AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of ratliver graft.METHODS: KCs were isolated at 0 (co...AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of ratliver graft.METHODS: KCs were isolated at 0 (control group), 2, 12,24 h (IR group) following IR in rat liver graft, mRNA expressionof TLR4 and MD-2 was detected by RT-PCR analysis, proteinexpression of TLR4/MD-2 was detected by flow cytometric(FCM) analysis, and tumor necrosis factor-(~ (TNF-(~) levelin supernatant was measured by ELISA. Then isolated KCswere incubated with anti-TLR4 polyclonal antibody (anti-TLR4 group), and TNF-(~ level was measured again.RESULTS: The mRNA and protein expression of TLR4/MD-2 and the level of TNF-(~ in IR group increased significantlyat 2 h following IR, and reached the maximum at 12 h, andslightly decreased at 24 h, but were still significantly higherthan those in the control group (P<O.01). The expression ofthese factors was markedly decreased after anti-TLR4antibody treatment as compared with the IR group (P<O.01).CONCLUSION: Lipopolysaccharide (LPS) following IR canup-regulate TLR4/MD-2 gene and protein expression inKCs, and synthesize cytokine TNF-(~. Anti TLR4 antibodycan inhibit the production of TNF-(~ induced by LPS. TLR4and its partner molecule MD-2 may play an important rolein Kupffer cell activation and IR injury.展开更多
Objective: To examine the changes in the express ion of mGluR4 after diffuse brain injury (DBI) and to determine the role of its specific agonist L-2-amino-4-phosphonobutyrate (L-AP4) in vivo. Methods: A total of 161 ...Objective: To examine the changes in the express ion of mGluR4 after diffuse brain injury (DBI) and to determine the role of its specific agonist L-2-amino-4-phosphonobutyrate (L-AP4) in vivo. Methods: A total of 161 male SD rats were randomized into the f ollowing groups. Group A included normal control,sham-operated control and DBI group. DBI was produced according to Marmarous diffuse head injury model. mRN A expression of mGluR4 was detected by hybridization in situ. Group B included D BI alone,DBI treated with normal saline and DBI treated with L-AP4. All DBI ra ts were trained in a series of performance tests,following which they were subj ected to DBI. At 1 and 12 hours,animals were injected intraventricularly with L -AP4 ( 100 mmol/L ,10 μl) or normal saline. Motor and cognitive performance s were tested at 1,3,7,14 days after injury and the damaged neurons were also detected. Results: There was no significant difference between normal con trol group and sham-operated group in the expression of mGluR4 ( P > 0.05 ) . The animals exposed to DBI showed significantly increased expression of mRNA o f mGluR4 compared with the sham-operated animals 1 h after injury ( P < 0.05 ). At 6 hours,the evolution of neuronal expression of mGluR4 in the trauma al one group was relatively static. Compared with saline-treated control animals,rats treated with L-AP4 showed an effective result of decreased number of damag ed neurons and better motor and cognitive performances.Conclusions: Increased expression of mGluR4 is important in the pathophysiological process of DBI and its specific agonist L-AP4 can provide r emarkable neuroprotection against DBI not only at the histopathological level bu t also in the motor and cognitive performance.展开更多
目的:基于"脑-肠-菌"轴探讨健脾益气针法对肥胖鼠肠道菌群及重要抗菌免疫分子Toll样受体4(TLR4)的调控机制。方法:复制营养性肥胖小鼠20只,随机均分为模型组与针刺组,另设正常组。针刺组取天枢、关元、后三里及三阴交等穴,施...目的:基于"脑-肠-菌"轴探讨健脾益气针法对肥胖鼠肠道菌群及重要抗菌免疫分子Toll样受体4(TLR4)的调控机制。方法:复制营养性肥胖小鼠20只,随机均分为模型组与针刺组,另设正常组。针刺组取天枢、关元、后三里及三阴交等穴,施以补法,得气后接通电针维持治疗,比较各组小鼠体质量及Lee’s指数差异;ELISA法测定脑与肠组织中炎性因子白细胞介素(IL)-6、IL-10、肿瘤坏死因子-α(TNF-α)的含量及血清中瘦素(LP)、脂联素(ADPN)及游离脂肪酸(FFA)的含量;免疫组化法检测脑-肠中TLR4蛋白分布变化;采用16S r DNA测序技术分析肠道菌群结构丰度变化。结果:模型组体质量、Lee’s指数及血清LP、ADPN及FFA含量显著高于正常组(P<0.01),与模型组比较,针刺组以上指标显著下降(P<0.01);针刺组脑-肠组织中IL-6、IL-10、TNF-α含量显著低于模型组(P<0.05,P<0.01)。针刺组脑及肠组织TLR4蛋白分布密度明显低于模型组(P<0.01);模型组厚壁菌门、绿弯菌门、芽单胞菌门显著升高(P<0.01),拟杆菌门显著降低(P<0.01),针刺组这些异常菌门显著恢复(P<0.05,P<0.01)。结论:应用健脾益气针法治疗肥胖,调节"脑-肠-菌"轴实现肠道菌群再平衡,达到减肥降脂消炎目的。展开更多
基金Supported by the National Natural Science Foundation of China,No.30300337,30200278,30170919
文摘AIM: To investigate the expression of toll-like receptor 4(TLR4) and MD-2 gene and protein in Kupffer cells (KCs)and their role in ischemia-reperfusion (IR) injury of ratliver graft.METHODS: KCs were isolated at 0 (control group), 2, 12,24 h (IR group) following IR in rat liver graft, mRNA expressionof TLR4 and MD-2 was detected by RT-PCR analysis, proteinexpression of TLR4/MD-2 was detected by flow cytometric(FCM) analysis, and tumor necrosis factor-(~ (TNF-(~) levelin supernatant was measured by ELISA. Then isolated KCswere incubated with anti-TLR4 polyclonal antibody (anti-TLR4 group), and TNF-(~ level was measured again.RESULTS: The mRNA and protein expression of TLR4/MD-2 and the level of TNF-(~ in IR group increased significantlyat 2 h following IR, and reached the maximum at 12 h, andslightly decreased at 24 h, but were still significantly higherthan those in the control group (P<O.01). The expression ofthese factors was markedly decreased after anti-TLR4antibody treatment as compared with the IR group (P<O.01).CONCLUSION: Lipopolysaccharide (LPS) following IR canup-regulate TLR4/MD-2 gene and protein expression inKCs, and synthesize cytokine TNF-(~. Anti TLR4 antibodycan inhibit the production of TNF-(~ induced by LPS. TLR4and its partner molecule MD-2 may play an important rolein Kupffer cell activation and IR injury.
文摘Objective: To examine the changes in the express ion of mGluR4 after diffuse brain injury (DBI) and to determine the role of its specific agonist L-2-amino-4-phosphonobutyrate (L-AP4) in vivo. Methods: A total of 161 male SD rats were randomized into the f ollowing groups. Group A included normal control,sham-operated control and DBI group. DBI was produced according to Marmarous diffuse head injury model. mRN A expression of mGluR4 was detected by hybridization in situ. Group B included D BI alone,DBI treated with normal saline and DBI treated with L-AP4. All DBI ra ts were trained in a series of performance tests,following which they were subj ected to DBI. At 1 and 12 hours,animals were injected intraventricularly with L -AP4 ( 100 mmol/L ,10 μl) or normal saline. Motor and cognitive performance s were tested at 1,3,7,14 days after injury and the damaged neurons were also detected. Results: There was no significant difference between normal con trol group and sham-operated group in the expression of mGluR4 ( P > 0.05 ) . The animals exposed to DBI showed significantly increased expression of mRNA o f mGluR4 compared with the sham-operated animals 1 h after injury ( P < 0.05 ). At 6 hours,the evolution of neuronal expression of mGluR4 in the trauma al one group was relatively static. Compared with saline-treated control animals,rats treated with L-AP4 showed an effective result of decreased number of damag ed neurons and better motor and cognitive performances.Conclusions: Increased expression of mGluR4 is important in the pathophysiological process of DBI and its specific agonist L-AP4 can provide r emarkable neuroprotection against DBI not only at the histopathological level bu t also in the motor and cognitive performance.
文摘目的:基于"脑-肠-菌"轴探讨健脾益气针法对肥胖鼠肠道菌群及重要抗菌免疫分子Toll样受体4(TLR4)的调控机制。方法:复制营养性肥胖小鼠20只,随机均分为模型组与针刺组,另设正常组。针刺组取天枢、关元、后三里及三阴交等穴,施以补法,得气后接通电针维持治疗,比较各组小鼠体质量及Lee’s指数差异;ELISA法测定脑与肠组织中炎性因子白细胞介素(IL)-6、IL-10、肿瘤坏死因子-α(TNF-α)的含量及血清中瘦素(LP)、脂联素(ADPN)及游离脂肪酸(FFA)的含量;免疫组化法检测脑-肠中TLR4蛋白分布变化;采用16S r DNA测序技术分析肠道菌群结构丰度变化。结果:模型组体质量、Lee’s指数及血清LP、ADPN及FFA含量显著高于正常组(P<0.01),与模型组比较,针刺组以上指标显著下降(P<0.01);针刺组脑-肠组织中IL-6、IL-10、TNF-α含量显著低于模型组(P<0.05,P<0.01)。针刺组脑及肠组织TLR4蛋白分布密度明显低于模型组(P<0.01);模型组厚壁菌门、绿弯菌门、芽单胞菌门显著升高(P<0.01),拟杆菌门显著降低(P<0.01),针刺组这些异常菌门显著恢复(P<0.05,P<0.01)。结论:应用健脾益气针法治疗肥胖,调节"脑-肠-菌"轴实现肠道菌群再平衡,达到减肥降脂消炎目的。
