Duhuo Jisheng decoction reduces apoptosis of human nucleus pulposus cells by regulating tunicamycin-induced endoplasmic reticulum stress
Objective:To study the effect and mechanism of Duhuo Jisheng decoction on endoplasmic reticulum stress and apoptosis of human nucleus pulposus cells induced by tunicamycin.Methods:Thirty SD rats of 10 weeks old were gavaged with Duhuo Jisheng decoction,and blood samples were collected through abdominal aorta after 14 days.Human nucleus pulposus cells were treated with tunicamycin and different concentrations of Duhuo Jisheng decoction,and CCK-8 was used to detect the activity of nucleus pulposus cells.Human nucleus pulposus cells were randomly divided into five groups:the Control group(not treated with tunicamycin and Duhuo Jisheng decoction-containing serum),the tunicamycin group(adding 10 μg/mL tunicamycin solu-tion),the tunicamycin+4-PBA group(adding 10 μg/mL tunicamycin solution and 5 mmol/mL 4-PBA),the tunicamycin+Duhuo Jisheng decoction group(adding 10 μg/mL tunicamycin solution and 10%medicated serum of Duhuo Jisheng decoction),the tunicamycin+normal saline group(adding 10 μg/mL tunicamycin solution and equal dose of normal saline).The expression levels of CHOP and GRP78 were detected by immunofluorescence.The apoptosis rate of nucleus pulposus cells was detected by TUNEL.The mRNA and protein expressions of CHOP,GRP78,endoplasmic reticulum stress and apoptosis-related pathways were detected by qRT-PCR and Western Blot,respectively.Results:Duhuo Jisheng decoction could enhance the activity of nucle-us pulposus cells,decrease the apoptosis rate of nucleus pulposus cells,and may have antagonistic effect on tunicamycin.Com-pared with the control group,protein and gene expressions of CHOP,GRP78,Cleaved-caspase3,Bax and apoptosis-related path-ways such as PERK/elF2α,IREI/JNK and Caspase12 were significantly increased after tunicamycin treatment,while protein and gene expressions of BCL-2 were significantly decreased.The difference was statistically significant(P<0.05).The expression of tunicamycin+saline group was similar to that of tunicamycin group,and the difference was not statistically significant.Protein and gene expressions of CHOP,GRP78,Cleaved-caspase3,Bax and apoptosis-related pathways such as PERK/elF2α,IREI/JNK and Caspase12,were significantly decreased in the tunicamycin+Duhuo jisheng decoction-containing serum group com-pared with the tunicamycin group.The expression of BCL-2 protein and gene was significantly increased,with statistical signifi-cance(P<0.05).The regulatory effect was similar to that of the positive control group,tunicamycin+4-PBA,and the difference was not statistically significant.Conclusion:Tunicamycin can induce endoplasmic reticulum stress in nucleus pulposus cells and promote apoptosis.Duhuo Jisheng decoction can regulate the tunicamycin-induced endoplasmic reticulum stress,inhibit the apopto-sis of nucleus pulposus cells,and delay the degeneration of intervertebral disc.
国家科技期刊平台
NSTL
万方数据
