Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line

Qiang Fu; Zhen-zhou Jiang; Lu-yong Zhang

doi:10.1007/s11655-012-1265-x

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Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line

Updated：2021-08-27

- Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line
- Chinese Journal of Integrative Medicine Vol. 19, Issue 9, Pages: 683-688(2013)
- Affiliations：
  
  1. Jiangsu Center for Drug Screening, China Pharmaceutical University,Nanjing,China
  2. Department of Pharmacology for Chinese Materia Medica, China Pharmaceutical University,Nanjing,China
  3. Ministry of Education, Key Laboratory of Drug Quality Control and Pharmacovigilance China Pharmaceutical University,Nanjing,China
- Author bio：
- Funds：
  
  Supported by the Special Fund of Traditional Chinese Medicine for Public Interest Research from the Ministry of Finance of China (No. 200707008) and Mega-projects of Science Research for the 11th Five-Year Plan (No. 2009ZX09302-002)
- DOI：10.1007/s11655-012-1265-x
  CLC：
- Published：2013，
  
  Published Online：10 April 2013，
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Fu, Q., Jiang, Zz. & Zhang, Ly. Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line., Chin. J. Integr. Med. 19, 683–688 (2013). https://doi.org/10.1007/s11655-012-1265-x

Qiang Fu, Zhen-zhou Jiang, Lu-yong Zhang. Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line. [J]. Chinese Journal of Integrative Medicine 19(9):683-688(2013)
Fu, Q., Jiang, Zz. & Zhang, Ly. Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line., Chin. J. Integr. Med. 19, 683–688 (2013). https://doi.org/10.1007/s11655-012-1265-x DOI：

Qiang Fu, Zhen-zhou Jiang, Lu-yong Zhang. Impairment of triptolide on liver mitochondria in isolated liver mitochondria and HL7702 cell line. [J]. Chinese Journal of Integrative Medicine 19(9):683-688(2013) DOI： 10.1007/s11655-012-1265-x.

摘要

To observe the impairing effects of triptolide on liver mitochondria in isolated rat-liver mitochondria and human normal liver HL7702 cell line. Rat-liver mitochondria were isolated from adult female Sprague-Dawley (SD) rats. Liver mitochondria were incubated with 0

1.25

2.5

5 and 10 μmol/L triptolide for detecting mitochondrial swelling and with 0

2.5

5 and 10 μmol/L triptolide for mitochondrial permeability transition pore (MPTP) activity. Mitochondrial swelling was estimated by measuring the apparent absorbance change during 600 s in the mitochondrial suspensions at 520 nm with a mitochondrial swelling examining kit. The effect of triptolide on MPTP was determined with a fluorescence detection kit by detecting the fluorescence intensity at an excitation wavelength of 488 nm emitted at 527 nm. Human normal liver HL7702 cells were treated without or with 0.02

0.1 and 0.5 μmol/L triptolide for 24 h for analyzing mitochondrial transmembrane potential (Δψm) and reactive oxygen species (ROS). Δψm was measured using the fluorescent probe 5

5′

6′-tetrachloro-1

1′

3′-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). ROS was measured using fluorescent probe 2′

7′-dichlorofluorescin diacetate (DCFH-DA). The cells were harvested and dyed with JC-1 and DCFH-DA

and analyzed by flow cytometry

respectively. Incubation of isolated mitochondria with triptolide results in swollen mitochondria in a concentration-dependent manner. Moreover

triptolide significantly activated mitochondrial permeability transition at 5 and 10 μmol/L (P<0.05 and P<0.01). When HL7702 cells were exposed to a various concentration triptolide for 24 h

mitochondrial membrane depolarization and increase of ROS were caused by triptolide in a concentration-dependent manner. Triptolide significantly induced the mitochondrial membrane depolarization at 0.1 and 0.5 μmol/L (P<0.05 and P<0.01) and the increase of ROS at 0.1 and 0.5 μmol/L (P<0.05 and P<0.01). Triptolide could induce mitochondrial impairment

which may be one of the mechanisms by which hepatotoxicity occurs.

Abstract

1.25

2.5

5 and 10 μmol/L triptolide for detecting mitochondrial swelling and with 0

2.5

0.1 and 0.5 μmol/L triptolide for 24 h for analyzing mitochondrial transmembrane potential (Δψm) and reactive oxygen species (ROS). Δψm was measured using the fluorescent probe 5

5′

6′-tetrachloro-1

1′

3′-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). ROS was measured using fluorescent probe 2′

7′-dichlorofluorescin diacetate (DCFH-DA). The cells were harvested and dyed with JC-1 and DCFH-DA

and analyzed by flow cytometry

respectively. Incubation of isolated mitochondria with triptolide results in swollen mitochondria in a concentration-dependent manner. Moreover

triptolide significantly activated mitochondrial permeability transition at 5 and 10 μmol/L (P<0.05 and P<0.01). When HL7702 cells were exposed to a various concentration triptolide for 24 h

which may be one of the mechanisms by which hepatotoxicity occurs.

关键词

Triptolidemitochondriahepatotoxicity

Keywords

Triptolidemitochondriahepatotoxicity

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