HIF-1α Was a Key Regulator to Improve hBMSCs to Secrete Vascular Endothelial Cytokine with Astragaloside in Hypoxia
American Journal of Clinical and Experimental Medicine
Volume 7, Issue 4, July 2019, Pages: 75-82
Received: Aug. 1, 2019;
Accepted: Sep. 6, 2019;
Published: Sep. 23, 2019
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Ji Hong Hu, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
Jin Hua, School of Chinese Medicine, Gansu University of Chinese Medicine, Lanzhou, China
Jia Jia, School of Chinese Medicine, Gansu University of Chinese Medicine, Lanzhou, China
Lu Juan, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
Jing Miao Zhao, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
Shu Xia Wang, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
Yun Wang, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
Li Mei Jin, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
Jin Juan Li, Center of Research and Experiment, Public Health School, Gansu University of Chinese Medicine, Lanzhou, China
This study aimed to assess the combined therapeutic efficacies with gene therapy and stem cell treatment and herb on the differentiation of hBMSCs into vascular endothelial cells under hypoxia condition. The third passage of hBMSCs were randomly divided into four groups, including control group (hBMSCs transfected with empty vectors), VEGF group (VEGF gene transfected to hBMSCs), AST group (cultured with AST), and VEGF + AST group (the intervention of VEGF group plus AST group). Each group was cultured at 37°C and 5% O2 for 2 weeks. Cell morphology was observed by inverted phase and were crowded and arranged irregularly in the control group and AST group, showing a fiber-like growth, while those in the VEGF group and VEGF plus AST group were mostly triangular or polygonal, exhibiting a colony-like growth, contrast microscope. CD31 was negative in the control group and AST group, while CD105 was positive in both groups, tested by flow cytometry assay. The positive rate of CD31 was significantly higher in the VEGF group than it in the VEGP + AST, and the positive rate of CD105 was lower in the VEGF group than it in the VEGF + AST group. The levels of VEGF and endothelial nitric oxide synthase (eNOS) by ELISA and the expression of endothelin and prostacyclin by west blot (WB) and RT-PCR were significantly higher in VEGF group and AST group and VEGF plus AST group, compared with control group. Further, the expression of endothelin and vWF and VEGFR-2 was highest in the VEGP + AST group and the expression of prostacyclin was lowest in VEGF group. AST can promote the secretion of VEGF from the differentiation of hBMSCs induced by VEGF gene transfected to hBMSCs by HIF-1α under hypoxia.
Ji Hong Hu,
Jing Miao Zhao,
Shu Xia Wang,
Li Mei Jin,
Jin Juan Li,
HIF-1α Was a Key Regulator to Improve hBMSCs to Secrete Vascular Endothelial Cytokine with Astragaloside in Hypoxia, American Journal of Clinical and Experimental Medicine.
Vol. 7, No. 4,
2019, pp. 75-82.
Hui Sui, Weiwei Chen, Wen Wang. "China cardiovascular disease report 2014". Chinese journal of hypertension. 2015; 23 (7): 627-629.
Chen X, Kata Kowskim, Li Y, et al. Human bone marrow stromal cell cultures conditioned by traumatic brain tissue extracts: growth factor production [J]. Neurosci Res. 2002; 69 (5): 687-691.
Tomita S, Mickle DA, Weisd RD, et al. Improved heart function withmyogenesis and angiogenesis after autologous porcine bone marrow stromalcell transplantation [J]. J Thora Cardiovasc Surg. 2002; 123 (6): 1132-1134.
Haiyan Feng, Ruifeng Liu, Kaiming Zhang, et al. In vitro isolation and culture of human bone marrow mesenchymal stem cells and differentiation into vascular endothelial cells [J]. Chinese journal of immunology. 2010; 26 (1): 70-74.
Jihong Hu, Jia Jia, Juan Lu, Qiuping Wang, Jingmiao Zhao. Differentiation of human bone marrow mesenchymal stem cells into vascular endothelioid cells transfected with vascular endothelial growth factor under hypoxia [J]. Tissue engineering research in China, 2017; 21 (9): 1352-1356.
DongSei, Shuqing Li, Donghua Li, et al. Effects of astragalus on improving cardiovascular function [J]. Chinese clinical rehabilitation. 2006; 10 (11): 21-23.
ZHANGJin-guo, GAO Dong-sheng, YANG Na, et al. Effect of astragalus injection on left ventricular remodeling and cardiac function in early morning patients with acute myocardial infarction [J]. Chinese Journal of Integrative Medicine. 2003; 9: 104-107.
LIU Yong-qi, LI Jing-ya, CAI Ling, et al. Differentiation of rat bone marrow mesenchymal stem cells induced by astragalus polysaccharide [J]. Chinese journal of traditional Chinese medicine. 2014; 21 (6): 60-64.
Shaoxian Xian, Zhongqi Yang, Zhaohui Wang, et al. In vitro induction of astragaloside iv into myocardial differentiation of bone marrow mesenchymal stem cells [J]. Journal of guangzhou university of traditional Chinese medicine. 2007; 24 (1): 37-40.
Lin Liu, Hai Hai, Cun bang Wang, et al. Effect of astragalus polysaccharide on proliferation and cytokine expression of bone marrow mesenchymal stem cells in patients with multiple myeloma [J]. Advances in modern biomedicine. 2013; 13 (10): 1873-1877.
Jihong Hu, Juan Lu, Yi Zhao, Limei Jin, Li Yan, Jia Jia, Qiuping Wang, Jingmiao Zhao. Effects of astragalus major active ingredients and extracts on proliferation of rat bone marrow mesenchymal stem cells [J]. Lishizhen Medicine and Materia Medica Research. 2016; 5: 1070-1072.
Muschler GF, Nakamoto C, Griffith LG. Engineering principles of clinical cell-based tissue engineering [J]. J Bone Joint Surg AM. 2004; 86: 1541-1558.
Junhua Song, Shuyun Wang. Effect of VEGF gene on ischemic heart disease and its influencing factors [J]. Journal of binzhou medical college. 2004; 27 (4): 281-284.
Xiaojuan Peng, Hai Bai, Cun-bang Wang, Rui Xi, et al. Effect of astragaloside iv on proliferation and cytokine expression of human bone marrow mesenchymal stem cells in vitro [J]. Advances in modern biology. 2014; 8: 1452-1455.
Xuelan LUO, Wei CHEN, Guojun MO, et al. The difference of eNOS expression/ activity and its metabolites in human bone marrow-derived mesenchymal stem cell-oriented vascular endothelial cells [J]. Journal of Xi'an Jiaotong University. 2017; (3): 386-391.
Irina Arutyunyan, TimurFatkhudinov, Evgeniya Kananykhina, et al. Role of VEGF-A in angiogenesis promoted by umbilical cord-derived mesenchymal stromal/stem cells: in vitro study [J]. Stem Cell Res Ther. 2016; 7.
Xiao Du, Xiaoping Chen, Kaiming Zhang, Haiyan Feng, Lei Wu. Experimental study on induced differentiation of human bone marrow mesenchymal stem cells into vascular endothelial cells in vitro. Chinese Journal of Integrative Medicine on Cardio-/Cerebrovascular Disease. 2009; 7 (8): 940-941.
Jin LI. Effect of heat shock culture on FGFRs and apoptosis-related proteins of bone marrow mesenchymal stem cells transfected with bFGF [J]. Health standard management in China. 2018; 1: 134-137.
Rong XU, Jinyong XU, Wei LIU. Experimental study on directional differentiation of Bone Marrow Mesenchymal Stem cells into Vascular Endothelial cells [J]. Journal of Biomedical Engineering. 2014; (2): 389-393.
YANG Li-Xin, XU Zhi-Yun, Zhang Bao-Ren, Huang Sheng-Dong, LIU Yan-Ling. In vitro induction of bone marrow stromal stem cells to differentiate into endothelial cells [J]. Acad J Secil Med Univ, 2003; 24 (12): 1300-1304.
Guan WANG, Jingya LI, Shengping YANG, Jianmin WANG. Induction of Astragalus Polysaccharide in differentiation of Bone Marrow Mesenchymal Stem cells into Neuron-like cells [J]. Research on Organization Engineering in China. 2015; (45) 7259-7262.
Wu JX, Xu BL, Huang WL. Advances in angiogenesis inhibitors [J]. Cancer. 2005; 24 (3): 376-384.
Bandara N, Gurusinghe S, Chen H, Chen S, Wang LX, Lim SY, Strappe P. Minicircle DNA-mediated endothelial nitric oxide synthase gene transfer enhances angiogenic responses of bone marrow-derived mesenchymal stemcells [J]. Stem Cell Res Ther. 2016; 7: 48doi: 10.1186/s13287-016-0307-2.
Baoming Zhao, Fengliang Li, Ximing Song, et al. Effects of glutathione on the secretion of prostaglandin and endothelin-1 in human umbilical vein endothelial cells in the glycochemical end-product environment [J]. Journal of Zhengzhou university (medical edition), 2011, 46 (4): 599-601.
Groop PH, Forsblom C, Thomas MC. Mechanisms of disease: Pathway-selective insulin resistance and microvascular complications of diabetes [J]. Nat Clin Pract Endocrinol Metab. 2005; 1 (2): 100.
Keliang Xie. Research progress of thrombosis and hemostasis test [J]. Medical information. 2014; 14: 655-656.
Shen ling. Intervention of astragalusmembranaceus on differentiation and migration of human bone marrow mesenchymal stem cells induced by vascular endothelial growth factor/stromal cell-derived factor-1 in hypoxic environment [D]. Beijing University of Chinese Medicine. 2010.
Argyris T, Paschalis N, Andreas K, et al. Expression of Hypoxia-Inducible Factor (HIF)-1a-Vascular Endothelial Growth Factor (VEGF)-Inhibitory Growth Factor (ING)-4- axis in sarcoidosis patients [J]. BMC Research Notes, 2012; 5 (1): 1-9.
Tao Y, Michael R, Soonpa M H, et al. Cardiac Engraftment of Genetically-Selected Parthenogenetic Stem Cell-Derived Cardiomyocytes [J]. PLOS ONE. 2015; 10 (6): e0131511.
Ikhapoh Attairu Izuagie, Christopher J Pelham, Devendra K Agrawal. Synergistic effect of angiotensin II on vascular endothelial growth factor-A-mediated differentiation of bone marrow-derived mesenchymal stem cells into endothelial cells [J]. Stem Cell Res Ther. 2015; 6.
Semenza GL. Hypoxia-inducible factors in physiology and medicine. Cell. 2012; 148 (3): 399–408.
Jingwen Si, Ning Wang, Huan Wang, Juan Xie, Jian Yang, Hui Yi, et al. HIF-1α signaling activation by post-ischemia treatment with Astragaloside IV attenuates myocardial ischemia-reperfusion injury. PLOS ONE. 2014; 9 (9): e107832.
Rissanen TT, Vajanto I, Hiltunen MO, et al. Expression of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 (KDR/Flk-1) in ischemic skeletal muscle and its regeneration. Am J Pathol. 2002; 160: 1393-403.
Tuomisto TT, Rissanen TT, Korkeela A, et al. HIF-VEGFVEGFR-2, TNF-a and IGF pathways are upregulated in critical human skeletal muscle ischemia as studied with DNA array. Atherosclerosis. 2004; 174: 111–20.