Research Progress in the Application of Chinese Herbal Medicines in Aquaculture： A Review

Due to increasing safety concerns regarding human consumption of fish products, an increasing number of medicinal chemicals are prohibited from use in aquaculture. As a result, Chinese herbal medicines are being increasingly used, coining the use of the term ＂green medicine.＂ Research shows that Chinese herbal medicines have many beneficial effects on fish, including growth promotion, enhancement of disease resistance, and improvement in meat quality. Many effective ingredients have been discovered in Chinese herbal medicines, which function to promote feed intake, improve meat flavor, and increase digestive enzyme activity. They also regulate and participate in processes that improve the specific and non-specific immunity of fish; however, the composition of Chinese herbal medicines is very complex and it is often difficult to identify the effective ingredients. This article reviews the latest research and application progress in Chinese herbal medicines regarding growth and feed utilization, immunity and disease resistance, and the meat quality of cultured fish. It also discusses research on the chemical constituents of classical Chinese medicinal herbs and problems with the application of Chinese herbal medicines in fish culture. This article concludes by proposing that future studies on Chinese herbal medicines should focus on how to cheaply refine and extract the effective ingredients in classical Chinese medicinal herbs, as well as how to use them efficiently in aquaculture.

Research Progress of Chinese Herbal Medicine Additives in Layer Breeding

Chinese herbal medicine have dual effects of medicinal value and nutrition regulation. In recent years, antibiotics have been widely forbidden in live- stock production, which are inevitably replaced by natural Chinese herbal medicine in livestock breeding. A brief summary of current research progress of Chinese herbal medicine in layer breeding is given in the paper.

Research and Development Procedures of Chinese Herbal-medicine Skin Care Products with Chinese Characteristics

As far as Chinese herbal medicine resources are concerned,China is one of the countries with the richest biodiversity in the world.As for the future development direction of Chinese skin care products,the development of special Chinese herbal skin care products is an important issue.The technical route for the research and development(R&D)of Chinese herbal-medicine skin care products should focus on“combining ancient and modern,combining Chinese and Western”,that is to say combining the ancient knowledge and wisdom with modern science and technology,and combining Chinese traditional therapy with western pharmacology.This article will elaborate the five stages of developing innovative Chinese herbal skin care products:pondering and thinking stage;extraction and purification stage;experimental evaluation stage;quality index establishment stage;product development stage.The key points of these five stages can provide a reference for the cosmetics R&D personnel.This project will be beneficial to the development of Chinese herbal-medicine skin care products and the formulation of an overall effective development strategy,which is helpful to actively respond to new challenges and continuously improve product competitiveness,so as to vigorously build a world-renowned high-end cosmetic brand in China,and strive to cultivate a world-class cosmetics talent team.

Regulating effect of Chinese herbal medicine on the peritoneal lymphatic stomata in enhancing ascites absorption of experimental hepatofibrotic mice

AIM: To observe the regulatory effect of Chinese herbal medicine on peritoneal lymphatic stomata and its significance in treating ascites in liver fibrosis model mice. METHODS: Two Chinese herbal composite prescriptions were used separately to treat the carbon tetrachloride-induced mouse model of liver fibrosis. The histo-pathologic changes of the liver sections (HE and VG stainings) were observed. The peritoneal lymphatic stomata was detected by scanning electron microscopy and computer image processing. The changes of urinary volume and sodium ion concentration were measured. RESULTS: In the model group, lots of fibrous tissue formed in liver and extended into the hepatic lobules to separate them incompletely. In the treated and prevention groups, the histo-pathologic changes of liver was rather milder, only showed much less fibrous tissue proliferation in the hepatic lobules. The peritoneal lymphatic stomata enlarged with increased density in the experimental groups (diameter: PA, 3.07 +/- 0.69 microm; PB, 2.82 +/- 0.37 microm; TA, 3.25 +/- 0.82 microm and TB, 2.82 +/- 0.56 microm; density: PA, 7.11 +/- 1.90 stomata.1000 microm(-2); PB, 8.76 +/- 1.45 stomata.1000 microm(-2); TA, 6.55 +/- 1.44 stomata.1000 microm(-2)and TB, 8.76+/-1.79 stomata.1000 microm(-2)), as compared with the model group (diameter: 2.00+/-0.52 microm density: 4.45+/-1.05 stomata.1000 microm(-2)). After treatment, the urinary volume and sodium ion excretion increased in the experimental groups (PA, 231.28+/-41.09 mmol.L(-1); PB, 171.69 +/- 27.48 mmol.L(-1) and TA, 231.44 +/- 34.12 mmol.L(-1)), which were significantly different with those in the model group (129.33 +/- 36.75 mmol.L(-1)). CONCLUSION: Chinese herbal medicine has marked effects in alleviating liver fibrosis, regulating peritoneal lymphatic stomata, improving the drainage of ascites from peritoneal cavity and causing increase of urinary volume and sodium ion excretion to reduce the water and sodium retention, and thus have favorable therapeutic effect in treating ascites.

Consensus statement on research and application of Chinese herbal medicine derived extracellular vesicles-like particles(2023 edition)

To promote the development of extracellular vesicles of herbal medicine especially the establishment of standardization,led by the National Expert Committee on Research and Application of Chinese Herbal Vesicles,research experts in the field of herbal medicine and extracellular vesicles were invited nationwide with the support of the Expert Committee on Research and Application of Chinese Herbal Vesicles,Professional Committee on Extracellular Vesicle Research and Application,Chinese Society of Research Hospitals and the Guangdong Engineering Research Center of Chinese Herbal Vesicles.Based on the collation of relevant literature,we have adopted the Delphi method,the consensus meeting method combined with the nominal group method to form a discussion draft of“Consensus statement on research and application of Chinese herbal medicine derived extracellular vesicles-like particles(2023)”.The first draft was discussed in online and offline meetings on October 12,14,November 2,2022 and April and May 2023 on the current status of research,nomenclature,isolation methods,quality standards and research applications of extracellular vesicles of Chinese herbal medicines,and 13 consensus opinions were finally formed.At the Third Academic Conference on Research and Application of Chinese Herbal Vesicles,held on May 26,2023,Kewei Zhao,convenor of the consensus,presented and read the consensus to the experts of the Expert Committee on Research and Application of Chinese Herbal Vesicles.The consensus highlights the characteristics and advantages of Chinese medicine,inherits the essence,and keeps the righteousness and innovation,aiming to provide a reference for colleagues engaged in research and application of Chinese herbal vesicles at home and abroad,decode the mystery behind Chinese herbal vesicles together,establish a safe,effective and controllable accurate Chinese herbal vesicle prevention and treatment system,and build a bridge for Chinese medicine to the world.

Chinese Medicine in the United States:Historical Development and Growth

Chinese medicine is growing rapidly in the United States both as a field of research and in the daily treatment of illnesses.The popularity of Chinese medicine,particularly acupuncture,as a healing mechanism demonstrates American people’s positive experience with Chinese medicine and their embracing it as an alternative to the mainstream Western biomedicine.Chinese medicine is an important part of the Alternative and Complementary Medicine or Integrated Medicine in the United States health system.It is increasingly used and covered by health insurance.Although Chinese medicine has been in the United States since colonial times,it was discriminated against by local authorities.However,things began to change in the 1970s when individual states gradually passed laws to legalize the practice of acupuncture as a healing mechanism.American public interest in Chinese medicine,particularly acupuncture,was aroused by the media’s report of the“magic”power of acupuncture during President Nixon’s visit to China.Non-Asians studied acupuncture in the following decades,which contributed to the establishment of acupuncture as a profession in the American health system.The history of Chinese medicine in the United States illustrates that Chinese and Asian immigrants promoted Chinese medicine in the communities,while Chinese American medical scientists who were trained in biomedicine played an important role in shaping the biomedical method of acupuncture research.This article investigates the changes of Chinese medicine in the United States,with a focus on acupuncture.Situating the discussion in a larger context of social and cultural changes,this study examines the contributions of Chinese immigrants and the popularity of acupuncture in the United States.It analyzes the development of acupuncture practice and research,the professional training in Chinese medicine,and American people’s interest in using Chinese medicine as a reliable healing mechanism to many health problems,including chronic pain,cancer,and drug addictions.

基于UPLC-Q-TOF-MS的参芪扶正注射液主要成分鉴定及其治疗三阴性乳腺癌的网络药理学研究

目的 应用超高效液相色谱串联四极杆飞行时间质谱联用(UPLC-Q-TOF-MS)技术和网络药理学方法,鉴定参芪扶正注射液的主要成分,探究其治疗三阴性乳腺癌(TNBC)的作用机制,为其药效物质基础研究提供依据。方法 (1)参芪扶正注射液主要成分鉴定:采用UPLC-Q-TOF-MS技术,结合天然产物高分辨质谱数据库及相关文献,明确参芪扶正注射液的主要成分。(2)网络药理学研究:基于中药系统药理学数据库与分析平台(TCMSP)、中草药化合物蛋白质靶标数据库(HIT)、有机小分子生物活性数据库(PubChem)、化合物靶点预测平台(Swiss Target Prediction)、本草组鉴数据库(HERB)获取参芪扶正注射液主要成分的潜在靶点,利用人类基因综合数据库(GeneCards)、在线人类孟德尔遗传数据库(OMIM)、治疗靶点数据库(TTD)获得TNBC疾病潜在靶点,二者相结合得到参芪扶正注射液治疗TNBC的潜在靶点;运用蛋白互作关系数据库(STRING)平台、Cytoscape 3.9.1软件构建蛋白互作网络图并进行网络拓扑分析,获得参芪扶正注射液治疗TNBC的关键靶点;运用R语言进行京都基因与基因组百科全书(KEGG)通路富集分析。(3)动物实验验证:制备异种移植瘤乳腺癌小鼠模型,分为模型组与参芪扶正注射液低、中、高剂量(20、40、60 mL/kg)组,干预4周后取肿瘤组织,运用Western blot方法检测肿瘤组织中磷酸化磷脂肌醇3激酶(p-PI3K)、磷脂肌醇3激酶(PI3K)、P-糖蛋白(P-gp)、乳腺癌耐药蛋白(ABCG2)表达情况,并对预测潜在靶点进行实验验证。结果 (1)参芪扶正注射液的主要成分为党参炔苷、鸟苷、毛蕊异黄酮苷、壬二酸、黄芪甲苷、美迪紫檀苷等。(2)网络药理学研究结果显示,参芪扶正注射液的主要成分主要作用于肿瘤蛋白p53(TP53)、白介素-2(IL-2)、白介素-6(IL-6)、白介素-10(IL-10)、己糖激酶2(HK2)、缺氧诱导因子1亚基α(HIF1A)、半胱氨酸天冬氨酸蛋白酶9(CASP9)、ABCG2、ATP结合盒转运体B1(ABCB1)、原癌基因(MYC)等关键靶点,调控磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)、缺氧诱导因子-1(HIF-1)、T细胞受体等信号通路。(3)动物实验验证结果显示,参芪扶正注射液降低了实验动物肿瘤组织中p-PI3K水平,抑制了P-gp和ABCG2的表达。结论 参芪扶正注射液中的主要成分为党参炔苷、鸟苷、毛蕊异黄酮苷、壬二酸、黄芪甲苷、美迪紫檀苷等,其抗TNBC的作用机制与其通过调控PI3K信号通路和调节耐药相关蛋白表达有关。

Research on Frequency of Application with Modern Chinese Herbal Medicine

Objective：To study the relationship between 500 kinds of commonly used Chinese herbal medicine and the classification of their efficacies in Chinese Materia Medica in relation to the common diseases listed in Internal Medicine.Methods：Database retrieval frequency of the quantitative statistical method was adopted.First,the 8 980 kinds of Chinese herbal medicine recorded in Chinese Materia Medica were used as the original search objects,and 4 493 kinds which were cited in more than five articles were picked out and then rechecked for further title citations.Second,as judged based on the Criterion,the numbers of articles which included the medicines in the line of standards were examined.As a result,500 species of Chinese herbal medicine were singled out based on their retrieval frequency and were then used for compilation of the classification statistics according to their efficacy and the common diseases in Internal Medicine.Results：From the classification of Chinese medicines,herbs with wide efficiency and a meek nature had higher frequencies, but those which were not appropriate as decoctions had relatively lower frequencies.However,according to the average frequency,the Chinese herbal medicine for nourishing qi and tonifying blood,at 36 346 times and 34 544 times,respectively,were the most commonly used.Analyzed from the frequency of application of the Chinese medicine in the treatment of common diseases,most of the top 10 kinds of Chinese herbal medicine with the highest frequencies generally coincided with the 500 selected medicines.In addition,the Chinese medicines with clear pharmacological efficiency were easily isolated and purified to be made into injections, although other forms are more commonly used.Conclusion：The results of the research objectively reflected the current applications of Chinese herbal medicine,and could be used as references in teaching,research,clinical applications,and in compiling and increasing the drugs in textbooks and Pharmacopoeia.

Challenges in Research and Development of Traditional Chinese Medicines

This review is briefly to recall the history of research and development (R&D) of Chinese materia medica (CMM) and to discuss the challenges of Chinese traditional and herbal medicines (CTHM) facing the modern science and technology. The R&D of CTHM is thought to be an important pathway for new drug discovery. Since1949, about 140 approved new drugs have been developed, among which about 80 originated directly or indirectly from medicinal plants. CTHM has gained interest from the international medical, biomedical, and pharmaceutical institutions as a valuable source of potential medicines. For the modernization of CMM and innovative research of CTHM, there are following challenges to be faced: (1) to evaluate the efficacy, pharmacological properties, action mechanism, and active chemical constituents; (2) to develop new methodologies for the quality and safety of CTHM; (3) to apply new '-omics' techniques to accelerate drug discoveries developed from CTHM; and (4) to apply international practices including good agricultural practice, good manufacturing practice, good laboratory practice, and good clinical practice in the R&D of CTHM.

Chinese herbal medicines for prostate cancer therapy: From experimental research to clinical practice

Prostate cancer remains the second most common malignancy in men worldwide, is a global health issue,and poses a huge health burden. Precision medicine provides more treatment options for prostate cancer patients, but its popularity, drug resistance, and adverse reactions still need to be focused on. Chinese herbal medicines(CHMs) have been widely accepted as an alternative therapy for cancer, with the advantages of multiple targets, multiple pathways, and low toxicity. We searched the experimental research and clinical practice of CHMs for prostate cancer treatment published in Pub Med, Embase, and Web of Science in the last five years. We found five CHM formulas and six single CHM extracts as well as 12 CHM-derived compounds, which showed induction of apoptosis, autophagy, and cell cycle arrest, suppression of angiogenesis, proliferation, and migration of prostate cancer cells, reversal of drug resistance,and enhancement of anti-tumor immunity. The mechanisms of action include the PI3K/Akt/mTOR, AR,EGFR and Wnt/β-catenin signaling pathways, which are commonly implicated in the development of prostate cancer. We also summarized the advantages of CHMs in patients with hormone-sensitive and castration-resistant prostate cancer and provided ideas for their further experimental design and application.

益神宁心方对斑马鱼睡眠剥夺模型行为学及神经递质的影响

目的观察益神宁心方对斑马鱼睡眠剥夺模型行为的影响,并对其机制进行初步探索。方法将受精后5 d(5 dpf)野生型AB系斑马鱼随机分为正常组、模型组(咖啡因300μmol/L)、褪黑素组(10μmol/L)、益神宁心方低剂量组(100 mg/L)、益神宁心方中剂量组(300 mg/L)和益神宁心方高剂量组(900 mg/L),咖啡因诱导建立斑马鱼睡眠剥夺模型。应用行为追踪系统定量监测自主活动1 h,明暗交替下连续测定斑马鱼的趋触性行为15 min,酶联免疫吸附试验(ELISA)法检测γ-氨基丁酸(GABA)、5-羟色胺(5-HT)和褪黑素(MT)的含量,实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测斑马鱼组织GABA A型受体a1亚单位基因(gabra1)、γ2亚单位基因(gabrg2)和5-HT受体1A类亚型基因(htr1aa)、1D类亚型基因(htr1bd)及MT受体1A类亚型基因(mtnr1aa)、1B类亚型基因(mtnr1ba)mRNA相对表达量。结果与正常组比较,模型组斑马鱼运动状态覆盖距离、运动状态持续时间及中运动时间均显著增加(P<0.05),静止状态持续时间显著减少(P<0.05);中心区域时间百分比及中心区域距离百分比均显著减少(P<0.05);GABA、MT含量及gabra1、gabrg2、mtnr1aa、mtnr1ba mRNA相对表达量均显著降低(P<0.05),5-HT含量及htr1aa、htr1bd mRNA相对表达量均显著升高(P<0.05)。与模型组比较,益神宁心方高、中剂量组运动距离、运动时间、中运动时间减少(P<0.05),静息时间增加(P<0.05);益神宁心方高剂量组中心区域时间百分比及中心区域距离百分比增加(P<0.05);益神宁心方高剂量组GABA、MT含量和gabra1、mtnr1aa、mtnr1ba mRNA相对表达量升高(P<0.05),5-HT含量和htr1aa mRNA相对表达量降低(P<0.05);益神宁心方中剂量组gabrg2、mtnr1ba mRNA相对表达量升高,5-HT含量和htr1bd mRNA相对表达量降低(P<0.05)。结论益神宁心方可以改善咖啡因造模斑马鱼的睡眠剥夺状态,改善焦虑行为,其机制可能与调节GABA、5-HT、MT相关紊乱的基因水平相关。

温经汤对大鼠子宫内膜异位症SPARC介导的上皮-间质转化的影响

目的探讨温经汤对子宫内膜异位症(EMT)大鼠模型富含半胱氨酸的酸性分泌蛋白(SPARC)介导的上皮-间质转化的影响。方法采用手术自体移植子宫内膜块法建立EMT大鼠模型,将建模成功的40只大鼠随机分为模型组(给予质量分数为0.9%的氯化钠溶液10 mL/kg)、孕三烯酮组(给予孕三烯酮溶液0.5 mg/kg)以及温经汤低、中、高剂量组(分别给予温经汤免煎颗粒剂8.19 g/kg、16.38 g/kg、32.76 g/kg),每组各8只,另取7只未手术大鼠作为正常对照组(给予质量分数为0.9%的氯化钠溶液10 mL/kg)。所有大鼠均连续灌胃给药3周后进行取材。腹主动脉取血,采用酶联免疫吸附试验(ELISA)法检测血清中SPARC水平。电子游标卡尺测量异位病灶体积,计算生长抑制率。取下异位病灶组织(正常对照组取下双侧子宫组织)采用免疫组织化学法检测内膜组织中SPARC、锌指转录因子(Snail)、E-钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)的表达,采用Western blot法和RT-qPCR法测定内膜组织中上述因子的蛋白和mRNA相对表达量。结果各治疗组异位病灶的生长抑制率与模型组比较差异均有统计学意义(P<0.05)。与正常对照组比较,模型组血清中SPARC水平显著升高(P<0.05),各治疗组血清中SPARC水平均明显低于模型组(P<0.05)。模型组异位内膜中SPARC、Snail、Vimentin呈高表达,E-cadherin呈低表达,经温经汤治疗后,异位内膜中SPARC、Snail、Vimentin的阳性表达减少,E-cadherin的阳性表达增加。温经汤中、高剂量组和孕三烯酮组异位内膜中SPARC、Snail、Vimentin的蛋白相对表达量均明显低于模型组(P<0.05),E-cadherin的蛋白相对表达量显著高于模型组(P<0.05)。温经汤高剂量组和孕三烯酮组异位内膜中Snail、Vimentin mRNA相对表达量均较模型组显著降低(P<0.05),E-cadherin mRNA相对表达量明显高于模型组(P<0.05)。结论温经汤可以降低EMT大鼠模型血清中SPARC水平,下调异位内膜中SPARC、Snail、Vimentin蛋白和mRNA表达,升高异位内膜中E-cadherin蛋白和mRNA表达,通过调控SPARC介导的上皮-间质转化,从而抑制异位内膜病灶的生长。

苍术健脾、燥湿的作用机制及研究进展

综述苍术健脾、燥湿的作用机制及研究进展。古代医家常用苍术治疗多种疾病的脾胃湿证。苍术有效成分能改善胃排空及小肠推进率;可增加胃泌素、胃动素等,促进胃酸分泌,增加胃肠动力;可降低血管活性肠肽水平,抑制肠蠕动亢进。苍术能双向调节胃肠功能紊乱,且还能改善胃黏膜的异常形态,减轻胃组织损伤,改善肠组织异常结构。苍术燥湿的主要有效成分为挥发油,可导致体液丢失,血黏度增加,影响唾液腺分泌,引起口干,降低肾脏水通道蛋白2的表达量。苍术及其提取物能降低大鼠胃、大肠、尿液中水通道蛋白含量,通过多脏器调节体内水液代谢。故苍术能从影响脏器形态、胃肠道激素水平、水通道蛋白表达等多方面达到健脾、燥湿的功效。

国家药品计划抽检品种麸炒薏苡仁质量问题分析及评价

目的:通过国家药品计划抽检工作,对药食两用中药材麸炒薏苡仁的质量进行考察,了解麸炒薏苡仁饮片质量现状及存在的问题,完善麸炒薏苡仁检验标准,为药品监管部门提供技术支持。方法:从全国24个省(市、自治区)的生产和使用环节抽取麸炒薏苡仁样品,按照《中华人民共和国药典》2020年版一部进行性状、水分、灰分、浸出物及含量测定项检验,分析检验数据以评价麸炒薏苡仁饮片的质量状况,并针对检验中发现的问题,开展探索性研究。结果与结论:本次抽验的麸炒薏苡仁样品共计98批次,按现行质量标准检验合格率均为100%。经探索性研究,发现麸炒薏苡仁存在炮制不到位的问题,33批次样品中4批次玉米赤霉烯酮超标;此外,甘油三油酸酯含量测定项限度偏低。通过探索性研究发现麸炒薏苡仁饮片和标准中的问题,结合标准提高工作完善麸炒薏苡仁检验标准。建议增加安全性控制指标及炮制控制指标,并提高含量测定限度。

越婢汤含药血清对阿霉素损伤的人脐静脉血管内皮细胞的干预作用

目的基于小窝蛋白-1(Cav-1)/内皮型一氧化氮合酶(eNOS)通路,探讨越婢汤含药血清对阿霉素(ADR)损伤的人脐静脉血管内皮细胞(HUVEC)模型的作用机制。方法制备越婢汤含药血清及空白血清,体外培养HUVEC,采用ADR诱导建立HUVEC损伤模型,将细胞分为对照组、模型组、越婢汤组(2.5%、5%、10%、20%含药血清),给予相应药物处理24 h。细胞增殖-毒性检测(CCK-8)法检测细胞存活率,划痕实验检测细胞迁移能力,Western blot法检测Cav-1/eNOS信号通路相关蛋白表达,免疫荧光法检测细胞Cav-1表达水平,酶联免疫吸附测定(ELISA)法检测细胞上清液Cav-1的含量。结果与对照组比较,模型组细胞增殖及迁移能力降低(P<0.05),细胞内Cav-1、磷酸化内皮型一氧化氮合酶(p-eNOS)、血管内皮细胞钙黏蛋白(VE-cadherin)表达下降(P<0.05),细胞上清液Cav-1含量上升(P<0.05)。与对照组比较,各浓度越婢汤含药血清对HUVEC细胞活力无明显影响(P>0.05)。与模型组比较,越婢汤各组能促进细胞迁移(P<0.05);2.5%浓度的含药血清对细胞Cav-1等蛋白表达无影响(P>0.05);5%、10%、20%浓度含药血清能增加细胞内Cav-1、p-eNOS、VE-cadherin蛋白的表达(P<0.05),下调细胞上清液Cav-1的含量(P<0.05)。结论越婢汤含药血清可以降低ADR对HUVEC细胞的损伤,促进HUVEC细胞的增殖、迁移,其保护作用可能与调控Cav-1/eNOS信号通路有关。

黄芩苷抗结直肠癌药理学作用研究进展

结直肠癌是常见的消化道肿瘤之一,目前其临床治疗手段中手术治疗占主要地位,但因其易复发及易侵袭转移,且常规治疗毒副作用较大等缺陷,治疗效果不佳。黄芩苷是从中药黄芩中提取出的重要有效成分,其生物活性显著,具有消炎、抗血栓形成、缓解哮喘、抗癌、降糖等药理学作用。通过梳理并总结相关文献,发现黄芩苷抗结直肠癌的药理学研究主要集中在调控癌细胞增殖、凋亡、侵袭转移、分化等方面,为黄芩苷在结直肠癌预防及临床中的应用提供参考。

银杏酮酯对小胶质细胞炎症诱导的PC12细胞损伤的保护作用

目的探讨银杏酮酯(GBE50)通过抑制小胶质细胞的炎症反应对神经元突触损伤的影响。方法①采用格里斯(Griess)试剂检测不同浓度GBE50(25~400 mg·L^(-1))干预对经脂多糖(LPS)刺激后BV2小胶质细胞培养液中亚硝酸盐含量的影响。②不同浓度GBE50(25~400 mg·L^(-1))干预经LPS刺激的BV2小胶质细胞,收集上清液作为条件培养基(CM)处理PC12细胞,采用四甲基偶氮唑盐比色(MTT)法检测BV2小胶质细胞和PC12细胞活力。③将对数生长的BV2和PC12细胞以2×106个/孔铺板于6孔板中,分为空白对照组(Ctrl组,仅培养液培养),对照加药组(Ctrl+GBE50组,GBE50干预浓度为200 mg·L^(-1)),模型组(LPS组,LPS刺激浓度为100μg·L^(-1)),模型加药组(LPS+GBE50组,GBE50预处理1 h后,加入LPS),各组进行相应干预。采用Western blot、实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测CM处理的PC12细胞N-甲基-D-天门冬氨酸受体(NMDAR)亚基蛋白及基因表达量。结果①Greiss法结果显示,GBE50干预可降低LPS刺激BV2细胞产生的一氧化氮(NO)水平(P<0.05)。②MTT法结果显示,GBE50对BV2细胞活力无影响;LPS刺激后的CM处理PC12细胞活力显著下降,该现象在GBE50干预的CM中有所改善,GBE50浓度在200 mg·L^(-1)时细胞活力已无明显下降(P<0.05)。③Western blot结果显示,与LPS组比较,LPS+GBE50组中NMDAR亚基NR1、NR2A和NR2B蛋白表达升高(P<0.05);RT-qPCR结果显示,与LPS组比较,LPS+GBE50组中NMDAR亚基基因表达量均增加(P<0.05)。结论GBE50可通过抑制激活小胶质细胞的炎症反应,减少NMDAR的损伤,对神经元突触起到保护作用。

基于网络药理学和分子对接探讨六味地黄丸异病同治老年性聋和阿尔茨海默病的作用机制

目的借助网络药理学和分子对接的方法探讨六味地黄丸异病同治老年性聋和阿尔茨海默病(AD)的共同分子机制,并采用细胞实验进行验证。方法利用中药系统药理学数据库与分析平台(TCMSP)、成分靶点数据库(SwissADME)获取六味地黄丸有效成分,并结合基因和化学物质相互作用预测数据库(STITCH)获取作用靶点。利用人类基因综合数据库(Genecard)、人类疾病相关基因与突变位点信息数据库(DisGeNET)获取老年性聋和AD靶点。使用Cytoscape 3.7.1软件构建“中药-成分-靶点-疾病”网络,借助蛋白互作网络分析数据库(STRING)构建蛋白-蛋白相互作用(PPI)网络,获取核心靶点。利用富集分析数据平台(Metascape)对共有靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析。采用AutoDock Vina 1.1.2软件对核心靶点与相关成分进行分子对接。采用过氧化氢(H2O2)致PC12细胞氧化损伤模型,观察六味地黄丸的作用。采用细胞计数(CCK-8)法观察细胞存活率,活性氧(ROS)检测试剂盒检测ROS水平,酶联免疫吸附测定(ELISA)法检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平,Western blot法检测基质金属蛋白酶-9(MMP-9)、蛋白激酶B(AKT)、磷酸化-蛋白激酶B(p-AKT)蛋白表达。结果共获得六味地黄丸有效成分86种,主要来自薯蓣皂素、视黄醇、丹皮酚、软脂酸等化合物,六味地黄丸防治老年性聋和AD共有靶点49个,核心靶点为AKT1、TNF、IL-6、MMP-9等。GO分析表明这些共有靶点主要参与对氧化应激的反应、对脂多糖的反应等生物进程。KEGG通路分析显示,共有靶点主要涉及缺氧诱导因子-1(HIF-1)、核因子-κB(NF-κB)等信号通路。分子对接结果显示,TNF-α、IL-6、AKT1、MMP-9与相关成分均存在结合可能。细胞实验结果发现,六味地黄丸含药血清可以增强细胞存活率,降低细胞ROS、TNF-α、IL-6水平,增加MMP-9蛋白表达水平,降低p-AKT蛋白表达水平。结论六味地黄丸异病同治老年性聋和AD具有物质基础,其共同的分子机制,可能是通过薯蓣皂素、视黄醇、丹皮酚等多种成分,作用于AKT、TNF-α、IL-6、MMP-9等核心靶点,抑制炎症反应和氧化应激,减少神经元死亡。

中药调控氧化应激治疗男性不育症实验研究进展

氧化应激(OS)是指机体遭受各种有害刺激时产生过多的活性氧自由基而无法及时清除,导致氧化系统和抗氧化系统平衡失调,引起组织或细胞功能紊乱和损伤的一种状态。OS与男性不育症(MI)关系密切,OS通过影响精子发生,损伤精子膜、脱氧核糖核酸(DNA)、线粒体,降低精子运动能力,加速细胞凋等途径使精子质量下降,导致MI。研究发现,单味中药的有效成分多糖类化合物(黄芪多糖、党参多糖、枸杞多糖、黄精多糖、山茱萸多糖、当归多糖、女贞子多糖、石斛多糖),黄酮类化合物(菟丝子总黄酮、槲皮素、紫云英苷、羊藿苷、金丝桃苷),皂苷类化合物(人参皂苷、蒺藜皂苷、薯蓣皂苷元),苯丙素类化合物(五味子甲素、姜黄素、白藜芦醇),环烯醚萜苷类化合物(桃叶珊瑚苷),氮苷类化合物(红景天苷),低聚糖类化合物(巴戟甲素),苯乙醇苷类化合物(肉苁蓉苯乙醇苷)多肽类化合物(牡蛎肽)具有抗氧化作用,可提升雄性动物MI模型的生殖功能。

经典名方中桂枝的关键信息考证与质量评价研究

目的通过本草考证和多批次道地产区桂枝药材的质量评价,为含桂枝的经典名方的研究提供依据与参考。方法对历代本草和经典医书进行考证,对经典名方中桂枝的药用部位、基原、产地与品质进行考证;并根据考证结果收集4个道地产地的15批次合格桂枝药材,建立适用于桂枝药材的高效液相色谱(HPLC)特征图谱方法并进行方法学验证;通过聚类分析、主成分分析和偏最小二乘-判别分析方法,结合特征图谱数据和桂皮醛含量,对古籍考证的道地产区桂枝药材的质量一致性进行综合评价。结果通过古籍考证,确定了经典名方中所用桂枝的药用部位(嫩枝)、基原(樟科植物肉桂)、产地(广东、广西);建立了通过方法学验证的HPLC特征图谱方法,形成的共有对照特征图谱共计10个共有特征峰,并指认出香豆素、肉桂酸、桂皮醛以及2-甲氧基肉桂醛4个色谱峰;3个化学识别分析结果均表明古籍考证的道地产区桂枝药材质量具有一致性。结论经典名方中桂枝选用樟科植物肉桂(Cinnamomum cassia Presl)的干燥嫩枝,产地可选用广西和广东两地;建立的桂枝特征图谱方法适用于桂枝的质量控制研究。