Effect of serum from overfatigue rats on JNK/c-Jun/HO-1 pathway in human umbilical vein endothelial cells and the intervening effect of Tongxinluo superfine powder.
OBJECTIVE
To cultivate human umbilical vein endothelial cells (HUVECs) in the serum of overfatigue rats with the intervention of Tongxinluo superfine powder (TXLSP). By examining the variation of the activity of JNK/c-Jun/HO-1 pathway, the possible mechanisms of vascular endothelial dysfunction under overfatigue conditions and the intervening effect of TXLSP were explored.


METHODS
The HUVECs were randomly divided into the normal control group, the model group, the SP600125 (a specific antagonist of JNK) group, the TXLSP group and the TXLSP + SP600125 group. The content of carboyhemoglobin (COHb) and the leak rate of lactic dehydrogenase (LDH) in different groups were measured. The mRNA and protein expression of JNK, c-Jun, HO-1 and the phosphorylation level of c-Jun (P-c-Jun) were detected using Western blot and PCR methods.


RESULTS
Compared with the normal control group, the COHb level in supernatant was increased significantly in the model group, and the expression of HO-1, JNK, c-Jun mRNA and corresponding proteins and P-c-Jun were also increased remarkably. The increases in these parameters were significantly decreased by SP600125. TXLSP showed remarkable up-regulation on the expression of JNK, c-Jun, P-c-Jun and HO-1 mRNA and their protein expression. Compared with the SP600125 group, the expressions of JNK, c-Jun, P-c-Jun and HO-1 mRNA and its protein in the TXLSP+SP600125 group were significantly increased at different time points (P<0.05, P<0.01).


CONCLUSIONS
The vascular endothelial dysfunction under overfatigue conditions is related to the activity of the JNK/c-Jun/HO-1 pathway. One of the mechanisms of TXLSP in improving the vascular endothelial function is to adjust the activity of the JNK/c-Jun/HO-1 pathway at gene and protein levels.
Liang
Jun-qing
JQ
Department of Pharmacology, Hebei Yiling Pharmaceutical Research Institute, Shijiazhuang, 050035, China.
Xu
Hai-bo
HB
Wu
Yi-ling
YL
Sun
Shi-ran
SR
Jia
Zhen-hua
ZH
Wei
Cong
C
You
Jia-Hua
JH
D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D019610:Cytoprotection / Q000187:drug effects / Q000235:genetics; D004353:Drug Evaluation, Preclinical; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005221:Fatigue / Q000097:blood* / Q000378:metabolism; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008297:Male; D010316:Particle Size; D011208:Powders / Q000008:administration & dosage / Q000494:pharmacology; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D044967:Serum / Q000378:metabolism / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D001798:Blood Proteins; D004365:Drugs, Chinese Herbal; D011208:Powders; D016755:Proto-Oncogene Proteins c-jun; C513941:tongxinluo; D006419:Heme Oxygenase (Decyclizing); C491523:Hmox1 protein, rat; D048031:JNK Mitogen-Activated Protein Kinases