Thymoquinone affects hypoxia-inducible factor-1αexpression in pancreatic cancer cells via HSP90 and PI3K/AKT/mTOR pathways

BACKGROUND Pancreatic cancer(PC)is associated with some of the worst prognoses of all major cancers.Thymoquinone(TQ)has a long history in traditional medical practice and is known for its anti-cancer,anti-inflammatory,anti-fibrosis and antioxidant pharmacological activities.Recent studies on hypoxia-inducible factor-1α(HIF-1α)and PC have shown that HIF-1αaffects the occurrence and development of PC in many aspects.In addition,TQ could inhibit the development of renal cancer by decreasing the expression of HIF-1α.Therefore,we speculate whether TQ affects HIF-1αexpression in PC cells and explore the mechanism.AIM To elucidate the effect of TQ in PC cells and the regulatory mechanism of HIF-1αexpression.METHODS Cell counting kit-8 assay,Transwell assay and flow cytometry were performed to detect the effects of TQ on the proliferative activity,migration and invasion ability and apoptosis of PANC-1 cells and normal pancreatic duct epithelial(hTERTHPNE)cells.Quantitative real-time polymerase chain reaction and western blot assay were performed to detect the expression of HIF-1αmRNA and protein in PC cells.The effects of TQ on the HIF-1αprotein initial expression pathway and ubiquitination degradation in PANC-1 cells were examined by western blot assay and co-immunoprecipitation.RESULTS TQ significantly inhibited proliferative activity,migration,and invasion ability and promoted apoptosis of PANC-1 cells;however,no significant effects on hTERT-HPNE cells were observed.TQ significantly reduced the mRNA and protein expression levels of HIF-1αin PANC-1,AsPC-1,and BxPC-3 cells.TQ significantly inhibited the expression of the HIF-1αinitial expression pathway(PI3K/AKT/mTOR)related proteins,and promoted the ubiquitination degradation of the HIF-1αprotein in PANC-1 cells.TQ had no effect on the hydroxylation and von Hippel Lindau protein mediated ubiquitination degradation of the HIF-1αprotein but affected the stability of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90,thus promoting its ubiquitination degradation.CONCLUSION The regulatory mechanism of TQ on HIF-1αprotein expression in PC cells was mainly to promote the ubiquitination degradation of the HIF-1αprotein by inhibiting the interaction between HIF-1αand HSP90;Secondly,TQ reduced the initial expression of HIF-1αprotein by inhibiting the PI3K/AKT/mTOR pathway.

Silencing of Jumonji domain-containing 1C inhibits the osteogenic differentiation of bone marrow mesenchymal stem cells via nuclear factor-κB signaling

BACKGROUND Osteoporosis is a common metabolic bone disorder induced by an imbalance between osteoclastic activity and osteogenic activity.During osteoporosis,bone mesenchymal stem cells(BMSCs)exhibit an increased ability to differentiate into adipocytes and a decreased ability to differentiate into osteoblasts,resulting in bone loss.Jumonji domain-containing 1C(JMJD1C)has been demonstrated to suppress osteoclastogenesis.AIM To examine the effect of JMJD1C on the osteogenesis of BMSCs and the potential underlying mechanism.METHODS BMSCs were isolated from mouse bone marrow tissues.Oil Red O staining,Alizarin red staining,alkaline phosphatase staining and the expression of adipo-genic and osteogenic-associated genes were assessed to determine the differen-tiation of BMSCs.Bone marrow-derived macrophages(BMMs)were incubated with receptor activator of nuclear factor-kappaΒligand to induce osteoclast differentiation,and osteoclast differen-tiation was confirmed by tartrate-resistant acid phosphatase staining.Other related genes were measured via reverse transcription coupled to the quantitative polymerase chain reaction and western blotting.Enzyme-linked immunosorbent assays were used to measure the levels of inflammatory cytokines,including tumor necrosis factor alpha,interleukin-6 and interleukin-1 beta.RESULTS The osteogenic and adipogenic differentiation potential of BMSCs isolated from mouse bone marrow samples was evaluated.JMJD1C mRNA and protein expression was upregulated in BMSCs after osteoblast induction,while p-nuclear factor-κB(NF-κB)and inflammatory cytokines were not significantly altered.Knockdown of JMJD1C repressed osteogenic differentiation and enhanced NF-κB activation and inflammatory cytokine release in BMSCs.Moreover,JMJD1C expression decreased during BMM osteoclast differentiation.CONCLUSION The JMJD1C/NF-κB signaling pathway is potentially involved in BMSC osteogenic differentiation and may play vital roles in the pathogenesis of osteoporosis.

Prognostic role of the stromal cell derived factor-1 in patients with hepatitis B virus-related acute-on-chronic liver failure

BACKGROUND Stromal cell derived factor-1(SDF-1)plays a pivotal role in the recruitment of stem cells to injured livers.However,the changes of SDF-l in patients with hepatitis B virus(HBV)-related acute-on-chronic liver failure(ACLF)have yet to be elucidated.AIM To study the SDF-1 changes in patients with HBV-related ACLF.METHODS 30 patients with HBV-related ACLF,27 patients with chronic hepatitis B and 20 healthy individuals are involved in our study.The SDF-l mRNA expression in liver tissue was detected by quantitative real-time polymerase chain reaction.Immunohistochemical staining was performed to illustrate the expression of SDFl,CXC receptor 4(CXCR4)and Ki67.The serum SDF-l concentrations were also detected by enzyme-linked immunosorbent assays.RESULTS The expression of SDF-1 mRNA from ACLF patients was remarkably higher than that from other patients(both P<0.05).The expression of SDF-l,CXCR4 and Ki67 from ACLF were the highest among the three groups(all P<0.01).The serum SDF-l levels in ACLF patients were significantly lower than that in other patients(both P<0.01).Moreover,in ACLF patients,the serum SDF-1 Levels were positively correlated with serum total bilirubin and international normalized ratio.In addition,the serum SDF-l levels in survival were significantly lower compared with the non-survivals(P<0.05).The area under the curve for the serum SDF-1 level in predicting 28-d mortality was 0.722(P<0.05).CONCLUSION This study provides the SDF-1 changes in patients with HBV-related ACLF.The SDF-1 Level at admission may serve as a promising prognostic marker for predicting short-term prognosis.

Analyze interleukin-1β,interleukin-6,and tumor necrosis factor-αlevels in dry eye and the therapeutic effect of cyclosporine A

BACKGROUND Dry eye is a common eye disease.Artificial tears supplements are widely used for the treatment of dry eyes.However,multiple adverse effects have been observed in patients receiving long-term treatment with artificial tears,which may affect the therapeutic effect.AIM To analyze the characteristics of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-alpha(TNF-α)levels in patients with dry eye and the therapeutic effect of artificial tears combined with cyclosporine A.METHODS A total of 124 dry eye patients treated at The First People’s Hospital of Xining from April 2020 to April 2022 were selected as the observation group,while 20 healthy individuals served as the control group during the same period.Levels of inflammatory markers,including IL-1β,IL-6,and TNF-α,were analyzed.The observation group was further divided into a study group and a control group,each consisting of 62 patients.The control group received artificial tears,whereas the study group received a combination of artificial tears and cyclosporine A.Inflammatory markers,Schirmer’s test(SIT),tear break-up time(TBUT),corneal fluorescein staining(CFS),National Eye Institute Visual Function Questionnaire-25(NEI-VFQ-25)scores,and adverse events(AEs)were compared between the two groups.RESULTS The observation group exhibited significantly elevated serum levels of IL-1β,IL-6,and TNF-αin comparison to the healthy group.Following treatment,the study group demonstrated substantial reductions in IL-1β,IL-6,and TNF-αlevels relative to the control group.Moreover,after treatment,the study group experienced a marked decrease in CFS scores and significant increases in both SIT and BUT levels when compared to the control group.Additionally,significant improvements were observed in the primary symptom of dry eye and secondary symptoms such as photophobia,foreign body sensation,fatigue,red eye,and burning sensation within the study group.Furthermore,post-treatment NEI-VFQ-25 scores across all dimensions exhibited significant enhancements in the study group compared to the control group(P<0.05).It is noteworthy that significant AEs were reported in both groups throughout the treatment period.CONCLUSION Cyclosporine A combined with artificial tears is effective in treating dry eye,yielding enhanced outcomes by improving SIT and TBUT levels,reducing CFS scores,and ameliorating vision-related quality of life.

Transforming growth factor-β1 and vascular endothelial growth factor levels in senile acute myeloid leukemia and correlation with prognosis

BACKGROUND Acute myeloid leukemia(AML)is a disease in which immature hematopoietic cells accumulate in the bone marrow and continuously expand,inhibiting hematopoiesis.The treatment and prognosis of this disease have always been unsatisfactory.AIM To investigate the correlation between vascular endothelial growth factor(VEGF)and transforming growth factor-β1(TGFβ1)expression and prognosis in older adults with AML.METHODS This study enrolled 80 patients with AML(AML group),including 36 with complete response(AML-CR),23 with partial response(AML-PR),and 21 with no response(AML-NR).The expression levels of VEGF and TGFβ1 were detected by reverse transcription polymerase chain reaction in bone marrow mononuclear cells isolated from 56 healthy controls.Kaplan-Meier analysis was performed to assess overall survival(OS)and progression-or disease-free survival(DFS).Prognostic risk factors were analyzed using a Cox proportional hazards model.RESULTS The AML group showed a VEGF level of 2.68±0.16.VEGF expression was lower in patients with AML-CR than those with AML-PR or AML-NR(P<0.05).TGFβ1 expression in the AML group was 0.33±0.05.Patients with AML-CR showed a higher TGFβ1 expression than those with AML-PR or AML-NR(P<0.05).VEGF and TGFβ1 expression in patients with AML was significantly correlated with the counts of leukocytes,platelets,hemoglobin,and peripheral blood immature cells(P<0.05);Kaplan-Meier survival analysis revealed that patients with high TGFβ1 expression had better OS and DFS than those with low TGFβ1 expression(P<0.05),whereas patients with low VEGF levels showed better OS and DFS than those with high VEGF levels(P<0.05).VEGF,TGFβ1,and platelet count were identified by the Cox proportional hazards model as independent risk factors for OS(P<0.05),while VEGF,TGFβ1,and white blood cell count were independent risk factors for DFS(P<0.05).CONCLUSION Decreased VEGF expression and increased TGFβ1 expression in patients with AML provide valuable references for determining and individualizing clinical treatment strategies.

Expression of transforming growth factor-β_1 and its typeⅠ receptor in different phases of post-burn hypertrophic scars

Objective: To analyze and compare the expression pattern of the transforming growth factor-β1(TGF-β1) and its type I receptor (TGF-β RI ) in nounal human skin and various phases of post-burn hypertrophic scars (HTS). Method: The immunohistochemical ABC method was employed. Results: In nounal human skin, no evident immunoreactivity of TGF-β1 and TGF-β R I was observed. In activation phase of post-burn HTS, TGF-β R I and TGF-β1 were highly expressed in most dermal fibroblasts which seemed to be the same subset. However, in remission phase, no staining was seen in der mal fibroblasts. Conclusion: The formation of all may involve the increase of TGF-β responsiveness in fibroblasts The ac cumulation at the wound site and failure of apoptosis of over-resposive fibroblasts may contribute to the formation of HTS.

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.

Expression of Mesenger RNA for Transforming Growth Factor-β_1 in Bovine Trabecular Meshwork

Purpose: To investigate the relationship between transforming growth factor-β1(TGF-β1) and primary open-angle glaucoma, we have determined whether trabec-ular tissues have the expression of messenger RNA for TGF-β1.Methods: Total RNA of 24 newborn bovine trabecular tissue were extracted byGuanidine isothiocyanate method. The TGF-β33 plasmid was brought into E. col-ibacillius HB101 and amplificated. After Bam HI endolase degradation and labelwith a-32p-dATP the RNA was hybridized with the cDNA (complementary DNA)probe and examined by autoradiography.Results: The presence of mRNA for TGF-β1 in bovine trabecular meshwork wasconfirmed.Conclusions: The TGF-β1 present in normal aqueous humor must be at least partlyderived from the trabecular meshwork. It offered a basis for understanding therelationship between abnormal synthesis, activation and clearance of TGF-β1 andthe pathogenesis of primary open-angle glaucoma (POAG) in molecular biology.Eye Science 1996; 12:1-4.

重组人内抑素对兔耳创面瘢痕组织血管内皮生长因子、转化生长因子-β_1和碱性成纤维细胞生长因子表达的影响

目的观察重组人内抑素(rh Endostatin)对兔耳创面瘢痕增生及血管内皮生长因子(VEGF)、转化生长因子β1(TGF-β1)和碱性成纤维细胞生长因子(b FGF)表达的影响,探讨rh Endostatin抑制瘢痕增生的分子机制。方法健康新西兰大耳白兔20只,均分为正常对照组、模型组、生理盐水(NS)对照组、rh Endostatin(5 g/L)治疗组和醋酸曲安奈德(TA,40 g/L)对照组;除正常对照组外其余各组均进行增生性瘢痕动物模型的复制,在兔耳腹侧面制作1cm×1cm大小创面。术后第28天,rh Endostatin治疗组瘢痕皮内注射相应浓度rh Endostatin(100μl),NS对照组注射等量生理盐水,隔日1次,共6次;TA对照组瘢痕块内注入相应浓度曲安奈德(100μl),每周1次,共2次;模型组术后不接受处理。术后第47天摄片并收集瘢痕及正常皮肤标本,应用免疫组织化学方法检测VEGF、TGF-β1和b FGF的表达。结果形态学观察结果显示,术后第47天rh Endostatin治疗组瘢痕皮肤色泽变浅,变软变平,体积减小,与模型组和NS对照组比较有明显差异;免疫组织化学检测结果可见,与模型组和NS对照组相比,rh Endostatin治疗组VEGF和TGF-β1蛋白表达减少,b FGF表达增加,差异均具有统计学意义(P<0.01)。结论 rh Endostatin能有效抑制兔耳创面瘢痕增生,其机制可能与rh Endostatin影响VEGF、TGF-β1和b FGF蛋白的表达有关。

TIMP-1mRNA表达在病毒性心肌炎小鼠心肌胶原重构中的作用及其与TGF-β_1的关系

目的探讨TIMP-1mRNA表达在病毒性心肌炎小鼠心肌胶原重构中的作用及其与转化生长因子(TGF-β1)的关系。方法4周龄雄性BALB/c鼠腹腔接种0.1ml100TCID50CVB3m,周龄、性别相同小鼠为对照,分别于接种后3、7、9、14、21、28、56d断脊处死小鼠,心肌标本经常规制片,VG染色观察心肌组织胶原的改变,原位杂交观察TIMP-1mRNA和TGF-β1mRNA的表达,免疫组织化学观察TGF-β1的表达。结果感染后28d小鼠心肌组织血管周围胶原明显沉积,感染后56d心肌组织血管周围及心肌细胞间隙胶原沉积均明显增加;感染后7d可见TIMP-1mRNA表达,14～21d最为明显,此后减弱,直到56d。感染后3d可见TGF-β1mRNA及TGF-β1表达,7～21d最为明显,此后减弱,持续至感染后56d;TIMP-1mRNA和TGF-β1表达等级间存在正相关关系(P<0.001)。结论TGF-β1表达增加及其上调TIMP-1mRNA的表达可能在病毒性心肌炎心肌胶原重构中起重要作用。

EMT、TGF-β_1、Ang Ⅱ与器官纤维化发生机制的研究进展

纤维化是大多数慢性炎症性疾病的病理转归,几乎能发生在身体的每个组织器官。纤维化以过多的细胞外基质沉积为特征,进一步发展可导致器官功能衰竭乃至死亡。关于器官纤维化的研究很多,但其确切机制目前尚不明确。近年来,上皮间质转化(EMT)、转化生长因子β1(TGF-β1)、血管紧张素Ⅱ(AngⅡ)在组织器官纤维化形成机制研究中备受关注。该文就EMT、TGF-β1、AngⅡ与各器官纤维化的相互关系及作用机制予以综述,以更全面地认识纤维化的发生机制。

肾小球系膜细胞金属蛋白酶1组织抑制剂对血管内皮生长因子及转化生长因子β_1表达的影响

目的:探讨高糖(葡萄糖25mmol/L)条件下大鼠肾小球系膜细胞(RMC)金属蛋白酶1组织抑制剂(TIMP-1)对血管内皮生长因子(VEGF)及转化生长因子β1(TGF-β1)表达的影响。方法:用高糖培养基体外培养未转染及分别用pcDNA3空载体、人反义TIMP-1基因重组真核表达载体转染的RMC24h,将细胞分为未转染组、空载体组及反义组,并设正常培养条件下的RMC作为正常对照组。RT-PCR检测各组RMC TIMP-1、VEGF及TGF-β1mRNA的表达。Western印迹检测胞质中TIMP-1、VEGF及TGF-β1蛋白的表达。免疫荧光检测各组RMC胞质中VEGF的表达。结果:高糖培养条件下未转染组及空载体组RMC大鼠TIMP-1、VEGF及TGF-β1mRNA表达均较对照组明显增加(P<0.01),而此两组间相应指标表达差异无统计学意义;反义组大鼠TIMP-1、VEGF mRNA表达较未转染组显著降低(P<0.01),而TGF-β1 mRNA表达则无变化。Western印迹也得到相同的结果。免疫荧光检测发现,未转染组及空载体组VEGF荧光表达较对照组显著增强,反义组荧光则较未转染组明显减弱。结论:高糖可促进RMC TIMP-1、VEGF及TGF-β1表达增加。在高糖条件下,TIMP-1可能位于VEGF上游,促进其表达;而TGF-β1表达并不受TIMP-1调控,高糖可能通过其他途径上调其表达。

Effect of porous titanium coated with IGF-1 and TGF-β_1 loaded gelatin microsphere on function of MG63 cells

Porous titanium with porosity of 60% was prepared by metal injection molding（MIM）,and coated with gelatin sustained-release microspheres which were made by improved emulsified cold condensation method.The effects of porous titanium coated with insulin-like growth factor-1（IGF-1） and transforming growth factor-β1（TGF-β1） gelatin microspheres on the function of MG63 cells were evaluated in vitro.The results show that porous titanium coated with gelatin sustained-release microspheres has no cytotoxicity.The IGF-1 and TGF-β1 loading concentrations are positively correlative with the proliferation and differentiation of MG63 after co-culturing with the concentrations of IGF-1 and TGF-β1 gelatin microspheres in the range of 0.1-10 ng/mg and 0.25-2.5 ng/mg,respectively.The MG63 cells exhibit the best proliferation and differentiation with the IGF-1 and TGF-β1 loading concentrations of 10 ng/mg and 2.5 ng/mg,respectively.The joint application of IGF-1 and TGF-β1 group,which promote adhesion,proliferation and differentiation of MG63 cells,is superior to a single application group.

Role of Hypoxia-inducible Factor-1α in Formation of Muttidrug Resistance Induced by Microenvironment in Hepatocellular Carcinoma

Objective： To explore the role of hypoxia-inducible factor-1α （HIF-1α） in formation of multidrug resistance （MDR） induced by microenvironment and to find a new and effective molecular target on preventing and reversing chemoresistance in hepatocellular carcinoma （HCC）. Methods： In HepG2 cells exposed to hypoxia, low glucose or transfected by plasmid pcDNA3/HBX, the expression of HIF-1α mRNA and protein was respectively detected using real-time fluorescent quantitative PCR and Westernblot technique and its expression localization was investigated by immunocytochemical technique. Plasmid pcDNA3/HIF-1α was transfected into HepG2 cells and then the expression of multidrug resistance related genes mdrl, multidrug resistance-associated protein 1 （MRP1） and lung resistance protein （LRP） in transfected cells was determined by the same methods. Results： In HepG2 cells respectively exposed to hypoxia, low glucose or transfected by plasmid pcDNA3/HBX, HIF-1α was overexpressed at mRNA and protein levels to varying degrees and translocated into nucleus. The gene expression levels of mdrl, MRP1 and LRP in HepG2 cells transfected by plasmid pcDNA3/HIF-1α were respectively increased by 2.4±0.2, 2.2±0.3 and 2.3±0.4 folds as compared with those in non-transfected HepG2 cells （all P〈0.01） and similar changes were observed in protein level. Conclusion： Microenvironmental factors around HCC could modulate the transcription of the MDR related genes by nuclear transcript factor HIF-1α, thereby conferred MDR of HCC. Up-regulation of HIF-1α expression could hold a central position in the formation of MDR of HCC induced by microenvironment. HIF-1α probably becomes a new and effective molecular target on preventing and reversing MDR in HCC.

GENE EXPRESSION OF TRANSFORMING GROWTH FACTOR β_1 TYPEII RECEPTOR IN HCC AND ITS CLINICAL SIGNIFICANCE

Objective: Transforming Growth Factor-β1 (TGF-β1)plays a central role in the process of . growth suppressionof the hepatocytes, and its type II receptor (TGF-β1R II)transfers the signal of growth suppression. In this study,the gene expression of TGF-β1R II in HCC and itsclinical significance was investigated. Methods: Theexpression of TGF-β1R II mRNA in 30 cases Of HCCtissue and the surrounding liver tissue was separatelydetected using reverse transcription-PCR. Results:The positive expression rate of TGF-β1R II mRNA wassignificantly lower in HCC tissue (11/30) than that in thesurrounding liver tissue (23/30) (P<0.01). Further, theless the cancer tissue expressed TGF-β1R II mRNA, themore poorly the tumoral hepatocyte differentiated(P<0.01) and the more portal vein cancer embolusexisted (p=0.0465). Conclusion: The decreaseexpression of TGF-β1 R II mRNA by tumoral hepatocyteresults in the defect of its negative growth regulation,and this may be one of the most important reasons forits carcinogenesis and uncontrolled growth.

荔枝核总黄酮对TGF-β_1诱导人肝星状细胞凋亡的影响及机制

目的探讨荔枝核总黄酮(TFL)对转化生长因子β_1(TGF-β_1)诱导的人肝星状细胞(HSC-LX2)凋亡的影响,探讨其相关的分子机制。方法将培养好的HSC-LX2随机分为正常组、TGF-β_1组及150、300、600 mg/L TFL组。正常组常规培养;其他四组接种于含10%FBS的DMEM+5μg/L TGF-β_1、培养皿中培养24 h后,150、300、600 mg/L TFL组分别加入150、300、600 mg/L TFL继续培养48 h。用Hoechst33258染色法观察各组细胞形态学变化;用流式细胞术检测细胞周期;用FITC Annexin V/PI双染色法检测细胞凋亡情况;用Western blotting法检测细胞中Toll样受体4(TLR4)、核因子κB(NF-κB)、白细胞介素1受体Ⅰ(IL-1RⅠ)蛋白表达。结果正常组、TGF-β_1组细胞核形态完整,TGF-β_1组细胞增殖活跃;150、300、600 mg/L TFL组可见细胞核固缩、裂解、凋亡小体形成等细胞凋亡形态学变化。随TFL浓度升高,TFL组G0/G1期细胞增多(P<0.05),S期细胞减少(P<0.05)。随TFL浓度升高,TFL对HSC-LX2凋亡具有促进作用,更倾向于促进细胞晚期凋亡。150、300、600 mg/L TFL组TLR4、NF-κB、IL-1RⅠ蛋白表达量低于TGF-β_1组(P均<0.05);且随TFL浓度升高,HSC-LX2内TLR4、NF-κB、IL-1RⅠ蛋白表达量逐渐降低(P均<0.05)。结论 TFL可促进TGF-β_1诱导的HSC-LX2细胞凋亡,尤其促进细胞晚期凋亡;其分子机制可能与降低细胞内TLR4、NF-κB、IL-1RⅠ的表达有关。

银杏叶提取物对TGF-β_1诱导的人Tenon's囊成纤维细胞增殖的影响及其机制的初步探讨

目的:观察银杏叶提取物(GBE)对转化生长因子-β1(TGF-β1)诱导的人Tenon's囊成纤维细胞(HTCFs)增殖的影响及可能机制。方法:体外培养HTCFs及鉴定;MTT法检测不同浓度的GBE(25、50、100、200mg/L)对TGF-β1(2ng/mL)诱导的HTCFs增殖的影响;流式细胞仪检测细胞周期;免疫细胞化学法观察α-平滑肌动蛋白(α-SMA)表达的情况。RT-PCR检测细胞结缔组织生长因子mRNA(CTGF mRNA)的表达。结果:TGF-β1可显著促进HTCFs的增殖,促进细胞由G0/G1期进入S期,上调α-SMA、CTGF在蛋白和mRNA水平的表达;GBE能够抑制TGF-β1的上述作用。结论:GBE具有抑制TGF-β1诱导的人Tenon's囊成纤维细胞增殖的作用,其机制可能是阻止细胞进入S期,下调α-SMA及CTGF的表达。

二黄汤对哮喘模型大鼠肺组织中TGF-β_1及体内IL-33的影响

目的观察二黄汤对哮喘模型大鼠肺组织中转化生长因子β1(TGF-β1)及体内白介素-33(IL-33)的影响。方法 50只SD大鼠随机均分为正常组、哮喘组、布地奈德组、高剂量二黄汤组(含生药量68 g/kg)和低剂量二黄汤组(含生药量17 g/kg)。以卵清白蛋白致敏与激发建立哮喘大鼠模型,随后分别用布地奈德、二黄汤干预治疗。肺组织切片HE染色观察病理变化并检测气道管壁厚度(Wat)及气道平滑肌厚度(Wam),用免疫组化法检测肺组织TGF-β1蛋白的表达,酶联免疫法检测血清及支气管肺泡灌洗液(BALF)中IL-33的含量。结果所有药物干预组较哮喘组炎症细胞浸润明显减轻;布地奈德组、高剂量二黄汤组和低剂量二黄汤组Wat均较哮喘组下降(μm2/μm:54.99±8.82、52.28±7.61、58.53±7.63 vs 79.50±5.64,P<0.05);布地奈德组、高剂量二黄汤组和低剂量二黄汤组Wam均较哮喘组下降(μm2/μm:22.74±2.73、20.63±1.72、21.20±4.50 vs 30.16±1.68,P<0.05);与正常组比较,哮喘组BALF、血清中IL-33的浓度增高,经药物干预后,布地奈德组、高剂量二黄汤组和低剂量二黄汤组低于哮喘组(P<0.05),但3干预组间差异无统计学意义;哮喘组TGF-β1高于正常组(IOD:12.60±2.25 vs 1.67±0.17),布地奈德组(5.51±2.48)、高剂量二黄汤组(5.22±2.52)和低剂量二黄汤组(6.92±2.18)均低于哮喘组(P<0.05),3干预组间差异无统计学意义。哮喘大鼠的气道壁厚度和平滑肌厚度与TGF-β1、IL-33呈正相关。结论二黄汤可在一定程度上干预哮喘大鼠气道重塑,其作用可能是通过调节TGF-β1和IL-33实现的。

Insulin-like growth factor-1 induces lymphangiogenesis and facilitates lymphatic metastasis in colorectal cancer

AIM:To investigate the expression of insulin-like growth factor-1(IGF-1)/insulin-like growth factor-1 receptor(IGF-1R)in colorectal cancer(CRC)tissues and to analyze their correlation with lymphangiogenesis and lymphatic metastasis.METHODS:Immunohistochemistry was used to evaluate IGF-1 and IGF-1R expression and lymphatic vessel density(LVD)in 40 CRC specimens.The correlation between IGF-1/IGF-1R and LVD was investigated.Effects of IGF-1 on migration and invasion of CRC cells were examined using transwell chamber assays.A LoVo cell xenograft model was established to further detect the role of IGF-1 in CRC lymphangiogenesis in vivo. RESULTS:Elevated IGF-1 and IGF-1R expression in CRC tissues was correlated with lymph node metastasis(r=0.715 and 0.569,respectively,P<0.05)and tumor TNM stage(r=0.731 and 0.609,P<0.05).A higher LVD was also found in CRC tissues and was correlated with lymphatic metastasis(r=0.405,P<0.05).A positive correlation was found between LVD and IGF-1R expression(r=0.437,P<0.05).Transwell assays revealed that IGF-1 increased the migration and invasion of CRC cells.In vivo mouse studies showed that IGF-1 also increased LVD in LoVo cell xenografts.CONCLUSION:IGF-1/IGF-1R signaling induces tumorassociated lymphangiogenesis and contributes to lymphatic metastasis of CRC.

Improvement in erectile dysfunction after insulin-like growth factor-1 gene therapy in diabetic rats

Aim： To determine whether adenoviral gene transfer of insulin like growth factor-1 （IGF-1） to the penis of streptozotocin （STZ）-induced diabetic rats could improve erectile capacity. Methods： The STZ diabetic rats were transfected with AdCMV-βgal or AdCMV-IGF-1. These rats underwent cavernous nerve stimulation to assess erectile function and their responses were compared with those of age-matched control rats 1 to 2 days after transfection. In control and transfected STZ diabetic rats, IGF-1 expression were examined by reverse transcription polymerase chain reaction （RT-PCR）, Western blot and histology. The penis β-galactosidase activity and localization of the STZ diabetic rats were also determined. Results： One to two days after transfection, the β-galactosidase was found in the smooth muscle cells of the diabetic rat penis transfected with AdCMV-βgal. One to 2 days after administration of AdCMV- IGF-1, the cavernosal pressure, as determined by the ratio of maximal intracavernous pressure-to-mean arterial pressure （ICP/MAP） and total intracavernous pressure （ICP）, was increased in response to cavernous nerve stimulation. Transgene expression was confirmed by RT-PCR, Western blot and histology. Conclusion： Gene transfer of IGF-1 significantly increased erectile function in the STZ diabetic rats. These results suggest that in vivo gene transfer of IGF- 1 might be a new therapeutic intervention for the treatment of erectile dysfunction （ED） in the STZ diabetic rats.