1.Protective effects of ferulic acid on mice bone marrow hematopoietic damage induced by irradiation
Shuai SHAO ; Mei TIAN ; Jianxiang LIU ; Qiao GOU ; Xuesong QI ; Chunyan WANG ; Gonglin QU ; Chen LI ; Xu SU
Chinese Journal of Radiological Medicine and Protection 2017;37(8):570-575
Objective To observe the effect and the mechanisms of ferulic acid on radiationinduced damage of mice peripheral blood and bone marrow hematopoietic function.Methods Ninety-six mice were randomly divided into sham irradiation group,irradiation group,positive drug group and 10,30,90 mg·kg-1 ·d-1 ferulic acid group,16 mice per group.Mice were exposed to 3.5 Gy γ-rays 24 h after first drug taken.Then,mice were given drugs for 7 d after irradiation.White blood cells in peripheral blood of 10 mice per group were counted 2 d before irradiation and 3,7,10,15 and 22 days after irradiation.The bone narrow of the other six mice was taken to detect the micronuclei frequency of polychromatic erythrocyte,the hematopoietic progenitor cell colony formation capacity,Thbd and HMGB1 protein expressions in mice bone marrow on the seventh day after irradiation.Results Compared with the irradiation alone group,the treatment of mice with ferulic acid 90 mg· kg-1 · d-1 increased the number of white blood cells in peripheral blood at 3,10,15 and 22 d after irradiation (t =2.267,2.399,1.945,2.828,P < 0.05).Treatment with mice with ferulic acid 90 mg· kg-1 · d 1 decreased the micronuclei rate of erythrocytes in irradiated bone marrow (t =4.013,P < 0.05),increased the clone numbers of CFU-E,BFU-E and CFU-GM of hematopoietic progenitor cells (t =2.366,2.953,3.115,P <0.05),improved the relative expression of the Thbd protein in bone marrow and the HMGB1 protein in nuclear (t =17.75,23.39,P < 0.01).Conclusions Ferulilc acid could protect the bone marrow hematopoietic of mice exposed to irradiation by regulating the expressions of Thbd and HMGB1 protein,and then accelerate the peripheral cells recovery.
2.Pentosan polysulfide sodium for chronic non-bacterial prostatitis in the rat model.
Wei-Ping LI ; Yang-Min WANG ; Bin ZHANG ; Ling-Min SONG ; Gou-Mei QIAO ; Xiao-Zheng LÜ
National Journal of Andrology 2011;17(4):310-313
OBJECTIVETo investigate the therapeutic effect of pentosan polysulfide sodium (PPS) on chronic non-bacterial prostatitis (CNP) in rats.
METHODSBased on Robinette's method, we established a CNP model in 80 male SD rats, aged 6 months and weighing 315 - 450 g, by castration followed by subcutaneous injection of estradiol at 0.25 mg / (kg x d) for 30 consecutive days. Then we randomly allocated the model rats into a placebo group (n = 40) and a PPS group (n = 40) to receive intragastric administration of normal saline and PPS, respectively. After 8 weeks of treatment, the pathological changes in the rat prostatic tissue were observed by HE staining.
RESULTSVaried degrees of chronic inflammation and inflammatory cell infiltration were seen in the prostatic tissues of both groups of rats before the treatment. The inflammation was significantly improved after the treatment in the PPS group but not in the placebo group.
CONCLUSIONPPS has some therapeutic effect on CNP in the rat, and its mechanism may be associated with the abilities of PPS to repair the damaged glycosaminoglycan layer and inhibit inflammation in the prostate.
Animals ; Chronic Disease ; Cystitis, Interstitial ; drug therapy ; Disease Models, Animal ; Male ; Pentosan Sulfuric Polyester ; therapeutic use ; Prostate ; pathology ; Prostatitis ; drug therapy ; pathology ; Rats ; Rats, Sprague-Dawley
3.Role of connexin 43 in X-ray induced apoptosis in human umbilical vein endothelial cells and its mechanism
Chen LI ; Mei TIAN ; Qiao GOU ; Jianxiang LIU ; Xuesong QI ; Chunyan WANG ; Xu SU
Chinese Journal of Radiological Medicine and Protection 2018;38(10):728-733
Objective To study the role of Cx43 in X-ray induced apoptosis of HUVEC cells and its mechanism.Methods Flow cytometry was used to detect the apoptosis of HUVEC cells at 48-96 h after 10 Gy X-ray irradiation and at 72 h after irradiation of different doses.Western blot was used to detect the protein expressions of Cx43 and cleaved caspase-3 in HUVEC cells at 72 h after 0,5,10 and 20 Gy irradiation.Small interfering RNA was transfected into HUVEC cells to silence Cx43 expression,the Cx43 bearing plasmid was transfected into cells to overexpress Cx43.The effect of Cx43 knockdown or overexpression on apoptosis induction and cleaved caspase-3 protein expression were detected by flow cytometry and Western blot,respectively.Results The apoptosis of HUVEC increased significantly from 48 h to 96 h after X-ray irradiation and in a dose-dependent manner at 72 h after irradiation.The expression of Cx43 protein was negatively correlated with the dose but the expression of cleaved caspase-3 was positively correlated with the dose in the range of 0-20 Gy.After Cx43 silencing,the proportion of early apoptosis and apoptosis combined with dead cells were significantly higher than that of the siRNA control group(t =3.674,6.375,P < 0.05).After Cx43 overexpression,the proportion of early apoptosis and apoptosis combined with dead cells were significantly lower than that of vector control group(t =9.399,11.190,P < 0.05).The expression of cleaved caspase-3 in the Cx43 silencing group was higher than that in the siRNA control group,but this protein in the Cx43 overexpressed group was lower than that in the vector control group.Conclusions Cx43 may protect X-ray irradiated HUVEC cells from apoptosis by down-regulating the activation of caspase-3.
4.Changes of connexin 43 in irradiated human vascular endothelial cells and its influence on cell stiffness
Chen LI ; Mei TIAN ; Qiao GOU ; Xuesong QI ; Xu SU
Chinese Journal of Radiological Medicine and Protection 2021;41(6):418-425
Objective:To investigate the changes of connexin 43 (Cx43) in human umbilical vein endothelial cells (HUVEC) after X-ray irradiation and its influence on the stiffness of irradiated cells.Methods:Western blot was used to detect the expression of Cx43 in HUVEC cells at different time points (0, 6, 12, 24 and 48 h) after different doses of X-ray irradiation (0, 2.5, 5, 10 and 20 Gy), and the phosphorylation levels of three phosphorylation sites (Ser279/282, Ser368 and Tyr265) of Cx43 at different time points (3, 6, 24 and 48 h) after 0, 5 and 10 Gy irradiation. The distribution of Cx43 protein in the irradiated HUVEC cells was detected by immunofluorescence. The stiffness changes of cells were detected by atomic force microscopy (AFM) at the depths of 50, 100 and 200 nm.Results:The expression of Cx43 in HUVEC cells was reduced at 6, 12, 24 and 48 h after 10 Gy X-ray irradiation( t=3.262, 3.708, 3.686, 6.825, P<0.05)and this decrease had a dose dependent manner at 24 h after 2.5, 5, 10 and 20 Gy irradiation ( t=3.034, 10.720, 13.130, 13.650, P<0.05). At 24 h after 5, 10 and 20 Gy X-ray irradiation, the distribution of Cx43 in HUVEC cells was transported from intercellular gap junctions to nucleus and perinuclear region. At 24-48 h after irradiation, the phosphorylation level of Ser368 at Cx43 increased and in a dose dependent manner. At 24 h after irradiation, the stiffness of the irradiated cells decreased significantly under the conditions of 100 and 200 nm ( t=3.362, 5.122, P<0.05), and recovered with overexpression of Cx43 ( t=2.674, 4.398, P<0.05). Conclusions:X-ray irradiation leads to the phosphorylation of Ser368 at Cx43, which promotes the degradation and nucleus/perinuclear translocation of Cx43 and reduces the stiffness of HUVEC. Increasing the expression level of Cx43 is helpful to the stiffness recovery of irradiated vascular endothelial cells, suggesting that Cx43 may be a potential target for regulating radiation injury of vascular endothelial cells.
5.Connexin43 Modulates X-Ray-Induced Pyroptosis in Human Umbilical Vein Endothelial Cells.
Chen LI ; Mei TIAN ; Qiao GOU ; Yong Rui JIA ; Xu SU
Biomedical and Environmental Sciences 2019;32(3):177-188
OBJECTIVE:
Pyroptosis is an inflammatory form of programmed cell death. This phenomenon has been recently reported to play an important role in radiation-induced normal tissue injury. Connexin43 (Cx43) is a gap junction protein that regulates cell growth and apoptosis. In this study, we investigated the effect of Cx43 on X-ray-induced pyroptosis in the human umbilical vein endothelial cells (HUVECs).
METHODS:
HUVECs, Cx43 overexpression, and Cx43 knockdown strains were irradiated with 10 Gy. Proteins were detected using western blot analysis. Cell pyroptosis was evaluated using the fluorescence-labeled inhibitor of caspase assay (FLICA) and propidium iodide staining through flow cytometry and confocal microscopy. Cell morphology and cytotoxicity were detected by scanning electron microscopy and lactate dehydrogenase release assay, respectively.
RESULTS:
Irradiation with 10 Gy X-ray induced pyroptosis in the HUVECs and reduced Cx43 expression. The pyroptosis in the HUVECs was significantly attenuated by overexpression of Cx43 as it decreased the level of active caspase-1. However, interference of Cx43 expression with siRNA significantly promoted pyroptosis by increasing the active caspase-1 level. Pannexin1 (Panx1), a gap junction protein regulates pyroptosis, and its cleaved form is used to evaluate channel opening and active state. The level of cleaved Panx1 in the HUVECs and Cx43 knockdown strains increased in the presence of X-ray, but decreased in the Cx43 overexpression strains. Furthermore, interference of Panx1 with siRNA alleviated the upregulation of pyroptosis caused by Cx43 knockdown.
CONCLUSION
Results suggest that single high-dose X-ray irradiation induces pyroptosis in the HUVECs. In addition, Cx43 regulates pyroptosis directly by activating caspase-1 or indirectly by cleaving Panx1.
Caspase 1
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genetics
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metabolism
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Connexin 43
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genetics
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metabolism
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Connexins
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genetics
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metabolism
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Gene Expression Regulation
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radiation effects
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Human Umbilical Vein Endothelial Cells
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physiology
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radiation effects
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Humans
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Nerve Tissue Proteins
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genetics
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metabolism
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Pyroptosis
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X-Rays
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adverse effects
6. Metabolic characteristics and radio-protective function of nano cerium oxide
Ning LI ; Chunyan WANG ; Peng TONG ; Liangliang YIN ; Shuai SHAO ; Chen LI ; Gonglin QU ; Qiao GOU ; Mei TIAN ; Xuesong QI
China Occupational Medicine 2017;44(03):331-335
OBJECTIVE: To observe the in vivo metabolism and distribution characteristics of nano-cerium oxide( nanoCeO_2) in rats,and to explore the radio-protective effect of nano-CeO_2. METHODS: i) A total of 18 specific pathogen free( SPF) SD rats were randomly divided into 3 groups. Rats of experiment group and CeO_2 blood group were gavaged with1. 0 g/kg body weight( bw) nano-CeO_2 suspension. Rats of control group were gavaged with double distilled water( DDW)in equal volume. At different time-points after treatment,venous blood was collected from the rats' eye socket in CeO_2 blood group,meanwhile urine and excrement of rats of experiment group were also collected. Organ and tissue samples of experiment group and control group were collected 24. 0 hours after treatment. The concentrations of cerium in biological samples were detected by inductively coupled plasma mass spectrometry. ii) A total of 72 SPF BALB/c mice were randomly divided into 6 groups. Mice of low-,medium-and high-dose groups were gavaged with 100,300 and 900 mg/kg bw nano-CeO_2 suspension respectively. Mice of negative control group,irradiation control group and drug positive control group were gavaged with DDW in equal volume once daily. After 14 days,mice of the other 5 groups were exposed by60Coγ-rays once with 3. 5 Gy( 1 Gy/min) except the negative control group. Mice of drug positive control group were given intraperitoneal injection with 200 mg/kg bw amifostine half an hour before irradiation. After exposure,mice were treated by the above gavages once daily. After 3 and 8 days,6 mice were randomly selected to collect the peripheral blood for the count of white blood cell( WBC) and lymph cell measuring. RESULTS: i) The cerium concentration in blood reached peak value in 4. 0 hours after exposure of nano-CeO_2,and the cerium concentration of urine and excrement reached maximum in8. 0 hours after exposure. After 24. 0 hours of exposure,the cerium concentration of brain in experiment group was higher than that of control group( P < 0. 05). Among the experiment group,the cerium concentrations of sternum,duodenum and brain were higher than that of kidney and heart( P < 0. 05),meanwhile the cerium concentrations of thymus and lung were higher than that of kidney( P < 0. 05). ii) There was no statistical difference in interactive effect of WBC count and lymph cell counts between nano-CeO_2 exposure ways and time( P > 0. 05). The WBC counts of the low-and medium-dose groups were lower than those of the negative control group and the drug positive control group( P < 0. 05). The WBC count of high-dose group was lower than those of irradiation control group,drug positive control group and medium-dose group( P <0. 05). The lymph cell counts of the 3 dose groups were lower than that of drug positive control group( P < 0. 05).CONCLUSION: The nano-CeO_2 is mainly cumulated in organs such as sternum,duodenum,brain,thymus and lung. After induced by radiation,nano-CeO_2 has a certain degree of promotion role in increasing the WBC counts.