradiation combined with pemetrexed can exacerbate pulmonary injury. Methods: Two groups of male Wister Rats were subjected to bilateral apex of lungs irradiation(a single dose of 12 Gy), with or without pemetrexed(20 mg/kg) by intraperitoneal injection at the same time; a third group of weightand age- matched animals were treated with pemetrexed alone, as the same dose scheme, time and root of injection. The fourth group served as control. The whole lung mounts were dissected to histological evaluation, while serum cytokine transforming growth factor-β1(TGF-β1) analysis were compared at 1, 7, 21, 35, 49 days post-irradiation after irradiation. Results: Histological examination showed a thickening of alveolar septal, accumulation of inflammatory cells. The irradiation treatment group and the radiation-chemo treatment group showed a statistically significant higher level of TGF-β1(P < 0.05) than other two groups, but there were no differences between these two irradiation groups. Conclusion: These results demonstrated that pemetrexed can not aggravate pulmonary injury and it could be safely used in concurrent or sequential radio-chemotherapy in lung adenocarcinoma.

Correlation between the Expression of TGF-β1,Rhoa,SOX9 and Renal Interstitial Fibrosis in Rats with Chronic Kidney Disease

Objective:To study the correlation between the expression of transforming growth factor-β1(TGF-β1),Rho A,SOX9 and renal interstitial fibrosis in rats with chronic kidney disease.Methods:Forty specific pathogen-free(SPF)male SD rats were randomly divided into study group and control group,with 20 cases in each group.The study group was given adenine suspension by gavage,while the control group was given the same amount of saline by gavage.Blood,urine and renal tissue specimens were collected from all rats at 3rd and 6th weeks after modeling.The kidney weight,kidney weight/body weight,renal function indexes,the expression of TGF-β1,Rho A and SOX9 m RNA in renal tissues,Masson staining and renal interstitial fibrosis score were compared between the two groups.Pearson correlation was used to analyze the relationship between the renal interstitial fibrosis score and the expression of TGF-β1,Rho A and SOX9 m RNA in renal tissues of rats with chronic kidney disease.Results:The kidney weight and kidney weight/body weight of rats in the study group were higher than those in the control group at 3rd and 6th weeks after modeling(P<0.05).The quantitative levels of creatinine,urea nitrogen and 24-hour urinary protein in the study group were higher than those in the control group at 3rd and 6th weeks after modeling(P<0.05).The expression levels of TGF-β1,Rho A and SOX9 m RNA in renal tissues of rats in the study group were higher than those in the control group at 3rd and 6th weeks after modeling(P<0.05).The renal interstitial fibrosis score in the study group was higher than that in the control group at 3rd and 6th weeks after modeling(P<0.05).Pearson correlation analysis confirmed that the renal interstitial fibrosis score in rats with chronic kidney disease was positively correlated with the expression of TGF-β1,Rho A and SOX9 m RNA in renal tissues(P<0.05).Conclusion:The expression of TGF-β1,Rho A and SOX9 was abnormally high in rats with chronic kidney disease and was closely related to renal interstitial fibrosis,which may play a promoting role in the process of renal interstitial fibrosis.

骨桥蛋白在急性肺损伤后肺纤维化大鼠肺组织中的表达及作用

目的：观察骨桥蛋白（ OPN）在脂多糖（ LPS）诱导的急性肺损伤（ ALI）后肺纤维化大鼠肺组织中的表达及作用。方法120只SD大鼠随机分为对照组、模型组、干预组，每组40只。对照组第0、1、2天腹腔注射生理盐水，模型组和干预组注入等量LPS，5 min后干预组再注入抗OPN抗体，而模型组和对照组用生理盐水代替，于24 h、7 d、14 d、28 d处死3组大鼠各10只。行HE及Masson染色观察肺泡炎及肺纤维化改变；免疫组化测定肺组织转化生长因子－β1（ TGF－β1）的表达；检测肺组织匀浆中OPN蛋白水平、羟脯氨酸含量及Ⅰ、Ⅲ型胶原mRNA的表达。结果模型组第24 h首先出现肺纤维化改变，第28天最明显；同一时间点，模型组肺纤维化评分、TGF－β1的表达、OPN水平、羟脯氨酸含量及Ⅰ、Ⅲ型胶原mRNA表达均显著高于对照组，而干预组上述指标较模型组均显著降低。结论 OPN在LPS诱导的ALI后肺纤维化大鼠肺组织中高表达，能加重LPS诱导的ALI后肺纤维化，机制可能与OPN促进TGF－β1及Ⅰ、Ⅲ型胶原的表达有关。

沙棘总黄酮干预兔耳增生性瘢痕组织块的消退

背景:沙棘总黄酮可抑制肾脏、肝脏、心肌纤维化,但其对增生性瘢痕纤维化的相关疗效鲜有报道。目的:对兔耳增生性瘢痕组织块局部注射中草药沙棘总黄酮,观察其对增生性瘢痕组织块消退的影响并探讨其作用机制。方法:选择8只新西兰大白兔,每只耳沿腹侧中线两侧各建立3个直径8 mm圆形创面,共96个创面。21 d创面上皮化后,随机分为5组:0.5,1.0,2.0 g/L沙棘总黄酮组,2只/组;二甲基亚砜组(药物溶剂对照)和空白对照组,1只/组。于组织块基底部注射相应药物,每间隔3 d注射1次,连续干预4周。通过苏木精-伊红染色和Masson染色对比各组增生性瘢痕组织块病理组织变化;实时荧光定量PCR检测各组组织块中Ⅰ、Ⅲ型胶原的mRNA表达;Western blot检测各组兔耳瘢痕组织块Ⅰ、Ⅲ型胶原、转化生长因子β1、α-平滑肌肌动蛋白、血管内皮生长因子的蛋白表达。结果与结论:①大体观察:不同质量浓度的沙棘总黄酮局部注射后瘢痕组织块明显软化、变平;②空白对照组大量炎性细胞浸润、血管生成、胶原纤维不规则排列;与空白对照组相比,各沙棘总黄酮组可见整齐的束状胶原纤维分布,新生血管较少,特别是2 g/L沙棘总黄酮组改善较为明显;不同质量浓度沙棘总黄酮组间瘢痕增生指数和胶原密度均低于空白对照组和二甲基亚砜组(P<0.05);③各沙棘总黄酮组和二甲基亚砜组瘢痕组织块的Ⅰ、Ⅲ型胶原mRNA表达水平均显著低于空白对照组,其中2 g/L沙棘总黄酮组的抑制作用最明显(P<0.05);④与空白对照组相比,二甲基亚砜一定程度上可以降低组织块中Ⅲ型胶原、转化生长因子β1、α-平滑肌肌动蛋白、血管内皮生长因子蛋白表达水平;但不同质量浓度沙棘总黄酮组均可明显抑制兔耳增生性瘢痕组织块Ⅰ、Ⅲ型胶原、转化生长因子β1、α-平滑肌肌动蛋白、血管内皮生长因子蛋白的表达水平,且呈剂量依赖性(P<0.05);⑤提示沙棘总黄酮可抑制瘢痕增生,使瘢痕组织块软化、变平,颜色变淡,降低兔耳增生性瘢痕组织块转化生长因子β1、Ⅰ、Ⅲ型胶原纤维的表达,抑制成纤维细胞向肌成纤维细胞转化,减少瘢痕组织内血管生成,从而达到治疗增生性瘢痕的作用。

Astragalus mongholicus ameliorates renal fibrosis by modulating HGF and TGF-β in rats with unilateral ureteral obstruction

Astragalus mongholicus （AM） derived from the dry root ofAstragalus membranaceus Bge. var. mongolicus （Bge.） Hsiao is a widely used traditional Chinese medicine. The present study investigated the potential role of AM on renal fibrosis on a rat model of unilateral ureteral obstruction （UUO）. We divided 48 Sprague-Dawley rats randomly into 4 groups： sham-operated group （Sham）, untreated UUO group, AM-treated （10 g/（kg.d）） UUO group, and losartan-treated （20 mg/（kg.d）） UUO group as positive control. Haematoxylin ＆ eosin （HE） and Masson staining were used to study the dynamic histological changes of the kidneys 7 and 14 d after operation. The expressions of fibronectin （FN）, type I collagen （coil）, hepatocyte growth factor （HGF）, transforming growth factor-β1 （TGF-β1）, and eL-smooth muscle actin （α-SMA） were analyzed by real-time polymerase chain reaction （PCR）, immunohistochemistry staining, and Western blot. Results show that, similar to losartan, AM alleviated the renal damage and decreased the deposition of FN and coil from UUO by reducing the expressions of TGF-β1 and α-SMA （P〈0.05）, whereas HGF increased greatly with AM treatment （P〈0.05）. Our findings reveal that AM could retard the progression of renal fibrosis. The renoprotective effect of AM might be related to inhibition ofmyofibroblast activation, inducing of HGF and reducing of TGF-β1 expression.

EXPRESSION AND SIGNIFICANCE OF TUMOR INFILTRATING DENDRITIC CELLS IN RENAL CELL CARCINOMA

Objective: To study the expression of dendritic cells in human renal cell carcinoma and explore the cause, so to reveal the mechanism of escaping immune surveillance in RCC. Methods: The expressions of CD83+DCS, CD1a+DCS,VEGF and TGF-β1 in tumoral, peritumoral and normal kidney tissues of RCC in 30 cases were detected by immunohistochemistry using streptavidin/peroxidese(SP) Results: CD83+DCS were mainly located in the peritumoral areas; whereas CD1a+DCS、were mainly retained within the cancer nests. The number of CD83+DCS was inversely correlated with the clinical stage(P<0.05); but there were no significant correlations between the number of CD1a+DCS、and the clinical stage(P>0.05). The expressions of CD83+DCS and CD1a+DCS have significant difference between the tumoral, peritumoral and normal kidney tissues(P<0.001). The expression of VEGF and TGF-β1 were significantly lower in samples with highly infiltrating CD83+DCS(P<0.05); Whereas CD1a+DCS were not (P>0.05). Conclusion: DC has the tendency to gathering in tumor, but because of the immunosuppressive cytokins, for example VEGF and TGF-β1, inhibits the maturation of DC, there are less mature TIDCS(CD83+TIDCS) in the tumoral tissues, they are mainly located in the peritumoral areas. This may contribute to the mechanism of escaping immune surveillance in RCC.

Hydraulic pressure inducing renal tubular epithelial-myofibroblast transdifferentiation in vitro

Objective： The effects of hydraulic pressure on renal tubular epithelial-myofibroblast transdifferentiation （TEMT） were investigated. Methods： We applied hydraulic pressure （50 cmH2O） to normal rat kidney tubular epithelial cells （NRK52E） for different durations. Furthermore, different pressure magnitudes were applied to cells. The morphology, cytoskeleton, and expression ofmyofibroblastic marker protein and transforming growth factor-β1 （TGF-β1） of NRK52E cells were examined. Results Disorganized actin filaments and formation of curling clusters in actin were seen in the cytoplasm of pressurized cells. We verified that de novo expression α-smooth muscle actin induced by pressure, which indicated TEMT, was dependent on both the magnitude and duration of pressure. TGF-β1 expression was significantly upregulated under certain conditions, which implies that the induction of TEMT by hydraulic pressure is related with TGF-β1. Conclusion： We illustrate for the first time that hydraulic pressure can induce TEMT in a pressure magnitude- and duration-dependent manner, and that this TEMT is accompanied by TGF-β1 secretion.

丹参多酚酸盐治疗老年糖尿病肾病的临床研究

目的通过研究丹参多酚酸盐对老年糖尿病肾病的疗效及对尿N-乙酰β-D-葡萄糖苷酶(NAG)、转化生长因子-β1(TGF-β1)、单核细胞趋化蛋白-1(MCP-1)的影响,探讨丹参多酚酸盐治疗糖尿病肾病的机制及糖尿病肾病发病过程中的关键因子。方法 90例糖尿病肾病患者随机分为2组,每组45例,对照组给予常规治疗,治疗组在对照组的基础上增加丹参多酚酸盐治疗,将200 mg注射用丹参多酚酸盐加入200 m L生理盐水中,静脉滴注,1次/d,4周为1个疗程。比较两组患者的疗效及NAG、尿微量白蛋白(m ALB)、β2-微球蛋白(β2-MG)、TGF-β1、MCP-1和生化指标的变化。结果治疗后对照组和治疗组的有效率分别为82.22%、91.11%,两组比较差异具有统计学意义(P<0.05)。治疗后,两组的尿白蛋白排泄率(UAER)、肌酐(Cr)、血尿素氮(BUN)均较治疗前明显好转,且差异具有统计学意义(P<0.01);与对照组治疗后比较,治疗组的UAER、纤维蛋白原(FIB)明显改善,且差异具有统计学意义(P<0.01)。治疗后,两组的NAG、m ALB、β2-MG、TGF-β1和MCP-1均明显好转,与治疗前相比差异具有统计学意义(P<0.01);与对照组治疗后比较,治疗组的NAG、β2-MG、TGF-β1和MCP-1显著改善,且差异具有统计学意义(P<0.01),两组的m ALB差异不具有统计学意义。结论丹参多酚酸盐对糖尿病肾病的治疗作用主要通过调节TGF-β1和MCP-1水平、延缓近端肾小管损伤实现。

Suppression of NLRP3 inflammasome by ivermectin ameliorates bleomycin-induced pulmonary fibrosis

Ivermectin is a US Food and Drug Administration(FDA)-approved antiparasitic agent with antiviral and anti-inflammatory properties.Although recent studies reported the possible anti-inflammatory activity of ivermectin in respiratory injuries,its potential therapeutic effect on pulmonary fibrosis(PF)has not been investigated.This study aimed to explore the ability of ivermectin(0.6 mg/kg)to alleviate bleomycin-induced biochemical derangements and histological changes in an experimental PF rat model.This can provide the means to validate the clinical utility of ivermectin as a treatment option for idiopathic PF.The results showed that ivermectin mitigated the bleomycin-evoked pulmonary injury,as manifested by the reduced infiltration of inflammatory cells,as well as decreased the inflammation and fibrosis scores.Intriguingly,ivermectin decreased collagen fiber deposition and suppressed transforming growth factor-β1(TGF-β1)and fibronectin protein expression,highlighting its anti-fibrotic activity.This study revealed for the first time that ivermectin can suppress the nucleotide-binding oligomerization domain(NOD)-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome,as manifested by the reduced gene expression of NLRP3 and the apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),with a subsequent decline in the interleukin-1β(IL-1β)level.In addition,ivermectin inhibited the expression of intracellular nuclear factor-κB(NF-κB)and hypoxia‑inducible factor‑1α(HIF-1α)proteins along with lowering the oxidative stress and apoptotic markers.Altogether,this study revealed that ivermectin could ameliorate pulmonary inflammation and fibrosis induced by bleomycin.These beneficial effects were mediated,at least partly,via the downregulation of TGF-β1 and fibronectin,as well as the suppression of NLRP3 inflammasome through modulating the expression of HIF‑1αand NF-κB.