黄芩苷、二甲双胍和氨基胍对D-半乳糖诱导大鼠体内蛋白非酶糖基化-氧化应激、醛糖还原酶活性及胰岛素抵抗作用

目的：观察黄芩苷、二甲双胍和氨基胍对D-半乳糖诱导大鼠体内蛋白非酶糖基化-氧化应激、醛糖还原酶活性及胰岛素抵抗作用。方法：清洁级健康SD大鼠90只,完全随机分为6组正常对照组、模型对照组、氨基胍组、二甲双胍组、黄芩苷高剂量和黄芩苷低剂量组,每组各15只。除空白对照组外,其余组大鼠均采用腹腔注射D-gal,灌胃;结束时观察各组大鼠晚期糖基化终末产物、糖化血红蛋白、胰岛素等,并计算胰岛素敏感指数,醛糖还原酶和超氧化物歧化酶的活性。结果：D-gal处理大鼠出现血糖升高、糖耐量减退、糖基化产物及胰岛素含量增高,SOD活性降低及红细胞内AR活性增强;Met,AG以及Bai高和低剂量均能改善D-gal诱导大鼠IGT和降低糖基化产物和胰岛素含量。结论：D-gal诱导能使大鼠体内糖基化反应、胰岛素抵抗和醛糖还原酶活性增强;Bai,Met和AG能抑制蛋白非酶糖基化反应和醛糖还原酶活性,改善高胰岛素血症。     

A Scutellaria baicalensis radix water extract inhibits morphine-induced conditioned place preference

Context: Scutellaria baicalensis Georgi (Lamiaceae) has been used as a traditional herbal preparation for the treatment of neuropsychiatric disorders in Asian countries for centuries.

Objective: To evaluate the effects of S. baicalensis on morphine-induced drug dependence in rats.

Materials and methods: In order to evaluate the effect of S. baicalensis and baicalin on morphine-induced dependence-like behavior, a water extract of S. baicalensis [500?mg/kg, intraperitoneally (i.p.)] or baicalin (50?mg/kg, i.p., a flavonoid found in S. baicalensis) was administered prior to morphine injection [5 and 2.5?mg/kg, respectively, subcutaneously (s.c.)] to rats for 8 and 4?d, respectively. Morphine-induced conditioned place preference was assessed by measuring the time spent in a drug-paired chamber. The effect of S. baicalensis on dopamine receptor supersensitivity (locomotor activity) and dopamine agonist-induced climbing behavior due to a single apomorphine treatment (2?mg/kg, s.c.) was also measured.

Results: At 50?mg/kg, a water extract of S. baicalensis decreased morphine (5?mg/kg)-induced conditioned place preference by 86% in rats. Apomorphine (2?mg/kg)-induced locomotor activity (dopamine receptor supersensitivity) in rats and climbing behavior in mice were attenuated after pretreatment with 500?mg/kg of S. baicalensis water extract by 41% and 56%, respectively. In addition, baicalin-reduced morphine-induced conditioned places preference by 86% in rats at 50?mg/kg.

Discussion and conclusion: These results suggest that S. baicalensis can ameliorate drug addiction-related behavior through functional regulation of dopamine receptors.     

黄芩苷调节IL-33/ST2信号通路对糖尿病视网膜病变大鼠视网膜新生血管生成的影响北大核心

目的探讨黄芩苷调节白细胞介素(IL)-33/基质裂解素2(ST2)信号通路对糖尿病视网膜病变(DR)大鼠视网膜新生血管生成的影响。方法SD大鼠随机分为对照组(正常大鼠,灌胃生理盐水)、DR模型组(大鼠建立DR模型后,灌胃生理盐水)和黄芩苷低、中、高剂量组(大鼠建立DR模型后,分别灌胃75 mg·kg^(-1)、150 mg·kg^(-1)、300 mg·kg^(-1)黄芩苷),所有大鼠均以右眼为实验眼。FFA检查各组大鼠视网膜血管生成情况,ELISA检测各组大鼠血清血管内皮生长因子(VEGF)、Ang-1、IL-6、IL-33、肿瘤坏死因子-α(TNF-α)水平,Western blot检测各组大鼠视网膜组织中IL-33、ST2蛋白表达水平。结果与对照组相比,DR模型组大鼠血清VEGF、Ang-1、IL-6、IL-33、TNF-α水平及视网膜组织中IL-33、ST2蛋白表达水平均显著升高(均为P<0.05),右眼眼底新生血管增多,荧光素渗漏明显;与DR模型组相比,黄芩苷低、中、高剂量组大鼠血清VEGF、Ang-1、IL-6、IL-33、TNF-α水平及视网膜组织中IL-33、ST2蛋白表达水平均显著降低(均为P<0.05),右眼视网膜新生血管生成减少,荧光素渗漏减少;随着黄芩苷剂量的升高,大鼠血清VEGF、Ang-1、IL-6、IL-33、TNF-α水平及视网膜组织中IL-33、ST2蛋白表达水平均逐渐降低(均为P<0.05)。结论黄芩苷能抑制IL-33/ST2信号通路激活,减轻DR大鼠体内炎症水平,减少视网膜新生血管生成。     

UPLC法测定痤疮凝胶中黄芩苷和蒙花苷的含量

[目的]建立痤疮凝胶中黄芩苷、蒙花苷含量测定方法。[方法]采用Acquity BEH C18(2.1 mm×50 mm,1.7μm)色谱柱;以乙腈-0.1%甲酸(20∶80)为流动相,流速0.3 m L/min,检测波长320 nm,柱温35℃。[结果]黄芩苷和蒙花苷分别在3.767~75.34μg和0.840 96~33.638 4μg范围内线性关系良好,平均回收率分别为99.04%和97.42%(n=6);RSD分别为1.97%和1.81%。[结论]该方法简便、准确、重现性良好,为痤疮凝胶的质量控制提供了理论依据。     

黄芩汤中黄芩苷的大鼠在体肠吸收特性研究

目的通过大鼠在体单向肠灌流实验研究黄芩汤中黄芩苷在体肠吸收特性。方法建立大鼠在体单向肠灌流模型,采用质量法计算黄芩苷吸收速率常数(Ka)和有效渗透系数(Peff)。结果黄芩汤中黄芩苷在十二指肠、空肠、回肠的吸收速率和程度都存在一定差异。其中黄芩苷在十二指肠的吸收速率显著高于空肠段,差异具有统计学意义(P<0.05),略高于回肠段但差异无统计学意义;黄芩苷在十二指肠的吸收程度明显高于空肠段且差异具有统计学意义(P<0.05),略高于回肠段但差异无统计学意义。结论黄芩汤中黄芩苷在十二指肠的吸收速度和吸收程度较好,表明黄芩汤中黄芩苷在体吸收具有肠道部位依赖特性。     

试剂盒法和福林酚法测定鹿瓜多肽注射液中多肽的比较研究

目的 筛选藿朴夏苓汤治疗早期新型冠状病毒肺炎（COVID-19）湿邪郁肺证的物质基础,预测其作用机制。方法 查阅文献及临床报道分析藿朴夏苓汤与早期COVID-19湿邪郁肺证的方证关系。运用TCMSP数据库筛选藿朴夏苓汤中潜在活性成分,将活性成分与新型冠状病毒（SARS-CoV-2）3CL水解酶、血管紧张素转化酶II（ACE2）进行分子对接,根据结合能筛选与两者均有较好结合作用的核心成分。借助Cytoscape软件构建关键成分靶点蛋白互作网络,筛选出核心靶点;通过STRING数据库进行核心靶点的GO分析,Cytoscape软件ClueGO插件进行Pathway、KEGG富集分析。结果 方证关系分析藿朴夏苓汤用于治疗早期COVID-19湿邪郁肺证,筛选出藿朴夏苓汤中潜在作用成分12个,核心靶点67个。其中通草中的通脱木皂苷元I,茯苓、猪苓中的过氧麦角甾醇,半夏中的黄芩苷与SARS-CoV-2 3CL水解酶、ACE2均具有较好的结合活性。GO、Pathway、KEGG富集分析结果显示藿朴夏苓汤中12个潜在作用成分参与调节刺激反应、信号转导、细胞死亡等生物过程以及白介素信号通路、癌症EGFR信号通路、酪氨酸激酶信号转导途径、编程性细胞死亡途径、MAPK信号通路等。结论 藿朴夏苓汤以化湿解毒、宣肺透邪治疗早期COVID-19湿邪郁肺证患者,菲酮、黄芩苷、酸枣仁皂苷、啤酒甾醇、常春藤皂苷元、过氧麦角甾醇、柠檬二烯醇、麦角甾-7,22-二烯-3-酮、通脱木皂苷元I、泽泻醇B-23-醋酸酯、泽泻醇B、新橙皮苷可能为其主要的物质基础,通过阻断SARS-CoV-2病毒蛋白合成,阻止病毒进入宿主细胞,通过调控白介素信号通路、MAPK信号通路、PI3K-Akt信号通路、T细胞受体信号通路、C型凝集素受体信号通路,抑制相关炎症因子的表达发挥作用。     

黄芩苷对变异链球菌UA159体外的抑制作用

目的探讨黄芩苷对变异链球菌国际标准株UA159的体外抑制作用。方法采用液体倍比稀释法结合酶标仪测OD600值测定黄芩苷对变异链球菌UA159的最低抑菌浓度(minimum inhibitory concentration,MIC)。利用酶标仪测定不同浓度的黄芩苷药液中变异链球菌UA159的OD600值,绘制生长曲线;计算黏附率及黏附抑制率;用结晶紫定量法测定黄芩苷对变异链球菌UA159生物膜形成的影响;用扫描电镜观察黄芩苷对变异链球菌UA159细菌总量的影响。结果黄芩苷对变异链球菌UA159最低抑菌浓度为12 mg/mL;随着黄芩苷药液浓度的增加,变异链球菌UA159生长速度缓慢,并且其黏附率下降,黏附抑制率增加(P<0.05)。结晶紫定量法结果显示,与菌液组相比,0、6、12 h时加入黄芩苷药液变异链球菌UA159生物膜形成量显著降低(P<0.001)。扫描电子显微镜下,0、6、12 h时加入黄芩苷后细菌总量均明显下降。结论黄芩苷能够抑制体外变异链球菌UA159的生长繁殖;同时其对变异链球菌UA159的黏附、生物膜形成均有抑制作用。     

基于模拟炮制法研究黄芩炒炭前后苷类成分的变化

目的:通过模拟炮制法探讨不同炒炭程度对黄芩饮片中苷类成分变化的影响。方法:采用Diamonsil C18(250 mm×4.6 mm,5μm)色谱柱;柱温箱温度25℃;进样量10μL;检测波长277 nm;流动相A为甲醇,B为0.4%磷酸,C为乙腈,使用梯度洗脱的方式,流速1 mL/min。通过模拟炮制的方法对黄芩苷与黄芩素、汉黄芩苷与汉黄芩素在炒炭温度下的转化进行初步研究。结果:黄芩炒炭后,黄芩苷以及汉黄芩苷的含量下降,而黄芩素以及汉黄芩素的含量均呈现先上升后下降的趋势。另外,在黄芩苷模拟样品中检出黄芩素,在汉黄芩苷模拟炮制品种检出汉黄芩素。结论:不同炒炭程度对黄芩中黄芩苷、汉黄芩苷、黄芩素、汉黄芩素含量有显著影响,模拟炮制法可较为真实地反映黄芩苷类成分在炒炭过程中的变化情况。     

黄芩苷对衣霉素诱导的内质网应激性心肌细胞损伤的影响

目的:观察黄芩苷(BC)对衣霉素(Tm)诱导的心肌细胞内质网应激(ERS)损伤的作用。方法:原代新生SD大鼠心肌细胞培养,随机分为对照组、BC组、Tm组、Tm+BC组。用MTT比色法检测心肌细胞的存活率,乳酸脱氢酶(LDH)释放法检测心肌细胞损伤,原位缺口末端标记法(TUNEL)检测心肌细胞凋亡,Western blot检测CCAAT增强子结合蛋白同源蛋白(CHOP)的水平。结果:①Tm能够损伤心肌细胞并呈现明显的时间依赖关系,各作用时间点心肌细胞的存活率差异显著(P0.05)。Tm可显著增加ERS的CHOP表达及细胞凋亡的水平。②相对与Tm组,50μmol/L的BC能够显著抑制Tm对心肌细胞的损伤,使心肌细胞存活率增加(P0.05)。同时,BC能够显著抑制Tm诱导的心肌ERS损伤,使CHOP表达及细胞凋亡的水平明显下调(P0.05)。结论:BC可通过抑制Tm诱导的心肌ERS及心肌细胞的凋亡,达到保护受损心肌细胞的作用。     

Antithrombotic effects of Huanglian Jiedu decoction in a rat model of ischaemia-reperfusion-induced cerebral stroke

ContextHuanglian Jiedu Decoction (HLJJD) has a variety of pharmacological activities, such as anti-inflammatory and neuroprotection against ischaemic brain injury.ObjectivesThis ex vivo study explores its antithrombosis activity and inhibition of platelet aggregation.Material and methodsTo study the antithrombosis activity of HLJJD ex vivo, saline, or HLJDD (100, 200, and 500 mg/kg) was treated prophylactically by gavage for 3 days in Wistar rats (n = 4). Based on the rat model of transient middle cerebral artery infarction (MCAO) or normal rats (n = 4), the antithrombotic activity in the normal group and HLJDD subgroups on prothrombin time, thrombus weight, platelet aggregation, and others was evaluated, followed by the antiplatelet aggregation of its main components (n = 4).ResultsThe weight of the thrombus increased significantly at 24 h after MCAO onset. HLJJD did not influence the change of PT, but significantly inhibited thrombosis by 12.5, 20.0, and 20.5% in reducing the dry weight of thrombus, and blocked collagen-induced platelet aggregation by 25.5, 39.0, and 42.7% and adhesion of blood platelet by 17.3, 26.2, and 27.3%. The IC₅₀ value of HLJJD on collagen-induced platelet aggregation was 670 mg/kg. Geniposide only facilitated antiplatelet aggregation induced by collagen, but not AA or ADP. Both baicalin and berberine showed markedly antiplatelet aggregation induced by all activators. The antithrombotic activity of baicalin was relatively higher than that of berberine (35.0–47.8% vs. 20.6–33.5%).ConclusionOur results indicated that HLJDD regulated blood circulation by inhibiting platelet aggregation and thrombosis, which might also extensively contribute to the clinical prevention and treatment of cerebrovascular diseases.