Chlorophyll a, an active anti-proliferative compound of Ludwigia octovalvis, activates the CD95 (APO-1/CD95) system and AMPK pathway in 3T3-L1 cells.
Ludwigia octovalvis is an aquatic plant widely distributed in Taiwan. It is traditionally used as a diuretic and is consumed as health drink. In this study, we evaluated the anti-proliferative activity of extracts and active constituent (chlorophyll a; CHL-a) of L. octovalvis in 3T3-L1 adipocytes; its mode of action on apoptosis was also investigated. Results showed that, among the different extracts and fractions, the ethylacetate layer (EAL) possessed the most potent anti-proliferative activity. Activity guided fractionation of the EAL obtained the bioactive constituent CHL-a (IC50: 24.10+/-0.83 nM). At concentrations 5-30 nM, CHL-a exhibited a dose-dependent accumulation of the Sub-G1 peak and caused cell cycle arrest at the G0/G1 phase. At 30 nM, it significantly reduced the cell viability, induced the appearance of DNA fragments, and enhanced the activation of caspase-3. Western blot data revealed that CHL-a decreased the level of Bcl-2, and increased the expression of CD95 (APO-1/CD95) and Bax. Furthermore, CHL-a up-regulated the AMPK and p-AMPK levels, and down-regulated the expression of PPAR-gamma. These results conclude that CHL-a possesses potent anti-proliferative activity, and its apoptotic effects on 3T3-L1 adipocytes are mediated through the activation of CD95 (APO-1/CD95) system and the AMPK signaling pathway.
Wu
Shu-Jing
SJ
Department of Nutritional Health, Chia-Nan University of Pharmacy and Science, Tainan, Taiwan. wsj268@mail.chna.edu.tw
Ng
Lean-Teik
LT
Wang
Gueu-Horng
GH
Huang
Yu-Jen
YJ
Chen
Jin-Liang
JL
Sun
Fang-Ming
FM
D041721:3T3-L1 Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000091162:AMP-Activated Protein Kinase Kinases; D017667:Adipocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D000818:Animals; D019440:Anti-Obesity Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D005591:Chemical Fractionation; D002734:Chlorophyll / Q000737:chemistry / Q000494:pharmacology*; D000077194:Chlorophyll A; D051379:Mice; D029594:Onagraceae / Q000737:chemistry*; D047495:PPAR gamma / Q000187:drug effects / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry; D011494:Protein Kinases / Q000187:drug effects* / Q000378:metabolism; D019014:fas Receptor / Q000187:drug effects* / Q000378:metabolism
D019440:Anti-Obesity Agents; D047495:PPAR gamma; D010936:Plant Extracts; D019014:fas Receptor; D002734:Chlorophyll; D011494:Protein Kinases; D000091162:AMP-Activated Protein Kinase Kinases; D000077194:Chlorophyll A