ABSTRACT
The apoptosis of mono-hepatocellular induced by the active ingredients of the Zanthoxyli Radix was investigated using laser Raman spectroscopy. Hepatoma cells (BEL-7404) were treated with 10 mg•L⁻¹ nitidine chloride and 3 g•L⁻¹ the extracts of Zanthoxyli Radix, respectively, then were divided into two parts, one for fluorescence staining, the other for determination of Raman spectroscopy. The acquired spectra were then processed by background elimination, smoothing, and normalization. Fluorescence staining results showed that the nucleuses from untreated group were uniformly stained, while those from the group treated for 48 hours were densely stained and broken. The spectra results revealed that the intensity of peaks associated with nucleic acid and protein decreased after the cells were incubated with the extracts of Zanthoxyli Radix for 12, 24, 36 and 48 hours. The intensity of peaks at 785,1 002,1 175,1 660 cm⁻¹ was decreased with the time of the cells were incubated by the extracts of Zanthoxyli Radix. The results indicated that the extracts of Zanthoxyli Radix could induce the apoptosis of hepatoma cells and reduce the amount of nucleic acid and protein in the cells. There is a certain relevance between the drug treatment time and the efficacy. The above results suggest that Raman spectra can provide abundant information about the changes in biological macromolecules within the cells after incubated by the extracts of Zanthoxyli Radix and serve as an effective method for the real time measurement of apoptosis.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of neurotoxicity induced by manganese, and observe the effects on the apoptosis of neurons in rat striatum.</p><p><b>METHODS</b>SD rats were divided into four groups, six rats each group. Three dose groups were exposed to high, middle, and low level of MnCl(2). At the end of experiment, all rats of the exposed groups and control group were decapitated, their striatums were removed and the Mn content of striatum, the apoptotic morphology, ratio and ultrastructural organization were analyzed.</p><p><b>RESULTS</b>The Mn content of striatum and apoptosis index of the three dose groups exposed to high, middle, and low level of Mn were significantly higher than control group (P < 0.05). The Mn content of striatum of the three dose groups exposed to high, middle, low level of MnCl(2) and control group were 2.98 +/- 0.52, 2.75 +/- 0.37, 2.61 +/- 0.73, 0.60 +/- 0.20 respectively. The apoptosis index of striatum of the three dose groups exposed to high, middle, low level of MnCl(2) and control group were 24.83 +/- 5.98, 17.00 +/- 5.33, 15.33 +/- 2.58, 2.83 +/- 0.41 respectively, and following higher level dose, the apoptosis index increased. The nucleus of neurons in striatum become smaller, condensed, etc, and these character showed apoptosis of neurons.</p><p><b>CONCLUSION</b>Mn can result in apoptotic morphology and increase level of apoptosis in striatum. The level of apoptos varies with Mn concentration.</p>
Subject(s)
Animals , Rats , Apoptosis , Corpus Striatum , Manganese , Neurons , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the change of free Ca(2+) in cytoplasma in the neurotoxicity of the manganese (Mn).</p><p><b>METHODS</b>The cortical neurons were separated from the neonatal Wistar rats and cultured in vitro. The neurons were grouped as the Mn-treated groups and the untreated group. The neurons in the Mn-added groups were incubated in the culture media containing lower, medium and high dosage manganese chloride (MnCl(2 x 4) H2O) with the concentration at 0.2, 0.6, 1.0 mmol/L respectively. Meanwhile, neurons in control were cultured in the normal culture media. All treatments stopped 24 h later. Neurons were labeled Ca(2+) sensitive prober, Fluo-3/AM. The fluorescence intensity of Fluo-3 combined with Ca(2+) was examined by LSCM (Laser scanning confocal microscope) and was treated by the picture analysis technique. The intensity was equal to the free Ca(2+) concentrations in cytoplasma of neurons.</p><p><b>RESULTS</b>MnCl(2) can induce free Ca(2+) overloaded in cytoplasma of neurons, but the increasing degree varied in MnCl(2) dosage. Cytoplasma Ca(2+) concentration in the moderate dosage The moderate dosage MnCl(2) group and the high dosage MnCl(2) group were significantly higher than that in the lower dosage MnCl(2) group and the control group (P < 0.05).</p><p><b>CONCLUSION</b>The Ca(2+) overload is involved in the neurotoxicity of manganese, and a dosage response relationship is found between the manganese chloride dose and Ca(2+) overload in cortical neurons.</p>