Effects of Ubiquitin on Human Umbilical Vein Endothelial Cells and Macrophages_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

 XingJuan ¹ , ZhangWei ¹ , ZhangZhenhuan ¹ , LiuWei ² , WangQian ²

1. Department of Criticial Care Medicine, Nanjing Benq Hospital, Nanjing, Jiangsu, 210019, China;
2. ;

Corresponding?author：

XingJuan, Email: medijine02@aliyun.com

Keywords：

Ubiquitin; Human umbilical vein endothelial cells; Human macrophages; Lipopolysaccharide

DOI：

10.7507/1671-6205.2015044

Video：

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Objective To compare the different effects of ubiquitin(UB) on human umbilical vein endothelial cells (HUVECs) and macrophages under normal circumstances,and analyze whether UB could protect HUVECs from lipopolysaccharide(LPS) induced injury. Methods The morphologic changes of HUVECs in vitro with up-rising concentrations of UB interventions were observed. HUVECs and human macrophages in vitro were divided into 4 groups according to UB concentration (0.01 μg/mL,0.1 μg/mL, 1 μg/mL, and 10 μg/mL). Supernatant and cells of each group were collected in 24 h after UB intervention. The levels of TNF-α and VCAM-1 in supernatant were measured by ELISA while NF-κB protein level in cells was detected by Western blot. HUVECs were divided into a LPS group(LPS 10 μg/mL) and an UB+LPS group(UB 0.1 μg/mL,LPS 10 μg/mL). The supernatant of the two groups were collected in 8,16 and 24 h after LPS and UB intervention. The levels of TNF-α and VCAM-1 in supernatant were measured by ELISA. Results The injury of HUVECs got worse with the ascending concentrations of UB.At the concentration of 50 μg/mL,UB induced HUVECs got ballooned and died massively. With the increase of UB concentration,the levels of TNF-α and VCAM-1 in HUVECs' supernatant ascended firstly and then descended,while those in human macrophages' supernatant ascended gradually. zHowever,the tendency of the NF-κB protein level in the two kinds of cells was similar when the concentration of UB increased.At the consentration of 0.1 μg/mL or 1 μg/mL,ubiquitin induced NF-κB protein level obviously increased.At the concentration of 0.01 μg/mL or 10 μg/mL,UB induced the protein level was similar with those of the control group and even decreased slightly. There was no significant difference in TNF-α or VCAM-1 levels at each time point between the LPS group and the UB+LPS group. Conclusions UB injuries HUVECs obviously at a low concentration but injuires human macrophages at much higher concentraton. UB can not protect HUVECs from LPS-induced injury in vitro.

Citation： XingJuan, ZhangWei, ZhangZhenhuan, LiuWei, WangQian. Effects of Ubiquitin on Human Umbilical Vein Endothelial Cells and Macrophages. Chinese Journal of Respiratory and Critical Care Medicine, 2015, 14(2): 173-177. doi: 10.7507/1671-6205.2015044 Copy

1.	?Hershko A,Ciechanover A. The ubiquitin system. Annu Rev Biochem,1998,67:425-479.
2.	Majetschak M. Extracellular ubiquitin:immune modulator and endogenous opponent of damage-associated molecular pattern molecules. J Leukoc Biol,2011,89:205-219.
3.	邢娟. 細胞外泛素作用的研究. 中國進修醫師雜志,2012,35:75-77.
4.	Daino H,Matsumura I,Takada K,et al. Induction of apoptosis by extracellular ubiquitin in human hematopoietic cells possible involvement of STAT3 degradation by proteasome pathway in interleukin 6-dependent hematopoietic cells. Blood,2000,95:2577-2585.
5.	Singh M,Roginskaya M,Dalal S,et al. Extracellular ubiquitin inhibits β-AR-stimulated apoptosis in cardiac myocytes:role of GSK-3β and mitochondrial pathways. Cardiovasc Res,2010,86:20-28.
6.	Nabika T,Terashima M,Momose I,et al. Synergistic effect of UB on lipopolysaccharide-induced TNF-α production in murine macrophage cell line RAW264.7 cells. Biochim Biophys Acta,1999,1450:25-34.
7.	Majetschak M,Krehmeier U,Bardenheuer M,et al. Extracellular ubiquitin inhibits the TNF-α response to endotoxin in peripheral blood mononuclear cells and regulates endotoxin hyporesponsiveness in critical illness. Blood,2003,101:1882-1890.
8.	Earle SA,Proctor KG,Patel MB,et al. Ubiquitin reduces fluid shifts after traumatic brain injury. Surgery,2005,138:431-438.
9.	Griebenow M,Casalis P,Woiciechowsky C,et al. Ubiquitin reduces contusion volume after controlled cortical impact injury in rats. J Neurotrauma,2007,24:1529-1535.
10.	邢娟,管向東,歐陽彬. 外源性泛素對致死型盲腸結扎穿孔術后小鼠早期肺損傷和血清硝酸鹽/亞硝酸鹽水平的影響. 中國呼吸與危重監護雜志,2009,8:432-435.
11.	Garcia-Covarrubias L,Manning EW 3rd,Sorell LT,et al. Ubiquitin enhances the TH2 cytokine response and attenuates ischemia-reperfusion injury in the lung. Crit Care Med,2008,36:979-982.
12.	邢娟,何振揚. 泛素對小鼠急性肺損傷肺組織細胞粘附分子及腫瘤壞死因子-α水平的影響. 實用醫學雜志,2010,15:567-569.
13.	Majetschak M,King DR,Krehmeier U,et al. Ubiquitin immunoreactivity in cerebrospinal fluid after traumatic brain injury:clinical and experimental findings. Crit Care Med,2005,33:1589-1594.
14.	Pirim I,Gumustekin K,Sahin Y. Serum ubiquitin concentration in patients with acute myocardial infarction. T Klin J Med Res,2000,18:1-5.
15.	Majetschak M,Sorell LT,Patricelli T,et al. Detection and possible role of proteasomes in the bronchoalveolar space of the injured lung. Physiol Res,2009,58:363-372.

1. ?Hershko A,Ciechanover A. The ubiquitin system. Annu Rev Biochem,1998,67:425-479.
2. Majetschak M. Extracellular ubiquitin:immune modulator and endogenous opponent of damage-associated molecular pattern molecules. J Leukoc Biol,2011,89:205-219.
3. 邢娟. 細胞外泛素作用的研究. 中國進修醫師雜志,2012,35:75-77.
4. Daino H,Matsumura I,Takada K,et al. Induction of apoptosis by extracellular ubiquitin in human hematopoietic cells possible involvement of STAT3 degradation by proteasome pathway in interleukin 6-dependent hematopoietic cells. Blood,2000,95:2577-2585.
5. Singh M,Roginskaya M,Dalal S,et al. Extracellular ubiquitin inhibits β-AR-stimulated apoptosis in cardiac myocytes:role of GSK-3β and mitochondrial pathways. Cardiovasc Res,2010,86:20-28.
6. Nabika T,Terashima M,Momose I,et al. Synergistic effect of UB on lipopolysaccharide-induced TNF-α production in murine macrophage cell line RAW264.7 cells. Biochim Biophys Acta,1999,1450:25-34.
7. Majetschak M,Krehmeier U,Bardenheuer M,et al. Extracellular ubiquitin inhibits the TNF-α response to endotoxin in peripheral blood mononuclear cells and regulates endotoxin hyporesponsiveness in critical illness. Blood,2003,101:1882-1890.
8. Earle SA,Proctor KG,Patel MB,et al. Ubiquitin reduces fluid shifts after traumatic brain injury. Surgery,2005,138:431-438.
9. Griebenow M,Casalis P,Woiciechowsky C,et al. Ubiquitin reduces contusion volume after controlled cortical impact injury in rats. J Neurotrauma,2007,24:1529-1535.
10. 邢娟,管向東,歐陽彬. 外源性泛素對致死型盲腸結扎穿孔術后小鼠早期肺損傷和血清硝酸鹽/亞硝酸鹽水平的影響. 中國呼吸與危重監護雜志,2009,8:432-435.
11. Garcia-Covarrubias L,Manning EW 3rd,Sorell LT,et al. Ubiquitin enhances the TH2 cytokine response and attenuates ischemia-reperfusion injury in the lung. Crit Care Med,2008,36:979-982.
12. 邢娟,何振揚. 泛素對小鼠急性肺損傷肺組織細胞粘附分子及腫瘤壞死因子-α水平的影響. 實用醫學雜志,2010,15:567-569.
13. Majetschak M,King DR,Krehmeier U,et al. Ubiquitin immunoreactivity in cerebrospinal fluid after traumatic brain injury:clinical and experimental findings. Crit Care Med,2005,33:1589-1594.
14. Pirim I,Gumustekin K,Sahin Y. Serum ubiquitin concentration in patients with acute myocardial infarction. T Klin J Med Res,2000,18:1-5.
15. Majetschak M,Sorell LT,Patricelli T,et al. Detection and possible role of proteasomes in the bronchoalveolar space of the injured lung. Physiol Res,2009,58:363-372.

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Effects of Ubiquitin on Human Umbilical Vein Endothelial Cells and Macrophages

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