Investigation The Molecular Mechanism Of HepG2 Apoptosis Induced By Niuhuangshen Containing Serum

Objective:NHS capsules containing serum induced Hep G2 apoptosis in vitro, and explored its molecular mechanism. Methods: Investigation the cellular apoptosis 、 Cyclin D1 m RNA 、 FAS expression and mitochondrial membrane △ψmt of Hep G2 that cultured with control serum、CTX or NHS。The process: Preparation of NHS capsules rats’ serum and culture Hep G2. CCK-8 method to detect the inhibitory ability of NHS on Hep G2. The nuclear was stained with DAPI and the morphology was detected with fluorescence microscopy after administration of the drug-containing serum 48 h.4. DNA ladder was investigated after administration of the drug-containing serum 48 h.5.Cyclin D1 m RNA was detected by real time RT-PCR after administration of the drug-containing serum 48 h.6.Fas protein expression was measured by taking advantage of Immunohistochemistry and flow cytometry after administration of the drug-containing serum 48 h.7.The mitochondrial membrane △ ψ mt of Hep G2 was detected after administration of the drug-containing serum 48 h. Results: 1. Various concentrations of NHS drug-containing serum and CTX group significantly inhibited the proliferation of Hep G2 cells. 2. The typical morphological changes in the serum containing group: the nucleus presents crease, corrugated, some chromatin appears condensed state by fluorescence microscopy.3. DNA Ladder was successfully formed in CTX group and NHS various concentrations of drug-containing serum group. 4. The Cyclin D1 m RNA expression was decreased companion with the increasing of NHS concentrations from 5% to 20%; compared with the control have significant difference in statistics. 5. The Fas protein expression increased in the CTX、5%NHS、10%NHS and 20% NHS group((6.54±0.98)%,(13.84±0.72)% and(27.03±2.01)%), respectively. 6. The mitochondrial membrane △ψmt of Hep G2 was decreased companion with the increasing of NHS concentrations from 5% to 20%; compared with the control have significant difference in statistics. Conclusion: The apoptosis of Hep G2 cells was induced by NHS, and the probably mechanism were increasing Fas expression and lowering the Cyclin D1 expression and mitochondrial membrane △ψmt in Hep G2.