The effect of folic acid on ovine fetuses in utero during late gestation

To investigate whether folic acid would have toxic effects on fetal cardiac, hepatic, and renal functions, this was the first in utero fetal study testing acute effects of folic acid at the last third of gestation. Folic acid (5?mg/day) or 0.9% saline as the control was intragastricly administrated into pregnant ewes. Both maternal and fetal blood were analyzed for pH, PO₂, PCO₂, SO₂%, hemoglobin, hematocrit, glucose, lactic acid, osmolality, Na⁺, and K⁺ concentrations. Maternal and fetal cardiovascular functions were assessed by examining cardiac enzymes and cardiovascular responses in vivo. Fetal hepatic and renal functions were examined by analysis of biochemistry index and renal excretion. Folic acid did not alter the blood values in both ewes and fetuses. Cardiac enzyme activities remained unchanged, and no alteration in cardiovascular responses was observed. Folic acid did not affect fetal urine volume, urine electrolytes, and osmolality. Enzyme activities related to hepatic and renal functions were not changed. In addition, maternal application of folic acid had no effect on maternal and fetal lipid profile. The results showed that folic acid used (5?mg/day) during the last third of gestation did not cause biochemical changes related to cardiac, hepatic, and renal functions in both maternal and fetal sheep, providing new information for use of folic acid during late pregnancy.     

Protective effects of tanshinone IIA derivative on myocardial ischemia/reperfusion injury in rats

Tanshinone IIA (TSIIA) is the major bioactive constituent of Salvia miltiorrhiza Bunge, a Chinese herbal medicine, which has protective effects on myocardial ischemia/reperfusion (MIR) injury. However, the cardioprotective effects of TSIIA as well as its clinical use were limited due to its poor water solubility. The objective of this study was to evaluate whether Tanshinone IIA derivative (TD), a new water soluble compound synthesized by TSIIA and N-Methyl-D-Glucamine, had protective effects on MIR injury and what the related mechanism was. The cardioprotective effects of TD were evaluated and compared with TSIIA in a rat MIR model. The results show that pretreatment with TD significantly alleviated inflammatory infiltration and exhibited antioxidant effect in MIR injury by reducing the activity of lactate dehydrogenase (LDH) and malondialdehyde (MDA), decreasing expression of nuclear factor-κ-gene binding (NF-κB) and upregulating expression of heme oxygenase (HO-1), but having no effect on the content of total superoxide dismutase (T-SOD) and mRNA expression of superoxide dismutase (SOD-1). Thus, our study reveals that TD exerted significant protective effects on MIR injury through attenuating oxidative stress and inflammatory responses.     

Continuous synthesis and anti-myocardial injury of tanshinone IIA derivatives

A series of tanshinone IIA derivatives were synthesized through sulfonation, slat-forming, chlorination, and amidation reactions. Meanwhile, anti-myocardial injury activity was evaluated in vitro. D8 and D9 exhibited a slightly higher anti-myocardial injury (5.78, 7.46 μM) activity compared with esmolol (8.12 μM). In addition, they also displayed a concentration-dependent inhibition on the anti-myocardial injury.     

Effects of tanshinone IIA on the hepatotoxicity and gene expression involved in alcoholic liver disease

Tanshinone IIA is one of the most abundant constituents of the root of Salvia miltiorrhiza BUNGE which exerts antioxidant and anti-inflammatory actions in many experimental disease models. In the present study, we demonstrated that the standardized fraction of S. miltiorrhiza (Sm-SF) was able to protect RAW 264.7 cells from ethanol-and lipopolysaccharide (LPS)-induced production of superoxide radical, activation of NADPH oxidase and subsequently death of the cells. Among four main components of Sm-SF, tanshinone IIA was the most potent in protecting cells from LPS-and ethanol-induced cytotoxicity. LPS or ethanol induced the expression of CD14, iNOS, and SCD1 and decreased RXR-alpha, which was completely reversed by tanshinone IIA. In H4IIEC3 cells, 10 microM tanshinone IIA effectively blocked ethanol-induced fat accumulation as evidenced by Nile Red binding assay. These results indicate that tanshinone IIA may have potential to inhibit alcoholic liver disease by reducing LPS-and ethanol-induced Kupffer cell sensitization, inhibiting synthesis of reactive oxygen/nitrogen species, inhibiting fatty acid synthesis and stimulating fatty acid oxidation.     

丹参酮IIA磺酸钠联合依达拉奉治疗急性脑梗死的疗效研究

目的探讨丹参酮ⅡA磺酸钠联合依达拉奉治疗急性脑梗死的临床疗效及安全性。方法选取100例急性脑梗死患者,随机数字表法分为两组,对照组患者（50例）给予依达拉奉治疗,观察组患者（50例）给予丹参酮ⅡA磺酸钠联合依达拉奉治疗,均治疗14 d。观察并记录两组患者治疗后1个月的疗效,治疗前后ESS评分、SF-36评分及治疗期间不良反应情况。结果治疗后,观察组有效率为90.0%,对照组有效率为72.0%,观察组治疗有效率明显高于对照组（P<0.05）。与治疗前相比,治疗后两组ESS评分均明显增加（P<0.05）,且观察组在治疗3、7、14 d的ESS评分均明显高于对照组（P<0.05）。治疗前两组SF-36各项得分相比,差异没有统计学意义;治疗后,两组SF-36各项得分均明显升高（P<0.05）,且观察组在生理功能、生理职能及精神健康上的评分高于对照组（P<0.05）。治疗期间,两组不良反应率无明显差异。结论丹参酮ⅡA磺酸钠联合依达拉奉对急性脑梗死具有较好的治疗作用,能改善患者神经功能,提高患者生活质量,用药具有安全性,值得临床推广使用。     

丹参酮ⅡA对异丙肾上腺素致心肌缺血损伤大鼠apelin及其受体的影响

目的:观察异丙肾上腺素(ISO)致心肌缺血损伤大鼠的心肌组织中apelin及其受体APJ的变化,探讨丹参酮ⅡA改善其心肌缺血损伤的作用机制.方法:雄性SD大鼠分为正常对照组、ISO组、不同剂量丹参酮ⅡA治疗组.皮下注射ISO建立大鼠心肌缺血损伤模型,ELISA分析检测血浆和心肌apelin和NO的含量,real-time PCR方法检测心肌中apelin及其受体APJmRNA表达,Western blot方法检测心肌组织中APJ、eNOS及其磷酸化蛋白水平.结果:与正常对照组比较,ISO组大鼠血浆和心肌组织apelin和NO含量均下降,心肌组织中apelin和APJ mRNA水平表达下调(P＜0.01),APJ、eNOS磷酸化蛋白水平降低;与ISO组相比,中、高浓度的丹参酮ⅡA可以明显增加大鼠血浆和心肌组织中apelin和NO含量,增加心肌组织中apelin和APJ mRNA表达水平,增加心肌组织中APJ和eNOS磷酸化蛋白水平.结论:丹参酮ⅡA抗大鼠心肌缺血损伤的机制可能与其上调心肌组织apelin/APJ mRNA的表达、提高apelin/APJ蛋白水平、增加eNOS磷酸化水平和NO生成有关.     

高效液相色谱法测定丹参中丹参酮IIA的含量

目的建立高效液相色谱法测定中药丹参不同部位中丹参酮IIA的含量。方法采用反相高效液相色谱法,选择色谱柱HypersilODSC18柱(4.6mm×250mm,5μm);流动相:甲醇∶水=75∶25;流速为1.2mL/min;柱温为30℃;检测波长为268～270nm。结果丹参酮IIA浓度在1.25～21.45μg/mL(r=0.9997)范围内线性关系良好,平均加样回收率为99.83%,RSD=2.04%(n=6)。结论采用反相高效液相色谱法测定丹参中丹参酮IIA的含量,该方法快速、灵敏、准确,可以为丹参的种植、品质鉴定、扩展药用部位以及控制丹参药材的质量提供测定依据。     

Simultaneous determination of danshensu, ferulic acid, cryptotanshinone and tanshinone IIA in rabbit plasma by HPLC and their pharmacokinetic application in danxiongfang

A selective and sensitive reversed-phase high performance liquid chromatography method was developed and validated for the simultaneous determination of danshensu, ferulic acid, cryptotanshinone, and tanshinone IIA in rabbit plasma using p-hydroxybenzoic acid as internal standard. Liquid–liquid extraction was used for sample preparation. Chromatographic separation was successfully achieved on an Agilent HC-C₁₈ column using a mobile phase composed of methanol–water (from 20:80 to 80:20, v/v) containing 0.5% (v/v) glacial acetic acid. The mobile phase was employing gradient elution at a flow rate of 1.0 ml/min. The method showed good linearity and no endogenous material interfered with the marked compounds and I.S. peaks. The limit of quantification of danshensu, ferulic acid, cryptotanshinone, and tanshinone IIA were 0.1, 0.03, 0.05, and 0.05 μg/ml, respectively. The average extract recoveries of the four compounds from rabbit plasma were all over 60%. The precisions determined from 5 days were all within 10%. The established method has been successfully applied in the pharmacokinetic study and drug interaction of danshensu, ferulic acid, cryptotanshinone, and tanshinone IIA in rabbits after intravenous administration of danxiongfang, a useful compound preparation of traditional Chinese medicine.     

不同浓度的安氟醚对肝肾功能的影响

目的 观察不同浓度的安氟醚对部分血生化指标的作用,探讨该药对肝肾功能的影响。方法24例 ASAⅠ～Ⅱ级全麻手术患者随机分为Ⅰ、Ⅱ、Ⅲ组,均为8例,分别吸入1％、2％、3％安氟醚。于吸入安氟醚前(T_0)、吸入安氟醚2h(T_1)、4h(T_2)、24h(T_3)和7d(T_4)取静脉血,测天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、血糖(GLu)、总胆固醇(CHOL)和总胆红素(TBIL)、血肌酐(CRE)及尿素氮(BUN)等生化指标。结果 Ⅰ组吸入安氟醚前后各生化指标无明显变化。其他两组吸入安氟醚后AST、ALT、GLu、CHOL、TBILL有不同程度升高(P<0.01或P<0.05),吸入浓度越大上述指标升高越显著,7d后回降至正常。三组吸入安氟醚前后CRE和BUN均无明显变化。结论 吸入较高浓度安氟醚可使AST、ALT、GLu、CHOL和TBIL出现不同程度的变化,而CRE、BUN无明显变化。吸入较高浓度对肝功能可能有一定损害。     

高效液相色谱法测定宁神补心片中丹参酮IIA含量

目的建立高效液相色谱测定宁神补心片中丹参酮IIA含量的方法。方法利用Lichrospher5C18色谱柱,流动相为甲醇水(73∶27);检测波长为270nm;流速为1.6mL·min-1;柱温为30℃。结果进样量在0.208～1.664μg范围内呈现良好的线性关系,回归方程为Y=2.735×107X+8.870×105,r=0.9996,平均回收率为98.54%,RSD为1.33%(n=5)。结论该方法快速简便,准确可靠,灵敏度高,可用于宁神补心片的质量控制。     

补心安泰片中丹参酮ⅡA的含量测定

目的：建立补心安泰片中丹参酮Ⅱ。的含量测定方法。方法：Themao C18柱（250mm×4．6mm,5μm）;流动相：甲醇-水（75：25）;流速：1．0mL·min^-1;检测波长：270nm;柱温：30℃结果：线形范围0．0790～0．6323μg;回归方程为Y=265732x-128293,R=0．9995;平均回收率为97．6％,RSD为0．77％（n=6）。结论：此方法简便、快速、准确,可用于补心安泰片中丹参酮ⅡA的含量测定。     

The effect of folic acid on ovine fetuses in utero during late gestation

To investigate whether folic acid would have toxic effects on fetal cardiac, hepatic, and renal functions, this was the first in utero fetal study testing acute effects of folic acid at the last third of gestation. Folic acid (5?mg/day) or 0.9% saline as the control was intragastricly administrated into pregnant ewes. Both maternal and fetal blood were analyzed for pH, PO(2), PCO(2), SO(2)%, hemoglobin, hematocrit, glucose, lactic acid, osmolality, Na(+), and K(+) concentrations. Maternal and fetal cardiovascular functions were assessed by examining cardiac enzymes and cardiovascular responses in vivo. Fetal hepatic and renal functions were examined by analysis of biochemistry index and renal excretion. Folic acid did not alter the blood values in both ewes and fetuses. Cardiac enzyme activities remained unchanged, and no alteration in cardiovascular responses was observed. Folic acid did not affect fetal urine volume, urine electrolytes, and osmolality. Enzyme activities related to hepatic and renal functions were not changed. In addition, maternal application of folic acid had no effect on maternal and fetal lipid profile. The results showed that folic acid used (5?mg/day) during the last third of gestation did not cause biochemical changes related to cardiac, hepatic, and renal functions in both maternal and fetal sheep, providing new information for use of folic acid during late pregnancy.     

丹参酮IIA磺酸钠对血管紧张素II诱导的心肌肥大及p-ERK表达的影响

目的:观察丹参酮IIA磺酸钠(STS)对血管紧张素II(Ang II)诱导的心肌肥大及p-ERK表达的影响。方法:培养新生大鼠心肌细胞,考马斯亮蓝法测定心肌细胞蛋白含量、[3H]-亮氨酸掺入法测定蛋白合成速率作为心肌肥大指标;用West-ern-blot测定p-ERK表达。结果:STS能显著降低Ang II诱导的心肌细胞总蛋白含量、蛋白合成速率上升,同时对p-ERK表达具有剂量、时间依赖性抑制作用。结论:STS可以抑制Ang II诱导的心肌肥大,机制与抑制p-ERK表达有关。     

丹参胶囊中丹参酮ⅡA及丹酚酸B含量测定方法研究

目的建立丹参胶囊中丹参酮ⅡA及丹酚酸B的高效液相色谱测定方法.方法 Diamonsil C18柱;丹参酮ⅡA测定:以甲醇-水(75∶25)为流动相,检测波长为270 nm;丹酚酸B测定:以甲醇-乙腈-甲酸-水(30∶10∶1∶59)为流动相,检测波长为286 nm;外标法计算.结果 10批丹参胶囊中丹参酮ⅡA的含量为0.189%～0.204%,平均为0.195%;10批丹参胶囊中丹酚酸B的含量为5.19%～5.97%,平均为5.55%.结论丹参胶囊中丹参酮ⅡA及丹酚酸B的含量应作为本品质量控制指标.     

RP-HPLC法测定大鼠血浆中丹参酮IIA浓度及其药代动力学研究RP-HPLC法测定大鼠血浆中丹参酮IIA浓度及其药代动力学研究

目的建立高效液相色谱法测定大鼠血浆中丹参酮IIA浓度的方法。方法血浆样品经液-液萃取后,用HPLC法进行分析。色谱柱为YMC C¹⁸(5 μm,ID 3.0 mm×150 mm);流动相为乙腈-水-冰醋酸(74∶26∶1);流速0.3 mL·min^-1;检测波长270 nm;内标为4-氯联苯。结果线性范围为0.05 mg·L^-1～6.40 mg·L^-1,最低检测浓度为0.05 mg·L^-1。高、中、低3种浓度的平均方法回收率分别为98.9%,102.1%和100.4%。日内、日间精密度(RSD)均小于5%。结论本方法稳定、简便、可靠,可用于丹参酮IIA的血药浓度分析及其药代动力学研究。     

Effects of the coexisting diterpenoid tanshinones on the pharmacokinetics of cryptotanshinone and tanshinone IIA in rat

Tanshinone II_A and cryptotanshinone are the main pharmacologically active components in the Chinese herb drug Salvia miltiorrhiza Bge. The objective of this study was to investigate the effect of coexisting tanshinones in liposoluble ethanol extract of S. miltiorrhiza Bge. on the rat pharmacokinetics of tanshinone II_A and cryptotanshinone after oral intra-gavage administration of the tanshinones extract. Rats were given the tanshinones extract 23.3 mg/kg (equivalent to 5.7 mg/kg cryptotanshinone and 8.0 mg/kg tanshinone II_A), cryptotanshinone 5.7 mg/kg, or tanshinone II_A 8.0 mg/kg orally under overnight fasted conditions. Blood samples were taken at predetermined sampling time interval and the concentrations of cryptotanshinone and tanshinone II_A were determined by a validated LC–MS/MS method. The peak plasma concentrations of cryptotanshinone and tanshinone II_A were considerably increased (about 8 and 10 folds) after oral administration of the extract in comparison with the equivalent dose of single component administration, respectively. The areas under the plasma concentration–time curve (AUC) of cryptotanshinone and tanshinone II_A were both significantly increased (P < 0.001) as well. Tanshinone II_A was also found after the administration of cryptotanshinone alone, and the fraction of metabolism of tanshinone II_A was 21.0 ± 4.1%. Therefore, the pharmacokinetics of cryptotanshinone and tanshinone II_A in rats after administration of the tanshinones extract were significantly affected by the coexisting tanshinones. In conclusion, the herb-drug interactions occurred between coexisting tanshinones and cryptotanshinone or tanshinone II_A affected their absorption, transformation and metabolism.     

丹参酮ⅡA对乳腺癌细胞株MDA-MB-231体外血管生成拟态的抑制作用

目的:研究丹参酮ⅡA对乳腺癌细胞株MDA-MB-231体外血管生成拟态的影响。方法:将培养的MDA-MB-231细胞分为丹参酮ⅡA组及对照组,应用MTT法检测细胞增殖活力的变化,体外管状形成试验检测管道形成能力的变化,通过RT-PCR和Western-blot技术检测细胞内与血管拟态形成相关基因表达的变化。结果:丹参酮ⅡA对MDA-MB-231细胞增殖有明显抑制作用,并抑制体外管道形成和肿瘤细胞VEGF的mRNA表达。结论:丹参酮ⅡA能抑制MDA-MB-231细胞的体外血管生成拟态的形成和细胞增殖,其机制可能是通过抑制VEGF的表达而实现。     

Preparation,preliminary pharmacokinetics and brain tissue distribution of Tanshinone IIA and Tetramethylpyrazine composite nanoemulsions

Objective: Tanshinone IIA (TSN) and Tetramethylpyrazine (TMP) were combined in a composite, oil-in-water nanoemulsions (TSN/TMP O/W NEs) was prepared to prolong in vitro and vivo circulation time, and enhance the bioavailability of TSN.

Material and methods: Physicochemical characterization of TSN/TMP O/W NEs was characterized systematically. The in vitro dissolution and in vivo pharmacokinetic experiments of TSN/TMP O/W NEs were also evaluated.

Result: A formulation was optimized, yielding a 32.5?nm average particle size, an encapsulation efficiency of over 95 %, and were spherical in shape as shown by TEM. TSN/TMP O/W NEs were shown to extend the release and availability in vitro compared to raw compounds. In pharmacokinetic study, the AUC_0→∞ and t_1/2 of the TSN/TMP O/W NEs were 481.50?mg/L*min and 346.39?min higher than TSN solution, respectively. Brain tissue concentration of TSN was enhanced with TSN/TMP O/W NEs over raw TSN and even TSN O/W NEs.

Conclusions: Therefore, nanoemulsions are an effective carrier to increase encapsulation efficiency of drugs, improve bioavailability and brain penetration for TSN – which is further enhanced by pairing with the co-delivery of TMP, providing a promising drug delivery.     

Tanshinone IIA attenuates atherosclerosis in ApoE mice through down-regulation of scavenger receptor expression

This study is designed to investigate the protection of tanshinone IIA (TSIIA) against atherosclerosis in apolipoprotein E deficient (ApoE^−/−) mice and to explore the mechanisms by focusing on the expressions of scavenger receptors, scavenger receptor-A (SR-A) and CD36. The in vivo study demonstrated that TSIIA (10–90 mg/kg) inhibited the atherosclerotic lesions, down-regulated the CD68 protein expression in lesion and decreased the contents of cholesterol in aortas of ApoE^−/− mice. In addition, TSIIA reduced the serum levels of oxidized LDL (oxLDL) and down-regulated the mRNA expression of CD36, SR-A and peroxisome proliferator-activated receptor gamma (PPARγ) in aortas. The in vitro study showed that TSIIA (0.1–10 μM) decreased cholesterol level and DiI-oxLDL uptake in mouse peritoneal macrophages treated with oxLDL (50 μg/ml). In addition, TSIIA down-regulated the mRNA and protein expression of CD36 but not that of SR-A in oxLDL treated macrophages. TSIIA also down-regulated the mRNA expression of PPARγ in oxLDL treated macrophages. Furthermore, TSIIA reduced the mRNA expression of CD36 in macrophages treated with PPARγ agonist 15d-PGJ₂ (2 μM) or troglitazone (50 μM), whereas both 15d-PGJ₂ (0.5–1.5 μM) and troglitazone (5–20 μM) dose-dependently abolished the down-regulation of CD36 expression by TSIIA in oxLDL treated macrophages. These results suggest that TSIIA attenuates the atherosclerotic lesion in ApoE^−/− mice, which might be attributed to the properties of both anti-oxidation and down-regulation of scavenger receptors. Furthermore, antagonism of PPARγ might be involved in the down-regulation of CD36 by TSIIA.