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Anti-fibrotic Effects and Mechanism of Shengmai Injection (生脉注射液) on Human Hepatic Stellate Cells LX-2

  • Yi Zhang
  • Li-tian Ma
  • Jie Li
  • Yu Qiao
  • Jun-ye Liu
  • Jin Wang
  • Qin-you Ren
  • Jin-tao Hu
  • Jin ZhengEmail author
Original Article
  • 14 Downloads

Abstract

Objective

To investigate the effects of Shengmai Injection (生脉注射液, SMI) on the proliferation, apoptosis and N-myc downstream-regulated gene 2 (NDRG2, a tumour suppressor gene) expression in varying densities of human hepatic stellate cells LX-2.

Methods

LX-2 cells were cultured in vitro. Then, cells were plated in 96-well plates at an approximate density of 2.5×104 cells/mL and cultured for 48, 72, 96 or 120 h followed by the application of different concentrations of SMI (0.6, 1.2, 2.4, 4.8 or 6 μL/mL). Cell proliferation was measured after an additional 24 or 48 h using the 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The effects of SMI on different cell growth states (cultured for 48, 72, 96, or 120 h) were observed by light microscopy at 24 h after treatment. When the cells reached 80% confluence, apoptosis was detected by flow cytometry after 24 h. Lastly, LX-2 cells were treated with different concentrations of SMI and extracted with protein lysis buffer. The levels of NDRG2 were measured by Western blot.

Results

When the LX-2 cells grew for 48, 72, 96 and 120 h, 4.8 and 6 μL/mL of SMI significantly inhibited cell proliferation at 24 and 48 h after treatment (P<0.05). And 2.4 μL/mL of SMI also inhibited cell proliferation at 24 h after treatment when cell growth for 48 h (P<0.05) and at 48 h after treatment when cell growth for 72, 96 and 120 h (P<0.05). The NDRG2 expression level in the LX-2 cell was significantly increased when treated with SMI at concentrations of 1.2, 2.4, 4.8 or 6 μL/mL (P<0.05).

Conclusions

The inhibitory effects of SMI on the proliferation of LX-2 cells were related to not only concentration dependent but also cell density. In addition, SMI (2.4, 4.8 and 6 μL/mL) could accelerate apoptosis in LX-2 cells, and the mechanism might be associated with NDRG2 over-expression.

Keywords

Shengmai Injection liver fibrosis N-myc downstream-regulated gene 2 LX-2 cell proliferation apoptosis Chinese medicine 

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Notes

Acknowledgement

We sincerely thank Prof. BIAN Hui-jie from the Fourth Military Medical University for donating LX-2 cell lines, and thank Dr. BAI Yang and Dr. ZHANG Ge from the Fourth Military Medical University for revising the manuscript. We also thank all of our colleagues for their generous support.

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Copyright information

© Chinese Association of the Integration of Traditional and Western Medicine 2018

Authors and Affiliations

  • Yi Zhang
    • 1
  • Li-tian Ma
    • 1
  • Jie Li
    • 2
  • Yu Qiao
    • 3
    • 4
  • Jun-ye Liu
    • 5
  • Jin Wang
    • 5
  • Qin-you Ren
    • 1
  • Jin-tao Hu
    • 6
  • Jin Zheng
    • 1
    Email author
  1. 1.Department of Traditional Chinese Medicine, Tangdu HospitalThe Fourth Military Medical UniversityXi’anChina
  2. 2.Department of EndocrinologyThe 986 Hospital of The People’s Liberation ArmyXi’anChina
  3. 3.Department of Anatomy and K.K. Leung Brain Research CenterThe Fourth Military Medical UniversityXi’anChina
  4. 4.Student BrigadeThe Fourth Military Medical UniversityXi’anChina
  5. 5.Department of Radiation MedicineThe Fourth Military Medical UniversityXi’anChina
  6. 6.Department of ImmunologyThe Fourth Military Medical UniversityXi’anChina

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