Evaluation of the famous classic formula Sanhua decoction based on network pharmacology and multi-component quantitative analysis

Background:Sanhua decoction has significant effects in the treatment of stroke.The study of the Sanhua decoction material benchmark was carried out to analyze the value transfer relationship between the Chinese herbal pieces and the substance benchmark.Methods:Network pharmacology was employed to investigate the potential active components and molecular mechanisms of Sanhua decoction in the treatment of stroke.15 batches of Sanhua decoction lyophilized powder were prepared using traditional formulas and subjected to high-performance liquid chromatography analysis to generate fingerprints of the Sanhua decoction substance benchmarks.Then,a multi-component quantitative analysis method was established,allowing for the simultaneous determination of ten components,to study the transfer of quantity values between pieces and substance benchmarks.Results:60 active ingredients were screened from Sanhua decoction by network pharmacology,of which gallic acid,magnolol honokiol,physcion,and aloe-emodin may have a greater effect than other active components.63 key targets and 134 pathways were predicted as the potential mechanism of Sanhua decoction in treating stroke.The fingerprint similarity of the Sanhua decoction substance benchmarks was found to be good among the 15 batches,confirming the 19 common peaks.The content of the 10 components was basically consistent.The components’transfer rates were within 30%of their respective means.Conclusions:This study provided a comprehensive and reliable strategy for the quality evaluation of Sanhua decoction substance benchmarks and held significant importance in improving its application value.

Establishment of Characteristic Chromatogram of Honeysuckle Stem Formula Granules Based on the Concept of Whole Process Quality Control

[Objectives]To establish a HPLC characteristic spectrum of honeysuckle stem formula granules,and to evaluate the quality consistency based on the standard decoction.[Methods]Using Agilent ZORBAX SB-C_(18)(5μm,250 mm×4.6 mm)chromatographic column;gradient elution with acetonitrile-0.4%phosphoric acid solution(0.2:99.8,V/V),column temperature at 25℃;detection wavelength at 236 nm;flow rate 1.0 mL/min;using"Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine",to evaluate the similarity of the characteristic chromatogram of crude drug,standard decoction and formula granules of honeysuckle stem.[Results]The similarity of characteristic chromatogram was higher than 0.94,and 6 components were identified from 10 characteristic peaks.[Conclusions]The established method can more comprehensively reflect the overall appearance of the chemical substances in the honeysuckle stem and the transmission law,and can provide an experimental basis for the consistency of the quality of the honeysuckle stem formula granules.

Mechanism of improving skin barrier function in a mouse model of atopic dermatitis using Mahuang Lianqiao Chixiaodou decoction and its disassembled formula

Objective: To reveal the mechanism of Mahuang Lianqiao Chixiaodou decoction and its disassembled formula for improving the skin barrier function in a mouse model of atopic dermatitis(AD).Methods: Sixty specific-pathogen free male BALB/c mice were randomly divided into the control group,model group, whole formula group(WF), exterior-releasing formula group(ERF), interior-clearing formula group(ICF), and positive control group(PC). A mouse model of AD was established using the semiantigen 2,4-dinitrofluorobenzene induction method. The lesion scores, transepidermal water loss and p H, and skin histopathology of mice in each group were observed. The expressions of filaggrin, loricrin,and involucrin were detected by the streptavidin peroxidase immunohistochemical method and western blotting, and their mRNA expressions were detected by quantitative polymerase chain reaction.Results: Mice in the WF, ERF, ICF, and PC groups showed reduced skin lesion performance, improved histopathology, decreased skin lesion score, transepidermal water loss and pH, and upregulated expressions of proteins including filaggrin, loricrin, and involucrin, and their mRNAs. The most obvious regulatory effect was observed in the WF group, followed by the ICF, ERF, and PC groups, accordingly.Conclusions: Mahuang Lianqiao Chixiaodou decoction and its disassembled formula can improve the skin barrier function in a mouse model of AD by upregulating filaggrin, loricrin, and involucrin, and their mRNA expressions, and the most optimal effect was noted in the WF group, followed by the ICF and ERF groups, which suggests that the effect of clearing heat and resolving dampness in improving the skin barrier function of AD is more obvious and is one of the key treatments for AD.

基于UPLC-Q-Exactive Orbitrap-MS的经典名方温胆汤化学成分分析

[目的]采用超高效液相色谱-四极杆静电场轨道阱高分辨质谱法(UPLC-Q-Exactive Orbitrap-MS)对温胆汤的化学成分进行快速识别和鉴定。[方法]采用Waters Acquity BEH C18(2.1×100 mm,1.7μm),以0.1%甲酸水溶液(A)-乙腈(B)作为流动相进行梯度洗脱,流速为0.3 mL/min、柱温30℃;采用加热电喷雾离子源(HESI),正、负离子模式同时检测,扫描模式为Full MS/dd-MS2,扫描范围m/z 100~1500。[结果]通过与对照品比对、精确相对分子质量、多级质谱碎片信息并结合文献报道、质谱数据库信息等对化合物进行鉴别。从温胆汤中共鉴别了121个成分,包括70个黄酮类成分,18个香豆素类成分,9个皂苷类成分,10个生物碱成分,3个苯丙素类成分及其他类化合物11个。其中15个成分通过与对照品比对后确认。[结论]利用UPLC-Q-Exactive Orbitrap-MS技术较为系统地阐明了温胆汤的物质基础,可为该复方药效物质基础的深入研究和质量标准的合理制定奠定基础。

Establishment of Specific Chromatogram of Spica Prunellae Formula Granules Based on Standard Decoction

[Objectives]This study aimed to establish HPLC chromatograms of decoction pieces,standard decoction and formula granules of Spica Prunellae.[Methods]The chromatographic conditions were as follows:column,SHISEIDO CAPCELL PAK C18 MGII column(4.6 mm×250 mm,5μm);mobile phase,acetonitrile-0.1%phosphoric acid;gradient elution;detection wavelength,280 nm;flow rate,1.0 mL/min;and column temperature,30℃.The correlation between the decoction pieces,standard decoction and formula granules of Spica Prunellae was analyzed by specific chromatograms.[Results]The fingerprint chromatograms of decoction pieces,standard decoction and formula granules of Spica Prunellae showed five common peaks,with good correlation.Among the five common peaks,four of them were tanshinol,protocatechuic acid,caffeic acid and rosmarinic acid.[Conclusions]The main chemical constituents of decoction pieces,standard decoction and formula granules of Spica Prunellae are basically the same.The HPLC specific chromatogram established can be used for the quality control of Spica Prunellae formula granules.

Qingchang decoction retention enema may induce clinical and mucosal remission in left-sided ulcerative colitis: A case report

BACKGROUND Ulcerative colitis(UC)is a chronic autoimmune disease characterized by relapsing-remitting abdominal pain,diarrhea,mucopurulent discharge and rectal bleeding.To date,the etiology of the disease remains unknown;therefore,medical therapy is not yet available.Left-sided UC is mainly treated with oral and topical mesalazine.However,due to its modest clinical effect,endoscopic mucosal remission is not achieved in all patients.CASE SUMMARY A 44-year-old man presented to Longhua Hospital with a history of left-sided UC for more than 6 years and slight bloody diarrhea over time.Endoscopy suggested hyperemia,edema,and erosive mucosa involving the rectum and sigmoid colon.The Traditional Chinese medicine Qingchang decoction(QCD)enema treatment was initiated once a day combined with a previous standard dose of mesalazine for 8 wk,and rectal bleeding ceased after 4 wk of treatment.Another QCD enema treatment was provided after symptom relapse due to drug withdrawal for nearly 6 mo.At the 2-mo follow-up,the colonoscopy results indicated mucosal healing with no erosion or ulcers.CONCLUSION The Chinese formula QCD retention enema represents a potential treatment for left-sided UC with predominant rectal bleeding to achieve clinical and mucosal remission.

基于UHPLC⁃Q⁃Exactive Orbitrap HRMS技术分析黄芪汤颗粒的化学成分与小鼠口服后的入血成分

目的运用超高效液相串联四极杆-静电场轨道阱高分辨质谱(UHPLC-Q-Exactive Orbitrap HRMS)技术,分析鉴定黄芪汤颗粒的化学成分及小鼠灌胃黄芪汤颗粒后的主要入血成分。方法色谱分离采用Acquity UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以甲醇-0.1%甲酸水溶液为流动相梯度洗脱。质谱采用电喷雾电离正离子模式(ESI+)和电喷雾电离负离子模式(ESI-)交替扫描,并自动触发二级质谱扫描功能(Full MS/dd-MS2)采集质谱数据。经Xcalibur 3.0软件获取色谱保留时间以及母离子和碎片离子信息,定性鉴别黄芪汤颗粒的化学成分及小鼠灌胃黄芪汤颗粒后的入血成分及可能的代谢途径。结果综合保留时间、准确分子量、二级碎片信息、对照品比对、文献参考等色谱质谱检测信息,从黄芪汤颗粒中共鉴定出167种化学成分,从小鼠灌胃黄芪汤颗粒后的血清中共检测到甘草素、毛蕊异黄酮、芒柄花素、黄芪甲苷等60种原型成分和43种代谢产物。结论完成了黄芪汤颗粒化学成分及主要入血成分快速鉴定,可为黄芪汤颗粒效应物质解析、质量标准研究及作用机制探讨等提供参考。

Study on therapeutic targets networks of xuefu-zhuyu decoction and gualou-xiebai-banxia decoction for the treatment of coronary artery disease

Aim This work is to provide a network approach to identify the potential therapeutic targets in molecular level for xuefu-zhuyu decoction （XZD） and gualou-xiebai-banxia decoction （GXBD） in treating Coronary artery dis- ease （CAD）. Methods The networks between the ingredients/drugs and relevant target proteins for XZD, GXBD, and modern anti-CAD drugs were constructed, respectively. A master network based on the three established networks was further generated. By comparing the similarities and the differences of the targets containing in the master net- work between the individual formula and the modern drugs, the potential anti-CAD targets for XZD and GXBD were i- dentified for further pharmacological investigations. Results Although the herbal formulations and the chemical con- stituents of XZD and GXBD were significant different, both formulas presented the great similarity on target proteins and with the Tanimoto coefficient of 0. 7225. Comparison the formula-specific targets to modem drugs targets, 50 mu- tual targets with higher possibility were modulated. Moreover, a total amount of 114 mutual targets between formulas derived from the master network were identified to be not yet related to those of the approved anti-CAD drugs. Among them, the top 10 targets were identified to be NOS3, PTPN1, GABRA1, PRKACA, CDK2, MAOB, ESR1, ADH1C, ADH1B and AKR1B1. The formula-specific targets of XZD or GXBD which were not yet covered by the current anti- CAD drugs provided the potential opportunities to discovery of the new drug candidates from the two formulas for CAD treatment. Conclusion The established method of network analysis provides a novel approach for screening of the potential therapeutic targets based on the chemical constituents in traditional Chinese medicines or formulas. It is cru- cial for this work to select relatively favorable therapeutic areas of traditional Chinese medicines, syndrome-oriented formulas and syndrome differentiation of same diseases. Meanwhile, this kind of work is helpful for unveiling the mo- lecular mechanism of TCM formulas.

Dissecting the underlying pharmaceutical mechanism of Danggui Buxue decoction acting on idiopathic pulmonary fibrosis with network pharmacology

Backgroud:Danggui Buxue decoction(DBD),a classical prescription in traditional Chinese medicine,has been found to have protective effect on bleomycin-induced pulmonary fibrosis in rats by reducing alveolar inflammation and fibrosis.However,the biological activity of individual chemical components and mechanism of action of whole formula are not clear.Methods:Potential targets of active ingredients of DBD were collected through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and SymMap database.Target genes related to idiopathic pulmonary fibrosis were obtained from the Online Mendelian Inheritance in Man database,Therapeutic Targets Database and Gkb database.Then,the common targets were obtained by overlapping the potential targets of active ingredients in DBD and diseases related targets.The selected targets were subjected to Kyoto Encyclopedia of Genes and Genomes signaling pathway and Gene Ontology analysis,and the network map of active component-target-pathway was established using Cytoscape 3.7.1 software.The active components of DBD with most targets were selected for fibrosis-related marker verification.The mRNA and protein expression of fibrosis markers,α-smooth muscle actin,collagen 1 and fibronectin,were detected in TGF-β1-induced fibroblast cell line after treatment with the active components.Results:The 14 active ingredients,such as quercetin and kaempferol,were screened from DBD.It acts on 26 targets like estrogen receptor 2 and prostaglandin-endoperoxide synthase 2,and mainly involves 38 signaling pathways such as cell inflammation and autophagy.Kaempferol and quercetin are the two compounds with the highest network regulation,which can inhibit the transformation of fibroblasts into myofibroblasts and reduce the expression of fibrosis markersα-smooth muscle actin,collagen 1 and fibronectin.Conclusion:The integration mode of multi-component,multi-target,multi-channel and mechanism of DBD in the treatment of idiopathic pulmonary fibrosis are predicted by means of network pharmacology.Our study could indicate the direction of further anti-fibrotic mechanism research.

Thoughts on Coronavirus Disease 2019 Based on JingFang Medicine (Classical Chinese Formula) Solutions for COVID-19

This article aims to provide some thoughts on the prevention and treatment of Coronaviras Disease 2019(COVTD-19)from the perspective of JingFang Medicine(Classical Chinese Formula).It is believed that the vague theoretical understanding of COVTD-19 in Traditional Chinese Medicine does not hinder the precise treatment of the disease by following the rale of“With this Zheng,prescribe this Fang.”According to the principle of“Fang-Zheng Correlation”and the knowledge gained from the thousands of years of experience in treating febrile diseases,Xiao Chai Hu Decoction(小柴胡汤)and its modifications are recommended with the emphasis on individualized treatment.As another form of practicing“Fang-Zheng Correlation,”generalized group treatment should also be paid attention to.Giving considerations to(ire historical medical data,Jing Fang Bai Du Powder(荆防败毒散)and Shi Shen Decoction(十神汤)are reconunended for group prevention treatment.Assisting the Zheng(Upright)Qi and using tonic formulas are two entirely different concepts.According to the principle of“Fang-Zheng Correlation,”tonics abuse should be avoided in the prevention of COVID-19,and the using of Huang Qi(黄芪Radix Astragali seu Hedysari)should also be very carefully done.

Thoughts on Coronavirus Disease 2019 Based on JingFang Medicine(Classical Chinese Formula)Solutions for COVID‑19

This article aims to provide some thoughts on the prevention and treatment of Coronavirus Disease 2019(COVID‑19)from the perspective of JingFang Medicine(Classical Chinese Formula).It is believed that the vague theoretical understanding of COVID‑19 in Traditional Chinese Medicine does not hinder the precise treatment of the disease by following the rule of“With this Zheng,prescribe this Fang.”According to the principle of“Fang‑Zheng Correlation”and the knowledge gained from the thousands of years of experience in treating febrile diseases,Xiao Chai Hu Decoction(小柴胡汤)and its modifications are recommended with the emphasis on individualized treatment.As another form of practicing“Fang‑Zheng Correlation,”generalized group treatment should also be paid attention to.Giving considerations to the historical medical data,Jing Fang Bai Du Powder(荆防败毒散)and Shi Shen Decoction(十神汤)are recommended for group prevention treatment.Assisting the Zheng(Upright)Qi and using tonic formulas are two entirely different concepts.According to the principle of“Fang‑Zheng Correlation,”tonics abuse should be avoided in the prevention of COVID‑19,and the using of Huang Qi(黄芪Radix Astragali seu Hedysari)should also be very carefully done.

真武汤联合血府逐瘀方治疗老年冠心病伴慢性心力衰竭患者的疗效观察

目的探讨真武汤联合血府逐瘀方治疗老年冠心病伴慢性心力衰竭(Chronic heart failure,CHF)患者的临床疗效。方法选取2020年5月—2023年6月期间六安市中医院收治的老年冠心病伴CHF患者106例作为研究对象,依据简单随机数字表法分为对照组和研究组,每组各53例。对照组采取常规治疗,研究组在对照组基础上采取真武汤合血府逐瘀方,均治疗4周。观察比较两组患者临床疗效、不良反应发生情况,治疗前后中医证候积分、心功能[左心室射血分数(Left ventricular ejection fraction,LVEF)、心输出量(Cardiac output,CO)、左心室舒张末期内径(Left ventricular end-diastolic diameter,LVEDD)、左心室收缩末期内径(Left ventricular end-systolic diameter,LVESD)]、血流动力学[肺动脉平均压(Mean pulmonary artery pressure,MPAP)、左心房平均压(Mean left atrial pressure,MLAP)、平均二尖瓣跨瓣压差(Mean mitral valve intervalve pressure difference,MMPG)]、心力衰竭标记物[血管内皮生长因子(Vascular endothelial growth factor,VEGF)、可溶性生长刺激表达基因2蛋白(Soluble growth stimulation expressed gene2,sST2)、脑钠肽(Brain natriuretic peptide,BNP)]水平。结果治疗后研究组临床总有效率94.34%(50/53)明显高于对照组81.13%(43/53),差异有统计学意义(P<0.05)。治疗后两组患者肢冷、心悸、气短、喘促及肢体浮肿、胸闷、盗汗、乏力分值均较治疗前降低,差异有统计学意义(P<0.05);且研究组中医证候积分明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者LVEF、CO指标均较治疗前升高,LVEDD、LVESD指标均较治疗前降低,差异有统计学意义(P<0.05);且研究组LVEF、CO指标明显高于对照组,LVEDD、LVESD指标明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者血流动力学MPAP、MLAP、MMPG水平均较治疗前降低,差异有统计学意义(P<0.05);且研究组血流动力学MPAP、MLAP、MMPG水平明显低于对照组,差异有统计学意义(P<0.05)。治疗后两组患者VEGF指标均较治疗前升高,sST2、BNP指标均较治疗前降低,差异有统计学意义(P<0.05);且研究组VEGF指标明显高于对照组,sST2、BNP指标均明显低于对照组,差异有统计学意义(P<0.05)。治疗期间,研究组不良反应发生率7.55%(4/53)与对照组3.77%(2/53)比较,差异无统计学意义(P>0.05)。结论常规基础上采取真武汤合血府逐瘀方治疗老年冠心病合并CHF可改善患者心功能,调节血流动力学状态及心力衰竭标记物水平,缓解临床症状,提升整体治疗效果,且具有安全性。

补肾活血膏方对肾虚血瘀证强直性脊柱炎患者的临床疗效

目的 探讨补肾活血膏方对肾虚血瘀证强直性脊柱炎患者的临床疗效。方法 167例患者随机分为对照组(常规治疗,55例)、暴露组(常规治疗+补肾活血汤剂,54例)、高暴露组(常规治疗+补肾活血汤剂+补肾活血膏方,58例),疗程2年。检测临床疗效、BASDAI评分、ASDAS-CRP、BASFI评分、脊柱痛评分、PGA评分、BASMI评分、ASQoL评分、SPARCC评分、肾虚血瘀证评分、ESR、CRP、IL-6、TNF-α、IL-17、IL-23、IL-35、NLR、PLR、安全性指标变化。结果 高暴露组ASAS40、ASAS5/6、BASDAI50例数多于暴露组和对照组(P<0.05)。治疗后,高暴露组BASDAI评分、ASDAS-CRP、BASFI评分、脊柱痛评分、PGA评分、BASMI评分、SPARCC评分、ASQoL评分、肾虚血瘀证评分、ESR、CRP、IL-6、TNF-α、IL-17、IL-23低于其他2组(P<0.05),IL-35更高(P<0.05)。经Logistic回归分析校正混杂因素后,补肾活血汤、补肾活血膏方降低BASDAI评分和ASDAS-CRP(P<0.05),提高临床疗效(P<0.05)。3组未发现严重不良反应。结论 补肾活血膏方可安全有效地抑制肾虚血瘀证强直性脊柱炎患者炎症,降低疾病活动度,减轻骨髓水肿,改善临床症状,提高关节功能和生活质量。

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。

《方剂学》教材商榷5则

《方剂学》教材中存在一些值得商榷的问题,如安神药服药时间、麻黄杏仁甘草石膏汤证是否兼有外邪、四逆散证有无外邪的方义解释、牡蛎散是否治疗盗汗、大秦艽汤的分类归属等。这些问题不单涉及到教学,更影响临床应用,对此作一探讨,以有利于中医教学与临床。

基于标准汤剂的鲜龙葵果配方颗粒质量标准研究

目的基于标准汤剂的质量评价方法,建立鲜龙葵果配方颗粒的质量标准。方法制备并计算10批鲜龙葵果标准汤剂及3批配方颗粒的出膏率;以鲜龙葵果药材为对照,采用薄层色谱法(TLC)鉴别鲜龙葵果配方颗粒的专属性;采用超高效液相色谱法(UPLC)测定鲜龙葵果配方颗粒中澳洲茄碱、澳洲茄边碱的含量,并建立鲜龙葵果配方颗粒的特征图谱,确定4个特征峰及其相对保留时间的规定范围。结果标准汤剂出膏率为(18.21±1.87)%,确定的配方颗粒的出膏率范围为12.75%~23.67%,3批配方颗粒的出膏率分别为15.90%、21.26%、18.01%。标准汤剂中澳洲茄碱含量为(24.54±1.24)mg·g^(-1),澳洲茄边碱的含量为(31.34±1.65)mg·g^(-1);3批配方颗粒中澳洲茄碱含量分别为19.3 mg·g^(-1)、18.5 mg·g^(-1)、20.4 mg·g^(-1);澳洲茄边碱含量分别为23.9 mg·g^(-1)、22.9 mg·g^(-1)、25.8 mg·g^(-1)。识别出4个共有特征峰,以澳洲茄碱为参照物峰,其余3个峰的相对保留时间规定值分别为1.13、1.70、1.85。结论本研究证明鲜龙葵果配方颗粒与标准汤剂具有质量一致性,为制定配方颗粒的质量标准提供了数据支持。

中药复方基于Wnt/β-catenin信号通路调控糖尿病肾病研究进展

糖尿病肾病是糖尿病最常见的微血管并发症之一,也是造成终末期肾病的主要原因。Wnt/β-catenin信号通路与糖尿病肾病肾小管间质纤维化密切相关,主要通过足细胞、肾小球系膜细胞、肾小管上皮细胞等来改善糖尿病肾病。中药复方基于调控Wnt/β-catenin信号通路来改善肾脏病变及纤维化,保护肾脏功能,进而延缓病程进展,在治疗中发挥激活自噬、减少炎症反应及减弱氧化应激等作用。目前,关于中药复方基于Wnt/β-catenin信号通路的研究仍存在以下不足之处:(1)在中药复方的研究中,能够得出中药复方可通过Wnt/β-catenin信号通路发挥作用,但由于中药复方成份复杂多样,大多数仅仅发现相关性,而没有分析其具体的活性成分;(2)Wnt/β-catenin信号通路的相关研究并未深入探索该信号通路在肾纤维化中的作用机制;(3)目前关于中医药的大部分实验为动物体内实验,缺乏体外细胞的相关实验探究。今后,需加强对中药复方活性成分的研究,并开展更多的体外细胞实验,进行深层次的作用机制及靶点研究。

橘皮竹茹汤对化疗性异食癖大鼠模型5-羟色胺3受体蛋白和mRNA表达的影响

目的观察橘皮竹茹汤对化疗性异食癖大鼠模型止呕作用及对5-羟色胺3受体(5-HT3R)蛋白和mRNA的影响。方法将36只Wistar雄性大鼠随机分为正常对照组、中药对照组(橘皮竹茹汤10.6 g/kg,不造模)、模型组、昂丹司琼组(2.6 mg/kg)、橘皮竹茹汤低剂量组(5.3 g/kg)和橘皮竹茹汤高剂量组(10.6 g/kg),每组6只。各组大鼠给予相应干预措施6 d(每日2次)后,采用腹腔注射顺铂(6 mg/kg)的方式诱导建立化疗性异食癖大鼠模型,观察并记录造模后24 h内大鼠体质量、摄食量和高岭土摄入量的变化。继续干预1 d后,苏木精-伊红(HE)染色观察各组大鼠胃窦、回肠组织形态学变化;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测延髓、回肠组织中5-HT3R、色氨酸羟化酶(TPH)、单胺氧化酶A(MAOA)mRNA表达;Western blot法检测延髓组织中5-HT3R蛋白的表达。结果①与正常对照组比较,模型组大鼠体质量、摄食量降低(P<0.05),高岭土摄入量显著增加(P<0.05);与模型组比较,昂丹司琼组和橘皮竹茹汤低、高剂量组大鼠高岭土摄入量显著降低(P<0.05)。②HE染色显示,与正常对照组比较,模型组胃窦组织上皮细胞受损,固有层腺体排列疏松、紊乱,回肠上皮部分缺失,腺体排列紊乱;与模型组比较,昂丹司琼组和橘皮竹茹汤低、高剂组胃窦和回肠组织病理改变减轻。③RT-qPCR结果显示,与正常对照组比较,模型组大鼠延髓、回肠5-HT3R mRNA表达水平显著升高(P<0.05),回肠MAOA mRNA表达水平显著降低(P<0.05);与模型组比较,橘皮竹茹汤高剂量组大鼠延髓5-HT3R mRNA表达水平明显降低(P<0.05),橘皮竹茹汤低剂量组回肠TPH1 mRNA、橘皮竹茹汤高剂量组延髓TPH2 mRNA表达水平明显降低(P<0.05),昂丹司琼组、橘皮竹茹汤低剂量组、橘皮竹茹汤高剂量组回肠MAOA mRNA表达水平明显升高(P<0.05)。④Western blot结果显示,与正常对照组比较,模型组大鼠延髓5⁃HT3R蛋白表达水平显著升高(P<0.05);与模型组比较,橘皮竹茹汤高剂量组大鼠延髓5-HT3R蛋白表达水平显著降低(P<0.05)。结论橘皮竹茹汤可以通过抑制延髓5-HT3R蛋白和mRNA的表达,调控5-羟色胺合成与代谢相关酶,进而改善化疗导致的恶心呕吐。

寒喘祖帕单煎配方与配伍合煎对16HBE细胞抗炎活性及非靶向代谢组学差异的研究

目的考察寒喘祖帕按照传统配伍合煎及单煎配方的抗炎活性,并采用UPLC-Q-TOF/MS研究非靶向代谢组学差异。方法通过体外脂多糖诱导16HBE炎症细胞,使用CCK-8法筛选出单煎配方组和配伍合煎组的剂量;采用逆转录酶聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)技术测定细胞中炎症因子IL-6、IL-8和TNF-α的mRNA表达水平,并采用UPLC-TOF/MS技术对细胞进行非靶向代谢组学分析:分为正常组(16HBE细胞+培养基)、模型组(16HBE细胞+培养基+0.1μg/mL LPS)、寒喘祖帕单煎配方组(16HBE细胞+培养基+0.1μg/mL LPS+50μg/mL单煎配方)、寒喘祖帕配伍合煎组(16HBE细胞+培养基+0.1μg/mL LPS+50μg/mL配伍合煎)。以此探究寒喘祖帕颗粒发挥抗炎活性的相关代谢通路,寻找寒喘祖帕颗粒抗炎活性相关的差异代谢物。结果与正常组比较,模型组在给药浓度为0.1μg/mL时细胞生长存活率未受到限制,差异无统计学意义(P>0.05);配伍合煎组在给药浓度为0~400μg/mL范围内时,细胞存活率未受影响,浓度>800μg/mL时,细胞生长存活率下降,差异具有统计学意义(P<0.01);而单煎配方组在浓度<50μg/mL时,细胞生长存活率未受到限制,浓度≥100μg/mL时,细胞生长存活率下降,差异具有统计学意义(P<0.01);RT-PCR实验结果显示:与正常组比较,模型组的IL-6、IL-8和TNF-αmRNA的表达上升,差异具有统计学意义(P<0.001),50μg/mL单煎配方较100μg/mL,IL-6、IL-8和TNF-αmRNA的表达下降,差异具有统计学意义(P<0.001);在相同药物浓度干预下,与正常组比较,模型组的IL-6、IL-8和TNF-αmRNA的表达上升,差异具有统计学意义(P<0.001),与模型组比较,单煎配方组和配伍合煎组可同时降低IL-6、IL-8和TNF-αmRNA的表达,差异具有统计学意义(P<0.001);配伍合煎组与单煎配方组比较,配伍合煎组IL-6、IL-8 mRNA的表达较单煎配方低,差异具有统计学意义(P<0.05)。代谢组学分析结果筛选出模型组与配伍合煎组间的内源性差异代谢物68个,主要与色氨酸代谢、甘油磷脂代谢和嘧啶代谢等有关;模型组与单煎配方组间识别出内源性差异代谢物62个,主要与甘油磷脂代谢、精氨酸和脯氨酸代谢等有关。结论寒喘祖帕单煎配方与配伍合煎对LPS诱导的炎症细胞模型抗炎效果存在差异,非靶向代谢组学分析结果表明两种方式通过不同的代谢通路发挥抗炎活性。

百合地黄汤本源考证及临床心得

百合地黄汤出自《金匮要略》,是医圣仲景诊治百合病的常用方剂。何庆勇教授临床运用百合地黄汤的关键有四:一是经方百合的入药品种应为食用百合,而不用卷丹;二是要重视百合地黄汤的主证,口干或口苦,小便黄赤,脉数;三是要注重古方原剂量,通过实测,原文“百合七枚”约(724.9±22.3)g鲜百合,“生地黄汁一升”约(418.8±11.3)g鲜地黄,故临床需重剂百合、地黄,何庆勇教授临证常用百合30~80 g、地黄30~80 g;四是应注重煎服法,煎药前须先浸泡百合,并去白沫。并举百合地黄汤医案一则,以资佐证。