1.Effects of statins on inflammatory reaction in chronic renal failure patients
Yuan YUAN ; Keliang SU ; Wei CHEN
Chinese Journal of Primary Medicine and Pharmacy 2008;15(8):1327-1328
Objective To observe the changes of serum CRP and plasma Fg in patients with chronic renal failure(CRF),to explore the effect of statins on inflammaory reaction in CRF patients.Methods 54 patients with CRF were randomly divided into non-statins group(routine therapy);atstina group(routine therapy plus simvastatin 20mg/d or pravastatin 20mg/d).Besides,a healthy control group consisted of 20 subjects was set up as control group.The changes of serum of CRP and plasma Fg of all groups before and four weeks after treatment were recorded.Results The serum CRP and plasma Fg levels increased in CRF patients,Which were significantly higher as compared to the control group.After treatment for four weeks,the level of CRP,Fg of matins group decreased significantly.The levels of CRP,Fg had no statistical changes in non-statins group.As compared to non-statins group,the differences of CRP,Fg levels after treatment in statins group were statistically significant respectively.Conclusions(1)Inilammaory reaction is a common condition in non-dialysis patients with CRF;(2)ststins show effects on decrease of CRP,Fg level in CRF patients,independently on the effect of decreasing hypedipemia.
2.Role of autophagy in activation of astrocytes in spinal cord of rats with neuropathic pain
Hongguang CHEN ; Yajun CHEN ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2015;35(11):1307-1309
Objective To evaluate the role of autophagy in activation of astrocytes in the spinal cord of rats with neuropathic pain (NP).Methods One hundred thirty-six male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 4 groups (n =34 each) using a random number table: sham operation group (group S), group NP, autophagy inhibitor 3-methyladenine (3-MA) group (group MA), and autophagy activator rapamycin group (group R).NP was induced by chronic constriction injury.In 3-MA and R groups, 3-MA 15 mg/kg and rapacymin 10 mg/kg were injected intraperitoneally, respectively, at 1 h before NP.Ten rats were randomly selected, and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before NP and at 1, 3, 7 and 14 days after NP.Before NP and at 1, 3 and 7 days after NP, 6 rats were randomly sacrificed, the L4-6 segments of the spinal cord was harvested to detect microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ), Beclin-1 and glial fibrillary acidic protein (GFAP) expression by Western blot.Results Compared with group S, the MWT was significantly decreased, the TWL was shortened, and the expression of LC3 Ⅱ , Beclin-1 and GFAP was up-regulated at each time point after NP in group NP (P< 0.05).Compared with group NP, the MWT was significantly decreased, the TWL was shortened, the expression of LC3 Ⅱ and Beclin-1 was down-regulated, and the expression of GFAP was up-regulated at each time point after NP in group MA, and the MWT was significantly increased, the TWL was prolonged, the expression of LC3 Ⅱ and Beclin-1 was up-regulated, and the expression of GFAP was down-regulated at each time point after NP in group R (P<0.05).Conclusion Autophagy is involved in the development and maintenance of NP through promoting the activation of astrocytes in the spinal cord of rats.
3.Effects of vitamin K 2 on sevoflurane-induced cognitive decline in aged mice
Yaxin WANG ; Lingling LIU ; Keliang XIE ; Jun CHEN
Chinese Journal of Anesthesiology 2021;41(2):165-168
Objective:To evaluate the effects of vitamin K 2 on sevoflurane-induced cognitive decline in aged mice. Methods:A total of 72 SPF healthy female C57BL/6J mice, aged 12 months, weighing 20-25 g, were divided into 4 groups ( n=18 each) using a random number table method: control+ corn oil group (group Con+ Oil), sevoflurane+ corn oil group (group Sevo+ Oil), control+ vitamin K 2 group (group Con+ K 2) and sevoflurane+ vitamin K 2 group (group Sevo+ K 2). The mice in Sevo+ Oil and Sevo+ K 2 groups were anesthetized with 2.5% sevoflurane+ 33% oxygen for 2 h. The mice in Con+ Oil and Con+ K 2 groups were treated with 33% oxygen only.The animals in Con+ Oil and Sevo+ Oil groups were intraperitoneally injected with corn oil 100 μl at 30 min before oxygen or sevoflurane inhalation.Vitamin K 2 (dissolved in corn oil, concentration 1 mg/ml) 100 mg/kg was injected intraperitoneally in Con+ K 2 and Sevo+ K 2 groups.At 24 h after sevoflurane inhalation, 8 mice from each group were randomly selected and sacrificed, and the hippocampal tissues were removed for determination of activity of ATPase, contents of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay) and the expression of AT8 and PHF1 (by Western blot). The remaining 10 mice in each group received standardized feeding, and the cognitive function was assessed using Y-maze at 1, 3, 5, 7 and 14 days after sevoflurane inhalation. Results:Compared with group Con+ Oil, the contents of IL-1β, IL-6 and TNF-α were significantly increased, expression of AT8 and PHF1 were up-regulated, activity of ATPase was decreased, and spontaneous alternation percentage was decreased at 1, 3, 5, 7 and 14 days after sevoflurane inhalation in group Sevo+ Oil ( P<0.05). Compared with group Sevo+ Oil, the contents of IL-1β, IL-6 and TNF-α were significantly decreased, expression of AT8 and PHF1 were down-regulated, activity of ATPase was increased, and spontaneous alternation percentage was increased at 1, 3, 5, 7 and 14 days in group Sevo+ K 2 ( P<0.05). There was no significant difference in the above indicators between group Con+ K 2 and group Sevo+ K 2 ( P>0.05). Conclusion:Vitamin K 2 can improve sevoflurane-induced cognitive decline in aged mice, the mechanism is related to increasing activity of ATPase and inhibiting the up-regulation of AT8 and PHF1 expression in hippocampus.
4.Propofol inhibits the metastasis of tumor cells via activating PI3K/Akt signaling pathway
Jun CHEN ; Wenhui ZHAO ; Lingling LIU ; Keliang XIE ; Zhangjun SONG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;(2):226-229
ABSTRACT:Objective To study the effects of propofol on the metastasis of tumor cells related PI3K/Akt signaling pathway.Methods The breast cancer model was established by transplanting human derived breast cancer cell lines into immunodeficient mice with naked gene.The mice,inoculated successfully,were randomly divided into 4 groups:control group (C group,n =6),propofol group (P group,n =6),propofol+PI3K inhibitor (BYL71 9)group (P+B group,n =6),and PI3K inhibitor group (BYL71 9)(B group,n =6).The expressions of PI3K,p-Akt and Akt were examined by Western blot at week 4 after administration;the gene levels of PI3KR1, Akt1 and Akt2 were detected by RT-PCR at week 4 after administration;the number of metastatic lung nodules from both lungs was also observed at week 4 after administration.Results Compared with those in C group,the expressions of PI3K and p-Akt were significantly higher in P group (P <0.05),the level of PI3KR1 mRNA but not Akt1 and Akt2 mRNA was significantly increased(P < 0.05 ),and metastatic lung nodules significantly decreased (P <0.05).In B group,the expressions of PI3K and p-Akt were significantly decreased (P <0.05 ),the levels of PI3KR1,Akt1 and Akt2 mRNA were not significantly increased (P >0.05),but metastatic lung nodules significantly increased (P < 0.05 ).Compared with those in B group,in P+ B group the expressions of PI3K and p-Akt were markedly higher (P <0.05),the level of PI3KR1 mRNA but not Akt1 and Akt2 mRNA was significantly increased (P <0.05),and metastatic lung nodules significantly decreased (P <0.05).Conclusion Propofol can inhibit the metastasis of tumor cells through the upregulated and activated PI3K/Akt signaling pathway.
5.Effect of hydrogen on expression of nuclear factor E2-related factor 2 during endotoxin-induced oxi-dative injury to macrophages
Gang LIU ; Hongguang CHEN ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2016;36(9):1150-1152
Objective To evaluate the effect of hydrogen on the expression of nuclear factor E2?related factor 2 ( Nrf2 ) during endotoxin?induced oxidative injury to macrophages. Methods Normally cultured Raw264.7 cells were divided into 4 groups ( n=36 each) using a random number table: control group (group C), hydrogen?rich saline group ( group H), endotoxin group ( group E) and endotoxin plus hydrogen?rich saline group (group EH). In E and EH groups, endotoxin was added with the final concentration of 1 μg∕ml. Hydrogen?rich saline ( hydrogen concentration 0. 06 mmol∕L) was added in H and EH groups. All the cells were incubated for 48 h. At 6, 24 and 48 h of incubation, cells were collect?ed to detect the activity of reactive oxygen species ( ROS) , and cells were collected and proteins extracted for determination of superoxide dismutase ( SOD) and catalase ( CAT) activities and Nrf2 expression by Western blot. Results Compared with group C, the ROS activity was significantly increased, the levels of SOD and CAT were significantly decreased, and the expression of Nrf2 was significantly up?regulated in E and EH groups (P<0.05), and no significant change was found in the parameters mentioned above in group H (P>0.05). Compared with group E, the ROS activity was significantly decreased, the levels of SOD and CAT were significantly increased, and the expression of Nrf2 was significantly up?regulated in group EH (P<0.05). Conclusion Hydrogen can attenuate endotoxin?induced oxidative injury to macro?phages, and the mechanism may be related to up?regulated expression of Nrf2.
6.Role of Nrf2∕ARE signaling pathway in inhibition of LPS-induced inflammatory factor release from macrophages by hydrogen
Zhiyong WANG ; Hongguang CHEN ; Lu WANG ; Yonghao YU ; Keliang XIE
Chinese Journal of Anesthesiology 2016;36(11):1393-1396
Objective To evaluate the role of nuclear factor erythroid 2?related factor 2 ( Nrf2)∕antioxidant response element( ARE) signaling pathway in inhibition of lipopolysaccharide ( LPS)?induced inflammatory factor release from macrophages by hydrogen. Methods RAW264. 7 macrophages of mice were cultured in 6?well plates (2×106 cells∕well) and were divided into 4 groups (n=24 each) using a random number table: control group ( group C); LPS group; hydrogen?rich saline+LPS group ( group LPS+H2); Nrf2 small interference RNA (siRNA)+LPS+hydrogen?rich saline group (siRNA+LPS+H2 group) . LPS 1 μg∕ml was added in group LPS. In group LPS+H2 , LPS 1μg∕ml was added, and the cul?ture medium was then replaced with the culture medium containing 0. 6 mmol∕L hydrogen?rich saline. In group siRNA+LPS+H2 , after Nrf2?siRN was successfully transfected into the cells, the cells were continu?ously incubated for 24 h, and the culture medium was then replaced with the culture medium containing 0.6 mmol∕L hydrogen?rich saline after LPS 1 μg∕ml was added. At 24 h of incubation, the supernatant was sep?arated for determination of the lactic dehydrogenase (LDH) activity (using colorimetric method) and for detection of the concentrations of tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) , high mobility group box?1 (HMGB1) and IL?6 (by ELISA). The cells were collected for measurement of the proliferation of cells ( by methyl thiazolyl tetrazolium assay) and for determination of the expression of Nrf2 and heme oxygenase?1 ( HO?1) in cells ( by Western blot) . Results Compared with group C, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly in?creased, the proliferation of cells was significantly decreased, and the expression of HO?1 in cells was sig?nificantly up?regulated in LPS and siRNA+LPS+H2 groups, and the expression of Nrf2 in cells was signifi?cantly up?regulated in LPS and LPS+H2 groups (P<0.05). Compared with group LPS, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly decreased, the proliferation of cells was significantly increased, and the expression of Nrf2 and HO?1 in cells was sig?nificantly up?regulated in group LPS+H2 , and the expression of Nrf2 and HO?1 in cells was significantly down?regulated in group siRNA+LPS+H2 ( P<0.05) . Compared with group LPS+H2 , the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly increased, the proliferation of cells was significantly decreased, and the expression of Nrf2 and HO?1 in cells was signifi?cantly down?regulated in group LPS+H2+siRNA ( P<0.05) . Conclusion The mechanism by which hydro?gen inhibits LPS?induced inflammatory factor release from macrophages is related to the activation of Nrf2∕ARE signaling pathway in mice.
7.The 4 year follow-up investigation of cognitive function in patients after acute stroke
Qingsong WANG ; Wenping GONG ; Keliang CHEN ; Qihao GUO
Chinese Journal of Geriatrics 2016;35(8):824-827
Objective To investigate the 4-year follow up of cognitive function outcomes and characteristics in patients after stroke.Methods Sixty three cases according with the diagnostic standard of acute unifocal subcortical stroke were consecutively collected in neurological ward from December 2009 to November 2010.They were followed up for average four years.Forty one out of them completed the neuropsychology tests identical to the baseline,which covered the general cognition function,attention,execution,memory,language,spatial,etc.According to the standard of clinical diagnosis,cognition function is divided into five degrees,including normal,VCI-ND,mild VaD,moderate VaD,and severe VaD.The improved group had 13 cases whose cognition function was improved by one or more ranks.The progressive group had 12 cases whose cognition function progressed by one or more ranks.The stable group had 16 cases whose cognition function remained the same as the baseline.Results According to qualitative analysis on the baseline versus 4-year follow-up outcome,in 13 improved cases,8 were VCI-ND and 5 were mild VaD.In 16 stable cases,11 were normal,4 were VCI-ND and 1 was mild VaD.In 12 progressive cases,3 were normal (change to mild VaD after follow-up),5 were VCI-ND (change to mild and moderate VaD after follow-up) and 4 were mild VaD (change to moderate VaD after 4-year follow-up).In the comparison of baseline cognition function among the improved,progressive and stable group,there was only one significantly different score (the right number of SCWT-A) in the improved and progressive group.The cognition function of improved group had significant differences in CFT-copy,right number of SCWT-C and the time of TMT-B before versus after follow-up.The cognition function of progressive group had significant differences in AVLT-Delay Recall and CFT-Recall.Conclusion Long-term cognitive function outcome after stroke is heterogenetic.The location of cognitive impairment or progression is not the same model for different cognitive outcome group.
8.Hydrogen-rich saline ameliorates the expressions of cytokines in a rat model of inflammatory pain through activating heme oxygenase-1
Guofeng ZHANG ; Hongguang CHEN ; Yonghao YU ; Guolin WANG ; Keliang XIE
Journal of Chinese Physician 2016;18(4):487-491
Objective To investigate whether hydrogen-rich saline could ameliorate the expressions of cytokines in a rat model of inflammatory pain through activating heme oxygenase-1 (HO-1).Methods Eighty male Sprague-Dawley (SD) rats,weighing 180 ~ 220 g,were randomly divided into five groups (n =16 in each group):Control group (Con),inflammation pain group (CFA),inflammation pain + hydrogen-rich saline group (CFA + H2),inflammation pain + HO-1 inhibitor Znpp-Ⅸ group (CFA + Znpp-Ⅸ),and inflammation pain + hydrogen-rich saline + HO-1inhibitor Znpp-Ⅸ group (CFA + H2 + Znpp-Ⅸ).The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were tested on days 1 (T1),2 (T2),3 (T3),5 (T4),7 (T5),and 14 (T6) after inflammation pain.The expressions of spinal HO-1 mRNA and protein were measured with real-time quantitative polymerase chain reaction (RT-PCR) and Western blot,and spinal inflammatory cytokines were measured with enzyme-linked immunosorbent assay (ELISA) on day 7 after inflammatory pain.Results Compared to Con group,MWT and TWL were significantly reduced;the spinal HO-1 mRNA level,protein expression and activity were increased;and the levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1β,IL-6 and IL-10 in spinal tissues were also increased in CFA group (P < 0.05).Compared to CFA group,MWT and TWL were significantly increased;the spinal HO-1 mRNA level,protein expression and activity were further increased;and the levels of TNF-α,IL-1β and IL-6 were decreased,while IL-10 was further increased in CFA + H2group (P < 0.05).Compared to CFA + H2 group,MWT and TWL were decreased;the spinal HO-1 mR-NA level,protein expression and activity were decreased;and the levels of TNF-α,IL-1β and IL-6 in spinal tissue were significantly increased,while IL-10 was decreased in CFA + H2 + Znpp-Ⅸ group (P <0.05).Conclusions Hydrogen-rich saline can ameliorate the mechanical and thermal allodynia in a rat model of inflammatory pain,and reduce the release of inflammatory cytokines via activating HO-1.
9.Effect of combination therapy with propofol and hydrogen-rich saline on organ damage and cytokines in a murine model of sepsis
Yunchuan HONG ; Hongguang CHEN ; Yonghao YU ; Keliang XIE
Chinese Critical Care Medicine 2017;29(4):316-320
Objective To investigate the effect of combining propofol with hydrogen on organ damage and inflammation of sepsis in cecal ligation and puncture (CLP) mice model.Methods One hundred and forty male C57BL/6 mice were randomly divided into groups (n = 28): sham group, CLP group, propofol group, H2 group, and propofol and H2 group. The sepsis was induced by CLP operation. Mice in sham group did the same operation with ligation and puncture. The mice of propofol group and propofol and H2 group were given 50 mg/kg propofol through tail vein at 1 hour and 6 hours after CLP and the mice of H2 group and propofol and H2 group were given 5 mL/kg H2-rich saline i.p. at 1 hour and 6 hours after CLP. The survival rates were observed during 7 days in twenty mice of each group. Inferior vena cava blood and part lung, liver and kidney tissue were collected for detection of the concentration of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and high mobility group box 1 (HMGB1) at 24 hours after CLP in the 40 animals left (eachn = 8). Then, the rest tissue of lung, liver and kidney tissue were harvested to test histopathology and histological score.Results The 1, 2, 3, 5, 7 days survival rate of septic mice were 80%, 40%, 20%, 10%, and 0%, respectively. The survival rate of animals increased significantly after propofol or hydrogen-rich treatment, and the combined treatment can further increase survival rate to 90%, 75%, 60%, 55%, and 55%, respectively. Compared with the sham group, inflammatory factors were significantly increased in blood and organ tissues, cell degeneration, necrosis, congestion and inflammatory cell infiltration in lung, liver and kidney, and tissues histological scores were significantly increased. The levels of inflammatory factors were reduced in blood and tissues, cell degeneration, necrosis, congestion and inflammatory cell infiltration were alleviated in lung, liver and kidney, and tissues histological scores were decreased after propofol or hydrogen-rich treatment compared with CLP group; these indicators were further improved in propofol and H2 group compared with propofol group or H2 group [2, 3, 5, 7-day survival rate: 75% vs. 60%, 65%; 60% vs. 50%, 50%; 55% vs. 45%, 40%; 55% vs. 40%, 40%; blood TNF-α (ng/L): 367±74 vs. 612±132, 588±117; blood IL-1β (ng/L): 321±68 vs. 502±95, 476±86; blood HMGB1 (μg/L): 4.6±0.9 vs. 7.0±1.4, 6.8±1.3; lung TNF-α(ng/g): 307±70 vs. 512±132, 488±102; lung IL-1β (ng/g): 367±77 vs. 571±108, 466±89; lung HMGB1 (μg/g):5.1±1.0 vs. 7.8±1.7, 7.1±1.5; liver TNF-α (ng/g): 247±57 vs. 431±112, 389±87; liver IL-1β (ng/g): 267±58 vs. 417±85, 399±76; liver HMGB1 (μg/g): 4.2±1.1 vs. 7.1±1.6, 6.6±1.2; kidney TNF-α (ng/g): 257±41 vs. 480±89, 448±82; kidney IL-1β (ng/g): 258±39 vs. 409±68, 411±66; kidney HMGB1 (μg/g): 3.9±0.7 vs. 6.8±1.2, 5.7±1.0; histological scores: lung: 1.22±0.28 vs. 2.61±0.49, 2.58±0.44; liver: 1.38±0.32 vs. 2.76±0.51, 2.62±0.46; kidney: 1.19±0.25 vs. 2.43±0.41, 2.36±0.40; allP < 0.05].Conclusions Both propofol and H2 can improve the survival rate of sepsis, reduce tissue damage and the release of cytokines, and combined application of the two treatment was better.
10.Role of autophagy in lung injury in septic mice
Aili DONG ; Hongguang CHEN ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2015;35(9):1124-1127
Objective To investigate the role of autophagy in the lung injury in the septic mice.Methods Thirty-six male C57BL/6 mice, aged 6 weeks, weighing 20-25 g, were randomly divided into 3 groups (n=12 each) using a random number table: sham operation group (group S);cecal ligation and puncture (CLP) group;CLP + autophagy inhibitor 3-methyladenine (3-MA) group (group CLP+3-MA).Sepsis was produced by CLP.In group CLP+3-MA, 3-MA 10 mg/kg was injected intraperitoneal at 1 h after operation.Arterial blood samples were taken at 24 h after operation for blood gas analysis, and the oxygenation index was calculated.The lungs were removed for microscopic examination of pathologic changes which were scored, and for determination of wet/dry lung weight ratio (W/D ratio) , myeloperoxidase (MPO) activity (using colorimetric method) and the expression of autophagy protein microtubule-associated protein 1 light chain 3 Ⅱ] (LC3 Ⅱ), Beclin-1 and lysosomes-associated protein Rab7 and lysosome-associated membrane protein-2 (LAMP2) (by Western blot).The lung was lavaged, and broncho-alveolar lavage fluid (BALF) was collected for determination of the total cell count and polymorphonuclear leukocyte (PMN) count.Results Compared with group S, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, and oxygenation index was decreased in CLP and CLP +3-MA groups, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was up-regulated in group CLP (P<0.05).Compared with group CLP, the pathological score, W/D ratio, MPO activity, and the total cell and PMN counts in BALF were significantly increased, the oxygenation index was decreased, and the expression of LC3 Ⅱ , Beclin-1, LAMP2 and Rab7 was down-regulated in group CLP+ 3-MA (P<0.05).Conclusion Autophagy is involved in the endogenous protective mechanism of acute lung injury in the septic mice.