The Effects of Acupuncture Serum on the Expression of Ca2+ and Caspase-12 Protein in Hippocampal Neurons of Neonatal Rats
LONG Xueping1, MA Yun2,YAN Fan1, WU Xu3, LIU Zhenbang3,ZHANG Hui1
(Anhui university of traditional Chinese medicine, clinical college of acupuncture and bone injury, Hefei 230038; 2 Wuhu city middle hospital, Wuhu 241002; Life sciences college, China university of science and technology, Hefei 230012)
Abstract: Objective: To observe the effect of acupuncture serum on the expression of endoplasmic reticulum and Caspase-12 protein in hippocampal neurons of neonatal rats induced by magnesium. Methods: SD rats were divided into acupuncture group and control group after intraperitoneal injection of amentazole induced acute convulsion. Acupuncture group choose "Baihui", "Dazhui" point, 1 day , a total of 7d. After the last acupuncture, rats inacupuncture group took serum as the acupuncture serum, and the control group as nonacupuncture serum. To magnesium induced primary cultured neonatal rat hippocampal neurons convulsions-like discharge model,rats were divided into normal extracellular fluid group (normal group), magnesium extracellular fluid group (hereinafter referred to as no magnesium group) and acupuncture group and the acupuncture serum groupaccording to different processing to the cultivation of 10 d hippocampal neurons. In the normal group, the serum-free medium was replaced by the extracellular fluid for 3 hours and then liquid-conditioned. The other groups were incubated with magnesium-free extracellular fluid for 3 hours and then liquid-conditioned. The endoplasmic reticulum Ca2 + -specific fluorescent probe combined with laser confocal microscopy was used to observe the changes of Ca2 + in the endoplasmic reticulum of hippocampal neurons. The expression of Caspase-12 protein in hippocampal neurons was observed by immunohistochemical method. Results: Compared with normal group, the fluorescence intensity of the neuron endoplasmic reticulum in the normal group was significantly lower than that in the normal group (p<0.05, p<0.01). Fluorescence intensity of endoplasmic reticulum in acupuncture group was significantly increased in 12h and 48h (p<0.05, p<0.01) compared with non-acupuncture group and non-acupuncture serum group. Compared with normal group, the expression of Caspase-12 protein increased significantly (P <0.05, P <0.01, P <0.01) in the group of magnesium-free neurons at three time points. The protein expression of Caspase-12 in acupuncture serum group decreased significantly at 12h and 48h (P <0.05, P <0.01 compared with non-acupuncture serum group). Conclusion: Acupuncture serum can stabilize Ca2+ in the endoplasmic reticulum of hippocampal neurons, which can significantly down-regulate Caspase-12 protein expression in hippocampal neurons.
Key words: convulsion, Ca2 +, Caspase -12 protein, Endoplasmic reticulum stress, Acupuncture serum