Neuroprotective Effects of Salidroside in the PC12 Cell Model Exposed to Hypoglycemia and Serum Limitation

  • Shu Yu
  • Mei Liu
  • Xiaosong Gu
  • Fei DingEmail author
Original Paper


The hypoglycemia and serum limitation-induced cell death in cultured PC12 cells represents a useful in vitro model for the study of brain ischemia and neurodegenerative disorders. Salidroside is a phenylpropanoid glycoside isolated from Rhodiola rosea L., a traditional Chinese medicinal plant, and has displayed a broad spectrum of pharmacological properties. In this study, MTT assay, Hoechst 33342 staining, and flow cytometry with annexin V/PI staining collectively showed that pretreatment with salidroside attenuated, in a dose-dependent manner, cell viability loss, and apoptotic cell death in cultured PC12 cells induced by hypoglycemia and serum limitation. RT-PCR, Western blot analysis, and enzymatic colorimetric assay indicated the changes in expression levels of Bcl-2, Bax, and caspase3 in PC12 cells on exposure to hypoglycemia and serum limitation with and without salidroside pretreatment, respectively. Rhodamine 123 staining and flow cytometry with 2′,7′-Dichlorofluorescin diacetate staining revealed the changes in the mitochondrial membrane potential and radical oxygen species (ROS) production in PC12 cells on exposure to hypoglycemia and serum limitation with and without salidroside pretreatment, respectively. The experimental results suggest that salidroside protects the PC12 cells against hypoglycemia and serum limitation-induced cytotoxicity possibly by the way of the modulation of apoptosis-related gene expression, the restoration of the mitochondrial membrane potential, and the inhibition of the intracellular ROS production. Our findings might raise a possibility of potential therapeutic applications of salidroside for preventing and treating cerebral ischemic and neurodegenerative diseases.


Salidroside Hypoglycemia and serum limitation Apoptosis PC12 cells Neuroprotection 



This study was supported by Hi-Tech Research and Development Program of China (973 Program, Grant no. 2003CB515306). We wish to thank Professor Jie Liu for assistance in the preparation of the manuscript.


  1. Bar-Am O, Weinreb O, Amit T, Youdim MB (2005) Regulation of Bcl-2 family proteins, neurotrophic factors, and APP processing in the neurorescue activity of propargylamine. FASEB J 19(13):1899–1901PubMedGoogle Scholar
  2. Bromont C, Marie C, Bralet J (1989) Increased lipid peroxidation in vulnerable brain regions after transient forebrain ischemia in rats. Stroke 20:918–924PubMedGoogle Scholar
  3. Cao LL, Du GH, Wang MW (2006) The effect of salidroside on cell damage induced by glutamate and intracellular free calcium in PC12 cells. J Asian Nat Prod Res 8:159–165PubMedCrossRefGoogle Scholar
  4. Choi JW, Yoo BK, Shin CY, Ryu MK, Ryu JH, el Kouni MH, Lee JC, Kim WK, Ko KH (2006) Uridine prevents the glucose deprivation-induced death of immunostimulated astrocytes via the action of uridine phosphorylase. Neurosci Res 56:111–118PubMedCrossRefGoogle Scholar
  5. Crompton M (2000) Bax, Bid and the permeabilization of the mitochondrial outer membrane in apoptosis. Curr Opin Cell Biol 12:414–419PubMedCrossRefGoogle Scholar
  6. Darbinyan V, Kteyan A, Panossian A, Gabrielian E, Wikman G, Wagner H (2000) Rhodiola rosea in stress induced fatigue-a double blind cross-over study of a standardized extract SHR-5 with a repeated low-dose regimen on the mental performance of healthy physicians during night duty. Phytomedicine 7:365–371PubMedGoogle Scholar
  7. De Sanctis R, De Bellis R, Scesa C, Mancini U, Cucchiarini L, Dacha M (2004) In vitro protective effect of Rhodiola rosea extract against hypochlorous acid-induced oxidative damage in human erythrocytes. Biofactors 20:147–159PubMedGoogle Scholar
  8. Diaz Lanza AM, Abad Martinez MJ, Fernandez Matellano L, Recuero Carretero C, Villaescusa Castillo L, Silvan Sen AM, Bermejo Benito P (2001) Lignan and phenylpropanoid glycosides from Phillyrea latifolia and their in vitro anti-inflammatory activity. Planta Med 67:219–223PubMedCrossRefGoogle Scholar
  9. Gross A, McDonnell JM, Korsmeyer SJ (1999) BCL-2 family members and the mitochondria in apoptosis. Genes Dev 13:1899–1911PubMedCrossRefGoogle Scholar
  10. Hall ED, Braughler JM (1989) Central nervous system trauma and stroke. II. Physiological and pharmacological evidence for involvement of oxygen radicals and lipid peroxidation. Free Radic Biol Med 6:303–313PubMedCrossRefGoogle Scholar
  11. Hillion JA, Takahashi K, Maric D, Ruetzler C, Barker JL, Hallenbeck JM (2005) Development of an ischemic tolerance model in a PC12 cell line. J Cereb Blood Flow Metab 25:154–162PubMedCrossRefGoogle Scholar
  12. Hoyer S (1993) Abnormalities in brain glucose utilization and its impact on cellular and molecular mechanisms in sporadic dementia of Alzheimer type. Ann N Y Acad Sci 695:77–80PubMedCrossRefGoogle Scholar
  13. Johnson EM Jr, Greenlund LJ, Akins PT, Hsu CY (1995) Neuronal apoptosis: current understanding of molecular mechanisms and potential role in ischemic brain injury. J Neurotrauma 12:843–852PubMedCrossRefGoogle Scholar
  14. Kostic V, Jackson-Lewis V, de Bilbao F, Dubois-Dauphin M, Przedborski S (1997) Bcl-2: prolonging life in a transgenic mouse model of familial amyotrophic lateral sclerosis. Science 277:559–562PubMedCrossRefGoogle Scholar
  15. Kucinskaite A, Briedis V, Savickas A (2004) Experimental analysis of therapeutic properties of Rhodiola rosea L. and its possible application in medicine. Medicina (Kaunas) 40:614–619Google Scholar
  16. Lazarova MB, Petkov VD, Markovska VL, Petkov VV, Mosharrof A (1986) Effects of meclofenoxate and Extr. Rhodiolae roseae L. on electroconvulsive shock-impaired learning and memory in rats. Methods Find Exp Clin Pharmacol 8:547–552PubMedGoogle Scholar
  17. Liu Y, Song XD, Liu W, Zhang TY, Zuo J (2003) Glucose deprivation induces mitochondrial dysfunction and oxidative stress in PC12 cell line. J Cell Mol Med 7:49–56PubMedCrossRefGoogle Scholar
  18. Love S (1999) Oxidative stress in brain ischemia. Brain Pathol 9:119–131PubMedGoogle Scholar
  19. Mattioli L, Perfumi M (2007) Rhodiola rosea L. extract reduces stress- and CRF-induced anorexia in rats. J Psychopharmacol 21:742–750PubMedCrossRefGoogle Scholar
  20. Ming DS, Hillhouse BJ, Guns ES, Eberding A, Xie S, Vimalanathan S, Towers GH (2005) Bioactive compounds from Rhodiola rosea (Crassulaceae). Phytother Res 19:740–743PubMedCrossRefGoogle Scholar
  21. Moley KH, Mueckler MM (2000) Glucose transport and apoptosis. Apoptosis 5:99–105PubMedCrossRefGoogle Scholar
  22. Oliver CN, Starke-Reed PE, Stadtman ER, Liu GJ, Carney JM, Floyd RA (1990) Oxidative damage to brain proteins, loss of glutamine synthetase activity, and production of free radicals during ischemia/reperfusion-induced injury to gerbil brain. Proc Natl Acad Sci USA 87:5144–5147PubMedCrossRefGoogle Scholar
  23. Perfumi M, Mattioli L (2007) Adaptogenic and central nervous system effects of single doses of 3% rosavin and 1% salidroside Rhodiola rosea L. extract in mice. Phytother Res 21:37–43PubMedCrossRefGoogle Scholar
  24. Petit PX, Susin SA, Zamzami N, Mignotte B, Kroemer G (1996) Mitochondria and programmed cell death: back to the future. FEBS Lett 396:7–13PubMedCrossRefGoogle Scholar
  25. Petkov VD, Yonkov D, Mosharoff A, Kambourova T, Alova L, Petkov VV, Todorov I (1986) Effects of alcohol aqueous extract from Rhodiola rosea L. roots on learning and memory. Acta Physiol Pharmacol Bulg 12:3–16Google Scholar
  26. Reimann-Philipp U, Ovase R, Weigel PH, Grammas P (2001) Mechanisms of cell death in primary cortical neurons and PC12 cells. J Neurosci Res 64:654–660PubMedCrossRefGoogle Scholar
  27. Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 27:612–616PubMedCrossRefGoogle Scholar
  28. Wise-Faberowski L, Raizada MK, Sumners C (2001) Oxygen and glucose deprivation-induced neuronal apoptosis is attenuated by halothane and isoflurane. Anesth Analg 93:1281–1287PubMedCrossRefGoogle Scholar
  29. Woronowicz A, Amith SR, Davis VW, Jayanth P, De Vusser K, Laroy W, Contreras R, Meakin SO, Szewczuk MR (2007) Trypanosome trans-sialidase mediates neuroprotection against oxidative stress, serum/glucose deprivation, and hypoxia-induced neurite retraction in Trk-expressing PC12 cells. Glycobiology 17(7):725–734PubMedCrossRefGoogle Scholar
  30. Yang E, Korsmeyer SJ (1996) Molecular thanatopsis: a discourse on the BCL2 family and cell death. Blood 88:386–401PubMedGoogle Scholar
  31. Zhang L, Yu H, Sun Y, Lin X, Chen B, Tan C, Cao G, Wang Z (2007) Protective effects of salidroside on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells. Eur J Pharm 564:18–25CrossRefGoogle Scholar
  32. Zhang WH, Wang X, Narayanan M, Zhang Y, Huo C, Reed JC, Robert M (2003) Fundamental role of the Rip2/caspase-1 pathway in hypoxia and ischemia-induced neuronal cell death. Proc Natl Acad Sci USA 100:16012–16017PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  1. 1.Jiangsu Key Laboratory of NeuroregenerationNantong UniversityNantongPeople’s Republic of China

Personalised recommendations