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23 December 2016 : Animal Research

Tetramethyl Pyrazine Protects Hippocampal Neurons Against Anoxia/Reoxygenation Injury Through Inhibiting Apoptosis Mediated by JNK/MARK Signal Pathway

Ming Zhong1ABCEF, Wuhua Ma1A*, Xiong Zhang1BC, Yong Wang1A, Xiaoqiu Gao1E

DOI: 10.12659/MSM.898921

Med Sci Monit 2016; 22:5082-5090

Abstract

BACKGROUND: Tetramethyl pyrazine (TMP) is a typical biologically active alkaloid isolated from the Chinese herb Ligusticum walliichi. It has been reported that TMP shows neuroprotective and stroke injury reductive properties in cerebral ischemia/reperfusion (I/R) animal models. In the present study we sought to investigate the effect and potential intervention mechanism of TMP in anoxia/reoxygenation (A/R) rat hippocampal neurons.

MATERIAL AND METHODS: After being cultured for 7 days, primary hippocampal neurons were randomly assigned into a normal control group (N), a TMP group (C: 0 ug/ml, L: 60 ug/ml, M: 200ug/ml and H: 800 ug/ml), and a JNK inhibitor group (S: SP600125, 10 μmol/L). A hypoxia/reoxygenation model were prepared 1 h after incubation. Hippocampal neurons were incubated in 90% N2 and 10% CO2 for 2 h, and then reoxygenated for 24 h in an incubator with 5%CO2 at the temperature of 37°C. The apoptosis rate, MKK4 and MKK7 mRNA and JNK kinase protein levels (C-fos, c-jun, and P-JNK) of hippocampal neurons were detected.

RESULTS: The apoptosis rates of hippocampal neurons induced by A/R showed significant reduction after being pre-treated with JNK inhibitor, TMP 60 µg/ml, 200 µg/ml, and 800 µg/ml. The JNK kinase MKK4mRNA and MKK7mRNA levels, as well as the expressions of C-fos, C-jun, and P-JNK protein levels, were also be reduced.

CONCLUSIONS: TMP may produce a protective effect in anoxia/reoxygenation-induced primary hippocampal neuronal injury by inhibiting the apoptosis of the hippocampal neurons; the possible mechanism may be inhibition of the JNK signal pathway.

Keywords: Anoxia, MAP Kinase Signaling System, Signal Transduction

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750