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.

Expression of Nerve Growth Factor and Hypoxia Inducible Factor-1α and Its Correlation with Angiogenesis in Non-Small Cell Lung Cancer

Summary： In order to investigate the expression of nerve growth factor （NGF） and hypoxia inducible factor-1α （HIF-1α） and its correlation with angiogenesis in non-small cell lung cancer （NSCLC）, paraffin-embedded tissue blocks from 20 patients with NSCLC were examined. Twenty corresponding para-cancerous lung tissue specimens were obtained to serve as a control. The expression of NGF, HIF-1α, and vascular endothelial growth factor （VEGF） in the NSCLC tissues was detected by using immunohistochemistry. The microvascular density （MVD） was determined by CD31 staining. The resuits showed that the expression levels ofNGF, HIF-1α and VEGF in the NSCLC tissues were remarkably higher than those in the para-cancerous lung tissues （P〈0.05）. There was significant difference in the MVD between the NSCLC tissues （9.19±1.43） and para-cancerous lung tissues （2.23±1.19） （P〈0.05）. There were positive correlations between NGF and VEGF, between HIF-1α and VEGF, and between NGF and HIF-1α in NSCLC tissues, with the spearman correlation coefficient being 0.588, 0.519 and 0.588, respectively. In NSCLC tissues, the MVD had a positive correlation with the three factors （P〈0.05）. Theses results suggest that NGF and HIF-1α are synergically involved in the angiogenesis of NSCLC.

Clinicopathological and Prognostic Significance of Hypoxia-inducible Factor-1 alpha in Lung Cancer: a Systematic Review with Meta-analysis

Hypoxia-inducible factor-1 alpha（HIF-1α） plays a vital role in the initiation, evaluation and prognosis in lung cancer. The prognostic value of HIF-1α reported in diverse study remains disputable. Accordingly, a meta-analysis was implemented to further understand the prognostic role of HIF-1α in lung cancer. The relationship between HIF-1α and the clinicopathological characteristics and prognosis of lung cancer were investigated by a meta-analysis. Pub Med and Embase were searched from their inception to January 2015 for observational studies. Fixed-effects or random-effects meta-analyses were used to calculate odds ratios and 95% confidence intervals of different comparisons. A total of 20 studies met the criteria. The results showed that HIF-1α expression in lung cancer tissues was significantly higher than that in normal lung tissues. Expression of HIF-1α in patients with squamous cell carcinoma was significantly higher than that of patients with adenocarcinomas. Similarly, non-small cell lung cancer（NSCLC） patients had higher HIF-1α expression than small cell lung cancer（SCLC） patients. Moreover, lymph node metastasized tissues had higher HIF-1α expression than non-lymph node metastasized tissues. A high level HIF-1α expression was well correlated with the expression of vascular endothelial growth factor and epidermal growth factor receptor in the NSCLC. Notably, NSCLC or SCLC patients with positive HIF-1α expression in tumor tissues had lower overall survival rate than patients with negative HIF-1α expression. It was suggested that HIF-1α expression may be a prognostic biomarker and a potential therapeutic target for lung cancer.

The Expression of Hypoxia Inducible Factor 1-alpha in Lung Cancer and Its Correlation with P53 and VEGF

To investigate the expression of hypoxia inducible factor 1-alpha (HIF-1α) and its correlation with P53 and vascular endothelial growth factor (VEGF), immunohistochemical technique was employed to detect the protein expressions of HIF-1α, P53 and VEGF in specimens from 57 patients with lung cancer. The results indicated that the total positive proportion of HIF-1α expression was 63 % and the HIF-1α expression was more frequent in bronchiole-alveolar carcinoma (86 %) than in other lung cancer. There was a strong association of HIF-1α with VEGF and P53 protein expressions. It is concluded that HIF-1α overexpression is a common event in lung cancer, which may be related to the up-regulation of the angiogenic factor VEGF and oncogene mutant P53 protein.

Chronic intermittent hypoxia increases β cell mass and activates the mammalian target of rapamycin/hypoxia inducible factor 1/vascular endothelial growth factor A pathway in mice pancreatic islet

Background Growing evidence from population and clinic based studies showed that obstructive sleep apnea （OSA） and its characterizing chronic intermittent hypoxia （IH） were independently associated with the development of type 2 diabetes mellitus.However,the pathogenesis by which OSA induces glucose metabolic disorders is not clear.We determined changes in pancreatic β cell mass and the mammalian target of rapamycin （mTOR）/hypoxia inducible factor 1 （HIF-1）/ vascular endothelial growth factor A （VEGF-A） pathway following IH exposure.Methods A controlled gas delivery system regulated the flow of nitrogen and oxygen into a customized cage housing mice during the experiment.Twenty-four male wild C57BL/6J mice were either exposed to IH （n=12） or intermittent air as a control （n=12） for 56 days.Mice were anaesthetized and sacrificed after exposure,pancreas samples were dissected for immunofluorescent staining.Insulin and DAPI staining labelled islet β cells.Insulin positive area and β cell number per islet were measured.P-S6,HIF-1α and VEGF-A staining were performed to detect the activation of mTOR/HIF-1NEGF-A pathway.Results After eight weeks of IH exposure,insulin positive area increased by an average of 18.5% （P 〈0.05）.The β cell number per islet increased （92 vs.55,respectively for IH and the control groups,P 〈0.05） with no change in the size of individual β cells.Islet expression of HIF-1α and VEGF-A were higher in IH group than control group,and percentage of p-S6 positive β cell also increased after IH exposure （16.8% vs.4.6% respectively for IH and the control groups,P 〈0.05）.Conclusion The number of pancreatic β cells increased as did the activity of the mTOR/HIF-1NEGF-A pathway after exposure to IH.

Hypoxia-inducible factor-1 alpha regulates the role of vascular endothelial growth factor on pulmonary arteries of rats with hypoxia-induced pulmonary hypertension

Background Hypoxia-inducible factor-1α (HIF-1α) is one of the pivotal mediators in the response of lungs to decreased oxygen availability, and increasingly has been implicated in the pathogenesis of pulmonary hypertension. Vascular endothelial growth factor (VEGF), a downstream target gene of HIF-1α, plays an important role in the pathogenesis of hypoxic pulmonary hypertension and hypoxic pulmonary artery remodelling. In this study, we investigated the dynamic expression of HIF-1α and VEGF in pulmonary artery of rats with hypoxia-induced pulmonary hypertension. Methods Forty male Wistar rats were exposed to hypoxia for 0, 3, 7, 14 or 21 days. Mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were estimated. Lungs were inflated and fixed for in situ hybridisation and immunohistochemistry. Results mPAP values were significantly higher than the control values after 7days of hypoxia [(18.4±0.4) mmHg, P<0.05]. RVHI developed significantly after 14 days of hypoxia. Expression of HIF-1α protein increased in pulmonary arterial tunica intima of all hypoxic rats. In pulmonary arterial tunica media, HIF-1α protein was markedly increased by day 3 (0.20±0.02, P<0.05), reached the peak by day 7, then declined after day 14 of hypoxia. HIF-1α mRNA increased significantly after day 14 of hypoxia (0.20±0.02, P<0.05). VEGF protein began to increase markedly after day 7 of hypoxia, reaching its peak around day 14 of hypoxia (0.15±0.02, P<0.05). VEGF mRNA began to increase after day 7 of hypoxia, then remained more or less stable from day 7 onwards. VEGF mRNA is located mainly in tunica intima and tunica media, whereas VEGF protein is located predominantly in tunica intima. Linear analysis showed that HIF-1α mRNA, VEGF and mPAP were correlated with hypoxic pulmonary artery remodelling. HIF-1α mRNA was positively correlated with VEGF mRNA and protein (P<0.01). Conclusion HIF-1α and VEGF are both involved in the pathogenesis of hypoxia-induced pulmonary hypertension in rats.

Expression levels and significance of hypoxia inducible factor-1 alpha and vascular endothelial growth factor in human colorectal adenocarcinoma

Background Hypoxia-inducible factor 1 (HIF-1), a transcription factor, is overexpressed in common human cancers and their metastases. This study aimed at determining the expression levels of HIF-1α and vascular endothelial growth factor (VEGF) in SW480 cells and in colorectal adenocarcinoma tissue and ascertaining whether HIF-1α and VEGF play important roles in tumor angiogenesis. Methods HIF-1α mRNA expression was analyzed using in situ hybridization and RT-PCR. HIF-1α and VEGF protein were detected in SW480 cells and colorectal adenomas and adenocarcinomas by immunohistochemistry using streptavidin/peroxidase (SP). Western blot was used to detect HIF-1α protein extracted from SW480 cells. Microvessel density (MVD) in colorectal carcinomas was determined by anti-CD_ 34 immunostaining in colorectal carcinomas. Results Optical density values representing HIF-1α mRNA expression levels were found to be significantly higher in SW480 cells in hypoxic conditions than in cells under normoxic conditions (P<0.05) or in hypoxic conditions but treated with genistein (P<0.05). The levels of HIF-1α and VEGF protein expression in SW480 cells were significantly higher in the hypoxia group than in the normoxia group (P<0.01, P<0.05, respectively) and hypoxia/genistein group (P<0.01, P<0.05, respectively). The positive expression rates of HIF-1α mRNA changed dramatically when comparing colorectal adenomas with adenocarcinomas of different Dukes’ stages (P<0.05). HIF-1α mRNA was also expressed at higher levels in adenocarcinomas than that in adenomas (P<0.01). HIF-1α protein expression correlated significantly with VEGF protein expression and MVD.Conclusions Hypoxia induces the expression of HIF-1α and VEGF in colorectal adenocarcinomas. HIF-1α may play an important role in angiogenesis and tumor progression by regulating the expression of VEGF in human colorectal carcinomas.

Effect of endothelial PAS domain protein 1 and hypoxia inducible factor 1~ on vascular endothelial growth factor expression in human pancreatic carcinoma

Background Transcription factors hypoxia inducible factor 1α （HIF 1α） and endothelial PAS domain protein 1 （EPAS1） promote the transcription of vascular endothelial growth factor （VEGF）. VEGF enhances angiogenesis and vascular permeability of tumours, which promotes tumour growth and facilitates entry of cancer cells into blood circulation and metastasizing. This study examined whether HIF 1α and EPAS1 stimulated angiogenesis through activation of VEGF in human pancreatic carcinoma. Methods Specimens from pancreatic carcinoma and healthy parts of same pancreas were taken from 60 patients. Real time quantitative reverse transcription polymerase chain reaction estimated expression of HIF 1α, EPAS1, and VEGF mRNAs. Western blotting and immunohistochemical, streptavidin peroxidase method assessed expression of HIF 1α, EPAS1, and VEGF proteins. Microvessel density （MVD） was assessed. Results Highly significant increases in expression of EPAS1, VEGF, and MVD were found in pancreatic carcinoma tissue but not in normal pancreatic tissue： VEGF at mRNA and protein levels （t=17.32, P=-0.0001; t=98.41, P=0.0001）; EPAS1 protein level （t=22.51, P=0.0001）. Expression of HIF la was similar in pancreatic carcinoma and normal pancreatic tissues at both mRNA and protein levels. Significant correlations were observed between EPAS1 and VEGF （r=0.736, P=0.0041）, between VEGF and MVD （r=0.858, P=0.0001）, and between EPAS1 and MVD （r=0.641, P=0.0003）. No significant correlations were observed between HIF la and VEGF, or between HIF 1α and MVD. MVD and expression of EPAS1 and VEGF were significantly related with TNM staging, so was EPASI and VEGF with size of tumour. Conclusions EPAS1 and VEGF, but not HIFla, are overexpressed in pancreatic carcinoma. The expression of EPAS1 is correlated with that of VEGF and MVD. EPAS1 may be involved in the angiogenesis of pancreatic carcinoma by upregulating the expression of VEGE Targeting EPAS1 may be a new method of antiangiogenic tumour therapy for pancreatic carcinoma.

Tongxinluo Inhibits Cyclooxygenase-2, Inducible Nitric Oxide Synthase, Hypoxia-inducible Factor-2α/Vascular Endothelial Growth Factor to Antagonize Injury in Hypoxia-stimulated Cardiac Microvascular Endothelial Cells

Background： Endothelial dysflinction is considered as the initiating process and pathological basis of cardiovascnlar disease. Cyclooxygenase-2 （COX-2） and prostacyclin synthase （PGIS）, inducible nitric oxide synthase （iNOS） and endothelial NOS （eNOS） are key enzymes with opposing actions in inflammation and oxidative stress, which are believed to be the major driver of endothelial dysfunction. And in hypoxia （Hx）, Hx-inducible factor （HIF）-1α and HIF-2α are predominantly induced to activate vascular endothelial growth factor （VEGF）, restllting in abnormal proliferation. Whether and how Tongxinluo （TXL） modulates COX-2, PGIS, iNOS, eNOS, HIF-1α, HIF-2α, and VEGF in Hx-stimulated human cardiac microvascular endothelial cells （HCM ECs） have not been clarified. Methods： HCMEC were treated with CoCl2 to mimic Hx and the mRNA expressions of COX-2, PGIS, iNOS, eNOS, HIF-1α, HIF-2α. and VEGF were first confirmed, and then their mRNA expression and protein content as well as the cell pathological alterations were evaluated for TXL treatment with different concentrations, In addition, the effector molecular of inflammation prostaglandin E2 （PGE2） and the oxidative marker nitrotyrosine （NT） was adopted to reflect HCMEC in.jury. Results： Hx could induce time-dependent increase of COX-2, iNOS, HIF-2α, and VEGF in HCMEC. Based on the Hx-induced increase, TXL could mainly decrease COX-2, iNOS, HIF-2α, and VEGF in a concentration-dependent manner, with limited effect on the increase of PGIS and eNOS. Their protein contents verified the mRNA expression changes, which was consistent with the cell morphological alterations. Furthermore, high dose TXL could inhibit the Hx-induced increase of PG E, and NT contents, attenuating the inflammatory and oxidative injury. Conclusions： TXL could inhibit inflammation-related COX-2, oxidative stress-related iNOS, and H IF-2α/VEGF to antagonize Hx-induced HCMEC injury.

电针调控HIF-1α/VEGF信号通路对类风湿性关节炎模型踝关节病理学改变的影响

目的探讨电针通过调控HIF-1α/VEGF信号通路活性对类风湿性关节炎(RA)大鼠踝关节病理损伤的影响。方法选取50只SD大鼠,随机分为Sham组(空白对照)、RA组(大鼠构建RA模型)、电针组(RA组大鼠给予电针治疗)、CAY10585组(RA大鼠腹腔内注射HIF-1α/VEGF信号通路抑制剂)、电针+CAY10585组(电针组大鼠腹腔注射HIF-1α/VEGF信号通路抑制剂),每组10只。模型建立后观察各组大鼠基本形态;8周后处死大鼠,比较治疗后大鼠踝关节直径;取大鼠踝关节行HE染色,观察整体病理形态及踝关节病理特征(滑膜细胞增殖、细胞侵蚀、血管翳形成、炎性细胞浸润、骨侵蚀)病理评分;酶联免疫吸附法(ELISA)检测5组大鼠血清炎性因子肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平;Western-blot检测大鼠踝关节组织内软骨基质因子Ⅱ型胶原(CollagenⅡ)、C端肽(CTXⅡ)、Ⅱ型胶原C前肽(CPⅡ)蛋白表达。结果与Sham组比较,RA模型建立可以上调大鼠踝关节直径,踝关节滑膜细胞增殖、细胞侵蚀、血管翳形成、炎性细胞浸润、骨侵蚀病理评分,血清炎性因子(TNF-α、IL-6)水平均明显提升(P<0.05),关节滑膜上皮复层出现增生、间质水肿以及炎性细胞浸润的现象;电针干预可以下调大鼠踝关节直径、踝关节滑膜细胞增殖、细胞侵蚀、血管翳形成、炎性细胞浸润、骨侵蚀病理评分,血清炎性因子(TNF-α、IL-6)水平(P<0.05),同时踝关节病理形态明显得到缓解;CAY10585干预再次增加RA大鼠病理形态,破骨细胞明显增加,骨质出现侵蚀、溶解骨层及炎性细胞浸润的现象,踝关节直径、踝关节滑膜细胞增殖、细胞侵蚀、血管翳形成、炎性细胞浸润、骨侵蚀病理评分,血清炎性因子(TNF-α、IL-6)水平再次提升(P<0.05);电针干预可以逆转CAY10585组大鼠趋势,再次下调大鼠踝关节直径、踝关节滑膜细胞增殖、细胞侵蚀、血管翳形成、炎性细胞浸润、骨侵蚀病理评分(P<0.05);与Sham组比较,RA组大鼠踝关节组织内CTXⅡ表达提升,CollagenⅡ、CPⅡ表达降低(P<0.05);电针干预可以下调CTXⅡ表达,上调CollagenⅡ、CPⅡ、HIF-1α、VEGF表达(P<0.05);CAY10585干预则体现出相反趋势,再次上调CTXⅡ表达,下调CollagenⅡ、CPⅡ、HIF-1α、VEGF表达(P<0.05);电针干预可以逆转CAY10585组大鼠软骨基质及HIF-1α、VEGF表达(P<0.05)。结论电针治疗可以改善RA大鼠踝关节的病理学表现,降低炎性反应,保护软骨损伤,其分子机制可能与激活HIF-1α/VEG信号通路有关。

Protective effects of piperine on the retina of mice with streptozotocin-induced diabetes by suppressing HIF-1/VEGFA pathway and promoting PEDF expression

AIM:To evaluate the protective mechanisms of piperine in the retina of mice with streptozotocin-induced diabetes.METHODS:In experiments in vitro,stimulation by chemical hypoxia was established in ARPE-19 cells.Then,the expression of hypoxia-inducible factor-1α(HIF-1α),vascular endothelial growth factor A(VEGFA),and pigment epithelium-derived factor(PEDF)was assessed at the m RNA and protein levels.In experiments in vivo,diabetes mellitus was established by intraperitoneally injecting 150 mg/kg streptozotocin once.After 3 wk of the onset of diabetes,15 mg/kg piperine was intraperitoneally injected once daily for 1 or 3 wk.Then,the retinal morphology and m RNA and protein expression were assessed.RESULTS:In hypoxia,1-100μmol/L piperine significantly decreased the expression of VEGFA m RNA and increased the expression of PEDF m RNA without affecting HIF-1αm RNA.Meanwhile,100μmol/L piperine substantially decreased the protein level of VEGFA and increased the protein level of PEDF.The HIF-1αprotein level was also hampered by piperine.In the diabetic retina of mice,the morphological damage was alleviated by piperine.Likewise,the retinal vascular leakage was substantially decreased by piperine.Further,the protein levels of HIF-1αand VEGFA were significantly reduced by piperine.Moreover,the level of the antiangiogenic factor of PEDF dramatically increased by piperine.CONCLUSION:Piperine may exert protective effects on the retina of mice with diabetes via regulating the pro-antiangiogenic homeostasis composed of HIF-1/VEGFA and PEDF.

MiR-181c-5p对卵巢癌干细胞样细胞肿瘤血管生成拟态的作用及其机制

目的探讨miR-181c-5p对卵巢癌干细胞样细胞(OCS-LCs)肿瘤血管生成拟态(VM)的作用及其机制。方法采用无血清悬浮培养法将人卵巢癌细胞系OVCAR3细胞诱导形成OCS-LCs。将OVCAR3细胞分为A~C组,各组分别转染NC-miR-181c-5p、siRNA-miR-181c-5p和pRNA-miR-181c-5p。通过成球实验评估A~C组细胞成球能力。采用实时荧光定量PCR(RT-qPCR)方法检测A~C组细胞miR-181c-5p的相对表达量,采用Western blot实验检测A~C组细胞Oct-4、Nanog、HIF-1α和VEGF蛋白相对表达量。采用CCK-8实验检测A~C组细胞的活性,采用三维立体培养实验检测A~C组的血管形成率。结果OVCAR3细胞成功被诱导形成OCS-LCs。RT-qPCR实验结果显示,B组细胞的miR-181c-5p相对表达量显著低于A组,C组高于A组(t=2.25、8.68,P<0.05)。成球实验结果显示,B组与A组、A组与C组相比,细胞的成球周期显著缩短,最大的细胞球直径显著增大,成球率显著增加(t=5.56~33.66,P<0.05)。Western blot实验结果表明,B组与A组、A组与C组相比,Oct-4、Nanog、HIF-1α和VEGF蛋白相对表达量显著升高(t=4.51~56.15,P<0.05)。CCK-8实验结果显示,B组的细胞活性高于A组,C组低于A组(F=97.70~281.80,P<0.05)。三维立体培养实验结果显示,B组与A组、A组与C组相比较,血管形成率显著性提高(t=3.70、18.67,P<0.05)。结论miR-181c-5p可能通过降低细胞中HIF-1α和VEGF蛋白的表达,从而抑制OCS-LCs的VM形成。

血清HIF-1α、HO-1和sFlt-1对妊娠期肝内胆汁淤积症患者胎儿宫内缺氧的诊断价值

目的 探讨血清缺氧诱导因子1α(HIF-1α)、血红素加氧酶-1(HO-1)和可溶性血管内皮生长因子受体-1(sFlt-1)对妊娠期肝内胆汁淤积症(ICP)患者胎儿宫内缺氧的诊断价值。方法 选择2020年1月至2022年12月在该院诊治的148例ICP患者纳入ICP组。选择同期该院66例健康孕妇纳入正常妊娠组。观察两组血清HIF-1α、HO-1和sFlt-1水平的变化,探讨宫内缺氧的影响因素,分析血清HIF-1α、HO-1和sFlt-1水平与ICP患者严重程度的关系,以及HIF-1α、HO-1和sFlt-1诊断ICP患者发生宫内缺氧的效能。根据ICP患者是否发生宫内缺氧分为宫内缺氧组和无宫内缺氧组,比较两组临床指标的差异。结果 ICP组血清HIF-1α和sFlt-1水平明显高于正常妊娠组(P<0.01),并且随着ICP疾病严重程度升高而升高(P<0.01),而血清HO-1水平明显低于正常妊娠组(P<0.01),随着ICP严重程度升高而降低(P<0.01)。宫内缺氧组的直接胆红素、间接胆红素、总胆汁酸、甘胆酸、HIF-1α和sFlt-1水平明显高于无宫内缺氧组(P<0.01),凝血酶原时间、活化部分凝血活酶时间长于无宫内缺氧组(P<0.01),而血清HO-1水平明显低于无宫内缺氧组(P<0.01)。多因素Logistic回归分析发现,直接胆红素、HIF-1α和sFlt-1水平升高,HO-1水平降低是宫内缺氧的独立危险因素(P<0.05)。血清HIF-1α、HO-1和sFlt-1对ICP患者发生宫内缺氧具有较高的诊断效能,联合检测的灵敏度为96.8%,特异度为88.4%,曲线下面积(AUC)为0.969,明显高于单项指标HIF-1α(Z=3.294,P<0.001)、HO-1(Z=4.841,P<0.001)和sFlt-1(Z=3.602,P<0.001)检测,而3项指标之间的AUC比较,差异无统计学意义(P>0.05)。结论 HIF-1α、HO-1和sFlt-1是反映ICP患者严重程度和宫内缺氧的指标,三者联合检测有利于提高对胎儿宫内缺氧的诊断效能。

白术内酯I调节HIF-1α/VEGF信号通路对急性心肌梗死大鼠心肌损伤的影响

目的:探讨白术内酯I(Atr-I)调节缺氧诱导因子1α(HIF-1α)/血管内皮生长因子(VEGF)信号通路对急性心肌梗死(AMI)大鼠心肌损伤的影响。方法:大鼠分为对照组、AMI组、Atr-I组、阿司匹林组、BAY87-2243组、Atr-I+BAY87-2243组,每组18只。除对照组外,其他组大鼠均采用结扎冠脉左前降支根部的方式构建AMI模型,建模1h后,开始处理,给药1次/d,持续7d。超声心动图监测左室短轴缩短率(FS)、左室射血分数(LVEF)的变化;2,3,5-三苯基氯化四氮唑(TTC)染色检测大鼠心肌梗死面积百分数;HE染色检测左心室心肌组织病理学变化;TUNEL染色检测心肌细胞凋亡;ELISA检测大鼠左心室心肌组织中肌红蛋白(Mb)、乳酸脱氢酶(LDH)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β含量;Western blot检测大鼠心肌组织匀浆中HIF-1α、VEGF蛋白表达。结果:与对照组相比,AMI组大鼠心肌损伤明显,FS、LVEF及HIF-1α、VEGF蛋白表达降低,心肌梗死面积百分数、Mb、LDH、TNF-α、IL-1β含量升高(P<0.05);与AMI组相比,Atr-I组、阿司匹林组大鼠心肌损伤有所改善,FS、LVEF及HIF-1α、VEGF蛋白表达升高,心肌梗死面积百分数、Mb、LDH、TNF-α、IL-1β含量降低,BAY87-2243组对应指标变化趋势与上述相反(P<0.05);与Atr-I组相比,Atr-I+BAY87-2243组大鼠心肌损伤加剧,FS、LVEF及HIF-1α、VEGF蛋白表达降低,心肌梗死面积百分数、Mb、LDH、TNF-α、IL-1β含量升高(P<0.05)。结论:Atr-I减轻AMI大鼠心肌损伤可能与激活HIF-1α/VEGF信号通路有关。

HIF-1α/HO-1/VEGF-A信号通路在自体动静脉内瘘成熟不良中的调控机制

目的 观察慢性肾脏病5期、行自体动静脉内瘘(AVF)患者在内瘘不同成熟结局条件下低氧诱导因子-1α/血红素加氧酶-1/血管内皮生长因子A(HIF-1α/HO-1/VEGF-A)信号通路的表达水平差异,探讨HIF-1α/HO-1/VEGF-A信号通路在AVF成熟过程中的调控机制。方法 选取慢性肾脏病5期、拟行AVF手术的45例患者作为研究对象,在内瘘手术过程中采集血管标本,依据术后8周内瘘成熟结局分为AVF成熟组(n=35)和AVF成熟不良组(n=10)。采用逆转录-聚合酶链反应(RT-PCR)检测HIF-1α/HO-1/VEGF-A信号通路HIF-1α、HO-1、VEGF-A的基因表达水平;采用蛋白免疫印迹法检测HIF-1α、HO-1、VEGF-A的蛋白表达水平;采用酶联免疫吸附测定(ELISA)检测下游炎症因子如白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)、转化生长因子-β(TGF-β)表达水平。结果 AVF成熟组HIF-1α mRNA、HO-1 mRNA、VEGF-A mRNA表达水平高于AVF成熟不良组,差异有统计学意义(P<0.01);AVF成熟组HIF-1α、HO-1、VEGF-A蛋白表达水平高于AVF成熟不良组,差异有统计学意义(P<0.01);AVF成熟组IL-1β、IL-8、TGF-β表达水平低于AVF成熟不良组,差异有统计学意义(P<0.01)。结论 HIF-1α/HO-1/VEGF-A信号通路表达水平的上调有利于内瘘的早期成熟。HIF-1α/HO-1/VEGF-A信号通路表达水平的测定对AVF的早期成熟结局有一定的预测意义。

全真一气汤通过HIF-1α/VEGF通路抑制COPD大鼠气道重塑的研究

目的:探讨全真一气汤抑制慢性阻塞性肺疾病(COPD)大鼠气道重塑的机制。方法:将39只SPF级雄性SD大鼠随机分为空白组(n=9)、模型组(n=10)、氨茶碱组(n=10)和全真一气汤组(n=10)。除空白组外,其余各组大鼠采用LPS联合烟熏的经典造模法进行COPD造模。验证造模成功后,各药物组大鼠给予相应药物灌胃,空白组和模型组大鼠给予等体积生理盐水灌胃,1次/d,连续给药28 d。处死各组大鼠,收集样本并行HE染色、透射电镜及免疫组化等相关检测。结果:HE染色结果显示:空白组大鼠肺泡结构完整,肺泡间隔无明显增厚;模型组大鼠可见肺泡腔扩张明显并且破裂、肺泡间隔明显增厚,单位面积内的肺泡数量明显减少;氨茶碱组大鼠可见肺泡腔轻度扩张并且部分破裂、肺泡间隔轻度增厚,单位面积内的肺泡数量稍有减少;全真一气汤组大鼠可见肺泡腔轻度扩张、肺泡间隔轻度增厚,单位面积内的肺泡数量稍有减少;与空白组比较,模型组大鼠单位面积内的肺泡数量明显减少(P<0.01);与模型组比较,氨茶碱组和全真一气汤组大鼠单位面积内的肺泡数量明显增加(P<0.01)。透射电镜结果显示:空白组线粒体(M)未见明显肿胀,大多结构尚可,基质较为均匀,嵴存在;全真一气汤组线粒体(M)未见明显肿胀,大多结构尚可,基质较为均匀,嵴存在;模型组线粒体(M)损伤严重,整体呈中重度肿胀,膜破损,基质较多溶解,嵴消失;氨茶碱组线粒体(M)损伤严重,整体呈中重度肿胀,膜破损,基质较多溶解,嵴消失。4组肺泡Ⅱ型上皮细胞存在一定程度差异性。模型组、氨茶碱组线粒体(M)损伤最为严重,整体呈中重度肿胀,膜破损,基质较多溶解,嵴消失;空白组、全真一气汤组线粒体(M)未见明显肿胀,大多结构尚可,基质较为均匀,嵴存在。4组细胞表面的微绒毛未见明显差异,数量较少,排列稀疏。免疫组化结果显示:与空白组比较,模型组大鼠肺组织HIF-1α、VEGF平均光密度值均明显升高(P<0.01),occludin、ZO-1平均光密度值均明显降低(P<0.05);与模型组比较,氨茶碱组大鼠肺组织HIF-1α平均光密度值明显降低(P<0.05),occludin、ZO-1平均光密度值均明显升高(P<0.05);与模型组比较,全真一气汤组大鼠肺组织HIF-1α、VEGF平均光密度值均明显降低(P<0.05或P<0.01),occludin、ZO-1平均光密度值均明显升高(P<0.05或P<0.01)。Western blotting结果显示:与空白组比较,模型组大鼠肺组织HIF-1α、VEGF蛋白相对表达量均明显升高(P<0.01);与模型组比较,氨茶碱组大鼠肺组织VEGF蛋白相对表达量明显降低(P<0.01),HIF-1α蛋白相对表达量与模型组比较,差异无统计学意义(P>0.05);全真一气汤组大鼠肺组织HIF-1α、VEGF蛋白相对表达量均明显降低(P<0.01)。结论:全真一气汤可能通过下调HIF-1α、VEGF基因及上调occloudin、ZO-1蛋白的表达,进而达到改善COPD大鼠气道重塑的作用。

经肝动脉灌注栓塞微球对比碘化油乳剂治疗VX2兔肝癌模型对VEGF、CTGF和HIFα-1的影响及疗效评估

目的:观察栓塞微球对比碘化油乳剂经肝动脉灌注化疗栓塞对VX2兔肝癌模型血管内皮生长因子(VEGF)、结缔组织生长因子(CTGF)和缺氧诱导因子(HIF)α-1的影响及疗效。方法:选取VX2兔肝癌模型24只,使用随机数字表法分为空白对照组、栓塞微球组、碘化油乳剂组,每组8只。经肝动脉分别灌注0.9%氯化钠溶液1 mL、栓塞微球0.5 mL+表柔比星溶液0.5 mL、碘化油乳剂0.5 mL+奥沙利铂0.5 mL,观察并比较各组VX2兔肝癌模型VEGF、CTGF和HIFα-1水平及病灶坏死情况。结果:行化疗药灌注及栓塞后,各组的VEGF、CTGF和HIFα-1水平均有不同程度升高。术后7、14 d,栓塞微球组、碘化油乳剂组的VEGF、CTGF和HIFα-1水平明显高于空白对照组,差异均有统计学意义(P<0.05)。术后28 d,栓塞微球组、碘化油乳剂组的Ⅱ—Ⅳ级病灶坏死率高于空白对照组。栓塞微球组与碘化油乳剂组病灶坏死率的差异无统计学意义(P>0.05)。结论:经肝动脉栓塞治疗后,因使用栓塞药物不同造成不同栓塞水平可能影响VEGF、CTGF和HIFα-1表达及病灶坏死率。

Effect of HIF-1αon VEGF-C Induced Lymphangiogenesis and Lymph Nodes Metastases of Pancreatic Cancer

The effect of hypoxia inducible factor-1 α （HIF-1 α） on vascular endothelial growth factor C （VEGF-C） and the correlation between HIF- 1 α and lymphangiogenesis and lymph nodes metastases （LNM） in pancreatic cancer were investigated. Immunohistochemical SP method was used to detect the protein expression of HIF-1 α and VEGF-C, and Lymphatic vessel density （LVD） was determined by stain of VEGFR-3, collagen type IV in 75 pancreatic head cancers from regional pancreatectomy （RP） during Dec. 2001 to Dec. 2003. The relationship between HIF-1α and VEGF-C, lymphangiogenesis, LNM was analyzed statistically. The results showed that the positive expression rate of HIF-1α and VEGF-C in pancreatic cancer tissues was 48.00 % （36/75） and 65.33 % （49/75） respectively. In positive group of HIF-1α, the positive rate of VEGF-C and LVD, and LVD rate was 80.56 % （29/36）, 13.22±3.76 and 88.89 % （32/36） respectively, and in negative group of HIF-10t, positive rate of VEGF-C and LVD was 51.28 % （20/39）, 5.98±2.17 and 66.67 % （26/39） respectively （P〈0.01 or P〈0.05）. It was suggested that HIF-1α could promote the expression of VEGF-C, lymphangiogenesis and LNM in pancreatic cancer.

半夏白术天麻汤加减治疗高血压合并颈动脉粥样硬化斑块患者的临床疗效及对其HIF-1α、VEGF、VEGFR2信号通路的影响

目的探讨半夏白术天麻汤加减治疗高血压合并颈动脉粥样硬化斑的疗效及对缺氧诱导因子、血管内皮生长因子、血管内皮生长因子受体(Hypoxia-inducible factor-1 alpha、vascular endothelial growth factor、vas-cular endothelial growth factor receptor 2,HIF-1α、VEGF、VEGFR2)信号通路的影响。方法选取2019年1月—2022年2月期间上海中医药大学附属曙光医院收治的高血压合并动脉粥样硬化斑块患者80例作为研究对象,采用随机数字表法分为对照组和研究组,每组各40例。对照组为常规西医治疗,研究组在对照组基础上联合半夏白术天麻汤加减治疗。连续治疗2个月后,观察比较两组患者治疗前后血压(收缩压和舒张压)、血脂[胆固醇(Cholesterol,TC)、甘油三酯(Triglyceride,TG)、低密度脂蛋白胆固醇(Low density lipoprotein cholesterol,LDL-C)以及高密度脂蛋白胆固醇(High density lipoprotein cholesterol,HDL-C)]、颈动脉内膜中层厚度(Intima-media thick-ness,IMT)、外周血缺氧诱导因子/血管内皮生长因子/血管内皮生长因子受体(Hypoxia-inducible factor-1 alpha、vascular endothelial growth factor、vascular endothelial growth factor receptor 2,HIF-1α、VEGF、VEGFR2)、中医症状积分及两组患者生活质量水平。结果治疗后两组患者收缩压和舒张压均较治疗前降低,差异有统计学意义(P<0.05);且研究组低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者血脂TC、TG、LDL-C指标均较治疗前降低,HDL-C指标较治疗前升高,差异有统计学意义(P<0.05);且研究组TC、TG、LDL-C指标低于对照组,HDL-C指标高于对照组,差异有统计学意义(P<0.05)。治疗后两组患者IMT和中医症状积分均较治疗前降低,差异有统计学意义(P<0.05);且研究组低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者外周血HIF-1α、VEGF、VEGFR2水平均较治疗前降低,差异有统计学意义(P<0.05);且研究组低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者心理功能、社会功能、躯体功能和物质生活状态等GQOLI-74评分均较治疗前升高,差异有统计学意义(P<0.05);且研究组高于对照组,差异有统计学意义(P<0.05)。结论对高血压合并颈动脉粥样硬化斑块患者采用半夏白术天麻汤治疗,有助于改善患者的血压及血脂水平,抑制HIF-1α/VEGF/VEGFR 2信号通路,改善患者临床症状,提升其生活质量水平,值得临床推广应用。

缺氧诱导因子1α基因修饰牙髓干细胞的体外成骨及成血管分化

背景:如何有效地修复和重建临界骨缺损是临床医学的难题。牙髓干细胞作为转基因靶细胞修复受损组织或器官具有一定优势。目的:探究缺氧诱导因子1α对牙髓干细胞体外成骨及成血管的作用。方法:选取临床上12-18岁患者因正畸治疗需要拔除的健康、完整双尖牙的牙髓组织分离、培养牙髓干细胞,实验组将缺氧诱导因子1α基因转染至牙髓干细胞,对照组不做任何处理,采用qPCR检测缺氧诱导因子1α的基因表达以及Western blot检测成骨及成血管因子的蛋白表达。结果与结论:(1)牙髓干细胞为短梭形的成纤维样细胞,细胞呈集落式生长。(2)缺氧诱导因子1α基因稳定转染至牙髓干细胞内,随着时间延长缺氧诱导因子1α基因表达逐渐增高,至14 d达到高峰,21 d明显降低。(3)与对照组相比,实验组的成血管相关因子表达水平更高。转染后1 d,血管内皮生成因子、基质细胞衍生因子1、促血管生成素2、胎盘生长因子蛋白表达明显增高,4 d达到高峰,随后实验组成血管相关蛋白表达虽略有降低,但与对照组相比明显增高。(4)与对照组相比,实验组的成骨相关因子表达水平更高。转染后1 d,人骨形态发生蛋白2、骨钙蛋白、Runt相关转录因子2蛋白表达明显增高,4 d达到高峰。(5)结果表明,缺氧诱导因子1α能够促进牙髓干细胞成骨及成血管因子的表达。