Study of Fufang Haishe capsule against cell apoptosis / 中国中药杂志

<p><b>OBJECTIVE</b>To study mechanismt of Fufang Haishe capsule for dementia by observing the effect of it on PC-12 cell apoptosis, which was induced by beta-amyloid protein (Abl-42).</p><p><b>METHOD</b>Nerve growth factor (NGF) was used to cultivate the PC-12 cells. Fufang Haishe capsule at different concentrations was added into the culture medium so as to identify the nontoxic concentrations with MTT. To analyze the PC-12 cell apoptosis respectively by MTT assay, Flow cytometry (FCM technique) with different concentrations of Fufang Haishe capsule (0.01, 0.1, 1, 5 mg x mL(-1)), adding Ab or not Western blot was used to detect apoptosis which was measured on the implementation of caspase-9 and caspase-3 activity.</p><p><b>RESULT</b>Fufang Haishe capsule could significantly inhibit the apoptosis of PC-12 cells induced by Abeta with increased colorimetric MTT asay ( compare among the control group and concentration 0, 0.01, 0.1, 1 and 5 mg x mL(-1) group, which is the same below: 1.75 +/- 0.12, 0.73 +/- 0.35, 0.79 +/- 0.11, 0.83 +/- 0.07, 1.31 +/- 0.07, 1.80 +/- 0.38, P < 0.01) and the decreased apoptosis rate of the cells which was analysed by flow cytometry (1.93 +/- 0.41)%, (46.17 +/- 4.08)%, (35.35 +/- 4.63)%, (28.62 +/- 3.81)%, (15.13 +/- 3.15)%, (7.84 +/- 1.76)%, P < 0.01. In addition, Fufang Haishe capsule inhibited the activity of caspase-9 and caspase-3 of PC-12 cells which was induced by Abeta.</p><p><b>CONCLUSION</b>Fufang Haishe capsule significantly inhibite apoptosis of PC-12 cells induced by Abeta. The mechanism might be that Fufang Haishe capsule decrease the activity of the apoptosis implementing protein,caspase-9 and caspase-3.</p>

Inhibitory effect of sinomenine on expression of cyclooxygenase-2 in lipopolysaccharide-induced PC-12 cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of sinomenine (Sin) on cell proliferation, intracellular expression of cyclooxygenase-2 (COX-2), and production of PGE2 in lipopolysaccharide-induced PC-12 cells, To explore the Sin's mechanism on nerve cell.</p><p><b>METHOD</b>PC-12 cells were cultured with nerve growing factors (NGF), and pretreated with Sin at various concentrations (0, 3 x 10(-6), 30 x 10(-6), 150 x 10(-6) mol x L(-1)) for 2 hours, then with or without stimulation of lipopolysaccharide (LPS). The proliferation activity of PC-12 cells was determined by 3H-TdR incorporation, and the production of PGE2 in culture supernatants of PC-12 cells was detected with competitive ELISA. Expression of COX-2 mRNA in PC-12 cells was analyzed by semi-quantitative RT-PCR, and expression of COX-2 protein was estimated by Western blot method and cellular enzyme immunoassay. Nuclear factor-kappa B (NF-kappaB) activity in whole-cell extract of PC-12 cells was also measured by an ELISA-based method.</p><p><b>RESULT</b>The data showed that Sin down-regulated the expression of COX-2 mRNA and protein, and reduced the production of PGE2 in the LPS-stimulated PC-12 cells which correlated with Sin's concentrations positively. In addition, NF-kappaB activity in LPS-stimulated cells was suppressed significantly by Sin. No inhibition of proliferation of PC-12 cells due to Sin treatment was observed.</p><p><b>CONCLUSION</b>Sin mediates the down-regulation of expression of COX-2 and production of induced PGE2 in PC-12 cells by suppressing the activity of NF-kappaB.</p>