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| Effect of hypoxia inducible factor on the expression of related proteins in apoptotic pathway of nucleus pulposus cells in SD rats |
| LI Xu1 YE Fang1 LIU Yi-xiu1 ZHAO Yu1 XIE Xin2 YANG Li-qun3 TU Guan-jun4▲ |
1.Department of Foot and Ankle Surgery,Shenyang Medical College Affiliated Central Hospital,Liaoning Province,Shenyang 110024,China;
2.Functional Experiment Center,Liaoning University of Traditional Chinese Medicine,Liaoning Province,Shenyang 110847,China;
3.Liaoning Research Institute of Family Planning,China Medical University,Liaoning Province,Shenyang 110031,China;
4.Department of Orthopedics,the First Hospital of China Medical University,Liaoning Province,Shenyang 110001,China |
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Abstract Objective To explore the effect of hypoxia inducible factor-1α (HIF-1α) on the expression of related proteins in apoptotic pathway of nucleus pulposus cells in SD rats.Methods The nucleus pulposus cells of SD rats were primary cultured.The expression of HIF-1α was induced in different degrees in different conditions.Group A (hypoxia condition group): O2 5%,CO2 5%,N2 90%;group B (over hypoxia condition group): O2 2%,CO2 5%,N2 93%;group C(hypoxia and CoCl2 treatment group): O2 5%,CO2 5%,N2 90%;group D (hypoxia and CAY10585 treatment group): O2 5%,CO2 5%,N2 90%.The apoptosis of each group was detected at the same time.Western Blot was used to detect the expression of HIF-1α,p53,Bax,caspase3 and Bcl-2 protein.The relationship between the changes of HIF-1α,p53,Bax,Bcl-2 and caspase3 and apoptosis rate was analyzed.Results HIF-1α protein expression was observed in all groups.The expression of HIF-1α protein in group B and group C was higher than that in group A,the expression of HIF-1α protein in group C was the highest,and the lowest in group D,the differences were statistically significant (P＜0.05).The expression trends of p53,Bax,and caspase3 proteins were the same,the highest in group C and higher than those in group A and B,the protein expression in group B was higher than that in group A,while the protein expression in group D was the lowest (P＜0.05).The expression trend of Bcl-2 and Bax was opposite,the lowest in group C,lower than that in group A,the highest in group D,higher than that in group A,the differences were statistically significant (P＜0.05).The results obtained by flow cytometry showed that the apoptotic rate of cells in group C was the highest,while that of group D was the lowest,there was a statistically significant difference between the two groups (P＜0.05).The changes of HIF-1α,p53,Bax,Bcl-2 and caspase3 were positively correlated with the apoptosis rate (P＜0.05).Conclusion The increase of HIF-1α expression can induce the apoptosis of nucleus pulposus cells.The expression of apoptosis related factors p53,Bax,Bcl-2 and caspase-3 is detected by HIF-1α expression in different degrees,which indicates the correlation between the expression of HIF-1α and the expression of apoptosis related proteins,thus reveals the effect of HIF-1α on the expression of apoptosis related factors in the mitochondrial apoptosis pathway of SD rat nucleus pulposus cell.
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|
| [10] |
Xia Y，Jiang L，Zhong T.The role of HIF-1α in chemo-/radioresistant tumors[J].Onco Targets Ther，2018，11：3003-3011.
|
| [1] |
Xiao B，Wang SY，Yang GS，et al.HIF-1α contributes to Hypoxia adaptation of the naked mole rat[J].Oncotarget，2017，8（66）：109 941-109 951.
|
| [2] |
Risbud MV，Guttapalli A，Stokes DG，et al.Nucleus pulposus cells express HIF-1 alpha under normoxic culture conditions：a metabolic adaptation to the intervertebral disc microenvironment[J].J Cell Biochem，2006，98（1）：152-159.
|
| [3] |
Ha KY，Koh IJ，Kirpalani PA，et al.The expression of hypoxia inducible factor-1α and apoptosis in herniated discs[J].Spine（Phila Pa 1976），2006，31（12）：1309-1313.
|
| [4] |
Lian CJ，Gao B，Wu ZZ，et al.Collagen type Ⅱis downregulated in the degenerative nucleus pulposus and contributes to the degeneration and apoptosis of human nucleus pulposus cells[J].Mol Med Rep，2017，16（4）：4730-4736.
|
| [5] |
Cheng A，He P，Zhang J，et al.Expression and correlation analysis of hypoxia inducible factor 1α and autophagy related molecules in rat nucleus pulposus cells under hypoxia[J].Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi，2020，34（3）：318-322.
|
| [6] |
Wang D，Zhu H，Cheng W，et al.Effects of hypoxia and ASIC3 on nucleus pulposus cells：From cell behavior to molecular mechanism[J].Biomed Pharmacotherapy，2019，117（1）：199 061.
|
| [7] |
Liu Y，Li Y，Huang ZN，et al.The effect of intervertebral disc degenerative change on biological characteristics of nucleus pulposus mesenchymal stem cell：an in vitro study in rats[J].Connect Tissue Res，2019，60（4）：376-388.
|
| [8] |
Bohuslavova R，Cerychova R，Papousek F，et al.HIF-1α is required for development of the sympathetic nervous system[J].Proc Natl Acad Sci U S A，2019，116（27）：13 414-13 423.
|
| [9] |
Kang Y，Roh MR，Rajadurai S，et al.Hypoxia and HIF-1α Regulate Collagen Production in Keloids[J].J Invest Dermatol，2020，140（11）：2157-2165.
|
| [11] |
Stegen S，Laperre K，Eelen G，et al.HIF-1α metabolically controls collagen synthesis and modification in chondrocytes[J].Nature，2019，565（7740）：511-515.
|
| [12] |
Liu Z，Li C，Meng X，et al.Hypoxia-inducible factor-lα mediates aggrecan and collagen Ⅱexpression via NOTCH1 signaling in nucleus pulposus cells during intervertebral disc degeneration[J].Biochem Biophys Res Commun，2017，488（3）：554-561.
|
| [13] |
Karaarslan N，Yilmaz I，Ozbek H，et al.Are Specific Gene Expressions of Extracellular Matrix and Nucleus Pulposus Affected by Primary CellCultures Prepared from Intact or Degenerative Intervertebral Disc Tissues?[J].Turk Neurosurg，2019，29（1）：43-52.
|
| [14] |
Yang SH，Hu MH，Lo WY，et al.The influence of oxygen concentration on the extracellular matrix production of human nucleus pulposuscells during isolation-expansion process[J].J Biomed Mater Res A，2017，105（6）：1575-1582.
|
| [15] |
Meng X，Zhuang L，Wang J，et al.Hypoxia-inducible factor（HIF）-1alpha knockout accelerates intervertebral disc degeneration in mice[J].Int J Clin Exp Pathol，2018，11（2）：548-557.
|
| [16] |
Huang Z，Cheng X，Zhao J，et al.Influence of simvastatin on the biological behavior of nucleus pulposus-derived mesenchymal stem cells[J].Iran J Basic Med Sci，2019，22（12）：1468-1475.
|
| [17] |
Moritz W，Meier F，Stroka DM，et al.Apoptosis in hypoxic human pancreatic islets correlates with HIF-1alpha expression[J].FASEB J，2002，16（7）：745-747.
|
| [18] |
Jin LZ，Lu JS，Gao JW.Silencing SUMO2 promotes protection against degradation and apoptosis of nucleus pulposus cells through p53 signaling pathway in intervertebral disc degeneration[J].Biosci Rep，2018，38（3）：BSR20171523.
|
| [19] |
Zhang M，Liu Q，Li L，et al.Cytoplasmic M-CSF facilitates apoptosis resistance by inhibiting the HIF-1α/BNIP3/Bax signalling pathway in MCF-7 cells[J].Oncol Rep，2019，41（3）：1807-1816.
|
| [20] |
Graeber TG，Osmanian C，Jacks T，et al.Hypoxia mediated selection of cells with diminished apoptotic potential in solid tumors[J].Nature，1996，379（6560）：88-91.
|
| [21] |
Greijer AE，van der Wall E.The role of hypoxia inducible factor 1 in hypoxia-induced apoptosis[J].J Clin Pathol，2004，57（10）：1009-1014.
|
|
|
|
