Carthamus tinctorius L. prevents LPS-induced TNFalpha signaling activation and cell apoptosis through JNK1/2-NFkappaB pathway inhibition in H9c2 cardiomyoblast cells.
OBJECTIVE
Severe and potentially fatal hypotension and cardiac contractile dysfunction are common symptoms in patients with sepsis. In our previous study, we found that estradiol and estrogen-receptor alpha have cardio-protective effects in myocardial cells exposed to LPS. Estradiol supplementation has been shown to induce breast and cervical cancers. Flos Carthami, the flower of Carthamus tinctorius L. (Compositae) is an important traditional Chinese medicine used for the treatment of heart disease and inflammation, and therefore might be a potential alternative to Estradiol in the prevention of heart damage. This study investigated the effect of Flos Carthami (FC(EtOH)) ethanolic extract on LPS-induced apoptosis in H9c2 cardiomyoblast cells.


METHODS
H9c2 cells induced apoptosis with LPS administration (1 microg/mL). H9c2 cells were divided into five groups: Control, LPS (1 microg/mL), and three FC(EtOH) (31.25, 62.5,and 125 microg/mL). We detected apoptosis using MTT, LDH, TUNEL assay. JC-1 staining and Western blot were used to detect pro-apoptosis proteins, anti-apoptosis proteins, MAPK proteins (JNK, ERK, and P38), and NFkappaB expression.


RESULTS
FC(EtOH) (62.5 microg/mL) inhibited LPS-induced apoptosis by suppressing JNK1/2 activity, which resulted in the reduction of both IkappaB degradation and NFkappaB activation. In addition, FC(EtOH) led to the activation of anti-apoptotic proteins, Bcl-2 and Bcl-xL, the stabilization of the mitochondria membrane and the down-regulation of extrinsic and intrinsic pro-apoptotic proteins, such as TNFalpha, active caspase-8, t-Bid, Bax, active caspases-9, and -3.


CONCLUSIONS
Carthamus tinctorius L. possesses the ability to suppress JNK activity and inhibit LPS-induced TNFalpha activation and apoptosis in H9c2 cardiomyoblast cells. Carthamus tinctorius L could potentially serve as a cardio-protective agent against LPS-induced apoptosis.
Tien
Yun-Chen
YC
Graduate Institute of Chinese Pharmaceutical Sciences, China Medical University, Taichung, Taiwan.
Lin
Jing-Ying
JY
Lai
Chao-Hung
CH
Kuo
Chia-Hua
CH
Lin
Wen-Yuan
WY
Tsai
Chang-Hai
CH
Tsai
Fuu-Jen
FJ
Cheng
Yi-Chang
YC
Peng
Wen-Huang
WH
Huang
Chih-Yang
CY
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002316:Cardiotonic Agents / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D031187:Carthamus tinctorius / Q000737:chemistry*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D035264:Flowers; D008070:Lipopolysaccharides; D008516:Medicine, Chinese Traditional; D048055:Mitogen-Activated Protein Kinase 8 / Q000378:metabolism; D048056:Mitogen-Activated Protein Kinase 9 / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects* / Q000378:metabolism
D002316:Cardiotonic Agents; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010936:Plant Extracts; D014409:Tumor Necrosis Factor-alpha; D048056:Mitogen-Activated Protein Kinase 9; D048055:Mitogen-Activated Protein Kinase 8