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Effect of total flavones of Clematis filamentosa Dunn on the oxLDL-induced injury of vascular smooth muscle cells by regulating miR-455-5p
Corresponding Author(s) : Qifang Liu
Cellular and Molecular Biology,
Vol. 66 No. 5: Issue 5
Abstract
This experiment was conducted to investigate whether total flavones of Clematis filamentosa Dunn affect the inflammatory response and apoptosis of vascular smooth muscle cells induced by oxidized low-density lipoprotein (oxLDL) by regulating microRNA-455-5p (miR-455-5p). 50 mg/mL oxLDL was performed to stimulate the injury of vascular smooth muscle cells, and the total flavones of Clematis filamentosa Dunn were added at concentrations of 75, 150, and 300 μg/mL. The expressions of inflammatory factors IL-1β and TNF-α were analyzed by ELISA, the apoptosis was evaluated by flow cytometry, the expression of Bcl-2 and Bax was determined by western blot, and the real-time fluorescence quantitative PCR (qRT-PCR) was applied to detect miR-455-5p expression. MiR-455-5p mimic was transfected into vascular smooth muscle cells and then induced injury with oxLDL; miR-455-5p inhibitor was transfected into vascular smooth muscle cells and treated with oxLDL and 300 μg/mL total flavones of Clematis filamentosa Dunn. The above methods were employed to investigate the inflammatory response and apoptosis of cells. The total flavones of Clematis filamentosa Dunn significantly inhibited the expression of IL-1β, TNF-α, apoptosis rate, Bax protein expression of oxLDL induced vascular smooth muscle cells, and remarkably promoted the expression of Bcl-2 protein and miR-455-5p, which all showed concentration dependence (p<0.05). Overexpression of miR-455-5p reduced IL-1β, TNF-α expression, apoptosis rate, Bax protein expression, and greatly increased Bcl-2 protein expression in oxLDL injured vascular smooth muscle cells (p<0.05). After interfering with the expression of miR-455-5p, the inhibitory effect of total flavones of Clematis filamentosa Dunn on the expression of IL-1β, TNF-α, apoptosis, Bax protein expression of oxLDL-induced vascular smooth muscle cells was reversed, and its promotion effect on Bcl-2 protein expression was also reversed. Total flavones of Clematis filamentosa Dunn can reduce oxLDL-induced vascular smooth muscle cell inflammation and inhibit its apoptosis. The mechanism of action is related to the up-regulation of miR-455-5p expression.
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- . Pi H, Wang Z, Liu M, Deng P, Yu Z, Zhou Z, Gao F. SCD1 activation impedes foam cell formation by inducing lipophagy in oxLDL"treated human vascular smooth muscle cells. J Cell Mol Med 2019; 23(8):5259-69.
- Zhang L, Cheng H, Yue Y, Li S, Zhang D, He R. H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL-stimulated vascular smooth muscle cells. J Biomed Sci 2018; 25(1):11.
- Durham AL, Speer MY, Scatena M, Giachelli CM, Shanahan CM. Role of smooth muscle cells in vascular calcification: implications in atherosclerosis and arterial stiffness. Cardiovasc Res 2018; 114(4):590-600.
- Li H, Pan Z, Chen Q, Yang Z, Zhang D. SMILR Aggravates the Progression of Atherosclerosis by Sponging miR-10b-3p to Regulate KLF5 Expression. Inflamm 2020; 1-4.
- Yang N, Bo L, Junfang Zh. Protective effect of total flavones of Clematis filamentosa Dunn on hypoxia/reoxygenation injury of primary cardiac myocytes in Suckling mice. Chin Arch Tradit Chin Med 2017; 35(7):1876-1878+1939.
- Gorur A, Celik A, Yildirim DD, Gundes A, Tamer L. Investigation of possible effects of microRNAs involved in regulation of lipid metabolism in the pathogenesis of atherosclerosis. Mol Biol Rep 2019; 46(1):909-20.
- Huang S, Tao W, Guo Z, Cao J, Huang X. Suppression of long noncoding RNA TTTY15 attenuates hypoxia-induced cardiomyocytes injury by targeting miR-455-5p. Gene 2019; 701:1-8.
- Torabi S, Tamaddon M, Asadolahi M, Shokri G, Tavakoli R, Tasharrofi N, Rezaei R, Tavakolpour V, Sazegar H, Kouhkan F. miR-455-5p downregulation promotes inflammation pathways in the relapse phase of relapsing-remitting multiple sclerosis disease. Immunogenet 2019; 71(2):87-95.
- Lu YH, Wang D, Jing HY. Effect and related mechanism on inflammatory response, proliferation and apoptosis of oxLDL-induced vascular smooth muscle cell of matrine. Chin J Immunol 2018; 34(4):537-43.
- Grootaert MO, Moulis M, Roth L, Martinet W, Vindis C, Bennett MR, De Meyer GR. Vascular smooth muscle cell death, autophagy and senescence in atherosclerosis. Cardiovasc Res 2018; 114(4):622-34.
- Moulis M, Grousset E, Faccini J, Richetin K, Thomas G, Vindis C. The Multifunctional Sorting Protein PACS-2 Controls Mitophagosome Formation in Human Vascular Smooth Muscle Cells through Mitochondria-ER Contact Sites. Cells 2019; 8(6):638.
- Shirai R, Sato K, Yamashita T, Yamaguchi M, Okano T, Watanabe"Kominato K, Watanabe R, Matsuyama TA, Ishibashi"Ueda H, Koba S, Kobayashi Y. Neopterin counters vascular inflammation and atherosclerosis. J Am Heart Assoc 2018; 7(3):e007359.
- Marchio P, Guerra-Ojeda S, Vila JM, Aldasoro M, Victor VM, Mauricio MD. Targeting early atherosclerosis: A focus on oxidative stress and inflammation. Oxid Med Cell Longev 2019.
- Arnett DK, Blumenthal RS, Albert MA, Buroker AB, Goldberger ZD, Hahn EJ, Himmelfarb CD, Khera A, Lloyd-Jones D, McEvoy JW, Michos ED. 2019 ACC/AHA guideline on the primary prevention of cardiovascular disease: executive summary: a report of the American College of Cardiology/American Heart Association Task Force on Clinical Practice Guidelines. J Am Coll Cardiol 2019; 74(10):1376-414.
- Yang N, Xinying Ch, Yanjun Y. Effects of total flavones of Clematis filamentosa Dunn against myocardial ischemia. Chin J Exp Tradit Med Formul 2014; 20(22):176-179.
- Haichao H, Yang N, Xiaohong Zh. Total flavones of Clematis filamentosa Dunn protects against H2O2-induced PC12 cell damage. Chin Tradit Pat Med 2015; 37(12):2585-2589.
- Jie Zh, Haichao H, Yang N. Inhibition of epirubicin myocardial toxicity by total flavones of Clematis filamentosa Dunn and its mechanism. J Chin Med 2017; 40(2):436-440.
- Nie Y, Ding L, Huang HC, Xu LK, Zhao J, Yang YJ. Effects of PI3K/Akt-eNOS signaling pathway on total flavones in Clematis filamentosa post-conditioning alleviated of myocardial ischemia-reperfusion injury in rats. Zhongguo Zhong Yao za zhi= Zhongguo Zhong Yao zazhi= Chin J Chin Mater Med 2018; 43(23):4692-7.
- Guo J, Zhang CD, An JX, Xiao YY, Shao S, Zhou NM, Dai DQ. Expression of miR"634 in gastric carcinoma and its effects on proliferation, migration, and invasion of gastric cancer cells. Cancer Med 2018; 7(3):776-87.
- Yao S, Tang B, Li G, Fan R, Cao F. miR-455 inhibits neuronal cell death by targeting TRAF3 in cerebral ischemic stroke. Neuropsychiatr Dis Treat 2016; 12:3083.
- . Chen P, Miao Y, Yan P, Wang XJ, Jiang C, Lei Y. MiR"455"5p ameliorates HG"induced apoptosis, oxidative stress and inflammatory via targeting SOCS3 in retinal pigment epithelial cells. J Cell Physiol 2019; 234(12):21915-24.
- Ghaheri M, Kahrizi D, Bahrami G, Mohammadi-Motlagh HR. Study of gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni under various mannitol concentrations. Mol Biol Rep 2019; 46(1): 7-16.
- Eruygur N, Ucar E, Akpulat HA, Shahsavari K, Safavi SM, Kahrizi D. In vitro antioxidant assessment, screening of enzyme inhibitory activities of methanol and water extracts and gene expression in Hypericum lydium. Mol Biol Rep 2019; 46(2): 2121-9.
- Esmaeili F, Ghaheri M, Kahrizi D, Mansouri M, Safavi SM, Ghorbani T, Muhammadi S, Rahmanian E, Vaziri S. Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 1-5.
- Akbari F, Arminian A, Kahrizi D, Fazeli A, Ghaheri M. Effect of nitrogen sources on gene expression of Stevia rebaudiana (Bertoni) under in vitro conditions. Cell Mol Biol, 64(2): 11-16.
- Ghaheri M, Adibrad E, Safavi SM, Kahrizi D, Soroush A, Muhammadi S, Ghorbani T, Sabzevari A, Ansarypour Z, Rahmanian E. Effects of life cycle and leaves location on gene expression and glycoside biosynthesis pathway in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 17-22.
- Kahrizi D, Ghaheri M, Yari Z, Yari K, Bahraminejad S. Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 23-27.
- Hashempoor S, Ghaheri M, Kahrizi D, Kazemi N, Muhammadi S, Safavi SM, Ghorbani T, Rahmanian E, Heshmatpanaah M. Effects of different concentrations of mannitol on gene expression in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 28-31.
- Akbarabadi A, Ismaili A, Kahrizi D, Firouzabadi FN. Validation of expression stability of reference genes in response to herbicide stress in wild oat (Avena ludoviciana). Cell Mol Biol 2018; 64(4). 113-118.
- Ghorbani T, Kahrizi D, Saeidi M, Arji I. Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression. Cell Mol Biol, 63(11): 32-36.
- Kahrizi D, Ghari SM, Ghaheri M, Fallah F, Ghorbani T, Beheshti AA, Kazemi E, Ansarypour Z. Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol, 63(7): 107-111.
- Fallah F, Nokhasi F, Ghaheri M, Kahrizi D, Beheshti Ale AA, Ghorbani T, Kazemi E, Ansarypour Z. Effect of salinity on gene expression, morphological and biochemical characteristics of Stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol 2017; 63(7): 102-106.
- Bordbar M, Darvishzadeh R, Pazhouhandeh M, Kahrizi D. An overview of genome editing methods based on endonucleases. Mod Genet J 2020; 15(2): 75-92.
References
. Pi H, Wang Z, Liu M, Deng P, Yu Z, Zhou Z, Gao F. SCD1 activation impedes foam cell formation by inducing lipophagy in oxLDL"treated human vascular smooth muscle cells. J Cell Mol Med 2019; 23(8):5259-69.
Zhang L, Cheng H, Yue Y, Li S, Zhang D, He R. H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL-stimulated vascular smooth muscle cells. J Biomed Sci 2018; 25(1):11.
Durham AL, Speer MY, Scatena M, Giachelli CM, Shanahan CM. Role of smooth muscle cells in vascular calcification: implications in atherosclerosis and arterial stiffness. Cardiovasc Res 2018; 114(4):590-600.
Li H, Pan Z, Chen Q, Yang Z, Zhang D. SMILR Aggravates the Progression of Atherosclerosis by Sponging miR-10b-3p to Regulate KLF5 Expression. Inflamm 2020; 1-4.
Yang N, Bo L, Junfang Zh. Protective effect of total flavones of Clematis filamentosa Dunn on hypoxia/reoxygenation injury of primary cardiac myocytes in Suckling mice. Chin Arch Tradit Chin Med 2017; 35(7):1876-1878+1939.
Gorur A, Celik A, Yildirim DD, Gundes A, Tamer L. Investigation of possible effects of microRNAs involved in regulation of lipid metabolism in the pathogenesis of atherosclerosis. Mol Biol Rep 2019; 46(1):909-20.
Huang S, Tao W, Guo Z, Cao J, Huang X. Suppression of long noncoding RNA TTTY15 attenuates hypoxia-induced cardiomyocytes injury by targeting miR-455-5p. Gene 2019; 701:1-8.
Torabi S, Tamaddon M, Asadolahi M, Shokri G, Tavakoli R, Tasharrofi N, Rezaei R, Tavakolpour V, Sazegar H, Kouhkan F. miR-455-5p downregulation promotes inflammation pathways in the relapse phase of relapsing-remitting multiple sclerosis disease. Immunogenet 2019; 71(2):87-95.
Lu YH, Wang D, Jing HY. Effect and related mechanism on inflammatory response, proliferation and apoptosis of oxLDL-induced vascular smooth muscle cell of matrine. Chin J Immunol 2018; 34(4):537-43.
Grootaert MO, Moulis M, Roth L, Martinet W, Vindis C, Bennett MR, De Meyer GR. Vascular smooth muscle cell death, autophagy and senescence in atherosclerosis. Cardiovasc Res 2018; 114(4):622-34.
Moulis M, Grousset E, Faccini J, Richetin K, Thomas G, Vindis C. The Multifunctional Sorting Protein PACS-2 Controls Mitophagosome Formation in Human Vascular Smooth Muscle Cells through Mitochondria-ER Contact Sites. Cells 2019; 8(6):638.
Shirai R, Sato K, Yamashita T, Yamaguchi M, Okano T, Watanabe"Kominato K, Watanabe R, Matsuyama TA, Ishibashi"Ueda H, Koba S, Kobayashi Y. Neopterin counters vascular inflammation and atherosclerosis. J Am Heart Assoc 2018; 7(3):e007359.
Marchio P, Guerra-Ojeda S, Vila JM, Aldasoro M, Victor VM, Mauricio MD. Targeting early atherosclerosis: A focus on oxidative stress and inflammation. Oxid Med Cell Longev 2019.
Arnett DK, Blumenthal RS, Albert MA, Buroker AB, Goldberger ZD, Hahn EJ, Himmelfarb CD, Khera A, Lloyd-Jones D, McEvoy JW, Michos ED. 2019 ACC/AHA guideline on the primary prevention of cardiovascular disease: executive summary: a report of the American College of Cardiology/American Heart Association Task Force on Clinical Practice Guidelines. J Am Coll Cardiol 2019; 74(10):1376-414.
Yang N, Xinying Ch, Yanjun Y. Effects of total flavones of Clematis filamentosa Dunn against myocardial ischemia. Chin J Exp Tradit Med Formul 2014; 20(22):176-179.
Haichao H, Yang N, Xiaohong Zh. Total flavones of Clematis filamentosa Dunn protects against H2O2-induced PC12 cell damage. Chin Tradit Pat Med 2015; 37(12):2585-2589.
Jie Zh, Haichao H, Yang N. Inhibition of epirubicin myocardial toxicity by total flavones of Clematis filamentosa Dunn and its mechanism. J Chin Med 2017; 40(2):436-440.
Nie Y, Ding L, Huang HC, Xu LK, Zhao J, Yang YJ. Effects of PI3K/Akt-eNOS signaling pathway on total flavones in Clematis filamentosa post-conditioning alleviated of myocardial ischemia-reperfusion injury in rats. Zhongguo Zhong Yao za zhi= Zhongguo Zhong Yao zazhi= Chin J Chin Mater Med 2018; 43(23):4692-7.
Guo J, Zhang CD, An JX, Xiao YY, Shao S, Zhou NM, Dai DQ. Expression of miR"634 in gastric carcinoma and its effects on proliferation, migration, and invasion of gastric cancer cells. Cancer Med 2018; 7(3):776-87.
Yao S, Tang B, Li G, Fan R, Cao F. miR-455 inhibits neuronal cell death by targeting TRAF3 in cerebral ischemic stroke. Neuropsychiatr Dis Treat 2016; 12:3083.
. Chen P, Miao Y, Yan P, Wang XJ, Jiang C, Lei Y. MiR"455"5p ameliorates HG"induced apoptosis, oxidative stress and inflammatory via targeting SOCS3 in retinal pigment epithelial cells. J Cell Physiol 2019; 234(12):21915-24.
Ghaheri M, Kahrizi D, Bahrami G, Mohammadi-Motlagh HR. Study of gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni under various mannitol concentrations. Mol Biol Rep 2019; 46(1): 7-16.
Eruygur N, Ucar E, Akpulat HA, Shahsavari K, Safavi SM, Kahrizi D. In vitro antioxidant assessment, screening of enzyme inhibitory activities of methanol and water extracts and gene expression in Hypericum lydium. Mol Biol Rep 2019; 46(2): 2121-9.
Esmaeili F, Ghaheri M, Kahrizi D, Mansouri M, Safavi SM, Ghorbani T, Muhammadi S, Rahmanian E, Vaziri S. Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 1-5.
Akbari F, Arminian A, Kahrizi D, Fazeli A, Ghaheri M. Effect of nitrogen sources on gene expression of Stevia rebaudiana (Bertoni) under in vitro conditions. Cell Mol Biol, 64(2): 11-16.
Ghaheri M, Adibrad E, Safavi SM, Kahrizi D, Soroush A, Muhammadi S, Ghorbani T, Sabzevari A, Ansarypour Z, Rahmanian E. Effects of life cycle and leaves location on gene expression and glycoside biosynthesis pathway in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 17-22.
Kahrizi D, Ghaheri M, Yari Z, Yari K, Bahraminejad S. Investigation of different concentrations of MS media effects on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 23-27.
Hashempoor S, Ghaheri M, Kahrizi D, Kazemi N, Muhammadi S, Safavi SM, Ghorbani T, Rahmanian E, Heshmatpanaah M. Effects of different concentrations of mannitol on gene expression in Stevia rebaudiana Bertoni. Cell Mol Biol, 64(2): 28-31.
Akbarabadi A, Ismaili A, Kahrizi D, Firouzabadi FN. Validation of expression stability of reference genes in response to herbicide stress in wild oat (Avena ludoviciana). Cell Mol Biol 2018; 64(4). 113-118.
Ghorbani T, Kahrizi D, Saeidi M, Arji I. Effect of sucrose concentrations on Stevia rebaudiana Bertoni tissue culture and gene expression. Cell Mol Biol, 63(11): 32-36.
Kahrizi D, Ghari SM, Ghaheri M, Fallah F, Ghorbani T, Beheshti AA, Kazemi E, Ansarypour Z. Effect of KH2PO4 on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol, 63(7): 107-111.
Fallah F, Nokhasi F, Ghaheri M, Kahrizi D, Beheshti Ale AA, Ghorbani T, Kazemi E, Ansarypour Z. Effect of salinity on gene expression, morphological and biochemical characteristics of Stevia rebaudiana Bertoni under in vitro conditions. Cell Mol Biol 2017; 63(7): 102-106.
Bordbar M, Darvishzadeh R, Pazhouhandeh M, Kahrizi D. An overview of genome editing methods based on endonucleases. Mod Genet J 2020; 15(2): 75-92.