Objective To explore the effect and mechanism of Chaihu Longgu Muli Decoction(柴胡龙骨牡蛎汤,CHLGMLD)in rats with temporal lobe epilepsy(TLE).Methods A total of 80 Sprague-Dawley(SD)male rats were randomized into cont...Objective To explore the effect and mechanism of Chaihu Longgu Muli Decoction(柴胡龙骨牡蛎汤,CHLGMLD)in rats with temporal lobe epilepsy(TLE).Methods A total of 80 Sprague-Dawley(SD)male rats were randomized into control(CON),model(MOD),carbamazepine(CBZ,0.1 g/kg),CHLGMLD low dose(CHLGMLD-L,12.5 g/kg),and high dose(CHLGMLD-H,25 g/kg)groups,with 16 rats in each group.TLE rat models were established in the four groups with the use of lithium-pilocarpine except for the CON group.After the successful establishment of TLE models,all drugs were administered through gavage,and distilled water was given to rats in the CON and MOD groups for four weeks.The frequency and duration of seizures before and after treatment were recorded for the evaluation of the alleviation degree.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-146a-3p and miR-146a-5p.The expression levels of toll-like receptor 4(TLR4),interleukin-1 receptor-associated kinase 1(IRAK1),tumor necrosis factor(TNF)receptor-associated factor 6(TRAF6),TAK1-binding protein(TAB),nuclear factor-kappa B(NF-κB),and interleukin-1 beta(IL-1β)in hippocampus were tested by immunofluorescence assay.Correlation analysis between the above factors and expressions of miR-146a-3p and miR-146a-5p were performed separately.Results CHLGMLD decreased the frequency(P<0.05)and duration(P<0.01)of seizures in rats.CHLGMLD down-regulated the expression levels of miR-146a-5p and miR-146a-3p(P<0.05),and inhibited the expression levels of TLR4,IRAK1,TRAF6,TAB,NF-κB,and IL-1β(P<0.01).The correlation analysis revealed that the expression levels of TLR4,IRAK1,TRAF6,TAB,NF-κB,and IL-1β were positively correlated with the expression levels of miR-146a-3p and miR-146a-5p detected by qRT-PCR,respectively(P<0.01).Conclusion CHLGMLD can inhibite the TLR4 signaling pathway by lowering the expression levels of miR-146a-3p and miR-146a-5p to alleviate hippocampal dentate gyrus inflammation in TLE rats,thus relieving seizures.展开更多
Chaihu plus Longgu Muli Decoction is the representative treatment method of harmonizing and releasing combined with easing for lesser yang disease proposed by famous physician Zhang Zhongjing.Through clinical cases,th...Chaihu plus Longgu Muli Decoction is the representative treatment method of harmonizing and releasing combined with easing for lesser yang disease proposed by famous physician Zhang Zhongjing.Through clinical cases,this paper firstly introduced the main framework of Chaihu plus Longgu Muli Decoction,to make the treatment method of harmonizing and releasing become more flexible in clinical application.Besides,it introduced the common changes in the learning and application of the decoction,to respond to various changes in the clinical diseases.展开更多
目的观察柴胡加龙骨牡蛎汤(CHLM)对心肌梗死合并焦虑大鼠肠道屏障、内毒素(lipopolysaccharides,LPS)/核因子κB(nuclear transcription factor-κB,NF-κB)介导的心肌炎性损伤、心脏收缩功能、行为学、血清炎症因子的影响。方法将24只...目的观察柴胡加龙骨牡蛎汤(CHLM)对心肌梗死合并焦虑大鼠肠道屏障、内毒素(lipopolysaccharides,LPS)/核因子κB(nuclear transcription factor-κB,NF-κB)介导的心肌炎性损伤、心脏收缩功能、行为学、血清炎症因子的影响。方法将24只雄性SD大鼠随机分为假手术组、模型组、中药组。其中模型组,中药组接受为期3周的焦虑造模,在第2周末模型组及中药组进行心梗造模。假手术组正常饮水,仅挂线,不结扎。术后第二天,中药组予柴胡加龙骨牡蛎汤13.6 g/kg灌胃。模型组及假手术组予等体积生理盐水灌胃。检测大鼠垂直运动分数、水平运动分数、开放臂进入次数百分比(the percentage of open arm entry,OE%)、开放臂进入时间百分比(the percentage of open arm time,OT%)等行为学指标,检测大鼠左室射血分数(left ventricular injection fraction,LVEF)、左室缩短分数(left ventricular systolic function,LVFS)、左心室舒张末期内径(left ventricular internal diameter at end-diastole,LVIDd)和左心室收缩末期内径(left ventricular internal diameter at end-systole,LVIDs)等超声指标。对心脏组织石蜡切片进行苏木精—伊红(hematoxylin-eosin,HE)染色后观察。用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)测定血清LPS及炎性因子水平,用蛋白免疫印迹法(Western-blot,WB)检测心肌组织NF-κB信号通路相关蛋白表达。结果(1)取样前,假手术组7只大鼠存活,模型组,中药组各6只大鼠存活。(2)肠道屏障指标:与模型组相比,中药组闭合蛋白(occludin)、闭锁连接蛋白-1(zonula occludens-1,ZO-1)表达升高,差异具有显著统计学意义。中药组紧密连接蛋白-1(claudin-1)表达相对模型组有升高趋势,但差异无统计学意义。(3)NF-κB相关蛋白:与模型组相比,中药组NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、磷酸化的核因子κB p65与核因子κB p65的比值(ratio of NF-κB pp65 to NF-κB p65,NF-κB pp65/p65)、白细胞介素1β(interleukin-1β,IL-1β)表达下调具有显著统计学意义。核因子κB抑制蛋白α(inhibitory subunit of NF-κB alpha,IκBα)表达上调具有统计学意义。(4)行为学指标:与模型组相比,中药组OE%、OT%、垂直运动评分、水平运动评分升高且具有统计学意义;(5)心功能指标:与模型组相比,中药组LVEF、LVFS升高且均具有统计学意义,LVIDs降低且具有统计学意义,LVIDd有下降趋势,但无统计学差异。(6)血清学指标:与模型组相比,中药组LPS、IL-1β、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)下降具有统计学意义,白细胞介素6(interleukin-6,IL-6)下降具有显著统计学意义。结论柴胡加龙骨牡蛎汤能够改善心肌梗死合并焦虑模型大鼠的肠道屏障损伤、心肌炎性损伤,增强左室收缩功能,减轻焦虑行为,其机制可能与肠道屏障及LPS/NF-κB信号介导的炎症反应相关。展开更多
基金National Natural Science Foundation of China(81874429)Natural Science Foundation of Hunan Province(2020JJ5294)+3 种基金Traditional Chinese Medicine Science&Research Project of Hunan Province(202145)Excellent Youth Program of Hunan Education Department(21B0081)Hunan Provincial Administration of Traditional Chinese Medicine(D2022027)Changsha Natural Science Foundation of China(KQ2202255).
文摘Objective To explore the effect and mechanism of Chaihu Longgu Muli Decoction(柴胡龙骨牡蛎汤,CHLGMLD)in rats with temporal lobe epilepsy(TLE).Methods A total of 80 Sprague-Dawley(SD)male rats were randomized into control(CON),model(MOD),carbamazepine(CBZ,0.1 g/kg),CHLGMLD low dose(CHLGMLD-L,12.5 g/kg),and high dose(CHLGMLD-H,25 g/kg)groups,with 16 rats in each group.TLE rat models were established in the four groups with the use of lithium-pilocarpine except for the CON group.After the successful establishment of TLE models,all drugs were administered through gavage,and distilled water was given to rats in the CON and MOD groups for four weeks.The frequency and duration of seizures before and after treatment were recorded for the evaluation of the alleviation degree.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-146a-3p and miR-146a-5p.The expression levels of toll-like receptor 4(TLR4),interleukin-1 receptor-associated kinase 1(IRAK1),tumor necrosis factor(TNF)receptor-associated factor 6(TRAF6),TAK1-binding protein(TAB),nuclear factor-kappa B(NF-κB),and interleukin-1 beta(IL-1β)in hippocampus were tested by immunofluorescence assay.Correlation analysis between the above factors and expressions of miR-146a-3p and miR-146a-5p were performed separately.Results CHLGMLD decreased the frequency(P<0.05)and duration(P<0.01)of seizures in rats.CHLGMLD down-regulated the expression levels of miR-146a-5p and miR-146a-3p(P<0.05),and inhibited the expression levels of TLR4,IRAK1,TRAF6,TAB,NF-κB,and IL-1β(P<0.01).The correlation analysis revealed that the expression levels of TLR4,IRAK1,TRAF6,TAB,NF-κB,and IL-1β were positively correlated with the expression levels of miR-146a-3p and miR-146a-5p detected by qRT-PCR,respectively(P<0.01).Conclusion CHLGMLD can inhibite the TLR4 signaling pathway by lowering the expression levels of miR-146a-3p and miR-146a-5p to alleviate hippocampal dentate gyrus inflammation in TLE rats,thus relieving seizures.
文摘Chaihu plus Longgu Muli Decoction is the representative treatment method of harmonizing and releasing combined with easing for lesser yang disease proposed by famous physician Zhang Zhongjing.Through clinical cases,this paper firstly introduced the main framework of Chaihu plus Longgu Muli Decoction,to make the treatment method of harmonizing and releasing become more flexible in clinical application.Besides,it introduced the common changes in the learning and application of the decoction,to respond to various changes in the clinical diseases.
文摘目的观察柴胡加龙骨牡蛎汤(CHLM)对心肌梗死合并焦虑大鼠肠道屏障、内毒素(lipopolysaccharides,LPS)/核因子κB(nuclear transcription factor-κB,NF-κB)介导的心肌炎性损伤、心脏收缩功能、行为学、血清炎症因子的影响。方法将24只雄性SD大鼠随机分为假手术组、模型组、中药组。其中模型组,中药组接受为期3周的焦虑造模,在第2周末模型组及中药组进行心梗造模。假手术组正常饮水,仅挂线,不结扎。术后第二天,中药组予柴胡加龙骨牡蛎汤13.6 g/kg灌胃。模型组及假手术组予等体积生理盐水灌胃。检测大鼠垂直运动分数、水平运动分数、开放臂进入次数百分比(the percentage of open arm entry,OE%)、开放臂进入时间百分比(the percentage of open arm time,OT%)等行为学指标,检测大鼠左室射血分数(left ventricular injection fraction,LVEF)、左室缩短分数(left ventricular systolic function,LVFS)、左心室舒张末期内径(left ventricular internal diameter at end-diastole,LVIDd)和左心室收缩末期内径(left ventricular internal diameter at end-systole,LVIDs)等超声指标。对心脏组织石蜡切片进行苏木精—伊红(hematoxylin-eosin,HE)染色后观察。用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)测定血清LPS及炎性因子水平,用蛋白免疫印迹法(Western-blot,WB)检测心肌组织NF-κB信号通路相关蛋白表达。结果(1)取样前,假手术组7只大鼠存活,模型组,中药组各6只大鼠存活。(2)肠道屏障指标:与模型组相比,中药组闭合蛋白(occludin)、闭锁连接蛋白-1(zonula occludens-1,ZO-1)表达升高,差异具有显著统计学意义。中药组紧密连接蛋白-1(claudin-1)表达相对模型组有升高趋势,但差异无统计学意义。(3)NF-κB相关蛋白:与模型组相比,中药组NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、磷酸化的核因子κB p65与核因子κB p65的比值(ratio of NF-κB pp65 to NF-κB p65,NF-κB pp65/p65)、白细胞介素1β(interleukin-1β,IL-1β)表达下调具有显著统计学意义。核因子κB抑制蛋白α(inhibitory subunit of NF-κB alpha,IκBα)表达上调具有统计学意义。(4)行为学指标:与模型组相比,中药组OE%、OT%、垂直运动评分、水平运动评分升高且具有统计学意义;(5)心功能指标:与模型组相比,中药组LVEF、LVFS升高且均具有统计学意义,LVIDs降低且具有统计学意义,LVIDd有下降趋势,但无统计学差异。(6)血清学指标:与模型组相比,中药组LPS、IL-1β、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)下降具有统计学意义,白细胞介素6(interleukin-6,IL-6)下降具有显著统计学意义。结论柴胡加龙骨牡蛎汤能够改善心肌梗死合并焦虑模型大鼠的肠道屏障损伤、心肌炎性损伤,增强左室收缩功能,减轻焦虑行为,其机制可能与肠道屏障及LPS/NF-κB信号介导的炎症反应相关。
