黄芪通过ACE2/Mas途径改善代谢综合征大鼠早期肾功能损害

探讨黄芪影响代谢综合征(MS)大鼠肾脏血管紧张素转换酶2(ACE2)以及血管紧张素1-7(Ang 1-7)受体Mas蛋白的表达及其抗氧化作用。将80只雄性SD大鼠分为:正常对照组、MS组、黄芪组、缬沙坦组。MS大鼠予以高脂肪高盐饲料,饮用10%果糖水制备代谢综合征动物模型,黄芪组予以黄芪6.0 g·kg^-1·d^-1灌胃,缬沙坦组予以缬沙坦30.0 mg·kg^-1·d^-1灌胃,MS组以及对照组予以同体积生理盐水灌胃,干预4周后行一般指标、生化指标以及血压测定,放免法检测血及肾组织血管紧张素Ⅱ(AngⅡ)、丙二醛(MDA)、一氧化氮(NO)和超氧化物歧化酶(SOD)水平,免疫蛋白印迹法检测各组大鼠肾组织Mas受体,ACE,ACE2,AT1R的表达。结果显示,与对照组比较,MS组、黄芪组大鼠收缩压、舒张压、体质量、空腹血糖、空腹胰岛素水平、甘油三酯、血清游离脂肪酸以及血浆、肾组织AngⅡ水平明显升高(P<0.05);缬沙坦组大鼠收缩压、舒张压与MS组比较明显降低;MS组大鼠肾组织NO含量以及SOD活性下降,经黄芪干预后较MS明显增高;黄芪治疗后较MS组大鼠肾脏组织中MDA水平明显降低(P<0.01)。与对照组比较,MS组肾组织中ACE和AT1R的表达明显升高,而ACE2和Mas受体的表达降低(均P<0.05);与MS组比较,MS+黄芪组肾组织中Mas受体的表达升高较显著,而MS+缬沙坦组AT1R的表达降低更明显(均P<0.05)。结论:黄芪可以提高肾脏组织Mas的表达,降低ACE表达,改变肾脏局部的AngⅡ,MDA,NO和SOD水平,从而对早期受损伤的肾组织起保护作用。     

“黄芪-莪术”药对通过PTEN与p-AKT对人乳腺癌细胞增殖的影响

目的：探讨“黄芪-莪术”药对通过PTEN与p-AKT对人乳腺癌细胞（MDA-MB-231）增殖的作用机制。方法：将MDA-MB-231细胞进行“黄芪-莪术”药对（药对组）、顺铂（顺铂组）、“黄芪-莪术”药对联合顺铂（药对+顺铂组）治疗,并设置空白对照组。MTT法检测细胞增殖,RT-qPCR检测PTEN与p-AKT的mRNA表达水平,Western Blotting检测p-AKT蛋白表达水平。结果：与空白对照组比较,药对组PTEN mRNA表达水平上升,OD值与p-AKT蛋白表达水平下降（P<0.05）,p-AKT mRNA蛋白表达水平变化差异无统计学意义（P>0.05）;顺铂组OD值与PTEN mRNA表达水平下降（P<0.05）,p-AKT mRNA与蛋白表达水平变化差异无统计学意义（P>0.05）;而药对+顺铂组在药对治疗的基础上能进一步抑制MDA-MB-231细胞增殖,促进PTEN mRNA的表达,抑制p-AKT mRNA与蛋白表达水平。结论：“黄芪-莪术”药对抑制MDA-MB-231细胞增殖的机制可能通过上调PTEN基因而直接抑制AKT蛋白水平。     

Enzyme-Linked Immunosorbent Assay and Immunochromatographic Strip for Rapid Detection of Atrazine in Three Medicinal Herbal Roots

Objectives: An enzyme?linked immunosorbent assay (ELISA) and colloidal gold?based immunochromatographic (ICG) strip assay will be developed for the rapid and high-throughput detection of atrazine (ATZ) in medicinal herbs. Methods: A monoclonal antibody against ATZ was obtained after the immunization of mice, cell fusion, and hybridoma screening, and the antibody was used to develop direct competitive ELISA (dcELISA) and the ICG strip assay. Results: Both dcELISA and ICG strip methods were established, optimized, and validated for the detection of ATZ in Salviae miltiorrhizae radix et rhizome, Astragali radix, and Isatidis radix. dcELISA had a half-maximum inhibition concentration of 10.56 ng/mL (Salviae miltiorrhizae radix et rhizome), 7.6 ng/mL (Astragali radix), and 8.15 ng/mL (Isatidis radix). The limit of detection (LOD) of the ICG strip assay was 12.5 ng/mL (Salviae miltiorrhizae radix et rhizome), 12.5 ng/mL (Astragali radix), and 6.25 ng/mL (Isatidis radix) in different herb matrices. Due to the recognition characteristics of the monoclonal antibody for the pesticides ATZ, propazine, sebuthylazine, and prometryn, the detection results of realsamples by the two immunoassays were confirmed by liquid chromatography–tandem mass spectrometry, which proved the accuracy and reliability of the established methods. Conclusion: The proposed dcELISA and ICG strip methods were suitable for the rapid, convenient, and high-throughput detection of ATZ in these medicinal herbs.     

黄芪治疗新生儿缺氧缺血性脑病的疗效观察和护理

目的 研究黄芪注射液辅助治疗新生儿缺氧缺血性脑病（HIE）的疗效和其对HIE患儿的神经功能保护作用，并探讨相应的护理措施。方法 HIE患儿50例，分治疗组和对照组，2组常规治疗方法相同，治疗组加用黄芪注射液，观察治疗前后临床症状及体征改善时间、新生儿行为神经测定（NBNA）评分。结果 治疗组临床症状恢复时间较对照组平均缩短1．4d，2组比较有显著性差异（t=2．06，P〈0．05）。NBNA评分在治疗后第7天治疗组优于对照组（t=2．18，P〈0．05），第14天优势更明显（t=2．79，P〈0．01）。结论 黄芪用于治疗新生儿缺氧缺血性脑病能缩短疗程，提高NBNA评分，促进患儿神经功能恢复。     

黄芪有效部位的复合型纯化工艺

目的:确定黄芪有效部位的复合型纯化工艺。方法:采用正交试验设计法,以多糖收率、多糖含量为指标,优选了多糖醇沉和除蛋白工艺;以单因素考察法优选了黄芪总皂苷的纯化工艺。结果:总多糖最佳醇沉工艺为醇沉前生药量与浓缩液体积比1∶1,醇沉温度40℃,加入3倍量95%的工业乙醇,醇沉静置18 h,粗多糖除蛋白的最佳工艺为配制质量浓度为15 g.L-1的粗多糖溶液,加入粗多糖溶液1/3体积的Sevage试剂,连续脱除2次;大孔树脂纯化黄芪总皂苷的最佳工艺为使用D101型树脂,树脂-药材比为5∶2,以4BV水洗除杂,再以4 BV 70%乙醇洗脱,收集乙醇洗脱液,蒸干即可。结论:此法优选出的工艺可以实现黄芪总多糖、总皂苷的次序提取。工艺流程科学、经济,可有效节约资源,指导工业生产。     

液相色谱-串联质谱法测定黄芪中119种农药残留

目的:建立液相色谱串联质谱法(LC-MS/MS)测定黄芪中119种农药多残留。方法:样品用乙腈提取,经PSA、C18、GCB净化后用HPLC分离。色谱柱:ZORBAX Eclipse plus C18(2.1 mm×100 mm,1.8μm);柱温:40℃;流动相:乙腈(含5%水,0.1%甲酸,5 mmol.L-1甲酸铵)-水(含0.1%甲酸,5 mmol.L-1甲酸铵)梯度洗脱;流速:0.4 mL.min-1;进样量:5μL;离子源:ESI;扫描模式:正离子;在动态MRM模式下,以保留时间和质荷比对分离的组分给予定性,外标法峰面积定量。结果:119种农药在0.012～0.75 ng的范围内线性关系良好(r>0.9930),样品最低检测限为0.003～22μg.kg-1,多数农药的加标回收率为70%～120%。结论:本方法简便准确,可用于黄芪中多种农药残留的检测。     

丹芪通片质量标准研究

目的:建立丹芪通片的质量标准。方法:采用薄层色谱(TLC)法鉴别方中丹参、黄芪;采用可见-紫外分光光度法和高效液相色谱法分别测定方中总皂苷和丹参酮ⅡA的含量。结果:丹参、黄芪的TLC斑点清晰、分离度好。总皂苷、丹参酮ⅡA的检测浓度分别在0.056～0.12mg.mL-1(r=0.9993)、0.002019～0.03213mg.mL-1(r=0.9999)范围内与各自峰面积积分值呈良好线性关系;平均回收率分别为97.61%、95.74%,RSD分别为2.10%、1.93%。结论:所建标准可用于丹芪通片的质量控制。     

黄芪三仙汤对去卵巢大鼠血清骨钙素的影响

目的：探讨黄芪三仙汤（由仙茅、淫羊藿、黄芪、三七等组成）对去卵巢大鼠血清骨钙素水平的影响。方法：将88只雌性大鼠随机分为对照组、假手术组、模型组、阳性组、黄芪三仙汤低剂量组、中剂量组、高剂量组。模型组、阳性组及受试药物各剂量组进行卵巢切除手术;假手术组动物进行开背,但不切除卵巢;对照组不进行手术。手术后7天开始给药,受试药物各剂量组用黄芪三仙汤灌胃,阳性组用尼尔雌醇灌胃,连续给药12周后观察各组血清钙离子、磷离子、骨钙素、碱性磷酸酶的浓度。结果：模型组大鼠血清钙离子、磷离子浓度降低。受试药物各剂量组大鼠血清钙离子、磷离子浓度升高,黄芪三仙汤中剂量组大鼠血清中骨钙素的浓度显著性升高。结论：黄芪三仙汤可能具有治疗去卵巢大鼠骨质疏松的作用。     

Effects of a Chinese Medicinal Plant Radix Astragali on the Ovariectomized Female Rats

Perimenopausal syndrome occurs during the transition to menopause. Complementary and alternative medicine, especially Chinese medicinal plants, has manifested significant effects in alleviating perimenopausal symptoms. However, little research has been focused on the effects of Chinese medicinal plant on the immune function of the perimenopausal women. The present study aimed to explore the effects of Radix Astragali (RA) on the sex hormone levels and the interleukins of the ovariectomized female rats. 24 female Sprague-Dawley (SD) rats were randomly divided into model control group (MOD group), sham-operation group (SHAM group), RA group and estrogen group (EST group). After all the treatment ended, the serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), interleukin-2 (IL-2), and interleukin-8 (IL-8) were measured using enzyme-linked immune-sorbent assay (ELISA) and the uterus was removed and weighed after blood exsanguinations immediately. In the MOD group, the serum levels of E2 were significantly lower, and the serum levels of FSH and LH were markedly higher than those of the RA group, EST group and SHAM group (P<0.05). In the RA group, the serum levels of E2 were significantly lower, and the serum levels of FSH were markedly higher than those of the SHAM group and EST group, respectively. In the MOD group, the serum levels of IL-2 and IL-8 were significantly lower than those of the RA group, EST group and SHAM group (P<0.05), and no marked differences existed among RA group, EST group and SHAM group in the serum levels of IL-2 and IL-8 (P>0.05). The uterine weight of the rats in the RA group, EST group and SHAM group were significantly higher than those of the rats in MOD group (P<0.05). There were no marked differences among the rats from RA group, EST group and SHAM group on the uterine weight (P>0.05). It is concluded that RA can significantly improve the immune functions of the ovariectomized female rats, although it cannot change the sex hormones levels as significantly as estrogen.     

山西省不同产地中药材黄芪HPLC指纹图谱研究

目的对山西省不同产地的黄芪药材建立HPLC指纹图谱,评定各类黄芪中所含化学物质的含量。方法采用HPLC法,乙腈-水二元梯度洗脱;流速0.8mL/min;检测波长208nm。结果山西省各个产地的黄芪药材中所含黄芪甲苷等成分均得到很好的分离,根据指纹图谱总结出了各个产地黄芪药材的质量区别。结论该法简便、快捷,为黄芪药材的质量评价提供依据。