Effects of Shenfu injection on calreticulin expression and neuronal apoptosis in the cerebral cortex of neonatal rats with hypoxic-ischemic brain damage
LIU Wen-Qiang, XU Yan, HAN Ai-Min, YANG Qian-Qian, WANG Jun
Department of Neonatology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu 221002, China
Abstract Objective To examine the expression of calreticulin (CRT) and the changes of intracellular free calcium and neuronal apoptosis in the cerebral cortex of neonatal rats with hypoxic-ischemic brain damage (HIBD), and to investigate the intervention effects of Shenfu injection. Methods Seven-day-old rats were randomly assigned to three groups: control, hypoxic-ischemia (HI) and Shenfu-treated. Each group (n=50) was subdivided into 5 groups sacrificed at 3, 6, 12, 24 and 72 hours. Rat models of HIBD were prepared according to the Rice's method. Rats in the control group only underwent the separation of right common carotidartery. Shenfu injection was administered by intraperitoneal injections right after HI insults and then once daily at a dosage of 10 mL/kg for 3 days in the Shenfu-treated group. The expression of CRT in the cerebral cortex was detected by RT-PCR and Western blot. The free calcium concentrations were determined under a fluorescent microscope. The apoptosis rate was measured by the flow cytometry. Results Compared with the control group, the expression levels of CRT in the HI and the Shenfu-treated groups were obviously up-regulated (P<0.05), and the expression levels of CRT in the Shenfu-treated group were notably higher than those in the HI group (P<0.05) at all time points. The concentrations of intracellular free calcium and the apoptosis rate of neurons in the cerebral cortex in the Shenfu-treated group were significantly reduced compared with those in the HI group (P<0.05), but increased significantly compared with those in the control group at all time points (P<0.05). Conclusions Shenfu injection may have neuroprotective effects against HIBD by up-regulation of CRT expression and relief of calcium overload.
About author:: 10.7499/j.issn.1008-8830.2015.03.017
Cite this article:
LIU Wen-Qiang,XU Yan,HAN Ai-Min et al. Effects of Shenfu injection on calreticulin expression and neuronal apoptosis in the cerebral cortex of neonatal rats with hypoxic-ischemic brain damage[J]. CJCP, 2015, 17(3): 281-286.
LIU Wen-Qiang,XU Yan,HAN Ai-Min et al. Effects of Shenfu injection on calreticulin expression and neuronal apoptosis in the cerebral cortex of neonatal rats with hypoxic-ischemic brain damage[J]. CJCP, 2015, 17(3): 281-286.
Cao G, Xing J, Xiao X, et al. Critical role of calpain I in mitochondrial release of apoptosis-inducing factor in ischemic neuronal injury[J]. J Neurosci, 2007, 27(35): 9278-9293.
Rice JE 3rd, Vannucci RC, Brierley JB. The influence of immaturity on hypoxic-ischemic brain damage in the rat[J]. Ann Neurol, 1981, 9(2): 131-141.
[6]
Liu H, Han F, Shi Y. Effect of calreticulin on Ca2+/CaM kinase II alpha and endoplasmic reticulum stress in hippocampal in a rat model of post-traumatic stress disorder[J]. Neurochem Res, 2013, 38(7): 1407-1414.
[7]
Li WH, Li YZ, Song DD, et al. Calreticulin protects rat microvascular endothelial cells against microwave radiationinduced injury by attenuating endoplasmic reticulum stress[J]. Microcirculation, 2014, 21(6): 506-515.
[8]
Lopez Sambrooks C, Carpio MA, Hallak ME. Arginylated calreticulin at plasma membrane increases susceptibility of cells to apoptosis[J]. J Biol Chem, 2012, 287(26): 22043-22054.
[9]
Shi Z, Hou W, Hua X, et al. Overexpression of calreticulin in pre-eclamptic placentas: effect on apoptosis, cell invasion and severity of pre-eclampsia[J]. Cell Biochem Biophys, 2012, 63(2): 183-189.
