A Clinical Study of Safflower Yellow Injection in Treating Coronary Heart Disease Angina Pectoris with Xin-Blood Stagnation Syndrome

Objective： To evaluate the clinical effect and safety of Safflower Yellow injection （SYI） in treating coronary heart disease angina pectoris （OHD-AP） with Xin-blood stagnation syndrome （XBSS）. Methods： Adopted was the multi-centered, randomized, positive parallel controlled method, 448 patients with CHD-AP-XBSS were enrolled and divided into two groups, 336 in the tested group treated with SYI and 112 in the control group treated with Salvia injection by intravenous dripping once a day for 14 days, so as to observe the conditions of angina, electrocardiogram, and therapeutic effect on traditinal Chinese medicine （TCM） symptoms as well as the safety of the treatment. Results： The significantly effective rate and total effective rate in the tested group were 60.06% （194/323） and 91.02 % （294/323） respectively; those in improvement of TOM symptoms were 40. 18% （129/321） and 75.23% （243/323） respectively, which were better than those in the control group （P〈0.01）. Conclusion： SYI Injection is effective and safe in treating OHD-AP-XBSS.

Inductive effect of hydroxyl safflower yellow-A on apoptosis in abnormal HUVEC via the mitochondrial pathway

Objective:To investigate the mechanism by which hydroxyl safflower yellow A,an active component of safflower (Carthamus tinctorius L.),promotes apoptosis in abnormal human umbilical vein endothelial cells (HUVECs).Methods:Supernatant of BGC-823 was used to stimulate HUVECs to establish a model of abnormal proliferation of HUVECs.After determining an ideal concentration of HSYA by MTT assay,apoptosis was detected with flow cytometry and TUNEL assay.Mechanism of apoptosis was assessed using quantitative real-time polymerase chain reaction,Western blot,and ELISA.Results:A range of concentrations of HSYA inhibited proliferation and promoted apoptosis of abnormal HUVECs.As the rate of apoptosis increased,mRNA expression of caspase-3 increased while expression of mutant p53 decreased.HSYA had no effect on Fas gene expression.Analogously,protein expression of Bax was increased while those of Bcl-2,Fas,and Fas-L were decreased.Conclusions:HSYA appears to induce apoptosis of HUVECs with the stimulation of the supematant of tumor cells.The mechanism of apoptosis by HSYA may involve activation of the mitochondrial apoptotic pathway and regulation of the expressions of Bcl-2,Bax,and p53.

Effect of safflower yellow pigment combined with cerebellin on neurological function in patients with ischemic stroke and its mechanism

Objective: To investigate the effects of safflower yellow pigment combined with cerebellin on neurological function in patients with ischemic stroke and analyze its possible mechanism. Methods: A total of 130 patients with ischemic stroke admitted to my hospital from May 2017 to August 2018 were enrolled. The patients were divided into the control group and the study group according to the random number table method, 65 cases in each group. The control group was treated with cerebroside carnosine. And the study group was treated with safflower yellow pigment on the basis of the control group. The changes of neurological function index, oxidative stress index, vascular endothelial function index and inflammatory factor were compared before and after treatment. Results: There were no significant differences in preoperative neurological parameters, oxidative stress, vascular endothelial function and inflammatory factors between the two groups (P>0.05). After treatment, the neurological function indexes S-100β and NSE were significantly lower in both groups, and NGF levels were significantly increased. The S-100β and NSE levels in the study group were (0.91±0.10) ng/L and (12.91±1.33) ng/L, respectively, which were significantly lower than control group. While the NGF level of the study group was (79.52±8.07) μg/L significantly higher than that of the control group (both P<0.05). The levels of Ox-LDL were significantly reduced and GSH-Px levels were significantly elevated in both groups after treatment. The level of oxidative stress index Ox-LDL was (563.51±57.10) μg/dL, which was significantly lower than that of the control group, and the GSH-Px level was (154.55±16.07) U/L, which was significantly higher than the control group (both P<0.05). The levels of vascular endothelial function NT-proBNP, TXB2 and ET-1 were significantly lower in the two groups after treatment. The NT-proBNP, TXB2 and ET-1 in the study group were (95.91±9.77) pg/mL, (245.69±25.06) pg/mL and (64.26±6.65) ng/L, respectively, which were significantly lower than the control group (both P<0.05). The levels of inflammatory factors MMP-9 and TNF-α were significantly lower in the two groups after treatment. The levels of MMP-9 and TNF-α in the study group were (60.64±6.12) ng/mL and (0.33±0.04) ng/mL, respectively, which were significantly lower than those in the control group (both P<0.05). Conclusion: Safflor yellow combined with cerebellin in the treatment of ischemic stroke has higher clinical efficacy and can significantly improve the neurological function of patients. The possible mechanism is related to the improvement of vascular endothelial function and stress response.

羟基红花黄色素A抑制糖氧剥夺/复糖复氧处理后神经元焦亡的作用

背景:羟基红花黄色素A具有抗缺血、抗氧化、抗血栓及抗炎等作用,其是否影响糖氧剥夺/复糖复氧处理后神经元焦亡,目前尚不清楚。目的:探讨羟基红花黄色素A对神经元焦亡的干预作用及机制。方法:取对数生长期的HT22细胞,随机分为5组:正常组、模型组、羟基红花黄色素A组、Colivelin组、Colivelin+羟基红花黄色素A组。利用糖氧剥夺/复糖复氧处理HT22细胞建立神经元焦亡模型,然后给予STAT3激动剂Colivelin、羟基红花黄色素A干预。干预后JC-1探针检测线粒体膜电位变化,活性氧试剂盒检测细胞内活性氧含量,GSDMD/TUNEL染色观察细胞焦亡情况,免疫荧光检测STAT3、GSDMD蛋白表达,RT-PCR检测STAT3、NLRP3、Caspase-1 mRNA表达,Western blot检测p-STAT3、NLRP3、GSDMD、Cleaved-caspase-1、白细胞介素1β蛋白表达。结果与结论:①与正常组相比,模型组焦亡细胞数量增多,p-STAT3、NLRP3、Cleaved-caspase-1、GSDMD、白细胞介素1β蛋白表达显著升高;与模型组相比,羟基红花黄色素A组焦亡细胞数量减少,焦亡相关蛋白的表达显著降低;②与模型组相比,Colivelin组细胞焦亡加剧,线粒体膜电位降低,活性氧含量增加,STAT3、NLRP3、Caspase-1 mRNA表达升高,p-STAT3、NLRP3、GSDMD、Cleaved-caspase-1、白细胞介素1β蛋白表达升高;与Colivelin组相比,Colivelin+羟基红花黄色素A组上述指标均有好转。结果表明:羟基红花黄色素A通过STAT3信号通路抑制糖氧剥夺/复糖复氧后HT22细胞焦亡发挥神经保护作用。

Safflower Yellow Inhibits Progression of Hepatocellular Carcinoma by Modulating Immunological Tolerance via FAK Pathway

Objective To explore the anti-tumor effect of safflower yellow(SY)against hepatocellular carcinoma(HCC)and the underlying potential mechanism.Methods An in vitro model was established by mixing Luc-Hepa1-6 cells and CD3^(+)CD8^(+)T cells,followed by adding programmed cell death protein 1(PD-1)antibody(Anti-mPD-1)with or without SY.The apoptosis was detected by flow cytometry and the level of inflammatory cytokines was determined by enzyme-linked immunosorbent assay.The protein levels of programmed cell death 1 ligand 1(PD-L1),chemokine ligand(CCL5),C-X-C motif chemokine ligand 10(CXCL10)were measured by Western blot.An in situ animal model was established in mice followed by treatment with anti-mPD-1 with or without SY.Bioluminescence imaging was monitored with an AniView 100 imaging system.To establish the FAK-overexpressed Luc-Hepa1-6 cells,cells were transfected with adenovirus containing pcDNA3.1-FAK for 48 h.Results The fluorescence intensity,apoptotic rate,release of inflammatory cytokines,and CCL5/CXCL10 secretion were dramatically facilitated by anti-mPD-1(P<0.01),accompanied by an inactivation of PD-1/PD-L1 axis,which were extremely further enhanced by SY(P<0.05 or P<0.01).Increased fluorescence intensity,elevated percentage of CD3+CD8+T cells,facilitated release of inflammatory cytokines,inactivated PD-1/PD-L1 axis,and increased CCL5/CXCL10 secretion were observed in Anti-mPD-1 treated mice(P<0.01),which were markedly enhanced by SY(P<0.05 or P<0.01).Furthermore,the enhanced effects of SY on inhibiting tumor cell growth,facilitating apoptosis and inflammatory cytokine releasing,suppressing the PD-1/PD-L1 axis,and inducing the CCL5/CXCL10 secretion in Anti-mPD-1 treated mixture of Luc-Hepa1-6 cells and CD3+CD8+T cells were abolished by FAK overexpression(P<0.01).Conclusion SY inhibited the progression of HCC by mediating immunological tolerance through inhibiting FAK.

红花黄色素对谷氨酸/过氧化氢诱导的SH-SY5Y细胞凋亡保护作用

为研究红花黄色素对谷氨酸/过氧化氢诱导SH-SY5Y细胞损伤的保护作用。将SH-SY5Y细胞随机分为正常组,模型组(谷氨酸/过氧化氢损伤),红花黄色素不同剂量组(0.01、0.1、1 g/L)。利用MTT法测定SH-SY5Y细胞存活率,Hoechst 33342染色法检测细胞凋亡情况,RT-PCR法测定Bax,Bcl-2和Bcl-x m RNA表达水平。MTT实验结果显示,谷氨酸/过氧化氢诱导后的细胞生存率分别下降至51.15%和63.08%(P<0.01),红花黄色素不同剂量组可以显著提高细胞存活率,且成浓度依赖性(P<0.05),红花黄色素高剂量组亦可以明显降低谷氨酸/过氧化氢损伤所诱导的细胞凋亡;与模型组相比,红花黄色素能够使促凋亡基因Bax的表达显著降低(P<0.05),而抗凋亡基因Bcl-2和Bcl-xl的表达显著增加(P<0.05)。由此可知,红花黄色素对谷氨酸/过氧化氢诱导的SH-SY5Y细胞损伤具有保护作用,其机制可能与调节凋亡相关基因的表达有关。

HSYA对骨关节炎大鼠膝关节软骨损伤的修复作用研究

目的探讨羟基红花黄色素A(HSYA)对骨关节炎(OA)大鼠膝关节及MMP-3、TGF-β1、Notch1的影响。方法将25只大鼠随机分为正常组(假手术)、OA组(模型大鼠)、低HSYA组(模型大鼠尾部注射2.5 mg/kg HSYA)、中HSYA组(模型大鼠尾部注射5.0 mg/kg HSYA)、高HSYA组(模型大鼠尾部注射10.0 mg/kg HSYA)。采用ELISA检测血清指标IL-1β、TNF-α,Pelletier评分评估膝关节软骨损伤情况,HE染色观察组织病理形态并以Mankin评分进行评估,免疫组化检测MMP-3阳性表达,Western blot检测TGF-β1、Notch1蛋白水平。结果OA组血清中IL-1β、TNF-α水平高于其余各组(P<0.05),高HSYA组IL-1β、TNF-α水平低于低HSYA组和中HSYA组(P<0.05)。各组大鼠Pelletier评分组间比较差异有统计学意义(P<0.05)。OA组软骨和滑膜组织Mankin评分高于其余各组(P<0.05),高HSYA组Mankin评分低于低HSYA组、中HSYA组(P<0.05)。各组大鼠关节组织MMP-3阳性细胞率组间比较差异有统计学意义(P<0.05)。各组大鼠关节组织TGF-β1、Notch1蛋白水平组间比较差异有统计学意义(P<0.05)。结论HSYA能够降低OA大鼠血清炎性因子IL-1β、TNF-α水平,缓解滑膜组织损伤,且存在浓度依赖性,其机制与抑制MMP-3和Notch1表达、提高TGF-β1水平相关。

HSYA与芍药苷联用对脑缺血再灌注损伤大鼠PI3K、Akt mRNA表达的影响

目的:探讨羟基红花黄色素A(HSYA)与芍药苷联合用药对急性局灶性脑缺血再灌注损伤大鼠PI3K/Akt信号通路中PI3K、Akt mRNA表达的影响。方法:SPF级雄性SD大鼠60只,随机分为6组,每组10只,分别为模型组、假手术组、HSYA组、芍药苷组、银杏内酯组、合用组。复制脑缺血再灌注模型。脑缺血1 h再灌注6 h后尾静脉注射相应的药物7天,每天1次。HSYA组灌胃HSYA溶液5.0 mg/(kg·d);芍药苷组灌胃芍药苷溶液5.0 mg/(kg·d);合用组是HSYA与芍药苷联合用药,灌胃HSYA溶液和芍药苷溶液各5.0 mg/(kg·d);银杏内酯组灌胃银杏内酯注射液5.0 mg/(kg·d);模型组和假手术组灌胃等量生理盐水。末次给药2 h后采集相应标本保存,qRT-PCR法检测大鼠海马组织PI3K、Akt mRNA的表达。结果:与假手术组比较,模型组大鼠PI3K mRNA表达量显著减少,差异有统计学意义(P<0.01)。与模型组比较,HSYA组、芍药苷组、合用组、银杏内酯组大鼠海马组织中PI3K mRNA的表达显著降低,差异均有统计学意义(P<0.05)。与合用组比较,HSYA组、银杏内酯组大鼠PI3K mRNA的表达量较高,差异均有统计学意义(P<0.05)。结论:HSYA与芍药苷联合用药对脑缺血再灌注损伤模型大鼠的保护机制可能与显著调节PI3K/Akt信号通路中PI3K mRNA过表达,微调Akt mRNA过表达有关。

Stability and degradation of hydroxysafflor yellow A and anhydrosafflor yellow B in the Safflower injection studied by HPLC-DAD-ESI-MS^n

Safflower is a popular Chinese medicinal plant and Safflower injection is extensively used for the clinical treatment of cerebrovascular and cardiovascular diseases. In this study, HPLC-DAD-ESI-MSn was utilized to study the stability and degradation of the two major but chemically unstable bioactive compounds hydroxysaffior yellow A and anhydrosaffior yellow B, in Safflower injection. The impact of light irradiation, temperature, and pH on the stability of these two compounds were studied. The results showed that hydroxysafflor yellow A and anhydrosafflor yellow B could degrade at high temperature （〉60 ℃） or extreme pHs （pH ≤ 3.0 or 〉7.0）, but not under light irradiation. The common degradation product was p-coumaric acid. Chemical structures of the other degradation products were characterized by LC-MS. Hypothetical degradation pathways were proposed. In addition, ADP-induced platelet aggregation tests showed that the degradation of anhydrosaffior yellow B could reduce the anticoagulation activities of Safflower injection. Our results suggest that temperature and pH are critically important for the preparation and storage of Safflower injection.

羟基红花黄色素调节Sirt 3改善心脏缺血再灌注损伤中线粒体功能障碍

目的探讨羟基红花黄色素对脂多糖诱导的H9c2心肌细胞损伤线粒体功能保护作用及其对Sirt3的调节作用。方法H9c2心肌细胞株培养24 h后随机分为对照组、脂多糖造模组、脂多糖造模+Sirt3抑制剂组,羟基红花黄色素给药+模型组,羟基红花黄色素给药+Sirt 3抑制剂+模型组,给予脂多糖建立氧化应激损伤心肌细胞模型。采用乳酸脱氢酶检测试剂盒测定细胞培养液中乳酸脱氢酶含量;测定细胞中ATP含量,测定细胞中细胞色素C含量及线粒体呼吸链复合物含量;Western blot法检测Sirt 3及Bax表达量。结果与对照组比较,脂多糖模型组细胞色素C含量升高,细胞中谷氨酸脱氢酶,ATP及呼吸链复合物含量降低,Sirt3表达量略升高,Bax表达上升(P<0.05)与模型组比较,脂多糖造模+Sirt3抑制组线粒体各项损失指标明显加剧,而羟基红花黄色素给药+模型组中线粒体损伤指标明显恢复(P<0.05),而羟基红花黄色素+Sirt 3抑制剂组中线粒体损伤程度低于脂多糖造模+Sirt3抑制剂组(P<0.05),Sirt3蛋白表达较抑制组明显上升(P<0.05)。结论羟基红花黄色素降低脂多糖诱导的H9c2心肌细胞线粒体氧化损伤,该作用可能与羟基红花黄色素提高受损心肌细胞内Sirt3的表达量有关。

基于TLR4/NF-κB信号通路探究红花黄色素对不同月龄APP/PS1小鼠学习记忆能力的影响

目的 采用3、6、9月龄APP/PS1转基因小鼠作为实验动物模型,明确红花黄色素(safflower yellow, SY)对不同疾病时期APP/PS1小鼠学习记忆能力的影响,探究SY抗阿尔茨海默病(Alzheimer’s disease, AD)的作用机制。方法 行为学实验观察SY对不同月龄APP/PS1小鼠学习记忆的影响;ELISA研究SY对不同月龄小鼠皮层炎症因子表达的影响,并采用Western blot检测小鼠小胶质细胞活化标志蛋白,进一步分析其作用机制。结果 SY提高了3、6、9月龄小鼠的学习记忆能力;降低脑组织中IL-6的含量,提高TGF-β1的含量;诱导各月龄小鼠脑组织中精氨酸酶-1(arginase-1, Arg-1)和髓样细胞触发受体2(triggering receptor expressed on myeloid cells 2,TREM2)的蛋白表达水平,同时下调诱导型一氧化氮合酶(inducible nitric oxide synthase, iNOS)、Toll样受体4(Toll-like receptors, TLR4)以及核因子-κB(nuclear factor-kappa B,NF-κB)的蛋白表达水平。结论 与3、9月龄小鼠相比,SY对6月龄APP/PS1小鼠的学习记忆能力改善效果最佳。SY通过诱导APP/PS1转基因小鼠脑组织中TREM2的表达,抑制TLR4/NF-κB通路激活,促进小胶质细胞表型向抗炎型转变,减轻慢性神经炎症反应,改善各月龄APP/PS1小鼠的学习记忆能力。

红花黄色素对糖尿病小鼠足溃疡创面愈合的影响及机制

目的探讨红花黄色素(SY)对糖尿病小鼠足溃疡(DFU)创面愈合的影响及分子机制。方法取45只C57BL/6小鼠腹腔注射链脲佐菌素建立糖尿病模型。将糖尿病小鼠随机分为假手术组、模型组、低剂量SY干预组、高剂量SY干预组及高剂量SY联合胰岛素样生长因子-1(IGF-1)组,每组9只。DFU模型制备前,模型组小鼠不给予干预处理,低剂量SY干预组和高剂量SY干预组分别腹腔注射5、20 mg·kg^(-1)SY,高剂量SY联合IGF-1组腹腔注射20 mg·kg^(-1)SY和0.03 mg·kg^(-1)IGF-1。除假手术组外,其余4组小鼠通过切开足背部皮肤制备DFU模型;假手术组不制作足背部皮肤伤口,余手术步骤与模型组一致。于造模后第14天,用电子秤测量各组小鼠的体质量,采集尾静脉血测定空腹血糖,并使用小型游标卡尺测量创面宽度,然后处死小鼠收集创面组织,采用苏木精-伊红染色检测各组小鼠创面组织病理学变化,反转录-实时荧光定量聚合酶链反应检测各组小鼠创面组织中血小板衍生生长因子(PDGF)、血管内皮生长因子(VEGF)、α-平滑肌肌动蛋白(α-SMA)和I型胶原蛋白(collagen I)、蛋白酪氨酸磷酸酶1B(PTP1B)、晚期糖基化终产物(AGEs)mRNA的相对表达量,蛋白质印迹法检测各组小鼠创面组织中增殖标志物Ki-67、增殖细胞核抗原(PCNA)、凋亡相关蛋白(caspase-3、caspase-6、caspase-7)、p85磷脂酰肌醇-3-激酶(PI3K)和磷酸化蛋白激酶B(p-AKT)蛋白的相对表达量,酶联免疫吸附试验检测各组小鼠创面组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β及IL-6水平。结果假手术组、模型组、低剂量SY干预组、高剂量SY干预组及高剂量SY联合IGF-1组小鼠血糖和体质量比较差异均无统计学意义(P>0.05)。高剂量SY干预组小鼠创面组织愈合率显著大于模型组、低剂量SY干预组及高剂量SY联合IGF-1组(P<0.01)。模型组、低剂量SY干预组及高剂量SY联合IGF-1组创面组织愈合率比较差异无统计学意义(P>0.05)。高剂量SY干预组小鼠创面组织中出现大量胶原纤维密集有序排列,并伴有大量新生血管;模型组、低剂量SY干预组及高剂量SY联合IGF-1组小鼠创面组织中胶原纤维稀疏,并伴有少量新生血管。高剂量SY干预组小鼠创面组织中PDGF、VEGF、α-SMA及collagen I mRNA相对表达量显著高于模型组、低剂量SY干预组、高剂量SY联合IGF-1组(P<0.01);高剂量SY干预组小鼠创面组织中PTP1B和AGEs mRNA相对表达量显著低于模型组、低剂量SY干预组、高剂量SY联合IGF-1组(P<0.01)。模型组、低剂量SY干预组、高剂量SY联合IGF-1组小鼠创面组织中PDGF、VEGF、α-SMA、collagen I、PTP1B及AGEs mRNA相对表达量比较差异无统计学意义(P>0.05)。高剂量SY干预组小鼠创面组织中Ki-67和PCNA蛋白相对表达量显著高于模型组、低剂量SY干预组、高剂量SY联合IGF-1组(P<0.01);caspase-3、caspase-6、caspase-7、p85 PI3K及p-AKT蛋白相对表达量显著低于模型组、低剂量SY干预组、高剂量SY联合IGF-1组(P<0.01)。模型组、低剂量SY干预组、高剂量SY联合IGF-1组中Ki-67、PCNA、caspase-3、caspase-6、caspase-7、p85 PI3K及p-AKT蛋白相对表达量比较差异无统计学意义(P>0.05)。高剂量SY干预组小鼠创面组织中TNF-α、IL-1β及IL-6水平显著低于模型组、低剂量SY干预组、高剂量SY联合IGF-1组(P<0.01)。模型组、低剂量SY干预组、高剂量SY联合IGF-1组小鼠创面组织中TNF-α、IL-1β及IL-6水平比较差异无统计学意义(P>0.05)。结论高剂量SY干预可通过提高血管生成、胶原形成及细胞增殖,降低胰岛素抵抗、炎症反应和细胞凋亡来促进小鼠DFU创面愈合,其机制可能与抑制PI3K/AKT通路有关。

Safflower yellow injection combined with conventional therapy in treating unstable angina pectoris:a meta-analysis

OBJECTIVE： To evaluate the clinical efficacy of safflower yellow injection combined with conventional therapy in treating unstable angina pectoris.METHODS： We searched online databases： Chinese journal full-text database, China National Knowledge Infrastructure, Wanfang database, Chinese journal full-text database, Pubmed, ScienceDirect,Embase, and the Cochrane Library with manual-screening of relevant literature. Eligible randomized controlled trials（RCT） on angina pectoris were included. We conducted meta-analysis using the RevMan 5.1 software from The Cochrane Collaboration. We treated the relief rate of angina symptoms and electrocardiograph（ECG） as evaluation.RESULTS： Seven articles, including in 1134 patients, were enrolled after the evaluation. Therewas no significant heterogeneity among the studies（χ2=1.08, df=6, P=0.98, I2=0%）. The safflower yellow injection with conventional therapy has a higher effective rate than the control group in relieving the symptoms of angina pectoris [odds ratio（OR）=2.95, 95%（CI）（1.81, 4.81）] and improving ischemic ECG [OR=2.85, 95% CI（1.67, 4.86）]. The difference was statistically significant in the ＂80 mg dosage＂ and ＂100 mg dosage＂ subgroups（P0.05） for improving clinical symptoms and ECG. The funnel graphic was nearly symmetrical. Sensitivity analysis suggested that the results were stable.CONCLUSION： Safflower yellow injection as an adjunct therapy with conventional drugs shows advantages in easing the clinical symptoms of unstable angina and improving ECG over basic therapy alone. However, the conclusions should be interpreted with care until more high-quality RCTs are reported.

穴位注射红花黄色素联合四逆酸枣仁合方在首发中风后不寐(肝郁血虚型)患者中的应用效果

目的 研究穴位注射红花黄色素联合四逆酸枣仁合方在首发中风后不寐(肝郁血虚型)患者中的应用效果。方法 选取158例首发中风后不寐(肝郁血虚型)患者作为研究对象,将其随机分为常规组、观察组,每组各79例。在常规西药治疗的基础上,常规组行穴位注射红花黄色素治疗,观察组行穴位注射红花黄色素联合四逆酸枣仁合方治疗,两组均治疗4周。比较两组治疗4周后的临床疗效和治疗前、治疗4周后中医证候积分、匹兹堡睡眠质量指数(PSQI)、脑卒中专用生活质量(SS-QOL)量表评分、血清神经递质水平、血清核转录因子-κB(NF-κB)、促食欲素A水平,以及不良反应发生率。结果 治疗4周后,观察组临床总有效率高于常规组(P<0.05);观察组中医证候积分、PSQI和血清去甲肾上腺素、P物质、NF-κB、促食欲素A水平均低于常规组,SS-QOL量表评分和血清多巴胺、5-羟色胺水平高于常规组(均P<0.05);两组不良反应总发生率差异无统计学意义(P>0.05)。结论 穴位注射红花黄色素联合四逆酸枣仁合方治疗首发中风后不寐(肝郁血虚型)患者疗效显著,可有效地缓解临床症状,调节神经递质水平,促进病情恢复。

补阳还五汤质量评价及其活性成分羟基红花黄色素A对内皮细胞损伤保护作用的研究

目的建立同时测定补阳还五汤中毛蕊异黄酮葡萄糖苷、阿魏酸、羟基红花黄色素A、芍药苷、苦杏仁苷5种成分的高效液相色谱(HPLC)分析方法,并研究羟基红花黄色素A对内皮细胞损伤的作用。方法采用HPLC法对补阳还五汤中的5种成分进行质量分数测定及方法学考察。细胞计数试剂(cell counting kit-8,CCK8)法检测羟基红花黄色素A不同浓度对EA.hy926细胞的细胞毒性;qRT-PCR法检测羟基红花黄色素A对TNF-a诱导的EA.hy926细胞损伤中TF、IL-6、IL-1β、MCP-1的mRNA水平。结果5种成分线性关系均良好,相关系数(R2)均在0.9950以上;平均加样回收率在93.80%~101.48%之间,RSD均小于3%,表明方法准确度良好;5种成分质量分数的RSD值在1.78%~2.59%之间。TNF-a(10 ng/mL)刺激EA.hy926细胞后,TF、IL-6、IL-1β、MCP-1的mRNA水平显著升高(P<0.05),而羟基红花黄色素A干预后呈浓度依赖性下调(P<0.05)。结论该方法专属性强、重复性好,稳定、可控,可为补阳还五汤的质量评价提供参考。其活性成分羟基红花黄色素A可改善TNF-a诱导的Ey.hA926内皮细胞损伤,为阐明补其药效物质基础提供参考。

红花黄色素对SCOP诱导小鼠认知功能损伤的保护作用

目的 研究红花黄色素(safflower yellow, SY)对氢溴酸东莨菪碱(scopolamine hydrobromide, SCOP)诱导的记忆障碍模型小鼠学习记忆的作用。方法 6月龄C57BL/6J小鼠随机分为Control组、Model组、SY-H组、SY-M组、SY-L组、石杉碱甲(huperzine-A)组,每组12只。采用SCOP建立记忆障碍模型,通过行为学实验评价记忆障碍模型小鼠的空间学习记忆和认知能力,ELISA试剂盒测定小鼠皮层胆碱能神经系统功能,尼氏染色观察脑组织病理变化情况,Western blot检测各组小鼠脑组织突触可塑性及BDNF/TrkB/CREB通路相关蛋白表达。结果 与Model组相比,各给药组小鼠学习记忆能力提高;海马和皮层神经元排列整齐,细胞形态趋于完整,正常神经元数量升高;皮层胆碱能神经系统功能显著改善,突触可塑性相关蛋白表达量升高,脑组织BDNF/TrkB/CREB信号通路激活。结论 SY能显著改善SCOP诱导的认知障碍小鼠的学习记忆能力,其作用机制可能是通过改善胆碱能神经系统功能,激活BDNF/TrkB/CREB信号通路,调节突触可塑性,改善神经元损伤,进而发挥神经保护作用。

高效液相色谱串联质谱法测定天然染料中的羟基红花黄色素A

为鉴别天然染料中羟基红花黄色素A区别于化学合成染料,采用高效液相色谱串联质谱法建立一种天然染料中羟基红花黄色素A的定量鉴别方法。采用25%甲醇水溶液提取、0.25 mL/min流动相流速、5μL进样体积、40℃柱温箱温度、以甲醇-10 mmol/L乙酸铵水溶液为流动相、梯度洗脱,在多反应监测模式下使用铂金埃尔默色谱柱C18(2.7μm,2.1mm×100 mm)对样品进行分析。结果表明:该方法的质量浓度在0.02~2.00 mg/L时具有良好的线性关系,且拟合度大于0.9990;最低检出限为0.125μg/L,定量限为0.42μg/L,回收率在不同质量浓度下为80%~95%;该方法能够为羟基红花黄色素A的判定提供高效灵敏的检测方案,也为天然染料染色纺织品的产品质量判定提供参考依据。

基于AHP-熵权TOPSIS混合加权法评价不同灭菌工艺对红花药材原粉质量的影响

目的采用“层次分析法”(AHP)-熵权“逼近理想解排序法”(TOPSIS)混合加权法考察并优选红花药材(Carthami Flos)原粉的灭菌工艺。方法以羟基红花黄色素A和山柰酚2种成分含量及灭菌率为评价指标,通过AHP-熵权TOPSIS混合加权法确定各指标权重系数,比较干热灭菌、湿热灭菌、紫外灭菌、辐照灭菌4种灭菌工艺对红花药材原粉灭菌前后质量的影响。结果AHP-熵权TOPSIS综合评价结果表明,4种灭菌工艺对红花药材原粉质量影响的优劣顺序依次为辐照灭菌、紫外灭菌、干热灭菌、湿热灭菌。其中辐照灭菌在9 kGy及以上剂量,与灭菌前原粉相比,其羟基红花黄色素A、山柰酚2种成分含量未见明显变化,且灭菌率≥98.79%;湿热灭菌条件下,红花药材原粉质量明显降低,且羟基红花黄色素A在灭菌后未能检出。结论相较于其他灭菌工艺,辐照灭菌更适于红花药材原粉,并确定辐照剂量为9 kGy,在确保灭菌效果的同时能最大程度地保留其有效成分,且优选得到的灭菌工艺重复性好、稳定可行、安全高效,可用于工业大生产。

红花黄色素对大鼠心梗后心肌重构的影响研究

目的观察红花黄色素(SY)对急性心肌梗死(AMI)后心肌重构的影响,并探究其与腺苷酸活化蛋白激酶(AMPK)/哺乳动物西罗莫司靶蛋白(mTOR)信号通路的关系。方法利用冠状动脉左前支结扎法构建AMI大鼠模型,随机分为Model组、氯沙坦组(10 mg/kg)、SY低剂量组(8 mg/kg)、SY高剂量组(32 mg/kg)、SY+Compound C组(32 mg/kg SY+10 mg/kg Compound C),并设假手术(Sham)组,每组12只。连续治疗4周后,使用超声心动图仪测定大鼠心功能指标;采用HE、Masson染色观察大鼠心肌组织变化,计算胶原容积积分(CVF)、测定大鼠心肌细胞直径;酶联免疫吸附法(ELISA)测定心肌组织乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)水平;Werstern blot法检测大鼠心肌组织Ⅰ、Ⅲ型胶原(COL-Ⅰ、COL-Ⅲ)、B型钠尿肽(BNP)、心房钠尿肽(ANP)、微管相关蛋白1轻链3Ⅱ/LC3-Ⅰ(LC3-Ⅱ/LC3-Ⅰ)、AMPK、自噬相关蛋白P62、Beclin-1、磷酸化AMPK(p-AMPK)、mTOR、磷酸化mTOR(p-mTOR)蛋白表达水平。结果与Sham组相比,Model组大鼠心肌细胞肥大,心肌纤维排列紊乱,左室短轴缩短率(LVFS)、左室射血分数(LVEF)及LC3-Ⅱ/LC3-I、Beclin-1、p-AMPK、p-mTOR蛋白表达水平及SOD水平降低(P<0.05),左室舒张末期内径(LVEDD)、左室收缩末期内径(LVESD)、心肌细胞直径、CVF、COL-Ⅰ、COL-Ⅲ、BNP、ANP及P62蛋白表达水平、LDH水平升高(P<0.05);与Model组相比,氯沙坦组、SY低、高剂量组大鼠心肌细胞有所缩小,心肌纤维排列趋于正常,LVFS、LVEF及LC3-Ⅱ/LC3-I、Beclin-1、p-AMPK、p-mTOR蛋白表达水平及SOD水平升高(P<0.05),LVEDD、LVESD、心肌细胞直径、CVF、COL-Ⅰ、COL-Ⅲ、BNP、ANP及P62蛋白表达水平、LDH水平降低(P<0.05);与SY高剂量组相比,SY+Compound C组大鼠心肌细胞肥大,心肌纤维排列紊乱,LVFS、LVEF及LC3-Ⅱ/LC3-I、Beclin-1、p-AMPK、p-mTOR蛋白表达水平及SOD水平降低(P<0.05),LVEDD、LVESD、心肌细胞直径、CVF、COL-Ⅰ、COL-Ⅲ、BNP、ANP及P62蛋白表达水平、LDH水平升高(P<0.05)。结论SY可提高AMI心肌组织抗氧化能力,降低心肌肥厚程度,缓解心肌纤维化进程,逆转心肌重构,可能与激活AMPK/mTOR信号通路,上调心肌自噬有关。

花黄色素调节AMPK/SIRT1/NF-κB信号通路对冠心病大鼠心肌损伤的影响

目的:探讨花黄色素(SY)调控腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子1(SIRT1)/核转录因子-κB(NF-κB)对冠心病(CHD)大鼠心肌损伤的影响。方法:将60只SD大鼠随机分为正常对照组(NC组,生理盐水)、模型组(Model组,生理盐水)、SY组(100 mg/kg SY)、EX-527组(47 mg/kg EX-527)、SY+EX-527组(100 mg/kg SY+47 mg/kg EX-527),每组12只,持续4周给予相应药物。试剂盒检测血清三酰甘油(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)水平和心肌白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)水平;苏木素-伊红(HE)染色法进行心肌组织病理学观察;末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测心肌细胞凋亡情况;蛋白免疫印迹(Western Blot)法检测心肌组织AMPK/SIRT1/NF-κB通路相关蛋白表达。结果:与NC组相比,Model组大鼠心肌细胞数量少且排列杂乱,细胞核皱缩,TC、TG、LDL-C、氧化修饰低密度脂蛋白(ox-LDL)、IL-6、TNF-α水平、心肌细胞凋亡率及NF-κB蛋白水平明显上升(P<0.05),SOD、GSH-Px水平及磷酸化-AMPK(p-AMPK)/AMPK、SIRT1蛋白水平明显下降(P<0.05)。与Model组比较,SY组大鼠心肌细胞数量增多且排列整齐有序,TC、TG、LDL-C、ox-LDL、IL-6、TNF-α水平、心肌细胞凋亡率及NF-κB蛋白水平明显下降(P<0.05),SOD、GSH-Px水平及p-AMPK/AMPK、SIRT1蛋白水平明显上升(P<0.05);而EX-527组以上指标呈现相反趋势。与SY组相比,SY+EX-527组大鼠心肌损伤加重,TC、TG、LDL-C、ox-LDL、IL-6、TNF-α水平、心肌细胞凋亡率及NF-κB蛋白水平明显上升(P<0.05),SOD、GSH-Px水平及p-AMPK/AMPK、SIRT1蛋白水平明显下降(P<0.05)。结论:SY可能通过调控AMPK/SIRT1/NF-κB通路降低氧化应激,减轻炎症反应,对冠心病大鼠心肌损伤起到一定改善作用。