Abstract
Objective
To observe the impairing effects of triptolide on liver mitochondria in isolated rat-liver mitochondria and human normal liver HL7702 cell line.
Methods
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′,6,6′-tetrachloro-1,1′,3,3′-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.
Results
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).
Conclusion
Triptolide could induce mitochondrial impairment, which may be one of the mechanisms by which hepatotoxicity occurs.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Tao X, Davis LS, Lipsky PE. Effect of an extract of the Chinese herbal remedy Tripterygium wilfordii Hook F on human immune responsiveness. Arthrit Rheum 1991;34:1274–1281.
Tao X, Lipsky PE. The Chinese anti-inflammatory and immunosuppressive herbal remedy Tripterygium wilfordii Hook F. Rheum Dis Clin North Am 2000;26:29–50, viii.
Takaishi Y, Ujita K, Tokuda H, Nishino H, Iwashima A, Fujita T. Inhibitory effects of dihydroagarofuran sesquiterpenes on Epstein-Barr virus activation. Cancer Lett 1992;65:19–26.
Li XY. Anti-inflammatory and immunosuppressive components of Tripterygium wilfordii Hook F. Int J Immunother 1993;9:181–187.
Yao W-C, Nian H-F. Medicated wine of Tripterygium wilfordii in treating rheumatoid arthritis in 392 patients. Pharmaceut Clin Res (Chin) 2004;23:35–37.
Shamon LA, Pezzuto JM, Graves JM, Mehta RR, Wangcharoentrakul S, Sangsuwan R, et al. Evaluation of the mutagenic, cytotoxic, and antitumor potential of triptolide, a highly oxygenated diterpene isolated from Tripterygium wilfordii. Cancer Lett 1997;112:113–117.
Zheng J. Screening of active anti-inflammatory, immunosuppressive and antifertility components from Tripterygium wilfordii. IV. A comparison of the male antifertility activities of 7 diterpene lactone epoxide compounds. Acta Acad Med Sin (Chin) 1991;13:398–403.
Zheng J. Screening of active anti-inflammatory, immunosuppressive and antifertility components of Tripterygium wilfordii. III. A comparison of the antiinflammatory and immunosuppressive activities of 7 diterpene lactone epoxide compounds in vivo. Acta Acad Med Sin (Chin) 1991;13:391–397.
Tao X, Cai JJ, Lipsky PE. The identity of immunosuppressive components of the ethyl acetate extract and chloroform methanol extract (T2) of Tripterygium wilfordii Hook F. J Pharmacol Exp Ther 1995;272:1305–1312.
Tao X, Schulze-Koops H, Ma L, Cai J, Mao Y, Lipsky PE. Effects of Tripterygium wilfordii Hook F extracts on induction of cyclooxygenase 2 activity and prostaglandin E2 production. Arthrit Rheum 1998;41:130–138.
Duan H, Takaishi Y, Momota H, Ohmoto Y, Taki T, Jia Y, et al. Immunosuppressive sesquiterpene alkaloids from Tripterygium wilfordii. J Nat Prod 2001;64:582–587.
Duan H, Takaishi Y, Momota H, Ohmoto Y, Taki T, Tori M, et al. Immunosuppressive terpenoids from extracts of Tripterygium wilfordii. Tetrahedron 2001;57:8413–8424.
Brinker AM, Ma J, Lipsky PE, Raskin I. Medicinal chemistry and pharmacology of genus Tripterygium (Celastraceae). Phytochemistry 2007;68:732–766.
Song HX, Gong J, Chen W. Effect of triptolide on urinary monocyte chemottractant protein-1 in patients with diabetic nephropathy. Chin J Integr Tradit West Med (Chin) 2005;25:416–418.
Peng A, Gu Y, Lin SY. Herbal treatment for renal diseases. Ann Acad Med Singapore 2005;34:44–51.
Wu SX, Guo NR. Clinical observation on effect of triptolide tablet in treating patients with psoriasis vulgaris. Chin J Integr Med 2005;11:147–148.
Yao J, Jiang Z, Duan W, Huang J, Zhang L, Hu L, et al. Involvement of mitochondrial pathway in triptolide-induced cytotoxicity in human normal liver L-02 cells. Biol Pharm Bull 2008;31:592–597.
Shu B, Duan W, Yao J, Huang J, Jiang Z, Zhang L. Caspase 3 is involved in the apoptosis induced by triptolide in HK-2 cells. Toxicol In Vitro 2009;23:598–602.
Liu L, Jiang Z, Liu J, Huang X, Wang T, Zhang Y, et al. Sex differences in subacute toxicity and hepatic microsomal metabolism of triptolide in rats. Toxicology 2010;271:57–63.
Ni B, Jiang Z, Huang X, Xu F, Zhang R, Zhang Z, et al. Male reproductive toxicity and toxicokinetics of triptolide in rats. Arzneimittelforschung 2008;58:673–680.
Ni B, Zhu T, Jiang Z, Zhang R, Zhang T, Zhang L. In vitro and in silico approaches for analyzing the toxicological effect of triptolide on cx43 in sertoli cells. Toxicol Mech Methods 2008;18:717–724.
Lee WM. Drug-induced hepatotoxicity. N Engl J Med 2003;349:474–485.
Pessayre D, Mansouri A, Berson A, Fromenty B. Mitochondrial involvement in drug-induced liver injury. Handb Exp Pharmacol 2010:311–365.
Fromenty B, Pessayre D. Inhibition of mitochondrial betaoxidation as a mechanism of hepatotoxicity. Pharmacol Ther 1995;67:101–154.
Pessayre D, Mansouri A, Haouzi D, Fromenty B. Hepatotoxicity due to mitochondrial dysfunction. Cell Biol Toxicol 1999;15:367–373.
Su Y, Yang S, Xiao Z, Wang W, Okunieff P, Zhang L. Triptolide alters mitochondrial functions. Adv Exp Med Biol 2007;599:139–146.
Author information
Authors and Affiliations
Corresponding author
Additional information
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)
Rights and permissions
About this article
Cite this article
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
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11655-012-1265-x