[Lingmao recipe inhibits starvation induced autophagy in HepG2.2.15 cell].
OBJECTIVE
To study the effects of Lingmao Recipe (LR) on starvation-induced autophagy in HepG 2.2.15 cell.


METHODS
Fifteen SD rats were selected to prepare LR drug serum. The Earle's balanced salt solution (EBSS) group, the EBSS + vehicle serum group, the EBSS + 5 times LR serum group, the EBSS + 10 times LR serum group, and the DMEM group were set up, 3 samples in each group. The HepG2.2.15 cells were cultured for 1, 2, 4, and 6 h respectively. The pEGFP-N1-LC3B eukaryotic expression vector was constructed and transfected with HepG2. 2.15 cell. The GFP-LC3B morphological changes were observed under fluorescent microscope. The ratio of the dotty GFP-LC3B number and the GFP-LC3B transfected HepG2.2.15 number was calculated. The intracytoplasmic autophagosome changes were observed using electronic transmission electron. The microtubule-associated protein 1 light chain 3 beta II/I was detected in HepG2.2.15 of each group using Western blot.


RESULTS
Two h after culture, when compared with the EBSS + vehicle serum group, GFP-LC3B changing from diffused distribution to dotted distribution was obviously inhibited in the EBSS +5 times LR serum group and the EBSS +10 times LR serum group (P<0.01). The electronic transmission electron showed that the formation of autophagosome was inhibited in the EBSS +5 times LR serum group and the EBSS +10 times LR serum group. Results of Western blot showed that microtubule-associated protein 1 light chain 3 beta II/I obviously decreased more in the EBSS +10 times LR serum group than in the EBSS +vehicle serum group (P<0.01).


CONCLUSIONS
LR could inhibit starvation-induced autophagy in HepG 2.2.15.
Department of Hepatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203.
Xiao-Jun
XJ
Zhu
Xue-Hua
XH
Sun
Shun-Qing
SQ
Liu
D000818:Animals; D001343:Autophagy / Q000187:drug effects*; D003470:Culture Media; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D056945:Hep G2 Cells; D006801:Humans; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D044967:Serum; D014162:Transfection
D003470:Culture Media; D004365:Drugs, Chinese Herbal