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ObjectiveTo investigate the effect of Chaihu Guizhitang on triple-negative breast cancer （TNBC） cells based on hypoxia-inducible factor-1α （HIF-1α）/vascular endothelial growth factor A （VEGFA） signaling pathway. MethodTNBC xenograft model was established and the cells were randomized into model group， capecitabine group （0.2 mg·kg-1）， Chaihu Guizhitang low-dose group， medium-dose group， and high-dose group （10.62， 21.23， 42.46 g·kg-1）， with 10 mice in each group. After 21 days of medication， the content of tumor necrosis factor-α （TNF-α） in serum was detected by enzyme-linked immunosorbent assay （ELISA）. The expression of HIF-1α mRNA was detected by real-time fluorogenic quantitative polymerase chain reaction （real-time PCR）. Immunohistochemistry （IHC） was employed to detect the expression of HIF-1α， TNF-α， and VEGFA in tumor tissues， and CD34 staining to examine the angiogenesis in tumor tissues. Microvessel density （MVD） was calculated， and the protein expression of HIF-1α， VEGFA， and epidermal growth factor receptor （EGFR） in tumor tissues was measured by Western blot. ResultCompared with the model group， the rest four groups showed low levels of TNF-α （P<0.01）， HIF-1α mRNA （P<0.01）， expression of HIF-1α， TNF-α， VEGFA， and CD34 in cells， and MVD （P<0.05， P<0.01）， and low protein levels of HIF-1α， VEGFA， and EGFR （P<0.01）. Compared with capecitabine group， medium-dose and high-dose Chaihu Guizhitang decreased the level of TNF-α （P<0.01）， HIF-1α mRNA （P<0.01）， expression of HIF-1α， TNF-α， and VEGFA in cells （P<0.01）， CD34 expression， MVD， and protein levels of HIF-1α， VEGFA， and EGFR （P<0.01）. ConclusionChaihu Guizhitang may inhibit the angiogenesis in TNBC cells by regulating the expression of HIF-1α/VEGFA signaling pathway， thus exerting anti-tumor effect.
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AIM:To investigate the mechanism of curcumin inhibiting the choroidal neovascularization(CNV)of brown Norway(BN)rats.METHODS: CNV model of 36 BN rats was established through laser photocoagulation induction, and they were divided into 6 groups with 6 rats in each group. Normal group was fed normally with no intervention, while 532nm laser photocoagulation was used to establish a experimental CNV model in BN rats. Rats after modeling were respectively intervened for 14d and divided into model group, ranibizumab group, curcumin low [100mg/(kg·d)], medium [200mg/(kg·d)], and high [400mg/(kg·d)] dose group. The model group was given intragastric administration of saline for 14d, ranibizumab(10mg/mL, 0.2mL/dose)was injected at 2d after photocoagulation with 5μL once for rats in ranibizumab group, and different concentrations of curcumin were intragastrically administrated to the rats in low, medium and high groups for 14d. Fundus photography, fundus fluorescein angiography(FFA)and indocyanine green angiography(ICGA)examination were performed at 14d after photocoagulation. Ocular histopathological specimens of rats with CNV were made, and the central thickness of CNV were observed by HE staining. Ocular histopathological specimens were made, and the expressions of AKT/p-AKT/HIF-1α/VEGF signaling pathway-related proteins were observed by immunohistochemistry. The mRNA relative expressions of AKT/HIF-1α/VEGF factor in CNV tissues were detected by RT-qPCR, and the protein expressions of AKT/p-AKT/HIF-1α/VEGF factor in CNV tissues were detected by Western-blot.RESULTS: CNV generation rates in the model group, the ranibizumab group, and the low, medium and high-dose curcumin groups were 78.18%, 73.21%, 77.19%, 75.86%, 74.55%, respectively, which were higher than 70%. The average absorbance were 182.12±6.59, 119.22±8.03, 166.45±8.33, 164.34±5.69, 149.22±6.45, respectively; the ranibizumab group was significantly lower than the model group(P&#x003C;0.05); the low-dose, medium-dose and high-dose groups were significantly higher than the ranibizumab group(P&#x003C;0.05), and the curcumin high-dose group was significantly lower than the model group(P&#x003C;0.05). HE staining showed that the retinal tissue structure of BN rats in normal group was clear and neatly arranged. The central thickness of CNV in the ranibizumab group was significantly reduced at 14d after photocoagulation compared with the model group(P&#x003C;0.05); While the curcumin high-dose group was significantly reduced compared with the model group(P&#x003C;0.05), but increased when compared with ranibizumab group(P&#x003C;0.05). Immunohistochemistry results showed that AKT, p-AKT, HIF-1α, and VEGF factors were negatively expressed in the retinal tissue structure of BN rats in the normal group, and no brown-yellow reactants were found. The expression of AKT, p-AKT, HIF-1α, and VEGF factors in the model group were higher than that in the normal group at 14d after photocoagulation(P&#x003C;0.05); the ranibizumab group was lower than the model group(P&#x003C;0.05). While the expression of the curcumin high-dose group was significantly decreased compared with the model group(P&#x003C;0.05), but significantly increased when compared with ranibizumab group(P&#x003C;0.05). The mRNA results showed that the relative expression levels of AKT, HIF-1α and VEGF mRNA in the model group at 14d after photocoagulation were higher than those of the normal group(P&#x003C;0.05); the ranibizumab group was lower than the model group(P&#x003C;0.05). While curcumin high-dose group was significantly decreased compared with the model group(P&#x003C;0.05), but significantly increased when compared with ranibizumab group(P&#x003C;0.05). Western-blot results showed that there was no significant difference in the relative expression of AKT protein among each experimental groups at 14d after photocoagulation. The relative expression of p-AKT protein in the model group was significantly higher than that in the normal group(P&#x003C;0.05); the ranibizumab group was significantly lower than the model group(P&#x003C;0.05); the curcumin high-dose group was significantly lower than the model group(P&#x003C;0.05). The relative expression levels of HIF-1α protein were significantly higher in the model group than in the normal group(P&#x003C;0.05), and the ranibizumab group was lower than in the model group(P&#x003C;0.05). The relative expression levels of HIF-1α protein was lower in the curcumin high-dose group than in the model group(P&#x003C;0.05)but higher than ranibizumab group(P&#x003C;0.05). The relative expression level of VEGF protein was significantly lower in the curcumin medium/high-dose group than in the model group(P&#x003C;0.05).CONCLUSION: Curcumin at 400mg/(kg·d)has an inhibitory effect on CNV in BN rats. The mechanism may be closely related to inhibiting the activation of AKT/p-AKT/HIF-1α/VEGF signaling pathways.
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ObjectiveTo explore the protective effect of Xielitang on ulcerative colitis （UC） mice induced by dextran sodium sulfate （DSS） and its possible mechanism. MethodSixty C57BL/6 mice were randomly divided into normal group， model group， sulfasalazine group and and low-， medium-， and high-dose Xielitang groups. Free drinking DSS solution to build the chronic UC model mice. Except for normal group， other groups were given 1.5% DSS for 3 cycles of drinking （days 1-7， days 22-28 and days 43-49） and distilled water for the rest of the time （days 8-21， days 29-42 and days 50-63）. After the first cycle， corresponding drugs were given for 42 days. The changes of general condition， body weight and disease activity index （DAI） score of mice were daily recorded during the experiment. At the end of the treatment， serum and colon tissue samples were collected， colon length was measured， intestinal weight index and colonic mucosal injury （CMDI） score were calculated. The pathological status of colon tissue was observed by hematoxylin-eosin （HE） staining. The levels of interleukin-6 （IL-6）， interleukin-10 （IL-10） and tumour necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. The gene and protein expressions of Toll like receptor 4 （TLR4）， nuclear transcription factor-κB （NF-κB） and hypoxia inducible factor-1α （HIF-1α） in colon tissue was detected by Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the normal group， the body weight， colon length and IL-10 content in the model group were significantly decreased （P<0.01）， DAI score， intestinal weight index， CMDI score， IL-6 and TNF-α contents， and mRNA and protein expression levels of TLR4， NF-κB and HIF-1α in the model group were significantly increased （P<0.01）. Moreover， the structure of colonic mucosa was destroyed and inflammatory cells infiltrated in the model group. Compared with model group， body weight， colon length and IL-10 content in each dose group of Xielitang were significantly increased （P<0.05， P<0.01）， DAI score， intestinal weight index and CMDI score， IL-6 and TNF-α contents， mRNA and protein expression levels of TLR4， NF-κB and HIF-1α were notably decreased （P<0.05， P<0.01）. The pathological injury of colon was obviously alleviated. ConclusionXielitang can significantly improve the inflammatory response of UC mice induced by DSS， and its mechanism may be related to the regulation of TLR4/NF-κB/HIF-1α signaling pathway.
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Accumulating evidence has confirmed the links between transfer RNA (tRNA) modifications and tumor progression. The present study is the first to explore the role of tRNA methyltransferase 5 (TRMT5), which catalyzes the m1G37 modification of mitochondrial tRNAs in hepatocellular carcinoma (HCC) progression. Here, based on bioinformatics and clinical analyses, we identified that TRMT5 expression was upregulated in HCC, which correlated with poor prognosis. Silencing TRMT5 attenuated HCC proliferation and metastasis both in vivo and in vitro, which may be partially explained by declined extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Mechanistically, we discovered that knockdown of TRMT5 inactivated the hypoxia-inducible factor-1 (HIF-1) signaling pathway by preventing HIF-1α stability through the enhancement of cellular oxygen content. Moreover, our data indicated that inhibition of TRMT5 sensitized HCC to doxorubicin by adjusting HIF-1α. In conclusion, our study revealed that targeting TRMT5 could inhibit HCC progression and increase the susceptibility of tumor cells to chemotherapy drugs. Thus, TRMT5 might be a carcinogenesis candidate gene that could serve as a potential target for HCC therapy.
Subject(s)
Humans , Carcinoma, Hepatocellular/pathology , Cell Hypoxia , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Liver Neoplasms/pathology , Signal Transduction/genetics , tRNA Methyltransferases/metabolismABSTRACT
Hypoxia, as an important hallmark of the tumor microenvironment, is a major cause of oxidative stress and plays a central role in various malignant tumors, including glioblastoma. Elevated reactive oxygen species (ROS) in a hypoxic microenvironment promote glioblastoma progression; however, the underlying mechanism has not been clarified. Herein, we found that hypoxia promoted ROS production, and the proliferation, migration, and invasion of glioblastoma cells, while this promotion was restrained by ROS scavengers N-acetyl-L-cysteine (NAC) and diphenyleneiodonium chloride (DPI). Hypoxia-induced ROS activated hypoxia-inducible factor-1α (HIF-1α) signaling, which enhanced cell migration and invasion by epithelial-mesenchymal transition (EMT). Furthermore, the induction of serine protease inhibitor family E member 1 (SERPINE1) was ROS-dependent under hypoxia, and HIF-1α mediated SERPINE1 increase induced by ROS via binding to the SERPINE1 promoter region, thereby facilitating glioblastoma migration and invasion. Taken together, our data revealed that hypoxia-induced ROS reinforce the hypoxic adaptation of glioblastoma by driving the HIF-1α-SERPINE1 signaling pathway, and that targeting ROS may be a promising therapeutic strategy for glioblastoma.
Subject(s)
Humans , Cell Hypoxia , Cell Line, Tumor , Glioblastoma/pathology , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Plasminogen Activator Inhibitor 1/metabolism , Reactive Oxygen Species/metabolism , Signal Transduction , Tumor Microenvironment , Brain Neoplasms/pathologyABSTRACT
ObjectiveTo investigate the mechanism of Xumingtang in Gu Jin Lu Yan （《古今录验》） in regulating cell pyroptosis through the hypoxia-inducible factor-1α （HIF-1α）/NOD-like receptor pyrin domain-containing protein 3 （NLRP3） pathway in ischemic stroke （IS）. MethodSD rats were randomly divided into a sham operation group， a model group， low- and high-dose Xumingtang groups， and a metformin group， with 20 rats in each group. Oral administration was performed for 3 days， and tissue samples were collected. Differential messenger RNA （mRNA） was screened using high-throughput sequencing， and Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analyses were performed on key differentially expressed genes. The modified neurological severity score （mNSS） and 2，3，5-triphenyltetrazolium chloride （TTC） staining were used to evaluate the effect of brain infarction. Hematoxylin-eosin （HE） staining was used for pathological morphological observation of brain tissue. Enzyme-linked immunosorbent assay （ELISA） was used to compare the levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in the ischemic cortical region. Double staining immunohistochemistry was used to detect the co-localization of HIF-1α and NLRP3. Real-time quantitative polymerase chain reaction （PCR） was performed to detect the mRNA expression of NLRP3， HIF-1α， Caspase-1 （CASP-1）， and gasdermin D （GSDMD）. Western blot was used to detect the protein expression of HIF-1α， NLRP3， CASP-1， and GSDMD. ResultA total of 5 705 differentially expressed genes （2 733 downregulated and 2 972 upregulated） were obtained by mRNA sequencing. After conversion to homologous genes and intersection with the pyroptosis gene set， 95 key differentially expressed pyroptosis genes were obtained. Compared with the sham operation group， the model group showed significantly increased mNSS scores， larger brain infarction areas （P<0.01）， diverse neuronal morphology， disordered arrangement， widened cell gaps， significantly increased levels of IL-1β and IL-18 in the ischemic cortical region （P<0.01）， enhanced co-localization fluorescence intensity， and significantly increased mRNA and protein expression levels of HIF-1α， NLRP3， CASP-1， and GSDMD （P<0.01）. Compared with the model group， the high-dose Xumingtang group showed the most significant improvement in neurological function scores and brain infarction areas （P<0.01）. The neuronal integrity and arrangement were more complete， and the cell gaps were narrower in all groups with drug treatment， with significantly reduced co-localization fluorescence intensity. Xumingtang could reduce the levels of IL-1β， IL-18， and the mRNA and protein expression of HIF-1α， NLRP3， CASP-1， and GSDMD （P<0.05， P<0.01）， with the high-dose Xumingtang group showing the most significant effect （P<0.01）. ConclusionXumingtang in Gu Jin Lu Yan can inhibit cell pyroptosis and promote neurological function recovery after IS， which may be related to the inhibition of the HIF-1α/NLRP3 pathway.
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AIM: To examine the effects of salidroside on choroidal thickness, hypoxia-inducible factor-1α(HIF-1α), dopamine(DA)and its D1 receptor expression in guinea pigs with lens-induced myopia(LIM).METHODS: A total of 18 two-week-old guinea pigs were randomly divided into the normal control(NC)group, the LIM group, and the LIM + salidroside(LIM+SA)group, with 6 guinea pigs in each group. The guinea pigs in the NC group were fed normally and intragastrically administered with 2 mL/d saline; those in the LIM group wore a -5D lens in front of their right eyes to establish a myopia model, then they were intragastrically administered with 2 mL/d saline. Finally, those in the LIM+SA group wore glasses along with intragastric administration of 2 mL/d salidroside at a dose of 100 mg/kg. The refraction, axial length, and choroidal thickness of guinea pigs in each group were measured 4wk following the establishment of the model. In addition, the relative mRNA expression and protein content of HIF-1α in the choroid and retina of guinea pigs in each group were detected by real-time quantitative PCR(qPCR)and immunohistochemistry(IHC). Finally, the DA concentration and its D1 receptor expression were detected by enzyme-linked immunosorbent assay(ELISA)and Western blot.RESULTS: At 4wk after model establishment, guinea pigs of LIM group and LIM+SA group exhibited increased negative refraction of the right eye, prolonged axial length, and decreased choroidal thickness compared to the NC group. The relative mRNA expression and protein content of HIF-1α in the choroid and retina of the guinea pigs increased. The concentration of DA and the expression of its D1 receptor both decreased. Moreover, compared to the LIM group, the diopter of the right eye of guinea pigs in LIM+SA group significantly reduced, the axial length was shorter, the thickness of choroid increased, the relative mRNA expression and protein content of HIF-1α in the choroid and retina decreased and the concentration of DA and the expression of its D1 receptor both increased.CONCLUSION: Salidroside can delay myopia progression in myopic guinea pigs by affecting choroidal thickness and the expression of HIF-1α, DA and its D1 receptor.
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ObjectiveTo observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α （HIF-1α）/nuclear factor-κB （NF-κB）/NOD-like receptor hot protein domain related protein 3 （NLRP3） signaling pathway in a rat model of high altitude hypoxia. MethodSixty male SPF SD rats were randomly divided into blank group， model group， dexamethasone group （5 mg·kg-1）， and high， middle， and low-dose groups of Huangqi Baihe granules （4.1， 2.05， 1.025 g·kg-1）. Among them， each Chinese medicine group was administrated orally for continuously 14 d， once a day， and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group. On the 15th d， the model group， dexamethasone group， and high， middle， and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude， low pressure， and low oxygen environment in the animal low-pressure simulation cabin， and the exposure lasted for 3 d. Blood was collected from the abdominal aorta and serum was separated， and the brain tissue was taken after being killed. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in brain tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in rat serum. Western blot was used to detect HIF-1α， NLRP3， phosphorylated nuclear factor-κB （p-NF-κB）， NF-κB， desquamation D （GSDMD）， and cysteine aspartate-specitis protein-1（Caspase-1） in rats of each group. The mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 were detected by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultThe results of HE staining showed that as compared with the normal group， the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder， with different sizes. Compared with the model group， the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules were reduced. The results of ELISA showed that as compared with the normal group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the model group was significantly higher （P<0.01）. Compared with the model group， the content of TNF-α， IL-6， and IL-1β in the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly （P<0.05， P<0.01）. The results of Western blot showed that as compared with the normal group， the relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly higher （P<0.01）. As compared with the model group， the relative expressions of HIF-1α， NLRP3， p-NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased （P<0.05， P<0.01）. The relative protein expression levels of HIF-1α， NLRP3， p-NF-κB p65， and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly （P<0.01）， and the relative protein expression of HIF-1α in the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced （P<0.05）. The Real-time PCR analysis showed that as compared with the normal group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of the model group were significantly increased （P<0.01）. As compared with the model group， the mRNA expression levels of HIF-1α， NLRP3， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased （P<0.01）. The mRNA expression levels of HIF-1α， NF-κB p65， GSDMD， and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly （P<0.01）. The mRNA expression levels of HIF-1α， NLRP3， and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased （P<0.05， P<0.01）. ConclusionThe protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway.
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Metabolic reprogramming is a common phenomenon in cancer, with aerobic glycolysis being one of its important characteristics. Hypoxia-inducible factor-1α (HIF1Α) is thought to play an important role in aerobic glycolysis. Meanwhile, naringin is a natural flavanone glycoside derived from grapefruits and many other citrus fruits. In this work, we identified glycolytic genes related to HIF1Α by analyzing the colon cancer database. The analysis of extracellular acidification rate and cell function verified the regulatory effects of HIF1Α overexpression on glycolysis, and the proliferation and migration of colon cancer cells. Moreover, naringin was used as an inhibitor of colon cancer cells to illustrate its effect on HIF1Α function. The results showed that the HIF1Α and enolase 2 (ENO2) levels in colon cancer tissues were highly correlated, and their high expression indicated a poor prognosis for colon cancer patients. Mechanistically, HIF1Α directly binds to the DNA promoter region and upregulates the transcription of ENO2; ectopic expression of ENO2 increased aerobic glycolysis in colon cancer cells. Most importantly, we found that the appropriate concentration of naringin inhibited the transcriptional activity of HIF1Α, which in turn decreased aerobic glycolysis in colon cancer cells. Generally, naringin reduces glycolysis in colon cancer cells by reducing the transcriptional activity of HIF1Α and the proliferation and invasion of colon cancer cells. This study helps to elucidate the relationship between colon cancer progression and glucose metabolism, and demonstrates the efficacy of naringin in the treatment of colon cancer.
Subject(s)
Humans , Glycolysis , Colonic Neoplasms/metabolism , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Phosphopyruvate Hydratase/metabolism , Flavanones/pharmacology , Cell Line, Tumor , Databases, Genetic , Cell Proliferation/drug effects , Transfection , Warburg Effect, OncologicABSTRACT
ObjectiveTo predict the underlying mechanism of Bushen Huoxuetang in treating osteoporosis related to endocrine therapy in breast cancer by network pharmacology and to verify the results through in vitro cell model. MethodThe main effective components and targets of Bushen Huoxuetang were screened out through network pharmacology， and the targets of osteoporosis related to endocrine therapy in breast cancer were further obtained. The intersected targets were analyzed by Gene Ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. Kaplan Meier plotter was used to analyze the survival of crucial targets. Finally， the inhibitory activity against cell proliferation was evaluated by in vitro methye thiazolye telrazlium（MTT） assay. The key targets and pathways were verified by Western blot， and the mRNA expression of the key targets was evaluated by real-time polymerase chain reaction（Real-time PCR）. ResultA total of 716 active components and 249 key targets of Bushen Huoxuetang were obtained from network pharmacology. There were 135 common targets， among which protein kinase B（Akt）1 and hypoxia-inducible factor-1α （HIF-1α） were two key targets. Additionally， 531 biological processes， 62 cellular components， 162 molecular functions， and 145 signaling pathways including breast cancer and endocrine resistance were involved. The key targets were effectively enriched in phosphatidylinositol 3-kinases（PI3K）/Akt and HIF-1 signaling pathways. According to the MTT assay， the cell proliferation rate and cell motility of MCF-7 and T47D cells in the luminal A cell line were reduced by Bushen Huoxuetang treatment （22.5， 45， 90 g·L-1， and 45， 90， 180 g·L-1） for 48 h as compared with the blank group. As revealed by Western blot， MCF-7 cells were treated with Bushen Huoxuetang （0， 15， 60 g·L-1） for 48 h， and the relative expression of p-PI3K， PI3K， p-Akt， Akt， and HIF-1α was decreased in a dose-dependent manner as compared with the blank group （P<0.05， P<0.01）. Real-time PCR was used to detect the mRNA expression of the key target HIF-1α. The results showed that the mRNA expression of HIF-1α in MCF-7 cells was decreased with the increase in the dose （P<0.01）， and the change was in a concentration-dependent manner. ConclusionThe mechanism of Bushen Huoxuetang in the treatment of osteoporosis related to endocrine therapy in breast cancer may be related to the key targets including Akt1 and HIF-1α through the PI3K/Akt/HIF-1α signaling pathway.
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ObjectiveTo investigate the effect and mechanism of Biejiajian Wan on liver fibrosis by regulating the polarization of macrophages. MethodRaw264.7 cells were cultured in vitro by serum pharmacological method， and the hypoxia model of RAW264.7 cells was established by stimulating RAW264.7 cells with cobalt chloride （CoCl2）. The cells were randomly divided into blank group， CoCl2 hypoxia model group （200 mmol·L-1）， Biejiajian Wan low-dose group （200 mmol·L-1+0.55 g·kg-1 Fuzheng Quyu capsules）， medium-dose group （200 mmol·L-1+1.1 g·kg-1 Biejiajian Wan）， and high-dose group （200 mmol·L-1+2.2 g·kg-1 Biejiajian Wan） and Fuzheng Quyu capsule group （200 mmol·L-1+0.56 g·kg-1 Biejiajian Wan）. Cell proliferation was detected by cell counting kit-8 （CCK-8）， and the gene expression of hypoxia inducible factor-1α （HIF-1α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6） in macrophages was detected by real time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of macrophage polarization-related protein and HIF-1α/nuclear factor-kappa B （NF-κB） signaling pathway-related protein was tested by Western blot， and the distribution and expression of NF-κB signaling pathway-related protein and HIF-1α were determined by cell immunofluorescence. ResultCompared with the conditions in the blank group， the proliferation of RAW264.7 cells was inhibited after CoCl2 stimulation for 24 hours （P<0.05）， the mRNA expression of HIF-1α， IL-1β and IL-6 in the model group were increased （P<0.05）， the protein expression of HIF-1α and M1 macrophage phenotypic proteins IL-6 and tumor necrosis factor-α （TNF-α） was boosted while that of M2 macrophage phenotypic protein interleukin-10 （IL-10） was reduced （P<0.05）， the protein expression of NF-κB p65， phosphorylation （p）-NF-κB p65， phosphorylated NF-κB inhibits protein kinase α/β （p-IKKα/β） and phosphorylated NF-κB inhibits protein α (p-IκBα） was elevated （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was promoted. Compared with the conditions in the model group， after 24 hours of treatment with corresponding drug-containing serum， each treatment group promoted the proliferation of RAW264.7 cells （P<0.05）， the mRNA expression levels of HIF-1α， IL-1β and IL-6 in macrophages were reduced （P<0.05）， the protein expression of HIF-1α， IL-6 and TNF-α was decreased， while that of CD163 and IL-10 was increased （P<0.05）， the protein expression of NF-κB p65， p-NF-κB p65， p-IKKα/β and p-IκBα was lowered （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was inhibited. ConclusionBiejiajian Wan could modulate the polarization of macrophages， attenuate the injury of macrophage-associated inflammatory response under hypoxia， and thus delay the progression of liver fibrosis， which might be related to its regulation of HIF-1α/NF-κB signaling pathway.
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ObjectiveTo investigate the effect and mechanism of Biejiajian Wan on liver fibrosis by regulating the polarization of macrophages. MethodRaw264.7 cells were cultured in vitro by serum pharmacological method， and the hypoxia model of RAW264.7 cells was established by stimulating RAW264.7 cells with cobalt chloride （CoCl2）. The cells were randomly divided into blank group， CoCl2 hypoxia model group （200 mmol·L-1）， Biejiajian Wan low-dose group （200 mmol·L-1+0.55 g·kg-1 Fuzheng Quyu capsules）， medium-dose group （200 mmol·L-1+1.1 g·kg-1 Biejiajian Wan）， and high-dose group （200 mmol·L-1+2.2 g·kg-1 Biejiajian Wan） and Fuzheng Quyu capsule group （200 mmol·L-1+0.56 g·kg-1 Biejiajian Wan）. Cell proliferation was detected by cell counting kit-8 （CCK-8）， and the gene expression of hypoxia inducible factor-1α （HIF-1α）， interleukin-1β （IL-1β） and interleukin-6 （IL-6） in macrophages was detected by real time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of macrophage polarization-related protein and HIF-1α/nuclear factor-kappa B （NF-κB） signaling pathway-related protein was tested by Western blot， and the distribution and expression of NF-κB signaling pathway-related protein and HIF-1α were determined by cell immunofluorescence. ResultCompared with the conditions in the blank group， the proliferation of RAW264.7 cells was inhibited after CoCl2 stimulation for 24 hours （P<0.05）， the mRNA expression of HIF-1α， IL-1β and IL-6 in the model group were increased （P<0.05）， the protein expression of HIF-1α and M1 macrophage phenotypic proteins IL-6 and tumor necrosis factor-α （TNF-α） was boosted while that of M2 macrophage phenotypic protein interleukin-10 （IL-10） was reduced （P<0.05）， the protein expression of NF-κB p65， phosphorylation （p）-NF-κB p65， phosphorylated NF-κB inhibits protein kinase α/β （p-IKKα/β） and phosphorylated NF-κB inhibits protein α (p-IκBα） was elevated （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was promoted. Compared with the conditions in the model group， after 24 hours of treatment with corresponding drug-containing serum， each treatment group promoted the proliferation of RAW264.7 cells （P<0.05）， the mRNA expression levels of HIF-1α， IL-1β and IL-6 in macrophages were reduced （P<0.05）， the protein expression of HIF-1α， IL-6 and TNF-α was decreased， while that of CD163 and IL-10 was increased （P<0.05）， the protein expression of NF-κB p65， p-NF-κB p65， p-IKKα/β and p-IκBα was lowered （P<0.05）， the nuclear expression of HIF-1α and NF-κB p65 was inhibited. ConclusionBiejiajian Wan could modulate the polarization of macrophages， attenuate the injury of macrophage-associated inflammatory response under hypoxia， and thus delay the progression of liver fibrosis， which might be related to its regulation of HIF-1α/NF-κB signaling pathway.
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It has been revealed that hypoxia is dynamic in hypertrophic scars; therefore, we considered that it may have different effects on hypoxia-inducible factor-1α (HIF-1α) and p53 expression. Herein, we aimed to confirm the presence of a teeterboard-like conversion between HIF-1α and p53, which is correlated with scar formation and regression. Thus, we obtained samples of normal skin and hypertrophic scars to identify the differences in HIF-1α and autophagy using immunohistochemistry and transmission electron microscopy. In addition, we used moderate hypoxia in vitro to simulate the proliferative scar, and silenced HIF-1α or p53 gene expression or triggered overexpression to investigate the changes of HIF-1α and p53 expression, autophagy, apoptosis, and cell proliferation under this condition. HIF-1α, p53, and autophagy-related proteins were assayed using western blotting and immunofluorescence, whereas apoptosis was detected using flow cytometry analysis, and cell proliferation was detected using cell counting kit-8 (CCK-8) and 5-bromo-2'-deoxyuridine (BrdU) staining. Furthermore, immunoprecipitation was performed to verify the binding of HIF-1α and p53 to transcription cofactor p300. Our results demonstrated that, in scar tissue, HIF-1α expression increased in parallel with autophagosome formation. Under hypoxia, HIF-1α expression and autophagy were upregulated, whereas p53 expression and apoptosis were downregulated in vitro. HIF-1α knockdown downregulated autophagy, proliferation, and p300-bound HIF-1α, and upregulated p53 expression, apoptosis, and p300-bound p53. Meanwhile, p53 knockdown induced the opposite effects and enhanced HIF-1α, whereas p53 overexpression resulted in the same effects and reduced HIF-1α. Our results suggest a teeterboard-like conversion between HIF-1α and p53, which is linked with scar hyperplasia and regression.
Subject(s)
Humans , Apoptosis , Autophagy , Cell Hypoxia , Fibroblasts/metabolism , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Tumor Suppressor Protein p53/metabolismABSTRACT
Objective:To explore the effect of hypoxia-inducible factor （HIF）-1<italic>α</italic> on T helper 17 （Th17）/regulatory T cell （Treg） balance in ulcerative colitis and the intervention mechanism of Shaoyaotang. Method:Forty-eight SD rats were randomly divided into normal group （normal saline）， model group， mesalazine group （0.42 g·kg<sup>-1</sup>）， Shaoyaotang group （11.1 g·kg<sup>-1</sup>）， inhibitor group ［2-methoxyestradiol （2ME<sub>2</sub>）， 0.015 g·kg<sup>-1</sup>］， and Shaoyaotang+inhibitor group. The ulcerative colitis model was induced by 2，4，6-trinitrobenzene sulfonic acid （TNBS）. The rats in all groups received corresponding treatments for 7 d， and the general condition and disease activity index （DAI） were observed. Hematoxylin-eosin （HE） staining was used to observe histopathological changes of the colon. Enzyme-linked immunosorbent assay （ELISA） was employed to detect serum levels of interleukin （IL）-10， IL-17， and IL-23 in rats. Western blot was used to detect the expression levels of forkhead box protein 3 （FoxP3）， retinoic acid-related orphan receptor <italic>γ</italic>t （ROR<italic>γ</italic>t）， and HIF-1<italic>α</italic> proteins in the colon tissue. Result:Compared with the normal group， the model group showed elevated disease activity index （DAI） score and pathological score for intestinal mucosa （<italic>P</italic><0.01）， reduced serum IL-10 level （<italic>P</italic><0.01）， up-regulated IL-17 and IL-23 levels （<italic>P</italic><0.01）， increased ROR<italic>γ</italic>t and HIF-1<italic>α</italic> expression （<italic>P</italic><0.01）， and decreased FoxP3 protein expression （<italic>P</italic><0.01）. Compared with the model group， the Shaoyaotang group displayed diminished DAI score and pathological score for intestinal mucosa （<italic>P</italic><0.05， <italic>P</italic><0.01）， increased serum IL-10 level （<italic>P</italic><0.01）， decreased IL-17 and IL-23 levels （<italic>P</italic><0.01）， dwindled protein levels of ROR<italic>γ</italic>t and HIF-1<italic>α </italic>（<italic>P</italic><0.01）， and up-regulated expression of FoxP3 （<italic>P</italic><0.01）. Compared with the inhibitor group， the Shaoyaotang group and the Shaoyaotang+inhibitor group exhibited significant differences in the expression of ROR<italic>γ</italic>t， FoxP3， and HIF-1<italic>α</italic> proteins （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:Shaoyaotang could effectively treat ulcerative colitis， and the underlying mechanism of action might be related to the regulation of Th17/Treg rebalance by inhibiting HIF-1<italic>α</italic>.
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Objective:To investigate the intervention effect of modified Shengjiangsan on hypoxia-inducible factor-1<italic>α </italic>（HIF-1<italic>α</italic>）/nicotinamide adenine dinucleotide phosphate oxidase 4 （NOX4） signaling pathway in membranous nephropathy （MN） rats and to explore its mechanism to reduce oxidative stress and apoptosis in renal tissues. Method:Cationized bovine serum albumin （C-BSA） was injected into the tail vein of rats to replicate the MN model. Rats were randomly divided into a model group， a modified Shengjiangsan group， and a benazepril group after modeling， and administered by gavage once a day accordingly. At the end of the 4<sup>th</sup> week， the 24-h urine total protein （UTP）， urea nitrogen （BUN）， and serum creatinine （SCr） levels of each group were detected. Enzyme-linked immunosorbent assay（ELISA）was used to detect the levels of superoxide dismutase （SOD）， malondialdehyde （MDA）， and reactive oxygen species （ROS） in renal tissues of rats. In situ end labeling（TUNEL） staining was used to detect the cell apoptosis rate. The mRNA and protein expression levels of HIF-1<italic>α</italic> and NOX4 were detected by real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）and Western blot， respectively. The immunohistochemistry method was used to detect the protein expression levels of B-cell lymphomas -2 （Bcl-2）， B-cell lymphomas xl （Bcl-xl）， Bcl-2 associated X protein （Bax）， Bcl-2 cell death regulator antibody （Bim）. Result:Compared with the normal group， the model group showed increased UTP （<italic>P</italic><0.05）， decreased SOD， elevated MDA and ROS （<italic>P</italic><0.05）， up-regulated mRNA and protein expression of HIF-1<italic>α</italic> and NOX4 （<italic>P</italic><0.05）， enhanced protein expression of Bax and Bim， declining protein expression of Bcl-xl and Bcl-2 （<italic>P</italic><0.05）， and increased cell apoptosis in renal tissues. Compared with the model group， the modified Shengjiangsan group and the benazepril group displayed declining UTP （<italic>P</italic><0.05）， up-regulated SOD， decreased MDA and ROS （<italic>P</italic><0.05）， down-regulated mRNA and protein expression of HIF-1<italic>α</italic> and NOX4 （<italic>P</italic><0.05）， diminished protein expression of Bax and Bim， elevated protein expression of Bcl-xl and Bcl-2 （<italic>P</italic><0.05）， and reduced cell apoptosis in renal tissues （<italic>P</italic><0.05）. Conclusion:The protective effect of modified Shengjiangsan on the kidney is presumedly achieved by reducing the oxidative stress and apoptosis in renal tissues of MN rats via inhibiting the HIF-1<italic>α</italic>/NOX4 signaling pathway.
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Objective:To observe the effects of Scutellariae Radix-Hedyotidis Herba on the proliferation of human lung adenocarcinoma A549 cells and the expression of interleukin-6（IL-6）， phosphatidylinositol 3-kinase （PI3K），protein kinase B （Akt）， p-protein kinase B （p-Akt）， mechanistic target of rapamycin （mTOR）， hypoxia-inducible factor-1<italic>α </italic>（HIF-1<italic>α</italic>）， and Cyclin D<sub>1</sub> at the cellular level， and to explore their molecular mechanism. Method:Following the set-up of the blank group （complete medium）， low-， moderate-，and high-dose （20， 40， and 60 mg·L<sup>-1</sup>） Scutellariae Radix-Hedyotidis Herba groups， and low-， moderate-， and high-dose （5， 10， and 20 mg·L<sup>-1</sup>） cisplatin groups， the cell were treated with the corresponding drugs for 24， 48， and 72 h for detecting their viability by tetrazolium bromide （MTT） colorimetry. A549 cells were then divided into the blank group， Scutellariae Radix-Hedyotidis Herba group， cisplatin group， and combined medication group and intervened with the<sup> </sup>complete medium， 40 mg·L<sup>-1 </sup>Scutellariae Radix-Hedyotidis Herba， 10 mg·L<sup>-1</sup> cisplatin， and 40 mg·L<sup>-1 </sup>Scutellariae Radix-Hedyotidis Herba + 10 mg·L<sup>-1 </sup>cisplatin， respectively， for 24， 48 and 72 h， followed by the measurement of inhibitory effects against the proliferation of A549 cells in each experimental group. The level of IL-6 in cell culture supernatant was determined by enzyme-linked immunosorbent assay （ELISA） after 72 h. The mRNA expression levels of HIF-1<italic>α</italic> and Cyclin D<sub>1</sub> in each group were assayed by real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of PI3K， Akt， p-Akt， mTOR， HIF-1<italic>α</italic>， and Cyclin D<sub>1</sub> by Western blot. Result:After 24 h intervention， Scutellariae Radix-Hedyotidis Herba did not significantly inhibit the proliferation of A549 cells. However， 48 h later， the inhibitory effect in Scutellariae Radix-Hedyotidis Herba groups were significantly enhanced in comparison with that in the blank group （<italic>P</italic><0.05）， exhibiting a time-dependent response. After 72 h of action， no significant change was present in the inhibitory effect of each Scutellariae Radix-Hedyotidis Herba group， so the optimal concentration of Scutellariae Radix-Hedyotidis Herba was set at 40 mg·L<sup>-1</sup> for follow-up experiments. As demonstrated by the comparison with the blank group， cisplatin at each concentration inhibited the cell proliferation in a time-dependent manner （<italic>P<</italic>0.05）. Considering the cell survival rate， the best concentration of cisplatin was set at 10 mg·L<sup>-1</sup>. Compared with the blank group， Scutellariae Radix-Hedyotidis Herba combined with cisplatin remarkably inhibited the proliferation of A549 cells in a time-dependent manner （<italic>P<</italic>0.05）， and the differences between the combined medication group and the other two groups became more significant after 72 h of medication （<italic>P<</italic>0.01）. The IL-6 level in each experimental group， especially in the combined medication group， significantly declined in contrast to that in the blank group （<italic>P<</italic>0.01）. The mRNA expression levels of HIF-1<italic>α</italic> and Cyclin D<sub>1</sub> in all experimental groups were obviously lower than those in the blank group， with the most significant changes observed in the combined medication group （<italic>P</italic><0.05，<italic>P</italic><0.01）. The protein expression levels of PI3K， p-Akt， mTOR， HIF-1<italic>α</italic>， and Cyclin D<sub>1</sub> in each experimental group was significantly down-regulated（<italic>P</italic><0.05，<italic>P</italic><0.01）， and the levels in the combined medication group were even lower than those in the cisplatin group （<italic>P<</italic>0.01）. Conclusion:Scutellariae Radix-Hedyotidis Herba has an inhibitory effect on the proliferation of A549 cells， which may be related to its inhibition against the expression and secretion of IL-6/PI3K/Akt/mTOR-HIF-1<italic>α</italic> axis.
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Objective:To observe the effects of artesunate （ART） on experimental choroidal neovascularization （CNV） and the expression of related proteins， and to explore the underlying mechanism. Method:Eighty BN rats were randomly divided into five groups： a normal group， a model group， a conbercept group， and low- （10.08 mg·kg<sup>-1</sup>·d<sup>-1</sup>） and high-dose （20.16 mg·kg<sup>-1</sup>·d<sup>-1</sup>） ART groups， with 16 rats in each group. A CNV model was established with 532 nm laser in rats of the groups except for the normal group. The rats in each group were treated with corresponding drugs by gavage for 14 days. The normal group， the model group， and the conbercept group received 1% CMC-Na solution at the same volume， while the conbercept group received an intravitreal injection （5 μL） once. On days 7 and 14， fundus fluorescein angiography （FFA） was used to evaluate the fluorescein leakage （gray value） of CNV. Hematoxylin-eosin （HE） staining was adopted to observe the histopathological changes of CNV. Western blot was employed to detect the protein expression levels of hypoxia-inducible factor-1<italic>α</italic> （HIF-1<italic>α</italic>） and vascular endothelial growth factor （VEGF） in the retina and choroid. Result:FFA results showed that compared with the normal group， other groups showed increased gray value on days 7 and 14 （<italic>P</italic><0.01）. On day 7， the gray value of the high-dose ART group and the conbercept group decreased compared with that in the model group （<italic>P</italic><0.01）. On day 14， the gray value of the ART groups and the conbercept group decreased in varying degrees compared with that in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. HE results showed that compared with the normal group， the model group showed increased thickness of CNV on days 7 and 14 （<italic>P</italic><0.01）. Compared with the model group， the ART groups and the conbercept group displayed decreased thickness of CNV （<italic>P</italic><0.01）. Western blot results revealed that the expression of HIF-1<italic>α</italic> and VEGF in the model group increased in varying degrees on the days 7 and 14 compared with that in the normal group （<italic>P</italic><0.05， <italic>P</italic><0.01）， while compared with the model group， the ART groups and the conbercept group showed decreased expression （<italic>P</italic><0.01）. Conclusion:ART can inhibit experimental CNV by down-regulating the expression of HIF-1<italic>α</italic> and VEGF in the early stage of experimental CNV formation.
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Objective To explore the predictive value of serum hypoxia-inducible factor-1α (HIF-1α) and interleukin-6 (IL-6) at admission on short-time 6-month prognosis of patients with severe traumatic brain injury (sTBI). Methods Seventy-two sTBI patients with Glasgow coma score (GCS) 3-8 points in our hospital were selected from September 2016 to January 2018 and divided into the group with good prognosis and group with poor prognosis according to Glasgow outcome score (GOS) after injury 6 months. Serum HIF-1α and IL-6 at admission were detected by using ELISA. The levels of plasma biochemistry indexes, acute physiology and chronic health evaluationⅡ(APACHEⅡ) scores and GCS scores were evaluated. Univariable and Multivariable COX proportional hazards models were performed to analyze the risk factors for short-time prognosis of patients with sTBI. Receiver operating characteristic (ROC) curve was built to analyze the predictive value of APACHEⅡ scores, HIF-1α and IL-6 on short-time prognosis of patients with sTBI. Results After 6-month followed up, there were 33 patients with good prognosis and 39 patients with poor prognosis. There was statistical difference of the baseline values of ages, serum HIF-1α and IL-6 at admission, APACHEⅡscores and GCS scores, the interval from injury to admission, the size of traumatic brain injury between two groups (t=2.312,14.132,16.628,3.172,3.644,3.073,4.284, P＜0.05). The serum HIF-1α [HR (95%CI)=2.645 (1.710-4.679), P＜0.05] and IL-6 [HR(95%CI)=1.821(1.674-2.957), P＜0.05] at admission, APACHEⅡscores [HR(95%CI)=1.789(1.105-2.928), P＜0.05] and the size of traumatic brain injury [HR (95%CI)=6.256 (1.727-10.834), P＜0.05] were the independent influence factors of short-time 6m prognosis of sTBI patients. The area under ROC curve and Youden＇s index of HIF-1α, IL-6 and APACHEⅡscores at admission on prediction of prognosis of sTBI patients were 0.94 (95% CI: 0.81-0.99) and 0.85, which was higher than separate predictive value of HIF-1α, IL-6 and APACHEⅡ scores. Conclusion The present study demontrated that serum HIF-1α and IL-6 at admission may be the early sensitive predictors of short-time prognosis in sTBI patients.
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Objective: To screen the anti-atherosclerosis (AS) activity of the compounds by using THP-1-HIF-1α-HER-Luciferase high-throughput model, and to verify the anti-AS function of the effective compounds. Methods: THP-1-HIF-1α-HER-Luciferase cells were pretreated with different concentrations of compounds (1, 10, and 100 μg/mL) for 2 h, then cultured under hypoxia for 24 h. Luciferase activity of cells was detected and compounds with anti-AS activity were screened by luciferase activity evaluation. THP-1 and U937 cells were pretreated with effective compounds, and then induced for 24 h by oxidized low density lipoprotein (OX-LDL). The formation of foam cells was observed by oil red staining. The mRNA level of hypoxia-inducible factor 1α (HIF-1α) was detected by real-time quantitative PCR (qPCR). HIF-1α protein expression was detected by Western blotting. Anti-AS activity of effective compounds were evaluated. Results: Among the 200 compounds, 11 compounds could significantly inhibit the increase of luciferase activity in THP-1-HIF-1α-HER-Luciferase cells induced by hypoxia (all P<0.05), and compound numbered 14 (C14) had the most significant inhibitory effect. THP-1 and U937 cells formed foam cells induced for 24 h by OX-LDL. However, cells were pretreated with C14 for 2 h, which could significantly inhibit the formation of foam cells induced by OX-LDL. Cells were induced for 24 h by OX-LDL, which could significantly increase the expression of HIF-1α mRNA and protein (all P<0.05), while cells pretreated with C14 could significantly inhibit the increase of HIF-1α mRNA and protein expression in a gradient-dependent manner (all P<0.05). Conclusion: THP-1-HIF-1α-HER-Luciferase high-throughput model can be reliability used in screening of compounds with anti-AS activity. C14 has the good anti-AS activity characteristics.
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Objective • To study the effect of inhibitor of differentiation 1 (ID1) on ocular neovascularization. Methods • The oxygen-induced retinal neovascularization (OIR), laser-induced choroidal neovascularization (CNV) and over-expression of vascular endothelial growth factor (VEGF) (Rho-VEGF) transgenic mice were established. The localization and mRNA level of ID1 in retina of OIR mice and Rho-VEGF transgenic mice were determined by immunofluorescence staining and quantitative real-time PCR. Mice deficient in ID1 (ID1-/-) were used to induce retinal neovascularization in accordance with the above three models, and to compare the changes of ID1 on the number of retinal, subretinal and choroidal neovascularization areas. In order to explore the role ID1 in neovascularization, the numbers and areas of retinal, subretinal and choroidal neovascularization in the mice models with or without ID1 deficiency were compared. Its effect on the related factors, i.e. hypoxia-inducible factor-1α (HIF-1α), VEGF and vascular endothelial growth factor receptor 1/2 (VEGFR1/2) were also observed. Results • Mice deficient in ID1 showed a significant reduction in the area of neovascularization in these three models(P<0.05). Mice lacking ID1 showed reduced levels of HIF-1α, VEGF and VEGFR 1. Conclusion • ID1 promotes the expression of HIF-1α, VEGF and VEGFR1 in the retina and choroidal neovascularization during hypoxia and oxidative injury.