Effects of Duhuo Jisheng Decoction on Interleukin-1β-Induced Human Nucleus Pulposus Cell Apoptosis and Endoplasmic Reticulum Stress
Objective:To study the effect of Duhuo Jisheng decoction drug-contained serum on interleukin 1β-induced hu-man nucleus pulposus cell apoptosis and endoplasmic reticulum stress,and to explore the mechanism of Duhuo Jisheng decoction in intervening intervertebral disc degenera-tion.Methods:The 40 ten-week-old SD rats were gavaged with Duhuo Jisheng decoction,and the abdominal aorta blood after 2 weeks continuous gavage was collected to obtain drug-contained serum.The nucleus pulposus cells were trea-ted by different concentrations of Duhuo Jisheng decoction for different times.The cell viability test kit(CCK-8)was used to detect the activity of nucleus pulposus cells.The nucleus pulposus cells were randomly divide into three groups:control group(without the treatment of IL-1β and Duhuo Jisheng decoction drug-contained serum),IL-1β group(adding 10 ng/mL IL-1β solution for 24 h),Duhuo Jisheng decoction group(adding 10 ng/mL IL-1β solution and Duhuo Jisheng decoction drug-contained serum for 24 h).The TUNEL method was used to analyze the apoptosis rate of nucleus pulposus cells.Immunofluorescence assay was used to detect the expressions of CHOP and GRP78 in each group;qRT-PCR was used to detect the mRNA expressions of CHOP,GRP78,Cleaved-caspase3,Bax,BCL-2,PERK,elF2α and ATF4 in each group;Western Blot was used to detect CHOP,GRP78,Cleaved-caspase3,Bax,BCL-2,PERK,elF2α,p-elF2α,p-PERK and ATF4 protein expressions.Results:Duhuo Jisheng decoction can significantly enhance the vitality and improve the apopto-sis of nucleus pulposus cells.Compared with the control group,the mRNA expression of CHOP,GRP78,Cleaved-caspase3,Bax,PERK,elF2α and ATF4 in IL-1β group was significantly decreased;the mRNA expression of BCL-2 was increased,and the difference was statistically significant(P＜0.05).Compared with the IL-1β group,the mRNA expres-sion of CHOP,GRP78,Cleaved-caspase3,Bax,PERK,elF2α and ATF4 in Duhuo Jisheng decoction group was significantly increased;the mRNA expression of BCL-2 was decreased,and the difference was statistically significant(P＜0.05).Com-pared with the control group,CHOP,GRP78,Cleaved-caspase3,Bax,PERK,elF2α,ATF4,p-PERK,p-elF2α protein expression in IL-1β group was significantly decreased;the protein expression of BCL-2 was increased,and the difference was statistically significant(P＜0.05).Compared with the IL-1β group,the CHOP,GRP78,Cleaved-caspase3,Bax,PERK,elF2 α,ATF4,p-PERK and p-elF2α protein expression in Juhuo Jisheng decoction group was significantly increased;the protein expression of BCL-2 was decreased,and the difference was statistically significant(P＜0.05).Con-clusion:The Duhuo Jisheng decoction drug-contained serum can inhibit endoplasmic reticulum stress and apoptosis in degenerated nucleus pulposus cells,thereby delaying intervertebral disc degeneration,which may be related to PERK/elF2α pathway.
NETL
NSTL
万方数据
