Role of JMJD3 in cisplatin-induced renal fibrosis following acute kidney injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of Jumonji domain-containing 3 (JMJD3) in cisplatin-induced renal fibrosis following acute kidney injury in mice.Methods:Forty-eight healthy C57BL/6 male mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=12 each) using a random number table method: control group (group CON), control plus JMJD3 inhibitor group (group CON-A), cisplatin group (group CIS), and cisplatin plus JMJD3 inhibitor group (group CIS-A). In group CIS and group CIS-A, cisplatin was intraperitoneally administered on 1st and 14th days, respectively, to develop a renal fibrosis model in mice with acute kidney injury, and the JMJD3 inhibitor GSKJ4 10 mg/kg and equal volume of PBS were intraperitoneally injected on 4th day, respectively, once every 3 days, 6 injections in total.The equal volume of PBS and GSKJ4 10 mg/kg were intraperitoneally injected at the corresponding time points in group CON and group CON-A, respectively.Six mice in each group were selected, and orbital blood samples were collected on 3rd day after the first injection of cisplatin to determine the concentrations of serum creatinine (Cr) and blood urea nitrogen (BUN), then the animals were sacrificed, and kidney tissues were obtained for microscopic examination of pathological changes after HE and PAS staining (with a light microscope), and the damage to kidneys was assessed and scored.Six mice were sacrificed on 28th day after the first injection of cisplatin, and kidney tissues were removed for determination of the area of renal fibrosis ( via Sirius red and Masson staining), expression of fibronectin (Fn), collagen type Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA) (by immunofluorescence), F4/80 + cell and CD3 + cell count (using immunohistochemical method), and expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), CXC chemokine ligand 16 (CXCL16), and monocyte chemoattractant protein1 (MCP-1) mRNA (by real-time polymerase chain reaction). Results:Compared with group CON, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly increased, the expression of Fn, Col Ⅰ and α-SMA was up-regulated, the F4/80 + cell and CD3 + cell count was increased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was up-regulated in group CIS ( P<0.05), and no significant change was found in the parameters mentioned above in group CON-A ( P>0.05). Compared with group CON-A, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly increased, the expression of Fn, Col Ⅰ and α-SMA was up-regulated, the F4/80 + cell and CD3 + cell count was increased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was up-regulated in group CIS-A ( P<0.05). Compared with group CIS, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly decreased, the expression of Fn, Col Ⅰ and α-SMA was down-regulated, the F4/80 + cell and CD3 + cell count was decreased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was down-regulated in group CIS-A ( P<0.05). Conclusions:JMJD3 is involved in the process of renal fibrosis following acute kidney injury in mice, and the mechanism may be related to promotion of inflammatory responses.

Role of spinal cord mitochondrial autophagy in alleviation of diabetic neuropathic pain by curcumin in mice / 中华麻醉学杂志

Objective:To evaluate the role of spinal cord mitochondrial autophagy in alleviation of diabetic neuropathic pain (DNP) by curcumin in mice.Methods:SPF healthy male C57BL/6 mice, aged 2 months, weighing 20-25 g, in which DNP model was established by intraperitoneal injection of streptozotocin (STZ) 130 mg/kg, were used in this study.A total of 36 mice with successfully established DNP model were divided into 3 groups ( n=12 each) using the random number table method: DNP group, DNP + curcumin group (DPR group), and DNP + curcumin + cyclosporine A group (DRC group). Another 12 C57BL/6 mice were selected and served as normal control group (NC group), and the equal volume of normal saline was intraperitoneally injected.In group DPR, curcumin 200 mg/kg was administered by intragastric gavage, once a day, for 7 consecutive days.In group DRC, the mitochondrial autophagy inhibitor cyclosporine A 10 mg/kg was intrathecally injected once a day for 7 consecutive days before each administration of curcumin.The equal volume of normal saline was administered by intragastric gavage at the same time point, once a day, for 7 consecutive days in group NC and group DNP.The mechanical paw withdrawal threshold (MWT) was measured before intragastric gavage and at 1, 3, 5 and 7 days after intragastric gavage.After the last behavioral testing, the L 4-6 spinal cord tissues were removed for determination of the mitochondrial membrane potential and ROS content (by JC-1 and DCFH-DA combined with flow cytometry), expression of microtubule-associated protein light chain 3 (LC3), Beclin1 and P62 (by Western blot), and mitochondrial autophagosomes (by transmission electron microscopy) and for microscopic examination of the co-expression of LC3-Ⅱwith mitochondrial translocase outer membrane protein 20 (TOM20) (using immunofluorescence double-labeling technique). Results:Compared with group NC, the MWT and mitochondrial membrane potential were significantly decreased, the ROS content and LC3-Ⅱ/Ⅰ ratio were increased, the expression of Beclin1 was up-regulated, the expression of P62 was down-regulated ( P<0.05), the number of mitophagosomes developed was increased, and the co-expression of LC3-Ⅱwith TOM20 was increased in group DNP.Compared with group DNP, the MWT and mitochondrial membrane potential were significantly increased, the ROS content was decreased, and LC3-Ⅱ/Ⅰ ratio was increased, the expression of Beclin1 was up-regulated, the expression of P62 was down-regulated ( P<0.05), the number of mitophagosomes developed was increased, and the co-expression of LC3-Ⅱwith TOM20 was increased in group DPR.Compared with group DPR, the MWT and mitochondrial membrane potential were significantly decreased, the ROS content was increased, LC3-Ⅱ/Ⅰ ratio was decreased, the expression of Beclin1 was down-regulated, the expression of P62 was up-regulated ( P<0.05), the number of mitophagosomes developed was decreased, and the co-expression of LC3-Ⅱ with TOM20 was decreased in group DRC. Conclusions:The mechanism by which curcumin reduces DNP may be related to the up-regulation of mitochondrial autophagy in the spinal cord and improvement in mitochondrial function in mice.

Relationship between CXCL16 and natural killer T cells during renal fibrosis in mice with acute kidney injury / 中华麻醉学杂志

Objective:To evaluate the relationship between chemokine CXC-ligand 16 (CXCL16) and natural killer T cells during renal fibrosis in mice with acute kidney injury (AKI).Methods:Twenty-four healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), AKI group, control+ rCXCL16 group (group C-rCXCL16) and AKI+ rCXCL16 group.In AKI-rCXCL16 and AKI groups, folic acid 250 mg/kg was intraperitoneally injected to induce AKI in anesthetized mice, and rCXCL16 0.1 mg/kg and the equal volume of solution were intraperitoneally injected, respectively, at 3, 6, 9 and 12 days after injection of folic acid.The equal volume of solution and rCXCL16 were intraperitoneally injected at the corresponding time points in group C and group C-rCXCL16, respectively.The orbital blood samples were taken on day 14 after injection of folic acid for determination of the serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The renal tissues were obtained for measurement of the renal fibrosis size (using Sirius red staining and Masson staining), for determination of the expression of fibronectin (FN), collagen-Ⅲ (Col-Ⅲ) and α-smooth muscle actin (α-SMA) (by immunofluorescence) and expression of interleukin-4 (IL-4), mannose receptor (CD206) and arginase 1 (Arg-1) mRNA (by real-time polymerase chain reaction), and for evaluation of the ratio of CD1d Tetramer + -IL-4 + cells (by flow cytometry). Results:Compared with group C, the serum BUN and Cr concentrations were significantly increased, the renal fibrosis size was increased, the expression of IL-4, CD206, Arg-1 mRNA, FN, Col-Ⅲ and α-SMA was up-regulated, and the ratio of CD1d Tetramer + -IL-4 + cells was increased in AKI and AKI-rCXCL16 groups ( P<0.05), and no significant change was found in the parameters mentioned above in group C-rCXCL16 ( P>0.05). Compared with group AKI, the serum BUN and Cr concentrations were significantly increased, the renal fibrosis size was increased, the expression of IL-4, CD206, Arg-1 mRNA, FN, Col-Ⅲ and α-SMA was up-regulated, and the ratio of CD1d Tetramer + -IL-4 + cells was increased in group AKI-rCXCL16 ( P<0.05). Conclusion:The mechanism by which CXCL16 is involved in the process of renal fibrosis is related to the recruitment of natural killer T cells secreting IL-4 which regulates macrophage M2 polarization in mice with AKI.

Role of natural killer T cells in renal fibrosis in mice with acute kidney injury / 中华麻醉学杂志

Objective:To evaluate the role of natural killer T cells in renal fibrosis in mice with acute kidney injury (AKI).Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), AKI group (group A), control plus CD1d antibody group (group C-MA), and AKI plus CD1d antibody group (group A-MA). The model of renal fibrosis in mice with AKI was established by intraperitoneal injection of folic acid 250 mg/kg.In group C, homotypic control antibody 20 mg/kg was injected via the tail vein.In group AKI, homotypic control antibody 20 mg/kg was injected via the tail vein at 24 h before establishing the model. In group C-MA, anti-CD1d monoclonal antibody 20 mg/kg was injected via the tail vein.In group A-MA, anti-CD1d monoclonal antibody 20 mg/kg was injected via the tail vein at 24 h before establishing the model.On the 14th day after folic acid injection, blood samples were taken from eyeballs to determine the concentrations of blood urea nitrogen (BUN) and creatinine (Cr) in serum.Then the mice were sacrificed, and the renal tissues were taken for Sirius red staining and HE staining to determine the area of renal fibrosis, and renal injury was scored.The expression of fibronectin (FN), type I collagen (Col-Ⅰ) and alpha-smooth muscle actin (α-SMA) in renal tissues was detected by immunofluorescence method.The expression of interleukin (IL)-4, IL-13, arginase-1 (Arg-1) and found in inflammatory zone 1 (FIZZ1) mRNA in renal tissues was detected by real-time polymerase chain reaction. Results:Compared with group C, the concentrations of BUN and Cr in serum, renal injury score, and area of renal fibrosis were significantly increased, the expression of FN, Col-Ⅰ and α-SMA and IL-4, IL-13, Arg-1 and FIZZ1 mRNA was up-regulated in A and A-MA groups ( P<0.05), and no significant change was found in the above indexes in group C-MA ( P>0.05). Compared with group A, the concentrations of BUN and Cr in serum, renal injury score, and area of renal fibrosis were significantly decreased, the expression of FN, Col-Ⅰ and α-SMA and IL-4, IL-13, Arg-1 and FIZZ1 mRNA was down-regulated in group A-MA ( P<0.05). Conclusion:Activation of natural killer T cells is involved in the process of renal fibrosis in mice with AKI, and the mechanism may be related to promoting the release of Th2 cytokines and M2 polarization of macrophages.

Role of CXCR6-mediated activation of natural killer T cells in renal fibrosis following acute kidney injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of CXC chemokine receptor 6 (CXCR6)-mediated activation of natural killer T (NKT) cells in renal fibrosis following acute kidney injury (AKI) in mice.Methods:Eighteen male wild-type C57BL/6 mice and 18 CXCR6 knockout C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 6 groups ( n=6 each) using a random number table method: wild-type mouse control group (group WT-CON), CXCR6 knockout mouse control group (group CXCR6 -/--CON), wild-type mouse with AKI group (group WT-AKI), CXCR6 knockout mouse with AKI group (group CXCR6 -/--AKI), wild-type mouse with AKI + NKT cell adoptive transfer group (group WT-AKI-NKT) and CXCR6 knockout mouse with AKI + NKT cell adoptive transfer group (group CXCR6 -/--AKI-NKT). Folic acid 250 mg/kg was intraperitoneally injected to establish the model of renal fibrosis in mice with AKI.NKT cellsuspension 250 μl(1×10 6 cells) was injected through the tail vein on the 4th and 9th days after folic acid injection in group WT-AKI-NKT and group CXCR6 -/--AKI-NKT, respectively.Blood samples were taken from orbital at day 14 after folic acid injection for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr). The animals were sacrificed, and renal tissues were obtained for observation of the area of renal fibrosis (by Sirius red staining) and renal injury (using H&E staining) which was scored and for determination of the proportion of CD1d Tetramer+ cells (by flow cytometry), the number of CD206 and α-smooth muscle actin (α-SMA) double positive (CD206 + -α-SMA + ) cells (by immunofluorescence) and expression of interleukin (IL)-4 and IL-13 mRNA (by real-time polymerase chain reaction). Results:Compared with group WT-CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI and WT-AKI-NKT ( P<0.05). Compared with group WT-AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group WT-AKI-NKT ( P<0.05), and the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly decreased, and the expression of IL-4 and IL-13 mRNA was down-regulated in group CXCR6 -/--AKI ( P<0.05). Compared with group CXCR6 -/--CON, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased in group CXCR6 -/--AKI and group CXCR6 -/--AKI-NKT ( P<0.05). Compared with group CXCR6 -/--AKI, the BUN and Cr levels, renal injury scores, area of renal fibrosis, proportion of CD1d Tetramer + cells and CD206 + -α-SMA + cell count were significantly increased, and the expression of IL-4 and IL-13 mRNA was up-regulated in group CXCR6 -/--AKI-NKT ( P<0.05). Conclusion:CXCR6-mediated activation of NKT cells is involved in renal fibrosis following AKI in mice, and the mechanism may be related to promoting Th2 cytokine-mediated M2 macrophage-myofibroblast transformation.

Effect of intrathecal insulin-like growth factor-1 on chemotherapy-induced neuropathic pain in mice / 中华麻醉学杂志

Objective:To evaluate the effect of intrathecal insulin-like growth factor-1 (IGF-1) on chemotherapy-induced neuropathic pain (NP) in mice.Methods:Forty clean-grade healthy male C57 mice, aged 7-9 weeks, weighing 22-24 g, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), chemotherapy-induced NP group (group CIPN), low-dose IGF-1 group (group I1) and high-dose IGF-1 group (group I2). In CIPN, I1 and I2 groups, oxaliplatin 5 mg/kg was intraperitoneally injected for 5 consecutive days to establish chemotherapy-induced NP model.Normal saline 0.2 ml was given in group C. After measurement of the pain threshold at 10 days after establishment of the model, IGF-1 0.5 and 1.0 μg were intrathecally injected in group I1 and group I2, respectively.Normal saline 5 μl was intrathecally injected in C and CINP groups.Mechanical withdrawal threshold (MWT) was measured at 3, 5, 8, 10, 11, 13 and 15 days after establishment of the model.After measurement of the pain threshold at 15 days after establishment of the model, the expression of spinal IGF-1, IGF-1receptor (IGF-1R), interleukin (IL)-17A, IL-1β and tumor necrosis factor (TNF)-α was detected, and IGF-1 positive cells were counted using immunofluorescence. Results:Compared with group C, MWT was significantly decreased, the expression of spinal IGF-1 was down-regulated, the count of IGF-1 positive cells was decreased, and expression of IL-17A, IL-1β and TNF-α was up-regulated at 3-25 days after establishment of the model in CINP, I1 and I2 groups ( P<0.05). Compared with group CIPN, MWT was significantly increased at 15 days after establishment of the model in group I1, and MWT was increased, the expression of spinal IGF-1 was up-regulated, the count of IGF-1 positive cells was increased, and expression of IL-17A, IL-1β and TNF-α was down-regulated at 13 and 15 days after establishment of the model in group I2 ( P<0.05). Compared with group I1, the count of IGF-1 positive cells in spinal dorsal horn was increased in group I2 ( P<0.05). There was no significant difference in the expression of spinal IGF-1R among the 4 groups ( P>0.05). Conclusion:Intrathecal IGF-1 can alleviate chemotherapy-induced NP, and the mechanism may be related to inhibiting the inflammatory responses in spinal cord of mice.

Role of interleukin-4 in renal fibrosis induced by acute kidney injury in mice / 中华麻醉学杂志

Objective:To evaluate the role of interleukin-4 (IL-4) in renal fibrosis induced by acute kidney injury (AKI) in mice.Methods:Twenty-four male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n =6 each) using a random number table method: control group (group CON), AKI group, control plus anti-IL-4 antibody group (group CON-A), and AKI plus anti-IL-4 antibody group (group AKI-A). In AKI-A and AKI groups, folic acid was intraperitoneally injected to induce AKI in anesthetized mice, and the anti-IL-4 antibody 40 mg/kg and the equal volume of PBS were intraperitoneally injected, respectively, at 3, 6, 9 and 12 days after injection. The equal volume of PBS and anti-IL-4 antibody was given at the corresponding time point in CON and CON-A groups, respectively.The orbital blood samples were taken on day 14 after injecting folic acid for determination of the serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The animals were then sacrificed, and renal tissues were obtained for examination of the pathological changes (using Sirius red staining and Masson staining) and for determination of the expression of fibronectin(FN), collagen-Ⅰ(Col-Ⅰ) and α-smooth muscle actin (α-SMA) (by Western blot) and expression of CD206, Arg-1 and FIZZ1 mRNA (by real-time polymerase chain reaction). The area of renal fibrosis was measured. Results:Compared with group CON, the serum BUN and Cr concentrations were significantly increased, the area of renal fibrosis was increased, and the expression of FN, Col-Ⅰ, α-SMA and CD206, Arg-1 and FIZZ1 mRNA in renal tissues was up-regulated in group AKI ( P<0.05), and no significant change was found in the parameters mentioned above in group CON-A( P>0.05). Compared with group AKI, the serum BUN and Cr concentrations were significantly decreased, the area of renal fibrosis was reduced, and the expression of FN, Col-Ⅰ, α-SMA and CD206, Arg-1 and FIZZ1 mRNA in renal tissues was down-regulated in group AKI-A ( P<0.05). Conclusion:IL-4 is involved in the process of renal fibrosis following AKI, and the mechanism may be associated with the regulation of macrophage M2 polarization in mice.

Effect of dexmedetomidine on hypoxia-inducible factor-1alpha signaling pathway during hypoxia in mice with lung cancer / 中华麻醉学杂志

Objective@#To evaluate the effect of dexmedetomidine on hypoxia-inducible factor-1alpha (HIF-1 α) signaling pathway during hypoxia in mice with lung cancer.@*Methods@#Eighteen clean-grade healthy adult male BALB/c nude mice, aged 8 weeks, weighing 20-30 g, were divided into 3 groups (n=6 each) using a random number table method: lung cancer group (group L), hypoxia+ lung cancer group (group HL), and hypoxia plus lung cancer plus dexmedetomidine group (group HLD). Human lung adenocarcinoma A549 cell suspension 1×106/150 μl was injected via the tail vein to establish the mouse model of lung cancer.After the model was established successfully, chronic intermittent hypoxia was performed as follows: the mice were placed in air-tight modular incubation chambers, and the atmosphere was controlled by a constant gas flow containing 10% O2 for 3 h once a day for 3 weeks.The mice in group HL were exposed to hypoxia.After the end of hypoxia exposure, the animals in group HLD were intraperitoneally injected with dexmedetomidine 25 μg/kg 3 times a week for 3 weeks in total.The mice in group L were placed in air-tight modular incubation chambers and the atmosphere was controlled by a constant gas flow containing 21% O2 for 3 h once a day for 3 weeks, and the equal volume of normal saline was injected intraperitoneally.The mice were sacrificed by cervical dislocation, and the lung tissues were removed for determination of lung cancer nodules count, HIF-1α expression (by immunohistochemistry), expression of HIF-1α, survivin and X chromosome linked inhibitor of apoptosis protein (XIAP) (by Western blot), and expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 (by real-time polymerase chain reaction).@*Results@#Compared with group L, the number of lung cancer nodules was significantly increased, and the expression of HIF-1α, survivin, XIAP, MMP-2 and MMP-9 was up-regulated in the other two groups (P<0.05). Compared with group HL, the number of lung cancer nodules was significantly increased, and the expression of HIF-1α, survivin, XIAP, MMP-2 and MMP-9 was up-regulated in group HLD (P<0.05). A small number of HIF-1α positive cells were found in group L, a medium number of HIF-1α positive cells in group HL, and a large number of HIF-1α positive cells in group HAD.@*Conclusion@#Dexmedetomidine can promote proliferation of tumor cells in mice with lung cancer during hypoxia, and the mechanism is related to activating HIF-1α signaling pathway.

Effects of dexmedetomidine on renal fibrosis in a mouse model of renal ischemia-reperfusion: the role of Akt / 中华麻醉学杂志

Objective@#To evaluate the effect of dexmedetomidine on renal fibrosis in a mouse model of renal ischemia-reperfusion (I/R) and the role of serine-threonine kinase (Akt).@*Methods@#Sixty male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 5 groups (n=12 each) using a random number table method: sham operation group (S group), renal I/R group (I/R group), renal I/R plus dexmedetomidine group (I/R + D group), renal I/R plus dexmedetomidine plus Akt agonist SC79 group (I/R + D + SC group), and renal I/R plus dexmedetomidine plus normal saline group (I/R+ D+ NS group). Renal I/R injury model was established by clamping the bilateral renal pedicle for 30 min followed by reperfusion.Dexmedetomidine was intraperitoneally injected at 30 min before surgery in I/R+ D, I/R+ D+ SC and I/R+ D+ NS groups.SC79 was intraperitoneally injected as a bolus of 0.04 mg/kg at 1 min of reperfusion, followed by an intraperitoneal injection of the same dose every 24 h until day 7.The serum blood urea nitrogen (BUN) and Scr concentrations were detected at 24 h of reperfusion.Renal tissues were taken, and the damage to the renal tubules was scored.Renal tissues were removed at 14 days of reperfusion to detect the degree of renal fibrosis and expression of collagen 1 (COL1), fibronectin (FN), and α-smooth actin (α-SMA) (by immunofluorescence and Western blot). The expression of phosphorylated Akt (p-Akt) in renal tissues was determined by Western blot at 24 h and 14 day of reperfusion.@*Results@#Compared with group S, the serum BUN and Scr concentrations, renal tubule damage score and degree of renal fibrosis were significantly increased, and the expression of COL1, FN, α-SMA and p-Akt was up-regulated in group I/R (P<0.05). Compared with I/R group, the serum BUN and Scr concentrations, renal tubular damage score and degree of renal fibrosis were significantly decreased, and the expression of COL1, FN, α-SMA and p-Akt was down-regulated in I/R+ D and I/R+ D+ NS groups (P<0.05). Compared with I/R+ D group, the serum BUN and Scr concentrations, renal tubule damage score and degree of renal fibrosis were significantly increased , and the expression of COL1, FN, α-SMA and p-Akt was up-regulated in I/R+ D+ SC group (P<0.05).@*Conclusion@#Dexmedetomidine can reduce the degree of renal fibrosis in a mouse model of renal I/R and the mechanism is related to inhibiting activation of Akt.

Efficacy of lumbar sympathetic ganglion pulsed radiofrequency in treatment of diabetic neuropathic pain / 中华麻醉学杂志

Objective To evaluate the efficacy of lumbar sympathetic ganglion pulsed radiofrequency (PRF) in treatment of diabetic neuropathic pain.Methods Forty-eight patients of both sexes with diabetic neuropathic pain,aged 45-75 yr,with the course of disease 4-7 yr,were divided into 2 groups (n =24 each) using a random number table method:control group (C group) and PRF group (P group).Group C was treated with conventional drugs.Bilateral lumbar sympathetic ganglion PRF was performed on the basis of conventional drug therapy in group P.The serum concentrations of tumor necrosis factor-alpha and interleukin-6 were measured by enzyme-linked immunosorbent assay.The visual analogue scale score was recorded before treatment and at 1,2 and 4 weeks of treatment.The degree of efficacy was recorded at 4 weeks of treatment,and the total effective rate was calculated.Results Compared with group C,the visual analogue scale score and serum concentrations of tumor necrosis factor-alpha and interleukin-6 were significantly decreased,and the total effective rate and degree of curative efficacy were increased in group P (P＜0.05).No pulsed radiofrequency-related complications or drugs-related adverse reactions were found in two groups.Conclusion Lumbar sympathetic ganglion PRF can effectively relieve diabetic neuropathic pain in the patients.

Effect of dexmedetomidine on hypoxia-inducible factor-1alpha signaling pathway during hypoxia in mice with lung cancer / 中华麻醉学杂志

Objective To evaluate the effect of dexmedetomidine on hypoxia-inducible factor1alpha (HIF-1 α) signaling pathway during hypoxia in mice with lung cancer.Methods Eighteen cleangrade healthy adult male BALB/c nude mice,aged 8 weeks,weighing 20-30 g,were divided into 3 groups (n=6 each).using a random number table method:lung cancer group (group L),hypoxia+lung cancer group (group HL),and hypoxia plus lung cancer plus dexmedetomidine group (group HLD).Human lung adenocarcinoma A549 cell suspension 1× 106/150 μl was injected via the tail vein to establish the mouse model of lung cancer.After the model was established successfully,chronic intermittent hypoxia was performed as follows:the mice were placed in air-tight modular incubation chambers,and the atmosphere was controlled by a constant gas flow containing 10％ O2 for 3 h once a day for 3 weeks.The mice in group HL were exposed to hypoxia.After the end of hypoxia exposure,the animals in group HLD were intraperitoneally injected with dexmedetomidine 25 μg/kg 3 times a week for 3 weeks in total.The mice in group L were placed in air-tight modular incubation chambers and the atmosphere was controlled by a constant gas flow containing 21％ O2 for 3 h once a day for 3 weeks,and the equal volume of normal saline was injected intraperitoneally.The mice were sacrificed by cervical dislocation,and the lung tissues were removed for determination of lung cancer nodules count,HIF-1α expression (by immunohistochemistry),expression of HIF-1α,survivin and X chromosome linked inhibitor of apoptosis protein (XIAP) (by Western blot),and expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 (by real-time polymerase chain reaction).Results Compared with group L,the number of lung cancer nodules was significantly increased,and the expression of HIF-1α,survivin,XIAP,MMP-2 and MMP-9 was up-regulated in the other two groups (P＜0.05).Compared with group HL,the number of lung cancer nodules was significantly increased,and the expression of HIF-1α,survivin,XIAP,MMP-2 and MMP-9 was up-regulated in group HLD (P＜0.05).A small number of HIF-1α positive cells were found in group L,a medium number of HIF-1α positive cells in group HL,and a large number of HIF-1α positive cells in group HAD.Conclusion Dexmedetomidine can promote proliferation of tumor cells in mice with lung cancer during hypoxia,and the mechanism is related to activating HIF-1α signaling pathway.

Effects of dexmedetomidine on renal fibrosis in a mouse model of renal ischemia-reperfusion: the role of Akt / 中华麻醉学杂志

Objective To evaluate the effect of dexmedetomidine on renal fibrosis in a mouse model of renal ischemia-reperfusion (I/R) and the role of serine-threonine kinase (Akt).Methods Sixty male C57BL/6 mice,aged 8 weeks,weighing 20-25 g,were divided into 5 groups (n=12 each) using a random number table method:sham operation group (S group),renal I/R group (I/R group),renal I/R plus dexmedetomidine group (I/R + D group),renal I/R plus dexmedetomidine plus Akt agonist SC79 group (I/R + D + SC group),and renal I/R plus dexmedetomidine plus normal saline group (I/R+D+NS group).Renal I/R injury model was established by clamping the bilateral renal pedicle for 30 min followed by reperfusion.Dexmedetomidine was intraperitoneally injected at 30 rmin before surgery in I/R+D,I/R+D+SC and I/R+D+NS groups.SC79 was intraperitoneally injected as a bolus of 0.04 mg/kg at 1 min of reperfusion,followed by an intraperitoneal injection of the same dose every 24 h until day 7.The serum blood urea nitrogen (BUN) and Scr concentrations were detected at 24 h of reperfusion.Renal tissues were taken,and the damage to the renal tubules was scored.Renal tissues were removed at 14 days of reperfusion to detect the degree of renal fibrosis and expression of collagen 1 (COL1),fibronectin (FN),and α-smooth actin (α-SMA) (by immunofluorescence and Western blot).The expression of phosphorylated Akt (p-Akt) in renal tissues was determined by Western blot at 24 h and 14 day of reperfusion.Results Compared with group S,the serum BUN and Scr concentrations,renal tubule damage score and degree of renal fibrosis were significantly increased,and the expression of COL1,FN,α-SMA and p-Akt was up-regulated in group I/R (P＜0.05).Compared with I/R group,the serum BUN and Scr concentrations,renal tubular damage score and degree of renal fibrosis were significantly decreased,and the expression of COL1,FN,α-SMA and pAkt was down-regulated in I/R+D and I/R+D+NS groups (P＜0.05).Compared with I/R+D group,the serum BUN and Scr concentrations,renal tubule damage score and degree of renal fibrosis were significantly increased,and the expression of COL1,FN,α-SMA and p-Akt was up-regulated in I/R+D+SC group (P＜0.05).Conclusion Dexmedetomidine can reduce the degree of renal fibrosis in a mouse model of renal I/R and the mechanism is related to inhibiting activation of Akt.

Modified strategy of anesthesia for laparoscopic radical resection of rectal carcinoma: ultrasound-guided anterior quadratus lumborum block combined with general anesthesia / 中华麻醉学杂志

Objective To evaluate the efficacy of ultrasound-guided anterior quadratus lumborum block combined with general anesthesia for laparoscopic radical resection of rectal carcinoma. Methods A total of 80 patients of both sexes, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 40-64 yr, scheduled for elective laparoscopic radical resection of rectal carcinoma, were divided into 2 groups ( n=40 each) using a random number table method: anterior quadratus lumborum block combined with general anesthesia group ( group QG) and general anesthesia group ( group G) . In group QG, anteri-or quadratus lumborum block was performed with 0. 33％ ropivacaine 25 ml and dexamethasone 5 mg under ultrasound guidance before operation, and the same procedure was performed on the other side. Combined intravenous-inhalational anesthesia was applied, propofol 3-5μg∕ml and remifentanil 3-5 ng∕ml were given by target-controlled infusion, and cisatracurium was intermittently injected in two groups. Patient-controlled intravenous analgesia with sufentanil 2μg∕kg was used for postoperative analgesia. The analgesic pump was set up to deliver a 2 ml bolus dose with a 15-min lockout interval. Bruggrmann comfort scale ( BCS) scores were recorded at 1, 6, 12, 24 and 48 h after operation ( T1-5 ) . Tramadol was used for rescue analgesic after operation. The consumption of remifentanil and sufentanil, requirement for tramadol, occurrence of adverse reactions and patients' satisfaction with postoperative analgesia were recorded. The emergence time, first ambulation time, time to first flatus∕poo and length of hospital stay were also recorded. The develop-ment of anterior quadratus lumborum block-related complications was recorded. Results Compared with group G, BCS scores were significantly increased at T4,5 , the consumption of remifentanil, requirement for tramadol and incidence of nausea and vomiting were decreased, patients' satisfaction with postoperative an-algesia was increased, and the emergence time, first ambulation time, time to first flatus∕poo and length of hospital stay were shortened in group QG (P<0. 05). Conclusion Ultrasound-guided anterior quadratus lumborum block combined with general anesthesia can reduce the consumption of opioids in the perioperative period and is helpful in improving outcomes when used for laparoscopic radical resection of rectal carcinoma.

Effect of pulsed radiofrequency on phenotypic transformation of lumbar sympathetic ganglion in rats with diabetic neuropathic pain / 中华麻醉学杂志

Objective To evaluate the effect of pulsed radiofrequency (PRF) on the phenotypic transformation of the lumbar sympathetic ganglion (LSG) in the rats with diabetic neuropathic pain (PDN).Methods Twenty-four clean-grade healthy adult male Sprague-Dawley rats,aged 2 months,weighing 180-220 g,were divided into 4 groups (n =6 each) according to the method of random number table:control group (group C),group PDN,group PRF,and PRF control group (group PC).The PDN model was established by intraperitoneal injection of streptozotocin 60 mg/kg in anesthetized rats.Citrate-sodium citrate buffer 6 ml/kg was intraperitoneally injected in group C.Group PC only received radiofrequency needle puncture.PRF was performed on the right L3 LSG in group PRF.The mechanical paw withdrawal threshold (MWT) to yon Frey filament stimulation was measured before intraperitoneal injection (baseline,T0),before PRF and at 1,3,5,7 and 14 days after PRF.The rats were then sacrificed,and ipsilateral L3 LSGs were removed for determination of the expression of tyrosine hydroxylase (TH) and vesicle glutamate transporter2 (VGLUT2) in LSGs (by double immunofluorescent staining) and for examination of pathological changes (with a light microscope).The number of neurons expressing VGLUT2 was counted.Results Compared with group C,the MWT was significantly decreased at T1-6,and the number of neurons expressing VGLUT2 was increased at T6 in PDN,PC and PRF groups (P＜0.05).Compared with PDN and PC groups,the MWT was significantly increased at T2-6,and the number of neurons expressing VGLUT2 was decreased at T6 in group PRF (P＜0.05).TH expression in LSGs was found,and no VGLUT2 expression in LSGs was observed in group C,the expression of TH and VGLUT2 in LSGs was found in the other three groups,especially in PDN and PC groups,and most of the neurons expressing VGLUT2 expressed TH simultaneously.Conclusion The mechanism by which PRF mitigates PDN is related to inhibiting the phenotypic transformation of LSGs in the rats.

Role of spinal cord TRAF6/NF-κB signaling pathway in development of diabetic neuropathic pain in rats / 中华麻醉学杂志

Objective To evaluate the role of spinal cord tumor necrosis factor receptor-associated factor 6 (TRAF6)/nuclear factor kappa B (NF-κB)signaling pathway in the development of diabetic neuropathic pain (DNP)in rats.Methods Clean-grade healthy adult male Sprague-Dawley rats,aged 2 months,weighing 180-230 g,in which IT catheters were implanted,were used in this study.Streptozotocin 60 mg/kg was intraperitoneally injected after IT catheterization to establish the model of DNP.Twelve DNP rats were divided into 2 groups (n =6 each) by a random number table method:DNP group and DNP plus TRAF6 inhibitor group (group DTR).Another 6 age-matched Sprague-Dawley rats were used as normal control group (group NC).The rats in group DC and group DTR received IT injection of dimethyl sulfoxide 10 μl and TRAF6 inhibition 10 μg,respectively,once a day for 7 consecutive days starting from day 21 after establishing the model.The mechanical paw withdrawal threshold (MWT) was determined before establishing the model (T1),on 7,14 and 21 days after establishing the model (T2-4),and on 1,4 and 7 days after IT injection (T5-7).The rats were sacrificed after the last MWT measurement,and the L3-5 segments of the spinal cord were removed for determination of the expression of TRAF6 and NFκB p65 by Western blot.Results Compared with group NC,the MWT at T3-7 in group DC and at T3-6 in group DTR was significantly decreased,and the expression of spinal TRAF6 and NF-κB p65 was up-regulated in DC and DTR groups (P＜0.05).Compared with group DC,the MWT was significantly increased at T6-7,and the expression of spinal TRAF6 and NF-κB p65 was down-regulated in group DTR (P ＜ 0.05).Conclusion Spinal cord TRAF6/NF-κB signaling pathway is involved in the development of DNP in rats.

Intrathecal infection of TRESK overexpression adenovirus to relieve neuropathic pain via inhibiting JNK activation and neuronal apoptosis / 实用医学杂志

Objective To observe the effect of intrathecal injection of TRESK overexpression adenoviruson phosphorylation of JNK and apoptosis of neurons in neuropathic pain rats.Methods Seventy-two male SD rats were randomly divided into six groups:groups C,S,NP,T,V,and NS,12 for each group.SNI was administrated to rats in groups NP,T,V and NS.TRESK adenovirus and negative virus were intrathecally injected after use of SNI in groups T and V,while equal volume of NS was injected to rats in group NS.MWT and TWL were measured at 1 day before operation(baseline,BL)and at 1,3,7 and 14 days after operation (days 1,3,7,and 14).Six rats in each group were sacrificed at D7 to determinate the expression of TRESK protein of DRG.The other rats were sacrificed at D14 to determinate neural apoptosis and the expressions of caspase3 and p-JNK of DRG.Results As compared with groups C,S and T,the expression of TRESK protein was significantly decreased at D7 in groups NP,NS and V (P＜0.05).Compared with groups C and S,MWT was significantly decreased at days 1,3,7 and 14 (P＜0.05),phosphorylation of JNK in DRG was significantly increased at D14 (P＜0.05),neuronal apoptosis rate and expressions of Caspase3 of DRG were significantly increased at D14 (P＜0.05) in groups NP,T,NS and V.Compared with groups NP,V and NS,MWT was significantly increased at time points of days 1,3,7 and 14 in group T (P＜0.05),phosphorylation of JNK of in DRG was significantly decreased at D14 in group T (P＜0.05),neuronal apoptosis rate and expression of Caspase3 of DRG were significantly decreased at D14 in group T (P＜0.05).Intrathecal injection ofpAd/CMV/VS-DEST-TRESK obviously reduced mechanical hyperalgesia,upregulated TRESK expression,and lowered JNK phosphorylation and NP in SNI rat.Conclusions Intrathecal injection of TRESK over expression adenovirus relieves NP via inhibiting JNK activation and neuronal apoptosis.

Role of interleukin-4 receptor in renal fibrosis following renal ischemia-reperfusion injury in mice / 中华麻醉学杂志

Objective To evaluate the role of interleukin-4 receptor (IL-4R) in renal fibrosis following renal ischemia-reperfusion (I/R) injury in mice.Methods Twelve male wild type BALB/C mice and 12 IL-4Rα gene-knockout mice,aged 8-10 weeks,weighing 20-30 g,were used in the study.The mice of either type were divided into 2 groups (n =6 each) using a random number table:sham operation group (group S) and group I/R.In group I/R,renal I/R was induced by occlusion of the right renal artery for 1 h with atraumatic microclips followed by 2 weeks of reperfusion.The right renal artery was only isolated in group S.At 2 weeks of reperfusion,blood samples were taken from the orbital vein for determination of the concentrations of serum blood urea nitrogen (BUN) and creatinine (Cr).The renal tissues were obtained,and the renal fibrosis area was measured by Sirius Red staining.The expression of fibronectin (FN),collagen Ⅰ (COL-Ⅰ) and α-smooth muscle actin (α-SMA) in renal tissues was detected by immunofluorescence.The expression of signal transducer and activator of transcription 6 (STAT6) and phospho-STAT6 in renal tissues was determined by Western blot.The ratio of phoshop-STAT6 to STAT6 was calculated to reflect the phosphorylation of STAT6.Results Compared with group S of wild type mice,the serum BUN and Cr concentrations and renal fibrosis area were significantly increased,the expression of FN,COL-Ⅰ and α-SMA in renal tissues was significantly up-regulated,and the phosphorylation of STAT6 in renal tissues was significantly increased in group I/R of wild type and IL-4Rα KO mice (P＜0.05).Compared with group I/R of wild type mice,the serum BUN and Cr concentrations and renal fibrosis area were significantly decreased,the expression of FN,COL-Ⅰ and α-SMA in renal tissues was significantly down-regulated,and the phosphorylation of STAT6 in renal tissues was significantly decreased in group I/R of IL-4RαKO mice (P＜0.05).Conclusion The mechanism of renal fibrosis following renal I/R injury is partially related to IL-4R,and IL-4R results in renal fibrosis through promoting activation of STAT6 signaling pathway in mice.

Effect of resveratrol on activity of NADPH oxidase during acute lung injury induced by intestinal ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To evaluate the effect of resveratrol on the activity of NADPH oxidase during acute lung injury induced by intestinal ischemia-reperfusion (I/R) in rats.Methods Thirty-two pathogenfree healthy female Sprague-Dawley rats,weighing 180-230 g,were divided into 4 groups (n =8 each) using a random number table:sham operation group (Sham group),intestinal I/R group (I/R group),vehicle group (Veh group) and resveratrol group (Res group).Intestinal I/R was produced by occlusion of the superior mesenteric artery for 75 min followed by 4 h of reperfusion.In group Res,resveratrol 15 mg/kg was intraperitoneally injected for 5 consecutive days before establishment of the model and at 15 min before ischemia.The equal volume of vehicle (0.5％ alcohol) was intraperitoneally injected in group Veh.The equal volume of normal saline was intraperitoneally injected in Sham and I/R groups.At the end of reperfusion,the rats were sacrificed and the lung was removed for microscopic examination of pathologic changes which were scored and for determination of wet/dry weight ratio (W/D ratio),malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (by colorimetric method) and expression of NADPH oxidase subunits (gp91phox and p47phox) in lung tissues (by Western blot).Results Compared with group Sham,pathologic scores,W/D ratio and MDA content were significantly increased,the expression of gp91phox and p47phox was up-regulated,and the activity of SOD was decreased in I/R and Veh groups (P＜0.05).Compared with I/R and Veh groups,pathologic scores,W/D ratio and MDA content were significantly decreased,the expression of gp91phox and p47phox was down-regulated,and the activity of SOD was increased in group Res (P＜0.05).Conclusion The mechanism by which resveratrol reduces intestinal I/R-induced acute lung injury is related to inhibiting NADPH oxidase-mediated oxidative stress response of rats.

Effect of intrathecal cardamonin on diabetic neuropathic pain in rats / 中华麻醉学杂志

Objective To evaluate the effect of intrathecal cardamonin on diabetic neuropathic pain (DNP) in rats.Methods Healthy adult male Sprague-Dawley rats,aged 2 months,weighing 180-220 g,were included in the study.Diabetes mellitus was produced by intraperitoneal injection of streptozotocin 60 mg/kg after intrathecal catheterization.When decrease in pain threshold＞50％ of the baseline at 21 days after diabetes mellitus was produced,the induction of DNP was considered successful.Twenty-four rats with DNP were divided into 4 groups (n=6 each) using a random number table:group DNP,rapamycin group (group R),cardamonin 50 μg group (group D50) and cardamonin 100 μg group (group D100).Another 6 healthy age-matched rats were used as normal control (group C).In DNP,R,D50 and D100 groups,dimethyl sulfoxide 10 μl,rapamycin 5 μg and cardamonin 50 μg and 100 μg were intrathecally injected,respectively,once a day for 7 consecutive days starting from 21 days after successful estabiishment of the model.Mechanical paw withdrawal threshold (MWT) was measured before establishment of the model,on 7,14 and 21 days after establishment of the model,and on 1,4 and 7 days after intrathecal injection.The rats were sacrificed after the last MWT measurement,and their L3-5 segments of the spinal cord were removed for determination of the expression of S6K,p-S6K and synapsin Ⅱ by Western blot.Results Compared with group C,MWT was significantly decreased,and the expression of spinal S6K,p-S6K and synapsin Ⅱ was up-regulated in group DNP (P＜0.05 or 0.01).Compared with group DNP,MWT was significantly increased,and the expression of spinal S6K,p-S6K and synapsin Ⅱ was down-regulated in R,D50 and D100 groups (P＜0.05 or 0.01).Compared with group D50,MWT was significantly increased,and the expression of spinal S6K,p-S6K and synapsin Ⅱ was down-regulated in group D100 (P＜0.05).Conclusion Intrathecal cardamonin can relieve DNP in rats,and the mechanism is related to inhibiting spinal mammalian target of rapamycin activity and down-regulating the expression of synapsin Ⅱ.

Changes in APPL1 expression during renal fibrosis in a mouse model of renal ischemia-reperfusion injury / 中华麻醉学杂志

Objective To evaluate the changes in the expression of adaptor protein containing pleck-strin homobgy domain, phosphotyrosine-binding domain and a leucine zipper motif 1(APPL1)during renal fibrosis in a mouse model of renal ischemia-reperfusion(I∕R)injury. Methods Twenty-four male C57BL∕6 mice, aged 8 weeks, weighing 20-25 g, were divided into 2 groups(n=12 each)using a random number table: sham operation group(S group)and renal I∕R group.The model of renal I∕R injury was established by clipping the bilateral renal pedicles for 30 min followed by reperfusion in group I∕R.Six mice were selected at 2 days of reperfusion, and venous blood samples were collected for determination of serum concentrations of blood urea nitrogen and creatinine.The animals were then sacrificed, the renal specimens were obtained for microscopic examination of tubular necrosis with a light microscope, and the damage to the renal tubules was scored using a semi-quantitative method.Six mice were sacrificed at 14 days of reperfusion, and the renal specimens were obtained for assessment of the degree of renal fibrosis(using picric acid-sirius red staining) and for determination of the expression of collagen type 1, fibronectin and α-smooth muscle actin in renal tis-sues(by Western blot or immunofluorescence method). At 2 and 14 days of reperfusion, the expression of APPL1 in renal tissues was detected by Western blot and the expression of APPL1 mRNA in renal tissues by real-time polymerase chain reaction. Results Compared with group S, the serum concentrations of blood u-rea nitrogen and creatinine, scores of renal tubular damage and degree of renal fibrosis were significantly in-creased at 2 days of reperfusion, the expression of collagen type 1, fibronectin and α-smooth muscle actin in renal tissues was up-regulated at 14 days of reperfusion, and the expression of APPL1 protein and mRNA was up-regulated at 2 and 14 days of reperfusion in group I∕R(P<0.05). Conclusion Up-regulated expression of APPL1 may be involved in the process of renal fibrosis in a mouse model of renal I∕R injury.