1.Effects of exogenous biliverdin on expression of Litaf in PC12 cells subjected to oxygen-glucose deprivation and restoration
Junjie LI ; Zhiyao ZOU ; Lijia PENG ; Jing LUO ; Li XIONG ; Jia LIU ; Tinghua WANG ; Jianlin SHAO
Chinese Journal of Anesthesiology 2020;40(3):347-349
Objective:To evaluate the effects of exogenous biliverdin (BV) on the expression of Litaf in PC12 cells subjected to oxygen-glucose deprivation and restoration (OGD/R).Methods:PC12 cells were seeded in a 96-well cell culture plate at a density of 1×10 4 cells/well for 3 days and were divided into 3 groups ( n=18 each) by a random number table method: control group (group C), OGD/R group, and biliverdin group (BV group). Group C was incubated in a 37 ℃ incubator (95% air+ 5%CO 2) for 6 h. To establish the OGD/R model, cells were incubated with sugar-free medium in a 37 ℃ incubator (95% air+ 5%CO 2) for 2 h, and the medium was then replaced with normal medium and cells were continuously incubated in a 37 ℃ incubator (95% N 2+ 5% CO 2). In BV group, 2 μg/ml biliverdin was added immediately after oxygen-glucose restoration.Cells in 6 wells in each group were selected at 6 h of restoration for determination of the expression of Litaf protein and mRNA (by real-time polymerase chain reaction) and tumor necrosis factor-alpha (TNF-α) concentration (by enzyme-linked immunosorbent assay). Results:Compared with group C, the expression of Litaf protein and mRNA was significantly up-regulated, and TNF-α concentration in supernatant was increased in group OGD/R ( P<0.05). Compared with group OGD/R, the expression of Litaf protein and mRNA was significantly down-regulated, and TNF-α concentration in supernatant was decreased in group BV ( P<0.05). Conclusion:The mechanism by which exogenous biliverdin reduces OGD/R damage to PC12 cells is related to inhibiting up-regulated expression of Litaf and alleviating the inflammatory responses.
2.Effect of propofol anesthesia on autophagy in hippocampal neurons of newborn rats
Cheng CHANG ; Lijia PENG ; Jing LUO ; Junjie LI ; Jia LIU ; Tinghua WANG ; Jianlin SHAO
Chinese Journal of Anesthesiology 2020;40(6):669-671
Objective:To evaluate the effect of propofol anesthesia on autophagy in hippocampal neurons of newborn rats.Methods:Thirty-nine healthy Sprague-Dawley rats, aged 7 days, weighing 10-12 g, were divided into 3 groups ( n=13 each) using a random number table method: control group (group C), fat emulsion group (group F) and propofol group (group P). Normal saline 8 ml/kg was intraperitoneally injected for 5 consecutive days in group C. Medium-/long-chain fatty emulsion injection 8 ml/kg was intraperitoneally injected for 5 consecutive days in group F. Medium-/long-chain propofol injection 80 mg/kg was intraperitoneally injected for 5 consecutive days in group P. Five rats were sacrificed on 1st day after the end of propofol anesthesia, and hippocampal tissues were taken for determination of the expression of microtubule-associated protein 1 light chain 3B (LC3B) and Beclin-1 (by Western blot). The remaining rats in each group underwent the Morris water maze test on 19th day after the end of propofol anesthesia (30 days after birth), and the escape latency, percentage of time of staying at the target quadrant and the number of crossing the original platform were recorded. Results:Compared with group C, no significant change was found in the expression of hippocampal LC3B and Beclin-1, escape latency, percentage of time of staying at the target quadrant, and the number of crossing the original platform in group F ( P>0.05), and the expression of hippocampal LC3B and Beclin-1 was significantly up-regulated, the escape latency was prolonged, percentage of time of staying at the target quadrant was decreased, and the number of crossing the original platform was decreased in group P ( P<0.05 or 0.01). Conclusion:The mechanism by which propofol anesthesia causes long-term cognitive dysfunction may be related to promoting autophagy in hippocampal neurons of newborn rats.
3.Effect of different treadmill training on cognitive function and TGF-β1 expression in rats
Xiujuan DONG ; Na LIN ; Hangping WANG ; Zhiwei CHEN ; Ruishen XIE ; Chaohui LIANG ; Shuhua SONG ; Jian LIU ; Tinghua WANG
Chinese Journal of Behavioral Medicine and Brain Science 2016;25(12):1063-1067
Objective To observe the effects of different modes treadmill training on cognitive func-tion and transforming growth factor β1 ( TGF-β1 ) expression in cerebral cortex of rats. Methods Two months old rats were divided into the control group,piecewise training group and intermittent training group ( n=10 in each group) . The training was performed five times a week for 6 weeks. Learning and memory a-bility of all rats was detected by water maze at 6 weeks after the training. TGF-β1 expression and localization in cerebral cortex was tested by QRT-PCR and immunofluorescence, respectively. Results The platform time in piecewise group ((30±28) s) and intermittent group ((25±23)s) was both significantly shorter than that in control group ((58±50)s). In the space exploration,the time around Ⅳ quadrant platform in piecewise group((23.6±3.9)s) and intermittent group ((24.3±8.9)s) was significantly higher than that in the control group((17.7±2.0)s). The expression of TGF-β1 mRNA in cerebral cortex in intermittent group (0.0067±0.0043)was obviously higher than that in piecewise group (0.0035±0.0006) and control group (0.0041±0.001). TGF-β1 was located in cell membrane and cytoplasm,and the relative optical density of intermittent group (0.0045±0.0017) was significantly higher than that of control group (0.0019±0.0004) and staging group (0.00175±0.00045). Conclusion (1)Learning and memory function both were im-proved after treadmill six weeks with piecewise and intermittent training models. ( 2) The level of TGF-β1 gene and protein was significantly increased after interval training in cortex of rats.
4.Bone marrow mononuclear cell transplantation for repair of traumatic brain injury via different approaches
Nan ZHAO ; Jun LIU ; Junyan LI ; Gang MA ; Jin LI ; Tinghua WANG ; Ping SU
Chinese Journal of Tissue Engineering Research 2015;(28):4531-4536
BACKGROUND:There are several routes for stem cel transplantation;however, it is stil unable to determine which one is the best. As for the different individuals with brain injury, the type of transplanted cel s, transplantation route and time wil affect the therapeutic effects.
OBJECTIVE:To investigate the effect of bone marrow mononuclear cel s transplanted via different approaches on neurological function of rats with traumatic brain injury.
METHODS:Bone marrow mononuclear cel s of rats were administered gradient centrifugation with Ficol lymphocyte separation medium, and were labeled with CFDA-SE in vitro as standby. Rat models of traumatic brain injury were established by the method of freefal . After successful establishment of rat models, bone marrow mononuclear cel s labeled with CFDA-SE were immediately transplanted into rats via injured area, lateral ventricle and internal carotid artery. One control group was designated for each transplantation route (bone marrow mononuclear cel s were replaced with the same volume of DMEM). The degree of neurological deficits was evaluated using mNSS scores at different time points after treatment. The brain tissue was harvested after the last neurobehavioral evaluation. The survival and migration of bone marrow mononuclear cel s in the injured area were observed under an inverted fluorescent microscope.
RESULTS AND CONCLUSION:At 7, 10, and 14 days after treatment, the mNSS scores of rats in al groups were al lower than those at 1 and 3 days (P<0.05). At 7 and 10 days, the mNSS scores of rats in the internal carotid artery transplantation group were significantly lower than those in the control group (P<0.05). At 14 days after treatment, the number of fluorescence-labeled cel s of rats in the internal carotid artery transplantation group was greater than that in the other groups (P<0.05) and these labeled cel s were widely distributed. The results demonstrate that the neurological function of rats can be improved by transplanting bone marrow mononuclear cel s via the internal carotid artery, and a large number of transplanted cel s can survive and migrate in the injured area.
5.Effect of Gold Belt combined with methylprednisolone on motor function and brain-derived neurotrophic factor expression in rats following traumatic spinal cord injury.
Cuiying LI ; Zhengguang XU ; Tinghua WANG
Journal of Southern Medical University 2015;35(2):276-280
OBJECTIVETo study the effects of a Gold Belt (GB, a traditional Chinese herbal medicine) combined with methyl-prednisolone (MP) on the motor function and brain-derived neurotrophic factor (BDNF) expression in rats with contusive spinal cord injury (SCI).
METHODSThirty adult female SD rats were randomly divided into 5 equal groups, namely the sham-operated group, SCI group, SCI with MP treatment group (MP group, with intramuscular injection of 50 mg/kg MP within 8 hours after SCI and then dosage reduced 10 mg/kg daily), SCI with GB treatment group (GB group, with intragastric gavage of GB 50 mg/kg once daily for 7 days), and combined GB and MP treatment group. The Basso, Beattie and Bresnahan (BBB) locomotor scale was used to evaluate the hindlimb motor function of the rats on days 1, 3, 7, 14, 21 and 28 after the injury. After the last evaluation the rats were sacrificed for immunohistochemistry to observe the localization of BDNF in the ventral and dorsal horn of spinal cord.
RESULTSBDNF were distributed mainly in neurons in the spinal cord grey matter ventral horn and dorsal horn of the rats. The number of BDNF-positive neurons and BBB scores in the combined treatment group were significantly higher than those in the other 4 groups (P<0.05).
CONCLUSIONGB combined with MP produces better therapeutic effects for treating SCI than GB or MP used alone, and such effects are probably related with enhanced BDNF expression in the spinal cord.
Animals ; Brain-Derived Neurotrophic Factor ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Female ; Immunohistochemistry ; Methylprednisolone ; pharmacology ; Neurons ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; drug therapy ; metabolism
6.Effect of Gold Belt combined with methylprednisolone on motor function and brain-derived neurotrophic factor expression in rats following traumatic spinal cord injury
Cuiying LI ; Zhengguang XU ; Tinghua WANG
Journal of Southern Medical University 2015;(2):276-280
Objective To study the effects of a Gold Belt (GB, a traditional Chinese herbal medicine) combined with methyl-prednisolone (MP) on the motor function and brain-derived neurotrophic factor (BDNF) expression in rats with contusive spinal cord injury (SCI). Methods Thirty adult female SD rats were randomly divided into 5 equal groups, namely the sham-operated group, SCI group, SCI with MP treatment group (MP group, with intramuscular injection of 50 mg/kg MP within 8 hours after SCI and then dosage reduced 10 mg/kg daily), SCI with GB treatment group (GB group, with intragastric gavage of GB 50 mg/kg once daily for 7 days), and combined GB and MP treatment group. The Basso, Beattie & Bresnahan (BBB) locomotor scale was used to evaluate the hindlimb motor function of the rats on days 1, 3, 7, 14, 21 and 28 after the injury. After the last evaluation the rats were sacrificed for immunohistochemistry to observe the localization of BDNF in the ventral and dorsal horn of spinal cord. Results BDNF were distributed mainly in neurons in the spinal cord grey matter ventral horn and dorsal horn of the rats. The number of BDNF-positive neurons and BBB scores in the combined treatment group were significantly higher than those in the other 4 groups (P<0.05). Conclusion GB combined with MP produces better therapeutic effects for treating SCI than GB or MP used alone, and such effects are probably related with enhanced BDNF expression in the spinal cord.
7.Effect of Gold Belt combined with methylprednisolone on motor function and brain-derived neurotrophic factor expression in rats following traumatic spinal cord injury
Cuiying LI ; Zhengguang XU ; Tinghua WANG
Journal of Southern Medical University 2015;(2):276-280
Objective To study the effects of a Gold Belt (GB, a traditional Chinese herbal medicine) combined with methyl-prednisolone (MP) on the motor function and brain-derived neurotrophic factor (BDNF) expression in rats with contusive spinal cord injury (SCI). Methods Thirty adult female SD rats were randomly divided into 5 equal groups, namely the sham-operated group, SCI group, SCI with MP treatment group (MP group, with intramuscular injection of 50 mg/kg MP within 8 hours after SCI and then dosage reduced 10 mg/kg daily), SCI with GB treatment group (GB group, with intragastric gavage of GB 50 mg/kg once daily for 7 days), and combined GB and MP treatment group. The Basso, Beattie & Bresnahan (BBB) locomotor scale was used to evaluate the hindlimb motor function of the rats on days 1, 3, 7, 14, 21 and 28 after the injury. After the last evaluation the rats were sacrificed for immunohistochemistry to observe the localization of BDNF in the ventral and dorsal horn of spinal cord. Results BDNF were distributed mainly in neurons in the spinal cord grey matter ventral horn and dorsal horn of the rats. The number of BDNF-positive neurons and BBB scores in the combined treatment group were significantly higher than those in the other 4 groups (P<0.05). Conclusion GB combined with MP produces better therapeutic effects for treating SCI than GB or MP used alone, and such effects are probably related with enhanced BDNF expression in the spinal cord.
8.CXCR4 and Nrf2 expressions in non-small cell lung cancer and their clinical implications.
Tinghua HU ; Yu YAO ; Shuo YU ; Hui GUO ; Tao TIAN ; Lili HAN ; Wenjuan WANG ; Qianqian GUO ; Jing WANG ; Kejun NAN ; Shuhong WANG
Journal of Southern Medical University 2014;34(2):153-158
OBJECTIVETo detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC.
METHODSWe investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immunohistochemistry and real-time PCR.
RESULTSThe expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis.
CONCLUSIONAbnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.
Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; NF-E2-Related Factor 2 ; metabolism ; Neoplasm Staging ; Receptors, CXCR4 ; metabolism ; Signal Transduction
9.CXCR4 and Nrf2 expressions in non-small cell lung cancer and their clinical implications
Tinghua HU ; Yu YAO ; Shuo YU ; Hui GUO ; Tao TIAN ; Lili HAN ; Wenjuan WANG ; Qianqian GUO ; Jing WANG ; Kejun NAN ; Shuhong WANG
Journal of Southern Medical University 2014;(2):153-158
Objective To detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC. Methods We investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immumohistochemistry and real-time PCR. Results The expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis. Conclusion Abnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.
10.CXCR4 and Nrf2 expressions in non-small cell lung cancer and their clinical implications
Tinghua HU ; Yu YAO ; Shuo YU ; Hui GUO ; Tao TIAN ; Lili HAN ; Wenjuan WANG ; Qianqian GUO ; Jing WANG ; Kejun NAN ; Shuhong WANG
Journal of Southern Medical University 2014;(2):153-158
Objective To detect the expressions of CXCR4 and Nrf2 in non-small cell lung cancer (NSCLC) tissues and analyze their association with the clinicopathological features of NSCLC. Methods We investigated the expressions of CXCR4 and Nrf2 in 66 NSCLC and corresponding distant normal tissue specimens using immumohistochemistry and real-time PCR. Results The expressions of CXCR4 protein and mRNA were significantly higher in NSCLC tissue specimens than in the distant normal tissues, while the expression of Nrf2 protein and mRNA increased significantly in NSCLC tissues compared to those in the distant normal tissues (P<0.01). A high expression level of CXCR4 was positively correlated with a large tumor size (P=0.048), poor differentiation (P=0.024), advanced TNM stage (P=0.018), lymph node metastasis (P=0.004), and distant metastasis (P=0.016). The expression of Nrf2 protein was positively correlated with a large tumor size (P=0.008), advanced TNM stage (P=0.028), lymph node metastasis (P=0.038), and distant metastasis (P=0.023). A strong correlation was found between CXCR4 and Nrf2 expressions in NSCLC tissues (r=0.324, P<0.01), and the co-expression of CXCR4 and Nrf2 was strongly correlated with lymph node metastasis and distant metastasis. Conclusion Abnormal expressions of CXCR4 and Nrf2 may contribute to the progression and malignant biological behavior of NSCLC.

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