Synthesis of a Novel Compound and Mediated Neuro-protection in Differentiated PC12 Cells via Mitochondria Related Pathway

doi:10.7503/cjcu20150723

Abstract

Abstract:

The neuro-protective effect of a new synthetic compound named 3h8b on differentiated PC12(DPC12) cells against neurotoxins 6-hydroxydopamine(6-OHDA)- and L-glutamic acid(L-Glu)-induced cell injury and potential mechanisms were investigated. Data show that 3h8b significantly enhanced cell viability, improved nucleus morphological apoptotic alterations, and relieved intracellular Ca²⁺ overload caused by neurotoxins in DPC12 cells. The abnormal alteration of mitochondria in neurotoxins-treated cells was strongly restored by 3h8b. Further experiments demonstrate that 3h8b reverses neurotoxins-suppressed levels of B-cell lymphoma 2(Bcl-2) and Bcl-xL. All these results indicate that mitochondria related pathway is essential for the neuro-protective effect of the novel synthetic compound 3h8b against neurotoxins-induce toxicity in DPC12 cells.

Key words: Neuroprotection, 6-Hydroxydopamine, L-glutamate, Mitochondria

CLC Number:

O629

TrendMD:

HU Shuang, TENG Lirong, ZHANG Junrong, MENG Jiatong, ZHUANG Zhe, DU Mengyan, WANG Di, LIAO Weiwei. Synthesis of a Novel Compound and Mediated Neuro-protection in Differentiated PC12 Cells via Mitochondria Related Pathway[J]. Chem. J. Chinese Universities, 2015, 36(11): 2322.

Figures/Tables 6

Scheme 1 Procedure for preparation of novel compound 3h8b

Fig.1 3h8b strongly enhanced cell viability of DPC12 cells exposed to 100 μmol/L 6-OHDA or 25 mmol/L L-Glu for 24 h Data are expressed as mean±standard deviation(SD)(n=6). ### P < 0.001 versus control(CTRL); * P < 0.05 versus 6-OHDA-exposed cells; ^^ P < 0.01 and ^^^ P < 0.001 versus L-Glu-exposed cells.

Fig.2 Nucleus morphological apoptotic alterations examined by Hoechst 33342 staining Cells were pre-treated with 3h8b at indicated doses for 3 h, followed by exposure to 100 μmol/L 6-OHDA or 25 mmol/L L-Glu for 24 h. The experiment was repeated three times. (A) Control; (B) 6-OHDA; (C) 6-OHDA+20 μmol/L 3h8b; (D) 6-OHDA-40 μmol/L 3h8b; (E) control; (F) L-Glu; (G) L-Glu+20 μmol/L 3h8b; (H) L-Glu+40 μmol/L 3h8b.

Fig.3 Overload of intracellular Ca2+ caused by 6-OHDA or L-Glu reversed after incubation with 3h8b Treated cells were stained by the probe Fluo-4-AM. The experiment was repeated three times. (A)—(H) are the same as Fig.2.

Fig.4 Disruption of MMP caused by 6-OHDA or L-Glu restored by 3h8b JC-1 staining was applied to determining MMP. Red fluorescence indicates healthy cells with high MMP, whereas green fluorescence indicates apoptotic or unhealthy cells with low MMP. The experiment was repeated three times. (A)—(H) are the same as Fig.2.

Fig.5 Enhancing effects of pretreatment with 3h8b for 3 h at doses of 20 and 40 μmol/L on the expression of Bcl-2 and Bcl-xL in the cells exposed to 6-OHDA(100 μmol/L)(A) and L-Glu(25 μmol/L)(B) for 24 h The average fold of band intensity compared to that of related non-treated cells was marked respectively(n=3).

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