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1.
该研究旨在探讨蚯蚓活性组分(EWAs)对衣霉素(tunicamycin,TM)诱导内质网应激(endoplasmic reticulum stress,ERS)所致肝细胞凋亡的保护作用。体外培养L-02细胞,采用CCK-8法检测细胞存活率,采用流式细胞仪检测细胞凋亡,分别采用Western和q PCR法检测相关蛋白和mRNA的表达情况。研究发现,TM能够以时间和剂量依赖的方式降低L-02细胞的存活率,与正常组相比,模型组中L-02细胞的凋亡率和ERS相关因子GRP78,PERK,e LF2α,ATF4,CHOP,Bax的mRNA和蛋白水平均显著升高(P0.05,P0.01),Bcl-2的mRNA和蛋白水平均显著下降(P0.05,P0.01)。与模型组相比,不同浓度的EWAs处理后,L-02细胞的存活率显著增加,细胞凋亡显著减少,ERS相关因子GRP78,PERK,e LF2α,ATF4,CHOP,Bax的mRNA和蛋白表达水平显著下降(P0.05,P0.01),Bcl-2的mRNA和蛋白水平明显升高(P0.05,P0.01),且呈剂量依赖性。这些结果表明,EWAs对TM诱导L-02细胞ERS所致的肝细胞凋亡具有显著的保护作用,其机制可能与下调ERS相关因子GRP78,PERK,ATF4,e LF2α,CHOP,Bax的表达,减轻L-02细胞的ERS,上调Bcl-2的表达,促进受损细胞增殖有关。  相似文献   

2.
该研究旨在探讨鬼针草乙酸乙酯提取物对内质网应激所致的肝细胞损伤的保护作用及机制。衣霉素诱导L02细胞建立内质网应激损伤模型;四甲基偶氮唑盐(MTT)比色法检测鬼针草乙酸乙酯提取物对内质网应激所致L02细胞损伤的存活率;Western blot法检测内质网应激信号通路蛋白葡萄糖调节蛋白78(GRP78)、蛋白激酶R样内质网激酶(PERK)、真核生物翻译起始因子2α(eIF2α)、转录因子激活蛋白4(ATF4)、C/EBP同源蛋白(CHOP)、B细胞淋巴瘤/白血病基因2(Bcl-2)、Bcl-2结合抗凋亡基因(Bax)的表达水平,同时检测加入内质网应激抑制剂(4-苯基丁酸)和shRNA沉默CHOP因子后上述表达情况;免疫荧光法检测L02细胞中GRP78、PERK、CHOP的表达水平。结果显示,鬼针草乙酸乙酯提取物显著提高内质网应激所致L02细胞损伤的存活率(P0.05或P0.01),并在一定范围内呈时间剂量依赖性。给药组的GRP78、PERK、eIF2α、ATF4、CHOP、Bax表达水平显著降低(P0.05或P0.01),Bcl-2表达水平显著升高(P0.01);加入内质网应激抑制剂和shRNA沉默CHOP因子后,GRP78、PERK、eIF2α、ATF4、CHOP、Bax表达水平显著降低(P0.01),Bcl-2表达水平显著升高(P0.01)。免疫荧光结果显示GRP78、PERK、CHOP表达情况与Western blot法检测结果一致。综上,鬼针草乙酸乙酯提取物对内质网应激所致的L02细胞损伤具有显著保护作用,其机制可能与抑制内质网应激,下调PERK/eIF2α/ATF4/CHOP信号通路所介导的细胞凋亡有关。  相似文献   

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目的研究对药酸枣仁-合欢花对抑郁模型大鼠海马CA3区神经元凋亡及葡萄糖调节蛋白78(GRP78)蛋白表达的影响,从内质网应激(ERS)角度探讨其抗抑郁作用机制。方法将雄性SD大鼠按随机数字表法分为正常对照组、模型组、对药酸枣仁-合欢花组、盐酸文拉法辛组;采用孤养加慢性不可预见性温和应激(CUMS)建立抑郁症大鼠模型,并用糖水消耗实验观察各组大鼠的行为学变化,用苏木素-伊红(HE)染色法观察海马CA3区神经细胞形态及数量。应用DNA原位末端缺口标记(TUNEL)法检测海马CA3区神经元凋亡情况,应用免疫组化法检测海马CA3区ERS通路的关键因子GRP78蛋白表达。结果与正常对照组比较,模型组大鼠糖水偏爱值下降(P 0.01),海马CA3区神经细胞数量减少(P 0.01)、凋亡细胞数和细胞凋亡率增加(P 0.01)、GRP78蛋白表达增加(P 0.01);与模型组比较,对药酸枣仁-合欢花组、盐酸文拉法辛组大鼠糖水偏爱值升高(P 0.05),海马CA3区神经细胞数量增加(P 0.05)、凋亡细胞数和细胞凋亡率降低(P 0.05或P 0.01)、GRP78蛋白表达增加(P 0.05)。结论对药酸枣仁-合欢花能显著改善抑郁模型大鼠的抑郁行为,具有抗抑郁功效,其机制可能与提高海马CA3区GRP78蛋白表达,抑制胱天蛋白酶-12(Caspase-12)表达,减缓ERS引起的细胞凋亡,维持内质网(ER)的稳定有关。  相似文献   

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目的研究对药酸枣仁-合欢花对抑郁模型大鼠海马CA3区神经元凋亡及葡萄糖调节蛋白78(GRP78)蛋白表达的影响,从内质网应激(ERS)角度探讨其抗抑郁作用机制。方法将雄性SD大鼠按随机数字表法分为正常对照组、模型组、对药酸枣仁-合欢花组、盐酸文拉法辛组;采用孤养加慢性不可预见性温和应激(CUMS)建立抑郁症大鼠模型,并用糖水消耗实验观察各组大鼠的行为学变化,用苏木素-伊红(HE)染色法观察海马CA3区神经细胞形态及数量。应用DNA原位末端缺口标记(TUNEL)法检测海马CA3区神经元凋亡情况,应用免疫组化法检测海马CA3区ERS通路的关键因子GRP78蛋白表达。结果与正常对照组比较,模型组大鼠糖水偏爱值下降(P 0.01),海马CA3区神经细胞数量减少(P 0.01)、凋亡细胞数和细胞凋亡率增加(P 0.01)、GRP78蛋白表达增加(P 0.01);与模型组比较,对药酸枣仁-合欢花组、盐酸文拉法辛组大鼠糖水偏爱值升高(P 0.05),海马CA3区神经细胞数量增加(P 0.05)、凋亡细胞数和细胞凋亡率降低(P 0.05或P 0.01)、GRP78蛋白表达增加(P 0.05)。结论对药酸枣仁-合欢花能显著改善抑郁模型大鼠的抑郁行为,具有抗抑郁功效,其机制可能与提高海马CA3区GRP78蛋白表达,抑制胱天蛋白酶-12(Caspase-12)表达,减缓ERS引起的细胞凋亡,维持内质网(ER)的稳定有关。  相似文献   

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目的:探讨斑蟊素致肝脏慢性损伤的可能机制。方法:用不同剂量斑蟊素持续对昆明小鼠灌胃14天,检测其肝功能、肝脏指数和肝脏病理变化,检测ATF-6、XBP1、GRP78、ATF-4、CHOP、BAX、Bcl-2、caspase3、caspase8和caspase9蛋白的表达。用不同浓度斑蟊素连续培养人肝细胞LO2系,显微观察生长,CCK-8检测活性,检测GRP78和CHOP的mRNA的表达,检测GRP78、ATF-4、CHOP、BAX、Bcl-2、caspase3、caspase8和caspase9蛋白的表达。结果:斑蟊素(≥1.0 mg/kg)能增高小鼠转氨酶和肝脏指数,肝细胞坏死及凋亡,且程度与其剂量正相关;ATF-6、XBP1、GRP78、ATF-4、CHOP、BAX、caspase3、caspase8和caspase9蛋白的表达上调,Bcl-2蛋白的表达下调。斑蟊素(≥0.392μg/ml)抑制LO2系细胞活性,增高GRP78和CHOP基因mRNA的表达,GRP78、ATF-4、CHOP、BAX、caspase3、caspase8和caspase9蛋白的表达上调,Bcl-2蛋白的表达下调。结论:斑蟊素可能主要通过内质网应激途径诱导肝细胞凋亡从而导致肝脏的慢性损伤。  相似文献   

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目的 观察茵黄合剂对雌激素诱导胆汁淤积大鼠肝脏组织内质网应激标志性蛋白CHOP、GRP78和PERK表达的影响,基于内质网应激信号通路,探讨茵黄合剂治疗妊娠期肝内胆汁淤积症的机制。方法 运用17-α乙炔雌二醇诱导形成肝内胆汁淤积症大鼠模型后,分别用高(259.2 g/kg/天)、中(129.6 g/kg/天)和低(64.8 g/kg/天)剂量的茵黄合剂干预7天,取血,检测血清总胆汁酸(total bile acid,TBA)、总胆红素(total bilirubin)等生化指标;取部分肝脏组织,分别在光学显微镜和透射电镜下观察大鼠肝脏形态和超微结构的改变;另取肝脏组织,置于液氮中保存,分别采用实时定量PCR法和蛋白免疫印迹法检测大鼠肝脏组织内质网应激标志性蛋白:C/EBP同源蛋白(C/EBP-homologous, CHOP)、葡萄糖调节蛋白78(glucose regulated protein, GRP78)和蛋白激酶R样ER激酶(protein kinase R-like ER kinase, PERK)mRNA与蛋白表达情况。结果 不同剂量的茵黄合剂干预均可显著降低胆汁淤积模型鼠肝脏组织内质网应激标志性蛋白CHOP、GRP78和PERK mRNA和蛋白表达,降低血清总胆汁酸和总胆红素水平,减轻肝脏组织形态和超微结构损伤,与模型组比较,差异具有统计学意义。结论 茵黄合剂可能通过减少雌激素诱导肝内胆汁淤积模型鼠肝脏CHOP、GRP78和PERK的表达,缓解内质网应激,从而降低血清总胆汁酸和总胆红素水平,发挥治疗肝内胆汁淤积的作用。  相似文献   

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目的:观察中药复方益气补肾调脂方对高脂高果糖饮食诱导的非酒精性脂肪性肝炎(NASH)小鼠的肝脏内质网应激相关基因m RNA的影响。方法:高脂高果糖饮食诱导16周,建立NASH小鼠模型,在造模第5周开始,分别予以益气补肾调脂方高、中、低三种剂量干预12周,在16周末处死小鼠。采用ELISA法检测血清游离脂肪酸(FFAs)的水平,real time-PCR法检测肝组织内质网应激相关基因mRNA的表达水平。结果:模型组血清FFAs、肝组织葡萄糖调节蛋白78(GRP78)、激活转录因子6(ATF6)、CCAAT增强子结合蛋白同源蛋白(CHOP)mRNA的表达水平较正常组升高(P0.05,P0.01),肝组织X-盒结合蛋白1(XBP1)mRNA的表达水平较正常组下降(P0.05)。经益气补肾调脂方干预后,小鼠血清FFAs水平、肝组织GRP78、ATF6、CHOPmRNA较模型组下降(P0.05),肝组织XBP1mRNA表达较模型组有所上升(P0.05)。结论:益气补肾调脂方可能通过改善肝脏内质网应激来发挥抗NASH的作用。  相似文献   

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目的基于IRE1-XBP1通路探究脊髓康(JSK)抑制内质网应激(ERS)诱导的PC12细胞凋亡的作用及机制。方法利用神经生长因子(NGF)诱导PC12细胞为神经元样细胞并通过流式细胞技术鉴定;诱导后细胞分为空白组(无干预措施)、模型组[2μmol/L毒胡萝卜素(TG)]、脊髓康组(2μmol/L TG及0.0521 g/mL JSK水提物冻干粉,合生药0.625 g/mL);CCK8法检测各组细胞24、48、72 h增殖活性,流式细胞技术检测各组细胞凋亡状态,Western Blot与q-PCR检测1型内质网转膜蛋白激酶(IRE1)、X盒结合蛋白1(XBP1)、葡萄糖调节蛋白78(Grp78)及C/EBP同源蛋白(CHOP)的蛋白及mRNA表达情况,透射电镜扫描各组细胞观察细胞及内质网形态学特征。结果与空白组比较,模型组NGF诱导的神经元样PC12细胞增殖活性显著降低(P0.05),晚期凋亡明显增高(P0.01),电镜扫描显示其细胞粗面内质网结构肿胀,囊泡化,呈典型的细胞凋亡形态;Grp78、CHOP蛋白表达显著升高(P0.05),XBP1及IRE1蛋白表达也有微微上调,但差异无统计学意义(P0.05);Grp78、CHOP、XBP1及IRE1 mRNA表达明显升高(P0.05,P0.01)。与模型组比较,JSK组24、48、72 h PC12细胞增殖活性升高(P0.05),晚期凋亡及早期凋亡均明显降低(P0.05,P0.01);Grp78、CHOP的蛋白及mRNA表达均降低(P0.05),XBP1及IRE1蛋白及mRNA表达明显上调(P0.05)。透射电镜观察JSK组内质网结构仍存在,存在完整的外膜,内质网膜破损较轻,可见部分凋亡形态学改变。结论 JSK可能通过活化IRE1-XBP1途径改善TG诱导的内质网应激状态,从而抑制ERS导致的神经元样PC12细胞凋亡,这可能是其有效改善脊髓损伤后神经元功能修复的机制。  相似文献   

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目的观察积雪草苷(AC)通过内质网应激(ERS)相关葡萄糖调节蛋白78(GRP78)/蛋白激酶R样内质网激酶(PERK)/转录因子C、EBP同源蛋白(CHOP)通路对急性心肌梗死(AMI)大鼠的心肌保护作用。方法选择SPF级8周龄级SD大鼠108只,结扎左冠状动脉前降支建立心肌梗死模型,随机分为假手术组,模型组,积雪草苷低、中、高剂量组,牛磺酸组,造模完成24 h后给药,每天早晚各1次,4周后测定左室质量和体质量,TCC染色法测定心肌梗死面积,TUNEL法检测心肌细胞凋亡率,HE染色观察心肌细胞形态,Westorn blotting检测GRP78、PERK、CHOP及凋亡相关蛋白Bax、Bcl-2、Caspase-12、Caspase-3的表达。结果与假手术组比较,模型组左室质量和左室质量/体质量的比值显著增加(P 0.05),心肌梗死面积显著增加,心肌细胞凋亡率显著增加;与模型组比较,积雪草苷各组和牛磺酸组左室质量和左室质量/体质量的比值显著下降(P 0.05),梗死面积下降,细胞凋亡比例显著减少(P 0.05),且积雪草苷各组呈浓度依赖性降低。HE染色显示假手术组心肌纤维排列整齐,横纹清晰,无水肿或坏死;模型组心肌纤维不规则,横纹不清或缺失,积雪草苷各组和牛磺酸组心肌纤维轻度扩张,心肌纤维局灶性丢失。Western blotting检测显示,与假手术组比较,模型组GRP78、PERK、CHOP、Bax、Caspase-12、Caspase-3蛋白表达水平显著升高(P 0.05),Bcl-2蛋白水平显著降低(P 0.05);与模型组比较,积雪草苷各组随着浓度的升高GRP78、p-PERK、CHOP、Bax、Caspase-12、Caspase-3蛋白表达逐渐降低(P 0.05),Bcl-2蛋白表达逐渐升高(P 0.05),均呈剂量依赖性。结论 AC可减少大鼠心肌梗死区,可通过抑制GRP78/PERK/CHOP通路发挥对急性心肌梗死大鼠的保护作用。  相似文献   

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汤卫国  王奇志  印敏  王鸣  陈雨  张建华  冯煦 《中草药》2014,45(15):2172-2177
目的建立微波消解-电感耦合等离子体质谱(ICP-MS)直接稀释测定脉络宁注射液中25种矿物质元素(Mg、Ca、Fe、Cu、Zn、Mn、Al、B、Ba、Co、Cr、K、Li、Mo、Na、Ni、P、Pb、Sr、Th、Ti、V、As、Cd和Hg)的方法。方法分别对微波消解条件和测试条件进行考察;样品经微波消解后,采用电感耦合质谱仪测定25种矿物质元素,并对测定方法学进行考察。结果确定最佳消解条件为3步缓慢升温:400 W 80℃升温10 min,保留5 min;600 W 120℃升温10 min,保留5 min;900 W 200℃升温20 min,保留20 min;25种矿物质元素在各自的线性范围内线性关系良好,r≥0.999 6,精密度、稳定性和重复性试验的RSD均符合定量分析要求;加标回收率为94.7%~106.1%,RSD在0.34%~2.79%。脉络宁注射液中检测出Mg、Ca、Fe、Cu、Zn、Mn、Al、B、Ba、Co、Cr、K、Li、Mo、Na、Ni、P、Pb、Sr、Th、Ti、V,未检出As、Cd和Hg。结论该方法简便、迅速、准确,适用于脉络宁注射液中25种矿物质元素的同时测定。  相似文献   

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The disturbance and instability of laterality are obstacles to diagnostics and therapy. Correction prior to starting therapy is required. They also predispose toward defined health problems and unspecific diseases. Numerous research activities provide evidence of the relevance of undisturbed laterality in diagnostics and therapy. Techniques of testing and therapeutic corrections will allow for optimized therapy success.  相似文献   

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The Journal of Acupuncture and Tuina Science, a journal with an international scope (IS SN 1672-3597, CN 31-1908/R, Bimonthly), is embodied by 'Springer Verlag' Database, Index Copernicus (IC) and Chinese Scientific and Technical Paper and Citations Data (CSTPCD). You can search full text on http://www, springerlink, com/content/1672 -3597.  相似文献   

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正Beverages based on Ilex paraguariensis A.St.-Hil.are used in the south Brazil and other Latin American countries located at the so-called southern cone.It is known as chimarro or mate in south and southeast Brazil,tererêin  相似文献   

16.
This paper deals with an interdisciplinary study covering historic, botanical, phytochemical, pharmacological and clinical aspects of rhubarb and related species, to lay stress on the correlation between plant phylogeny, chemical constituents and purgative activity.

It was found that the official rhubarbs were exclusively restricted in the Sect. Palmata e.g. Rheum palmatum R. palmatum var. tanguticum R. officinale; the following criteria may serve as their standard, viz., the presence of sennoside derivatives and rhein, the occurrence of the reduced form of rhein and aloe-emodin, the leaves with any kind of palmate incision. Comprehensive multivariate analyses showed that there is a very close relationship between the leaf incision, existence of sennosides or rhein and purgative activity.  相似文献   


17.

Background

At present, the state-of-the-art medication in treating idiopathic taste disorder (gustatory dysfunction) is zinc. If zinc medication was unsuccessful, not tolerated or if it is contraindicated the persons affected can hardly be helped.

Objectives

In a randomized placebo-controlled clinical trial an attempt was made to determine the efficacy of acupuncture.

Methods

The treatment group was treated with acupuncture needles, the placebo group with an inactive acupuncture-laser. Endpoints of the study were changes in the taste test, the quality of life and the severity of symptoms of depression (BDI) as well as mood changes (ZMS). Satisfaction with the success of the respective therapy was also evaluated. Data were collected before and after therapy and again for the treatment group 10 weeks after completion of the treatment period.

Findings

Treatment with real acupuncture showed a significant improvement in gustatory function as well as a significant improvement in psychological symptoms. Greater satisfaction with the treatment results was evident in the treatment group. The treatment results of acupuncture remained stable over a period of ten weeks after completion of the treatment.

Conclusion

Acupuncture was effective in the therapy of idiopathic taste disorders and therefore it is an effective alternative when zinc medication is not tolerated or unsuccessful. But it is also suitable as a therapy of first choice. Acupuncture can be considered as an extension of the therapeutic spectrum in the difficult treatment of taste disorders.  相似文献   

18.

Background

Experimental and clinical studies postulate a neurobiological mechanism of acupuncture by modulating central pain systems, neurohormones and -transmitters. It seems to be quite likely that the autonomic nervous system (ANS) plays an important mediating role in this neuromodulation.

Aim

In measuring heart rate variability (HRV), as a non-invasive approach to the ANS-function, the object of this study was to monitor probable acupuncture-induced changes of autonomic balance. The innovative technology of laser-needle acupuncture offers for the first time a stimulation method which fulfils the demands on a placebo-controlled acupuncture trial not requiring controversial sham acupuncture.

Method

19 healthy voluntary subjects underwent 3 treatments in randomized sequence, consisting of placebo-, 1-point- (PC 6) and simultaneous 3-point- (PC 6, LR 3, SI 3) laser-needle-stimulation over 20 minutes. Before, during and after placebo-/verum laser-needle acupuncture, HRV-data for normalized HF(high frequency)- and LF(low frequency)-Power (nHF, nLF) and their quotient nLF/nHF obtained by power spectrum analysis (FFT), were measured and analyzed statistically at 5 different time points (ANOVA repeated measures, p < 0,05).

Results

During both verum applications, the 1-point- (PC 6) as well as the simultaneous 3-point- (PC 6, LR 3, SI 3) laser-needle stimulation, significant changes of the ANS-activity were found in comparison to the placebo application. The most significant modification of HRV was an increase of nHF (p < 0,05 (PC 6), p < 0,0001 (PC 6, LR 3, SI 3)) as an expression of growing vagal activity during acupuncture.

Conclusion

Laser-needle acupuncture causes in 1-point-stimulation (PC 6) as well as in simultaneous 3-point-stimulation (PC 6, LR 3, SI 3) significant changes in HRV in comparison to placebo application. HRV-monitoring seems to be a suitable approach in exploration of acupuncture-induced changes of ANS and could possibly be established in combination with laser needle acupuncture as a standard for further scientific and clinical acupuncture investigations which are greatly needed.  相似文献   

19.
瑞香科植物广泛分布于世界热带和温带地区,其中多种植物可供药用。在对该科植物的研究中发现,黄酮类、木脂素和香豆素等酚性化合物的低聚体为其重要的特征性成分,药理活性主要表现在抗肿瘤、抗炎、抗氧化、抗菌和抗病毒等方面。对已发现的酚性聚合体的化学结构、植物来源及药理活性进行综述,为瑞香科植物今后的研究和开发利用提供参考。  相似文献   

20.
目的采用HPLC梯度洗脱法同时测定沉香化滞丸中沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚11种成分。方法采用Thermo Syncronis C18色谱柱(250 mm×4.6 mm,5μm),流动相为水-乙腈,梯度洗脱:0~10 min,20%乙腈;10~20 min,20%~40%乙腈;20~24 min,40%乙腈;24~26 min,40%~52%乙腈;26~30 min,52%乙腈;30~31 min,52%~90%乙腈;31~35 min,90%乙腈;35~40 min,90%~100%乙腈;40~43min,100%乙腈;43~45min,100%~20%乙腈;检测波长215nm,体积流量1.0m L/min,柱温30℃,进样量20μL。结果各成分在43 min内分离良好,沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚的线性范围分别为1.4~13.6、10.0~200.0、31.5~315.0、1.0~120.1、1.8~50.6、0.93~10.1、1.8~30.0、0.2~40.3、1.8~18.1、1.7~25.0、0.45~10.70μg/mL;样品中各成分的平均回收率均在98.90%~100.87%;11种成分精密度RSD在0.55%~1.54%;供试品溶液在30 h内稳定性良好,RSD在0.75%~1.94%;重复性RSD在0.39%~1.73%。6批次样品中沉香四醇、柚皮苷、橙皮苷、新橙皮苷、和厚朴酚、大黄素、厚朴酚、木香烃内酯、去氢木香内酯、大黄酚、大黄素甲醚质量分数分别为92.0~201.0、511.5~9 033.0、5 475.0~12 635.5、54.5~5 095.5、192.0~2 137.5、117.0~391.5、106.5~1 281.5、13.0~136.5、93.5~199.0、177.0~1 207.0、33.5~251.5μg/g。结论本方法准确、快速、简便,重复性好,精密度高,适用于沉香化滞丸中多种活性成分的定量分析。  相似文献   

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