, Volume 41, Issue 4, pp 1229–1237 | Cite as

Geniposide Attenuates LPS-Induced Injury via Up-Regulation of miR-145 in H9c2 Cells

  • Qiang Su
  • Junjing Yao
  • Cunjian ShengEmail author


Myocarditis is a cardiomyopathy associated with inflammatory response. It has been reported that geniposide (GEN), a traditional Chinese herb extract from Gardenia jasminoides Ellis, possesses an anti-inflammatory effect and a protective effect on cardiomyocytes. The present study aimed to explore the protective role of GEN and the underlying mechanism in LPS-injured H9c2 cells. H9c2 cells were treated with LPS to induce cell injury and then we investigated the effect of GEN. miR-145 expression was inhibited by transfection with miR-145 inhibitor and its expression was measured by RT-PCR. Cell viability and apoptotic cells were measured by CCK-8 assay and flow cytometry analysis. The levels of pro-inflammatory factors (IL-6, TNF-α, and MCP-1) were assessed by western blot and RT-PCR. Western blot was performed to detect the expression of the MEK/ERK pathway-related factors. LPS exposure reduced cell viability, increased apoptotic cells, and promoted the expression of pro-inflammatory factors in H9c2 cells. However, GEN pretreatment significantly reduced LPS-induced cell injury, as increased cell viability, reduced apoptotic cells, and inhibited the expression of pro-inflammatory factors. Moreover, we found that miR-145 expression was down-regulated by LPS exposure but was up-regulated by GEN pretreatment. The protective effect of GEN on LPS-injured H9c2 cells was blocked by miR-145 inhibitor. In addition, GEN inhibited the MEK/ERK pathway through up-regulating miR-145. Our results suggested that GEN exerted a protective role in LPS-injured H9c2 cells. The GEN-associated regulation might be related to its regulation on miR-145 and the MEK/ERK signaling pathway.


geniposide (GEN) inflammation miR-145 MEK/ERK pathway cardiomyocytes 











American Type Culture Collection


Dulbecco’s modified Eagle’s medium


Fetal bovine serum


Phosphate buffered saline


Cell Counting Kit-8


Propidium iodide


Fluorescein isothiocynate


Sodium dodecyl sulfate-polyacrylamide gel electrophoresis


Polyvinylidene fluoride


Bovine serum albumin


Toll-like receptor 4


Myocardial infarction


  1. 1.
    Banerjee, Aditya, Aryadeep Roychoudhury, and S. Krishnamoorthi. 2016. Emerging techniques to decipher microRNAs (miRNAs) and their regulatory role in conferring abiotic stress tolerance of plants. Plant Biotechnology Reports 10 (4): 185–205.CrossRefGoogle Scholar
  2. 2.
    Basso, C., F. Calabrese, A. Angelini, E. Carturan, and G. Thiene. 2013. Classification and histological, immunohistochemical, and molecular diagnosis of inflammatory myocardial disease. Heart Failure Reviews 18 (6): 673–681.CrossRefPubMedGoogle Scholar
  3. 3.
    Chiyomaru, T., H. Enokida, S. Tatarano, K. Kawahara, Y. Uchida, K. Nishiyama, L. Fujimura, N. Kikkawa, N. Seki, and M. Nakagawa. 2010. miR-145 and miR-133a function as tumour suppressors and directly regulate FSCN1 expression in bladder cancer. British Journal of Cancer 102 (5): 883–891. Scholar
  4. 4.
    Cho, W.C., A.S. Chow, and J.S. Au. 2009. Restoration of tumour suppressor hsa-miR-145 inhibits cancer cell growth in lung adenocarcinoma patients with epidermal growth factor receptor mutation. European Journal of Cancer 45 (12): 2197–2206. Scholar
  5. 5.
    Coggins, M., and A. Rosenzweig. 2012. The fire within: cardiac inflammatory signaling in health and disease. Circulation Research 110 (1): 116–125.CrossRefPubMedGoogle Scholar
  6. 6.
    Dong, Maolong, Nan Hu, Yinan Hua, Xihui Xu, Machender R. Kandadi, Rui Guo, Shasha Jiang, Sreejayan Nair, Dahai Hu, and Jun Ren. 2013. Chronic Akt activation attenuated lipopolysaccharide-induced cardiac dysfunction via Akt/GSK3β-dependent inhibition of apoptosis and ER stress. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease 1832 (6): 848–863. Scholar
  7. 7.
    Higashi, K., Y. Yamada, S. Minatoguchi, S. Baba, M. Iwasa, H. Kanamori, M. Kawasaki, K. Nishigaki, G. Takemura, M. Kumazaki, Y. Akao, and S. Minatoguchi. 2015. MicroRNA-145 repairs infarcted myocardium by accelerating cardiomyocyte autophagy. American Journal of Physiology. Heart and Circulatory Physiology 309 (11): H1813–H1826. CrossRefPubMedGoogle Scholar
  8. 8.
    Hua, Fang, Tuanzhu Ha, Jing Ma, Yan Li, Jim Kelley, Xiang Gao, I. William Browder, Race L. Kao, David L. Williams, and Chuanfu Li. 2007. Protection against myocardial ischemia/reperfusion injury in TLR4-deficient mice is mediated through a phosphoinositide 3-kinase-dependent mechanism. The Journal of Immunology 178 (11): 7317–7324. Scholar
  9. 9.
    Jiang, Y.Q., G.L. Chang, Y. Wang, D.Y. Zhang, L. Cao, and J. Liu. 2016. Geniposide prevents hypoxia/reoxygenation-induced apoptosis in H9c2 cells: improvement of mitochondrial dysfunction and activation of GLP-1R and the PI3K/AKT signaling pathway. Cellular Physiology and Biochemistry 39 (1): 407–421. Scholar
  10. 10.
    Kano, M., N. Seki, N. Kikkawa, L. Fujimura, I. Hoshino, Y. Akutsu, T. Chiyomaru, H. Enokida, M. Nakagawa, and H. Matsubara. 2010. miR-145, miR-133a and miR-133b: tumor-suppressive miRNAs target FSCN1 in esophageal squamous cell carcinoma. International Journal of Cancer 127 (12): 2804–2814. Scholar
  11. 11.
    Li, D.W., J.P. Liu, Y.W. Mao, H. Xiang, J. Wang, W.Y. Ma, Z. Dong, H.M. Pike, R.E. Brown, and J.C. Reed. 2005. Calcium-activated RAF/MEK/ERK signaling pathway mediates p53-dependent apoptosis and is abrogated by alpha B-crystallin through inhibition of RAS activation. Molecular Biology of the Cell 16 (9): 4437–4453.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Li, F., W. Li, X. Li, F. Li, L. Zhang, B. Wang, G. Huang, X. Guo, L. Wan, Y. Liu, S. Zhang, S. Kang, and J. Ma. 2016. Geniposide attenuates inflammatory response by suppressing P2Y14 receptor and downstream ERK1/2 signaling pathway in oxygen and glucose deprivation-induced brain microvascular endothelial cells. Journal of Ethnopharmacology 185: 77–86. CrossRefPubMedGoogle Scholar
  13. 13.
    Li, R., G. Yan, Q. Li, H. Sun, Y. Hu, J. Sun, and B. Xu. 2012. MicroRNA-145 protects cardiomyocytes against hydrogen peroxide (H2O2)-induced apoptosis through targeting the mitochondria apoptotic pathway. PLoS One 7 (9): e44907.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Li, R., G. Yan, Q. Zhang, Y. Jiang, H. Sun, Y. Hu, J. Sun, and B. Xu. 2013. miR-145 inhibits isoproterenol-induced cardiomyocyte hypertrophy by targeting the expression and localization of GATA6. FEBS Letters 587 (12): 1754–1761. Scholar
  15. 15.
    Lin, Chien-Hung, and Chung-Ching Lin. 2016. Sitagliptin attenuates inflammatory responses in lipopolysaccharide-stimulated cardiomyocytes via nuclear factor-κB pathway inhibition. Experimental and Therapeutic Medicine 11 (6): 2609–2615. Scholar
  16. 16.
    Liu, J.H., F. Yin, L.X. Guo, X.H. Deng, and Y.H. Hu. 2009. Neuroprotection of geniposide against hydrogen peroxide induced PC12 cells injury: involvement of PI3 kinase signal pathway. Acta Pharmacologica Sinica 30 (2): 159–165. Scholar
  17. 17.
    Livak, K.J., and T.D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25 (4): 402–408.CrossRefPubMedGoogle Scholar
  18. 18.
    Maddahi, A., and L. Edvinsson. 2010. Cerebral ischemia induces microvascular pro-inflammatory cytokine expression via the MEK/ERK pathway. Journal of Neuroinflammation 7: 14. Scholar
  19. 19.
    Pan, Li-Long, Xin-Hua Liu, Qi-Hai Gong, and Yi-Zhun Zhu. 2011. S-Propargyl-cysteine (SPRC) attenuated lipopolysaccharide-induced inflammatory response in H9c2 cells involved in a hydrogen sulfide-dependent mechanism. Amino Acids 41 (1): 205–215. Scholar
  20. 20.
    Parthasarathy, G., and M.T. Philipp. 2014. The MEK/ERK pathway is the primary conduit for Borrelia burgdorferi-induced inflammation and P53-mediated apoptosis in oligodendrocytes. Apoptosis 19 (1): 76–89. Scholar
  21. 21.
    Peng, C.H., C.N. Huang, S.P. Hsu, and C.J. Wang. 2007. Penta-acetyl geniposide-induced apoptosis involving transcription of NGF/p75 via MAPK-mediated AP-1 activation in C6 glioma cells. Toxicology 238 (2–3): 130–139. Scholar
  22. 22.
    Pollack, A., A.R. Kontorovich, V. Fuster, and G.W. Dec. 2015. Viral myocarditis—diagnosis, treatment options, and current controversies. Nature Reviews Cardiology 12 (11): 670–680.CrossRefPubMedGoogle Scholar
  23. 23.
    Robinson, Emma, Roslyn S. Cassidy, Mitchel Tate, Youyou Zhao, Samuel Lockhart, Danielle Calderwood, Rachel Church, et al. 2015. Exendin-4 protects against post-myocardial infarction remodelling via specific actions on inflammation and the extracellular matrix. Basic Research in Cardiology 110 (2): 20. Scholar
  24. 24.
    Rong, Y.P., H.T. Huang, J.S. Liu, and L. Wei. 2017. Protective effects of geniposide on hepatic ischemia/reperfusion injury. Transplantation Proceedings 49 (6): 1455–1460. Scholar
  25. 25.
    Sachdeva, M., S. Zhu, F. Wu, H. Wu, V. Walia, S. Kumar, R. Elble, K. Watabe, and Y.Y. Mo. 2009. p53 represses c-Myc through induction of the tumor suppressor miR-145. Proceedings of the National Academy of Sciences of the United States of America 106 (9): 3207–3212.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Sandeep, Sagar, Peter P. Liu, and Leslie T. Cooper. 2012. Myocarditis. Lancet 379 (9817): 738–747.CrossRefGoogle Scholar
  27. 27.
    Song, X., W. Zhang, T. Wang, H. Jiang, Z. Zhang, Y. Fu, Z. Yang, Y. Cao, and N. Zhang. 2014. Geniposide plays an anti-inflammatory role via regulating TLR4 and downstream signaling pathways in lipopolysaccharide-induced mastitis in mice. Inflammation 37 (5): 1588–1598. Scholar
  28. 28.
    Wang, R., and H. Wu. 2017. Antiinflammation effects and mechanisms study of geniposide on rats with collagen-induced arthritis. 31 (4):631–637. doi: 10.1002/ptr.5775.Google Scholar
  29. 29.
    Wang, Z., X. Zhang, Z. Yang, H. Du, Z. Wu, J. Gong, J. Yan, and Q. Zheng. 2012. MiR-145 regulates PAK4 via the MAPK pathway and exhibits an antitumor effect in human colon cells. Biochemical & Biophysical Research Communications 427 (3): 444–449.CrossRefGoogle Scholar
  30. 30.
    Wu, S.Y., G.F. Wang, Z.Q. Liu, J.J. Rao, L. Lu, W. Xu, S.G. Wu, and J.J. Zhang. 2009. Effect of geniposide, a hypoglycemic glucoside, on hepatic regulating enzymes in diabetic mice induced by a high-fat diet and streptozotocin. Acta Pharmacologica Sinica 30 (2): 202–208. Scholar
  31. 31.
    Yong, L.U., Michael Chopp, Xuguang Zheng, Mark Katakowski, Benjamin Buller, and Feng Jiang. 2013. MiR-145 reduces ADAM17 expression and inhibits in vitro migration and invasion of glioma cells. Oncology Reports 29 (1): 67–72.CrossRefGoogle Scholar
  32. 32.
    Yuan, Ming, Liwei Zhang, Fei You, Jingyu Zhou, Yongjiang Ma, Feifei Yang, and Ling Tao. 2017. MiR-145-5p regulates hypoxia-induced inflammatory response and apoptosis in cardiomyocytes by targeting CD40. Molecular and Cellular Biochemistry 431 (1): 123–131. Scholar
  33. 33.
    Zhang, T., T. Yan, J. Du, S. Wang, and H. Yang. 2015. Apigenin attenuates heart injury in lipopolysaccharide-induced endotoxemic model by suppressing sphingosine kinase 1/sphingosine 1-phosphate signaling pathway. Chemico-Biological Interactions 233: 46–55. Scholar
  34. 34.
    Zhang, Z., Z. Wang, H. Ren, M. Yue, K. Huang, H. Gu, M. Liu, B. Du, and M. Qian. 2011. P2Y(6) agonist uridine 5′-diphosphate promotes host defense against bacterial infection via monocyte chemoattractant protein-1-mediated monocytes/macrophages recruitment. Journal of Immunology 186 (9): 5376–5387. Scholar
  35. 35.
    Zhou, H., Y. Yuan, Y. Liu, J. Ni, W. Deng, Z.Y. Bian, J. Dai, and Q.Z. Tang. 2015. Icariin protects H9c2 cardiomyocytes from lipopolysaccharide-induced injury via inhibition of the reactive oxygen species-dependent c-Jun N-terminal kinases/nuclear factor-κB pathway. Molecular Medicine Reports 11 (6): 4327–4332.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of EmergencyAffiliated Hospital of Jining Medical UniversityJiningChina
  2. 2.Department of Cardiac SurgeryJining No. 1 People’s HospitalJiningChina

Personalised recommendations