Mechanism of Yanghe Decoction against subcutaneous tumor in pulmonary metastasis from breast cancer through HIF-1α signaling pathway regulating glycolysis:based on network pharmacology and animal experiment / 中国中药杂志

This study aims to explore the mechanism of Yanghe Decoction(YHD) against subcutaneous tumor in pulmonary metastasis from breast cancer, which is expected to lay a basis for the treatment of breast carcinoma with YHD. The chemical components of medicinals in YHD, and the targets of the components were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and SwissTargetPrediction. The disease-related targets were searched from GeneCards and Online Mendelian Inheritance in Man(OMIM). Excel was employed to screen the common targets and plot the Venn diagram. The protein-protein interaction network was constructed. R language was used for Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. A total of 53 female SPF Bablc/6 mice were randomized into normal group(same volume of normal saline, ig), model group(same volume of normal saline, ig), and low-dose and high-dose YHD groups(YHD, ig, 30 days), with 8 mice in normal group and 15 mice in each of the other groups. Body weight and tumor size was measured every day. Curves for body weight variation and growth of tumor in situ were plotted. In the end, the subcutaneous tumor sample was collected and observed based on hematoxylin and eosin(HE) staining. The mRNA and protein levels of hypoxia inducible factor-1α(HIF-1α), pyruvate kinase M2(PKM2), lactate dehydrogenase A(LDHA), and glucose transporter type 1(GLUT1) were detected by PCR and Western blot. A total of 213 active components of YHD and 185 targets against the disease were screened out. The hypothesis that YHD may regulate glycolysis through HIF-1α signaling pathway to intervene in breast cancer was proposed. Animal experiment confirmed that the mRNA and protein levels of HIF-1α, PKM2, LDHA, and GLUT1 in the high-and low-dose YHD groups were lower than those in the model group. YHD has certain inhibitory effect on subcutaneous tumor in pulmonary metastasis from breast cancer in the early stage, which may intervene pulmonary metastasis from breast cancer by regulating glycolysis through HIF-1α signaling pathway.

Exploring the common mechanism of Yindan Xinnaotong soft capsule in the treatment of stroke and coronary heart disease through HIF1α-MMP9-mediated HIF1α signaling pathway / 药学学报

Coronary heart disease (CHD) and stroke are the most well-known cardiovascular diseases, which share many common pathological basis. Yindan Xinnaotong soft capsule (YDXNT) is a commonly used Chinese patent medicine in the treatment of stroke and CHD. However, its action of mechanism of co-treatment for stroke and CHD is still unclear. The aim of this study was to explore the common mechanism of YDXNT in co-treatment of CHD and stroke using network pharmacology, experimental verification and molecular docking. An integrated literature mining and databases of IPA, ETCM, HERB, Swiss Target Prediction, OMIM and GeneCards were used to screen and predict active ingredients and potential targets of YDXNT in co-treatment of CHD and stroke. The protein-protein interaction network, GO analysis and pathway analysis were analyzed by IPA software. The effect of YDXNT on core targets was verified by immunofluorescence. UPLC-QTOF/MS and molecular docking were used to screen and predict the main active constituents of YDXNT and their interactions with core targets. A total of 151 potential targets are predicted for YDXNT in co-treatment of CHD and stroke. Hypoxia-inducible factor-1α (HIF1α)-matrix metalloproteinase-9 (MMP9)-mediated HIF1α signaling pathway serves as one of the common mechanisms. YDXNT could reduce the increase of mitochondrial fluorescence intensity and the protein expression of HIF1α and MMP9 in HL-1 and HA induced by oxygen and glucose deprivation/reperfusion (OGD/R) in a dose-dependent manner. Baicalin may be the material basis for treating stroke and CHD with YDXNT. In conclusion, the HIF1α signaling pathway is one of the common key mechanisms of YDXNT in the co-treatment of stroke and CHD. The study provides support and basis for the in-depth scientific connotation of the traditional Chinese medicine theory of "same treatment to different diseases".

To Explore the Mechanism of Xintongtai Medicated Serum on Inhibiting Apoptosis of Rabbit Aortic Vascular Smooth Muscle Cell by Interfering PI3K/Akt/HIF-1α Signaling Pathway / 世界科学技术-中医药现代化

Objective Bioinformatics analysis combined with cell experiment to explore the effect of Xintongtai medicated serum on apoptosis of rabbit aortic vascular smooth muscle cells by down-regulating PI3K/Akt/HIF-1α.Methods ox-LDL was used to induce the apoptosis of VSMC so as to establish the atherosclerotic cell model.The CCK8 method was used to select the optimal concentration of Xintongtai medicated serum.The main chemical components of Xintongtai were collected by TCMSP,the information of active compounds was collected by PubChem,the targets of active compounds were predicted by SwissTargetPrediction,the targets of"atherosclerosis"and"apoptosis"were collected by genecards and disgenet,and the protein-protein interaction(PPI)network was constructed by string platform.David online analysis gene ontology(GO)enrichment analysis function and Kyoto Encyclopedia of genes and genomes(KEGG)enrichment analysis.VSMC was divided into blank serum group,model group,Xintongtai medicated serum group and phosphatidylinositol 3 kinase(PI3K)inhibitor(LY294002)group,and Xintongtai medicated serum+ LY294002 group.The apoptosis of VSMC was detected by TUNEL and flow cytometry,and the apoptosis rate was calculated.The mRNA expression of PI3K,Akt,HIF-1α,caspase-3,caspase-9 were determined by polymerase chain reaction(PCR).Protein expression of p-PI3K/PI3K,p-Akt/Akt,HIF-1α,cleaved caspase-3,cleaved caspase-9 were determined by Western blot.α-SMA(Contractive VSMC specific marker)Fluorescence quantification of VSMC was determined by cellular immunofluorescence.Results The optimal concentration selected by CCK-8 was 20%middle dose Xintongtai medicated serum.Compared with the blank group,the VSMC early apoptosis rate,late apoptosis rate and total apoptosis rate in model group were increased(P<0.01),the mRNA expression of PI3K,Akt,HIF-1α,caspase-3,caspase-9 of the model group were up-regulated(P<0.01),the protein expression of p-PI3K/PI3K,p-Akt/Akt,HIF-1α,cleaved caspase-3,cleaved caspase-9 of the model group were up-regulated(P<0.01).Compared with the model group,the VSMC early apoptosis rate,late apoptosis rate and total apoptosis rate in Xintongtai medicated serum group,LY294002 group,and Xintongtai medicated serum+LY294002 group decreased(P<0.01 or P<0.05),and the mRNA expression of PI3K,Akt,HIF-1α,caspase-3,caspase-9 of Xintongtai medicated serum group,LY294002 group,and Xintongtai medicated serum+LY294002 group were down regulated(P<0.01 or P<0.05),the protein expression of p-PI3K/PI3K,p-Akt/Akt,HIF-1α,cleaved caspase-3,cleaved caspase-9 of Xintongtai medicated serum group,LY294002 group,and Xintongtai medicated serum+LY294002 group were down regulated(P<0.01 or P<0.05),the α-SMA increased significantly(P<0.01 or P<0.05).Compared with the Xintongtai medicated serum group,the VSMC early apoptosis rate,late apoptosis rate and total apoptosis rate were no difference(P>0.05),the mRNA expression of PI3K,Akt,HIF-1α,caspase-3,caspase-9 were no difference(P>0.05),the protein expression of p-PI3K/PI3K,p-Akt/Akt,HIF-1α,cleaved caspase-3,cleaved caspase-9 were no difference(P>0.05),the α-SMA was no difference(P>0.05).Conclusion Xintongtai medicated serum may down regulate PI3K/Akt/HIF-1α Signal pathway and downstream apoptosis related factors to alleviate the ox-LDL induced VSMC apoptosis,so as to stabilize vulnerable arterial plaque.

Hypoxic three-dimensional culture microenvironment promotes proliferation of bone marrow mesenchymal stem cells through HIF-1α signaling pathway / 中国胸心血管外科临床杂志

@#Objective To investigate the effects of hypoxic three-dimensional culture microenvironment on the proliferation of bone marrow mesenchymal stem cells and its mechanism. Methods P5 generation mouse bone marrow mesenchymal stem cells and P (3HB-co-4HB) were co-cultured under normoxic three-dimensional (20%) and hypoxic three-dimensional microenvironment (4%) respectively. After 24 hours, the proliferation of the two groups was determined by CCK-8 method. The expression of HIF-1α gene was detected by real-time quantitative PCR after 12 hours. Western blotting was used to detect the expression of HIF-1α protein after 24 hours. Results After 24 hours, the CCK-8 method showed that the OD value of the hypoxia group was significantly higher than that of the normoxia group (0.455±0.027 vs. 0.352±0.090, n=12, P<0.05). After 12 hours of hypoxic culture, the expression level of HIF-1α mRNA in the hypoxia group was significantly higher than that in the normoxia group (P<0.05). Compared with the normoxia group (0.47± 0.05), the relative expression level of HIF-1α protein in the hypoxia group (0.63±0.06) significantly increased in the Western blotting after 24 hours (n=3, P<0.05). Conclusion The hypoxic three-dimensional microenvironment can promote the proliferation of bone marrow mesenchymal stem cells, which may be related to the activation of HIF-1α signaling pathway.

Electroacupuncture for myocardial ischemia injury in rats via AMPK-HDAC5-HIF-1α signaling / 中国针灸

OBJECTIVE@#To explore the impact of electroacupuncture (EA) on the AMPKα-HDAC5-HIF-1α signaling in the heart of the rats with myocardial ischemia (MI) via detecting the expressions of AMP-activated protein kinase α (AMPKα), histone deacetylase 5 (HDAC5), hypoxia inducible factor (HIF-1α) and vascular endothelial growth factor (VEGF).@*METHODS@#Thirty-six healthy male SD rats were randomized into a sham operation group (6 rats), a sham + EA group (6 rats), a model group (12 rats) and an EA group (12 rats). We ligated the left anterior descending artery (LAD) for MI model, and exposed the heart of rats after opening the chest without ligation for the rats in the sham operation gorup and the sham + EA group. On the 2nd day after LAD ligation, EA was applied at "Neiguan" (PC 6) with 2 Hz/15 Hz and 1.5-2 mA for 30 min in the EA group and sham+EA group, once a day for 4 days. The same fixation was used in the sham operation group and the model group, without EA. Myocardial infarction area was observed by TTC staining and serum cardiac troponin T (cTnT) was detected by radioimmunoassay. The expression of VEGF mRNA was detected by real time PCR. The protein expressions of AMPKα, HDAC5, HIF-1α and VEGF were detected by western blot.@*RESULTS@#Compared with the sham operation group, 4 days after LAD ligation, the myocardial infarction was obvious and the expression of serum cTnT increased in the model group (0.05). After EA for 4 days, the myocardial infarction area and cTnT expression decreased in the EA group (both <0.01); the VEGF mRNA and protein expressions and AMPKα, HDAC5, HIF-1α protein expressions increased (<0.05, <0.01).@*CONCLUSION@#EA could regulate the AMPKα-HDAC5-HIF-1α signaling in myocardial tissue, which may activate VEGF expression for angiogenesis signaling, reduce myocardial infarction area so as to achieve cardioprotective effect.