Effects of traditional herbal medicine, Hwaotang, on atherosclerosis using the spontaneous familial hypercholesterolemia model, Kurosawa and Kusanagi-hypercholesterolemic rabbits and the venous thrombosis rats

Hwaotang (HOT), a traditional Korean medicinal formulation, is a dried decoctum of a mixture of seven herbal medicines, consisting of Angelica gigantis Radix, Rehmanniae Radix, Paeoniae Radix, Ciniamomi Cortex, Cnidii Rhizoma, Persicae Semen and Carthami Flos. In the present study, the inhibitory effects and anti thrombic properties of HOT on the progression of atherosclerotic lesions were studied using the spontaneous familial hypercholesterolemia (FH) model, Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits and rats. Changes in blood chemistry, pathology and low-density lipoprotein (LDL) oxidation were measured in a control and HOT group. In the control group, the area of atheromatous plaques of the aorta progressed between week 12 (36.65%) and week 14 (46.22%). This progression of atherosclerotic lesions did not occur in the HOT-treated group after 12 (24.24%) and 14 (23.34%) weeks. Antioxidative effects on LDL were seen in the HOT in weeks 12 and 14. HOT improved the hypercholesterolemia in the KHC rabbits. On the other hand, HOT and five of the seven herbs, except Cnidii Rhizoma and Carthami Flos, inhibited the endotoxin-induced hepatic venous thrombosis in high cholesterol diet-treated rats. However, Ciniamomi Cortex showed a very weak inhibitory effect on the endotoxin-induced hepatic venous thrombosis. The extract also inhibited the endotoxin-induced decrease in blood platelets and fibrinogen, and endotoxin-induced increase in fibrin degradation products (FDP) on disseminated intravascular coagulation in normal rats. In conclusion, these results suggest that HOT has inhibitory effects on the development of atheromatous plaque formation in spontaneous FH rabbits. It is also suggested that the antioxidative effects of HOT on LDL led to the beneficial effects observed in this study. The protection by HOT and its herbs on the artificially induced ischemic infarction might be related to their inhibitory effects on disseminated intravascular coagulation, platelet coagulation and thrombotic action.     

药对“蛇床子一补骨脂”与乳腺癌骨转移裸鼠体重变化与骨代谢的量效关系研究

目的 从调节破骨细胞分化的主要途径护骨素(OPG)/细胞核因子κB受体活化因子配体(RANKL)/细胞核因子κB受体活化因子(RANK)系统研究蛇床子—补骨脂温肾药对抑制乳腺癌骨转移的作用机制.方法 将42只乳腺癌骨转移裸鼠随机分为模型组、唑来磷酸组、蛇床子组、补骨脂组、2:2配比组、1:3配比组、3:1配比组,每组6只;另备正常裸鼠6只作为正常组.分别给予相应的药液灌胃,唑来磷酸组皮下注射,正常组给予生理盐水灌胃,持续6周.观察裸鼠的体重、骨转移灶病理和OPG、RANKL mRNA表达水平变化.结果 3:1配比组和2:2配比组体重下降率明显低于其他配比组,而2:2配比组低于3:1配比组;不同比例配伍组的骨抗酒石酸酸性磷酶(TRAP)(+)细胞数差异均有统计学意义,2:2配比组优于1:3配比组(P＜0.05);不同配比的蛇床子—补骨脂可不同程度地上调OPG基因表达,下调RANKL基因表达,从基因方面调节OPG/RANKL的分泌比例,其中2:2配比组的疗效优于其他配比组.结论 不同配比的蛇床子—补骨脂均具有一定的抑制乳腺癌骨转移作用,其中2:2配比组疗效最突出,其作用机制可能是调节了OPG/RANKL基因分泌比例.     

基于数据挖掘与实验验证的抗湿疹中药筛选及活性研究

目的 通过数据挖掘与实验相结合,筛选抗湿疹中药并评价其活性。方法 收集具有抗湿疹活性的中药方剂,筛选后录入Excel软件建立数据库,运用中医传承计算平台V3.0对其高频药物、复方配伍进行关联规则分析,挖掘核心药物及其组合,并对核心药物组合及单味药的抑菌作用进行测定。将40只昆明种小鼠随机分为空白组、模型组、基质组、阳性药组及核心药物组合凝胶组,并采用2,4-二硝基氯苯建立小鼠湿疹模型,验证核心药物组合的抗湿疹药效。应用超高效液相色谱-串联质谱代谢组学技术检测核心药物组合治疗湿疹小鼠前后血清中内源性代谢物,并采用主成分分析及正交偏最小二乘法判别分析等多元统计分析,结合人类代谢数据库（HMDB）等数据库和相关文献指认、鉴定差异代谢物,通过MetaboAnalyst平台富集相关代谢通路。结果 筛选得到255首符合标准的抗湿疹中药方剂,包含中药347味,使用频次由高到低分别是黄柏、苦参、蛇床子、白鲜皮、枯矾等清热解毒、清热燥湿中药。通过用药模式分析,并结合《已使用化妆品原料目录（2021年版）》,得到“苦参-蛇床子”核心药物组合。抑菌实验结果表明,苦参抑菌活性优于蛇床子,而二者组合物的抗菌活性有所提高。皮肤形态学观察显示,苦参-蛇床子凝胶组能明显减轻皮损的严重程度,减少皮肤组织炎性细胞的浸润情况。从血清中筛选出与核心药物组合治疗湿疹相关的12种内源性差异代谢物,主要涉及甘油磷脂代谢、谷胱甘肽代谢、甘油酯代谢等代谢通路。结论 基于数据挖掘方法从抗湿疹中医方剂中筛选得到14味核心药物,在此基础上进一步结合《已使用化妆品原料目录（2021年版）》确定的苦参-蛇床子组合物呈现较好的抗菌和抗湿疹活性,其抗湿疹药效与调节甘油磷脂代谢、谷胱甘肽代谢、甘油酯代谢等有关,这为其进一步研究开发奠定了基础。     

蛇床子总香豆素的抗炎作用

蛇床子总香豆素（ＴＣＣＭ）对角叉菜胶和鸡蛋清引起的大鼠足跖肿均有明显的抑制作用。ＴＣＣＭ还能减轻二甲苯引起的耳壳肿胀，明显抑制大鼠滤纸片肉芽肿。     

Effects of a Korean herbal formulation, Silsosangami, consisting of seven medicinal herbs, and its seven herbs on endotoxin-induced experimental thrombosis in rats

A traditional Korean medicine, Silsosangami (SSG), consisting of seven different herbs of Typhae Pollen, Pteropi Faeces, Paeoniae Radicis rubra, Cnidii Rhizoma, Persicae Semen, Carthami Flos and Curcumae Tuber, has been reported to have a hypolipidemic effect in human subjects. In the present study, the inhibitory effects of SSG on a thrombosis in rats, induced by endotoxin treatment were examined. The anti-thrombic properties of SSG were also investigated with respect to blood parameters. The extracts of SSG and five of the seven herbs, except Cnidii Rhizoma and Carthami Flos, inhibited both endotoxin-induced disseminated intravascular coagulation (DIC) and thrombosis in rats. The extract also inhibited the endotoxin-induced decrease in blood platelets and fibrinogen, and the endotoxin-induced increase in fibrin degradation products (FDP) on disseminated intravascular coagulation in normal rats. In conclusion, the artificially induced, protective effects of SSG on ischemic infarction might be related to their inhibitory effects on DIC, platelet coagulation and thrombotic action.     

Effect of a Korean Traditional Formulation, Hwaotang, on Superoxide Generation in Human Neutrophils, Platelet Aggregation in Human Blood, and Nitric Oxide, Prostaglandin E2 Production and Paw Oedema Induced by Carrageenan in Mice

Hwaotang, a traditional Korean medicinal formulation, is a dried decoctum of a mixture of 7 herbal medicines, consisting of Angelica gigantis Radix, Rehmanniae radix, Paeoniae radix, Ciniamomi cortex, Cnidii rhizoma, Persicae semen and Carthami flos. We have investigated that Hwaotang water extract (HOT) has various effects on stimulus-induced superoxide generation in human neutrophils. The effects of HOT on superoxide generation in human neutrophils were investigated. HOT significantly inhibited N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced superoxide generation in a concentration-dependent manner, but not that induced by arachidonic acid (AA). On the other hand, HOT enhanced superoxide generation induced by phorbol 12-myristate 13-acetate (PMA) in a concentration-dependent manner. The superoxide generation induced by PMA with HOT was suppressed by staurosporine, an inhibitor of protein kinase C, but was not suppressed by genistein, an inhibitor of protein tyrosine kinase. Tyrosyl phosphorylation of a 58 kDa protein, which was increased by fMLP, was inhibited by HOT. HOT also inhibited the generation of a 47 kDa protein and platelet aggregation in human blood. The results suggest that protein tyrosine kinase participates in fMLP-mediated superoxide generation by HOT-treated human neutrophils. HOT inhibited neutrophil functions, including degranulation, superoxide generation, and leukotriene B4 production, without any effect on 5-lipoxygenase activity. HOT reduced nitric oxide (NO) and prostaglandin E2 production in mouse peritoneal macrophages stimulated with lipopolysaccharide, whereas no influence on the activity of iNOS, COX-2 or COX-1 was observed. HOT significantly reduced mouse paw oedema induced by carrageenan. Western blot analysis showed that HOT reduced the expression of iNOS and COX-2. The results indicate that HOT exerts anti-inflammatory effects related to the inhibition of neutrophil functions and of NO and prostaglandin E2 production, which could be due to a decreased expression of iNOS and COX-2.     

祛瘀止痛酊的薄层定性鉴别

目的建立祛瘀止痛酊的定性鉴别标准,从而控制产品质量。方法;采用薄层色谱法,对本品所含樟脑、三七、蛇床子进行定性鉴别研究。结果采用薄层色谱法鉴别专属性强。结论该方法可用于祛瘀止痛酊的定性鉴别标准。     

蛇床子总香豆素对新生大鼠成骨细胞的作用

目的：研究蛇床子总香豆素（TCFC）在体外对新生大鼠颅盖骨成骨细胞增殖及成骨作用的影响。方法：酶消化法分离培养新生大鼠颅盖骨成骨细胞;^3H-胸腺嘧啶和^3H-脯氨酸掺入法分别测定细胞增殖和胶原合成;磷酸苯二钠法测定细胞内骨碱性磷酸酶（ALP）活性;并以抗骨质疏松药物异丙黄酮（ipriflavone,IP)作为阳性对照药物。结果：TCFC对新生大鼠成骨细胞的增殖,ALP活性,骨胶原合成具有显的促进作用。结论：TCFC抗骨质惊讶松的作用与其增加成骨细胞的数量,促进细胞胶魇蛋白及ALP的活性有关。     

HPLC法同时测定蛇床子中蛇床子素和欧前胡素含量

目的用高效液相色谱法同时测定中药蛇床子中蛇床子素和欧前胡素的含量。方法采用KromasilC8(2 5 0mm× 4 6mm ,5 μm)为色谱柱 ,以甲醇 水 (体积比 6 5∶35 )为流动相的HPLC法 ,检测波长为 310nm ,流速为 0 8mL·min-1,柱温为室温。结果以醋酸氟轻松为内标 ,蛇床子素和欧前胡素分别在 13～ 2 0 8mg·L-1和 8～ 12 8mg·L-1内 ,线性关系良好 (r≥ 0 9998) ;平均回收率分别为 98 2 %和 97 5 % ;RSD分别为 1 7%和 2 0 % (n =9)。结论本法简便快速 ,回收率及重现性良好 ,适用于中药蛇床子中蛇床子素和欧前胡素的定量分析     

正交实验优化蛇床子素的提取工艺

目的:探讨蛇床子中蛇床子素的最佳提取工艺。方法:采用正交实验,以提取物中的蛇床子素含量为指标,优化提取工艺。结果:蛇床子中提取蛇床子素的最佳工艺为A1B2C2(A:乙醇浓度;B:乙醇体积;C:提取时间)。结论:蛇床子素最佳提取工艺为:加入8倍体积的65％乙醇,提取3次,每次1 h。