Objective:To examine the therapeutic effect of Fangji Fuling Decoction(FFD) on sepsis through network pharmacological analysis combined with in vitro and in vivo experiments.Methods:A sepsis mouse model was constructe...Objective:To examine the therapeutic effect of Fangji Fuling Decoction(FFD) on sepsis through network pharmacological analysis combined with in vitro and in vivo experiments.Methods:A sepsis mouse model was constructed through intraperitoneal injection of 20 mg/kg lipopolysaccharide(LPS).RAW264.7 cells were stimulated by 250 ng/m L LPS to establish an in vitro cell model.Network pharmacology analysis identified the key molecular pathway associated with FFD in sepsis.Through ectopic expression and depletion experiments,the effect of FFD on multiple organ damage in septic mice,as well as on cell proliferation and apoptosis in relation to the mitogen-activated protein kinase 14/Forkhead Box O 3A(MAPK14/FOXO3A) signaling pathway,was analyzed.Results:FFD reduced organ damage and inflammation in LPS-induced septic mice and suppressed LPS-induced macrophage apoptosis and inflammation in vitro(P<0.05).Network pharmacology analysis showed that FFD could regulate the MAPK14/FOXO signaling pathway during sepsis.As confirmed by in vitro cell experiments,FFD inhibited the MAPK14 signaling pathway or FOXO3A expression to relieve LPS-induced macrophage apoptosis and inflammation(P<0.05).Furthermore,FFD inhibited the MAPK14/FOXO3A signaling pathway to inhibit LPS-induced macrophage apoptosis in the lung tissue of septic mice(P<0.05).Conclusion:FFD could ameliorate the LPS-induced inflammatory response in septic mice by inhibiting the MAPK14/FOXO3A signaling pathway.展开更多
目的研究利拉鲁肽对骨质疏松大鼠叉头框蛋白O3a(FoxO3a)/Wnt信号通路及椎骨密度的影响。方法将雌性Sprague-dawley大鼠根据随机数字表法分成假手术组、模型组、利拉鲁肽干预组,每组10只,模型组、干预组均采用双侧卵巢切除术建立骨质疏...目的研究利拉鲁肽对骨质疏松大鼠叉头框蛋白O3a(FoxO3a)/Wnt信号通路及椎骨密度的影响。方法将雌性Sprague-dawley大鼠根据随机数字表法分成假手术组、模型组、利拉鲁肽干预组,每组10只,模型组、干预组均采用双侧卵巢切除术建立骨质疏松症模型。干预组每天分别皮下注射利拉鲁肽,假手术组、模型组给予等体积的生理盐水。连续给药12周后,检测骨密度、骨生物力学,采用酶联免疫吸附法血清骨保护素、抗酒石酸酸性磷酸酶及骨钙素水平,采用Real-time PCR技术检测O3a/Wnt途径中相关mRNA表达水平,采用免疫印迹法检测O3a/Wnt途径中相关蛋白表达水平。结果造模成功大鼠的L4~5(0.33±0.04 vs 0.18±0.03)及左、右股骨骨密度(0.37±0.05 vs 0.23±0.04,0.35±0.04 vs 0.24±0.03)水平明显低于假手术组(P<0.05)。治疗12周后3组大鼠骨最大载荷、三点弯曲应力、骨密度及弹性模量差异有统计学意义,其中假手术组各指标水平最高(36.53±5.23,154.13±6.27,0.34±0.04,3102.34±160.44),其次为依次干预组(19.37±4.32,141.54±6.58,0.18±0.03,2270.18±145.53)、模型组(26.17±4.68,147.56±5.84,0.28±0.03,2804.24±140.42)(P<0.05)。3组大鼠血清骨保护素、抗酒石酸酸性磷酸酶及骨钙素水平差异有统计学意义,其中假手术组的骨保护素(假手术组vs模型组vs干预组:7.53±0.63 vs 4.57±0.42 vs 6.15±0.61)明显高于干预组、模型组,抗酒石酸酸性磷酸酶(假手术组vs模型组vs干预组:14.34±2.87 vs 19.53±3.52 vs 15.96±3.14)、骨钙素(假手术组vs模型组vs干预组:0.84±0.09 vs 1.13±0.12 vs 0.95±0.08)明显低于干预组、模型组(P<0.05)。3组大鼠FoxO3a、Wnt2及β-catenin mRNA和蛋白表达水平差异有统计学意义,其中假手术组的Wnt2、β-catenin明显高于干预组、模型组,FoxO3a明显低于干预组、模型组(P<0.05)。结论利拉鲁肽通过活化O3a/Wnt信号通路而增加骨密度,改善骨生物力学状态,从而有效治疗骨质疏松。展开更多
文摘Objective:To examine the therapeutic effect of Fangji Fuling Decoction(FFD) on sepsis through network pharmacological analysis combined with in vitro and in vivo experiments.Methods:A sepsis mouse model was constructed through intraperitoneal injection of 20 mg/kg lipopolysaccharide(LPS).RAW264.7 cells were stimulated by 250 ng/m L LPS to establish an in vitro cell model.Network pharmacology analysis identified the key molecular pathway associated with FFD in sepsis.Through ectopic expression and depletion experiments,the effect of FFD on multiple organ damage in septic mice,as well as on cell proliferation and apoptosis in relation to the mitogen-activated protein kinase 14/Forkhead Box O 3A(MAPK14/FOXO3A) signaling pathway,was analyzed.Results:FFD reduced organ damage and inflammation in LPS-induced septic mice and suppressed LPS-induced macrophage apoptosis and inflammation in vitro(P<0.05).Network pharmacology analysis showed that FFD could regulate the MAPK14/FOXO signaling pathway during sepsis.As confirmed by in vitro cell experiments,FFD inhibited the MAPK14 signaling pathway or FOXO3A expression to relieve LPS-induced macrophage apoptosis and inflammation(P<0.05).Furthermore,FFD inhibited the MAPK14/FOXO3A signaling pathway to inhibit LPS-induced macrophage apoptosis in the lung tissue of septic mice(P<0.05).Conclusion:FFD could ameliorate the LPS-induced inflammatory response in septic mice by inhibiting the MAPK14/FOXO3A signaling pathway.
文摘目的研究利拉鲁肽对骨质疏松大鼠叉头框蛋白O3a(FoxO3a)/Wnt信号通路及椎骨密度的影响。方法将雌性Sprague-dawley大鼠根据随机数字表法分成假手术组、模型组、利拉鲁肽干预组,每组10只,模型组、干预组均采用双侧卵巢切除术建立骨质疏松症模型。干预组每天分别皮下注射利拉鲁肽,假手术组、模型组给予等体积的生理盐水。连续给药12周后,检测骨密度、骨生物力学,采用酶联免疫吸附法血清骨保护素、抗酒石酸酸性磷酸酶及骨钙素水平,采用Real-time PCR技术检测O3a/Wnt途径中相关mRNA表达水平,采用免疫印迹法检测O3a/Wnt途径中相关蛋白表达水平。结果造模成功大鼠的L4~5(0.33±0.04 vs 0.18±0.03)及左、右股骨骨密度(0.37±0.05 vs 0.23±0.04,0.35±0.04 vs 0.24±0.03)水平明显低于假手术组(P<0.05)。治疗12周后3组大鼠骨最大载荷、三点弯曲应力、骨密度及弹性模量差异有统计学意义,其中假手术组各指标水平最高(36.53±5.23,154.13±6.27,0.34±0.04,3102.34±160.44),其次为依次干预组(19.37±4.32,141.54±6.58,0.18±0.03,2270.18±145.53)、模型组(26.17±4.68,147.56±5.84,0.28±0.03,2804.24±140.42)(P<0.05)。3组大鼠血清骨保护素、抗酒石酸酸性磷酸酶及骨钙素水平差异有统计学意义,其中假手术组的骨保护素(假手术组vs模型组vs干预组:7.53±0.63 vs 4.57±0.42 vs 6.15±0.61)明显高于干预组、模型组,抗酒石酸酸性磷酸酶(假手术组vs模型组vs干预组:14.34±2.87 vs 19.53±3.52 vs 15.96±3.14)、骨钙素(假手术组vs模型组vs干预组:0.84±0.09 vs 1.13±0.12 vs 0.95±0.08)明显低于干预组、模型组(P<0.05)。3组大鼠FoxO3a、Wnt2及β-catenin mRNA和蛋白表达水平差异有统计学意义,其中假手术组的Wnt2、β-catenin明显高于干预组、模型组,FoxO3a明显低于干预组、模型组(P<0.05)。结论利拉鲁肽通过活化O3a/Wnt信号通路而增加骨密度,改善骨生物力学状态,从而有效治疗骨质疏松。
