Thapsigargin induces endoplasmic reticulum stress in rat cortical neurons and intervention of Danhong injection

Abstract

Abstract: AIM: To study the apoptosis induced by thapsigargin on rat cortical neurons and the intervention of Danhong injection. METHODS: Primary cortial neurons were cultured in vitro, and NSE-positive cells were detected by immunohistological chemistry and immunofluorescence staining.Protein levels of GRP78, bcl-2, cytochrome c(cyt c), active caspase-12 were assessed by immunoblotting cell extracts with specific antibodies.The percentage of apoptotic(annexin V positive and propidium iodide negative)cells was determined by flow cytometric analysis.Protein expression of active caspase-3, caspase-9, caspase-8, caspase-7 was measured by flow cytometric(FCM)analysis cytometry;Intracellular calcium concentrations([ Ca2+] i) were measured by fluorescence spectrophotometer with calcium sensitive fluorescence indicator Fura-2/AM. RESULTS: As expected, GRP78 and caspase-12 protein levels elevated in cortical neurons in response to thapsigargin(2 μmol L, 24 h and 48 h)but not to vehicle.The rate of apoptosis induced by thapsigargin 24 h and 48 h was 17.88% and 21.38%, and was depressed to 6.30% and 6.11% after treated by Danhong injection (8 mL L) respectively.Activated caspase-3,-9,-8 was detected at 24 h and 48 h after thapsigargin contribution and was reduced by Danhong injection.The enhancement of [ Ca2 + ] i in neurons were induced by thapsigargin and were depressed by Danhong injection. CONCLUSION: Thapsigargin activates ER-initiated apoptosis which can be inhibited by Danhong injection in in rat cortical neurons.

Key words: thapsigargin, apotosis, endoplasmic reticulum stress, caspase, Danhong injection

CLC Number:

R965

ZHANG Hong, LIU Xue-qiu, ZHANG Xiao-qiao, ZHENG Li-fang, XU Jiang-hua, MEI Yuan-wu. Thapsigargin induces endoplasmic reticulum stress in rat cortical neurons and intervention of Danhong injection[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(3): 259-265.

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