Investigation on the mechanism of YQHX against cerebral ischemic injury based on network pharmacology and molecular docking

Cerebral ischemia(CI)is the world’s second-largest lethal disease,with a high recurrence and teratogenic rate.Traditional Chinese medicine(TCM)YQHX(a pharmaceutical preparation from herbs)has a certain effect in the clinical treatment of CI,while its underlying mechanism remains largely undetermined.To explore the potential mechanism,we used network pharmacology and molecular docking in the present study.TCMSP and CNKI databases were used to explore the active ingredients of YQHX;the Pharmmapper database was used to get the ingredient targets;the OMIM,GeneCards,and DisGeNET databases were used to obtain the disease targets;the Venn diagram was used to obtain the intersection targets,the Cytoscape was used to visualize results and plug-in MCODE to obtain core targets;the Metascape database was used to perform GO and KEGG pathway enrichment analyses on core targets.The top 20 KEGG pathway enrichment pathways were used to construct the“ingredient-target-pathway”network by Cytoscape;the top 10 ingredients and the top five protein targets were used for molecular docking with AutoDock Vina software,and PyMoL and Ligplus software were used to visualize the results.A total of 83 active ingredients were screened from YQHX.Moreover,432 corresponding targets,2005 disease-related targets,and 140 drug-disease intersection targets were obtained.GO biological function and KEGG pathway enrichment analyses yielded 507 biological function entries and 141 signaling pathways.KEGG pathway enrichment was mainly involved in cell proliferation,adhesion,migration,and other processes.Molecular docking results showed that the key ingredients and core targets screened had a strong binding activity,including EGFR,MAP2K1,and KDR.The combination of miltionone I and miltiodiol was relatively stable.The main biological mechanism of YQHX in the treatment of CI might play a role through the signaling pathway related to the tyrosine kinase receptor,which was also the improvement of the theory of“benefiting qi and activating blood circulation for promoting the production of blood and blood vessels”.

Yiqi Huoxue prescription can prevent and treat post-MI myocardial remodeling through promoting the expression of AMPK signal pathway

Objective:To investigate how Yiqi Huoxue (YQHX) prescription regulates mitochondrial biosynthesis and ATP synthesis via AMP-activated protein kinase (AMPK) and to reveal its molecular mechanism in preventing and treating post-MI myocardial remodeling.Methods:The MI animal model of myocardial infarction were established by ligating Sprague Dawley (SD) rats' left anterior descending coronary arteries;the animals were randomly divided into MI group,YQHX prescription group and perindopril group,and a sham operation group was set at the same time.Related drug intervention was administered on the 2nd day after surgery,the YQHX group was given Astragalus,angelica,ginseng,Ligusticum wallichii and pseudoginseng,provided by the Dongzhimen Hospital of Beijing University of Chinese Medicine,once per day,at a dose of 21 g/kg body weight/day (the clinical equivalent dose based on a previous study),and the changes in relevant indicators were observed at 1 week and 4 weeks.Echocardiography (ECG) was used to observe the changes in rat cardiac structure and functions;the morphology-based technique was used to observe the changes in myocardial cells and mitochondria;the expression of AMPK signal pathway-related proteins and mRNA was detected using western blotting and real-time fluorescence quantification respectively,while fluorescence enzyme-labeled method was used for detecting ATP synthesis.Results:Cardiac structure and functions:Compared with the MI group at 1 week,the YQHX prescription group and the perindopril group exhibited increased LVEF and LVFS (Pall <.05);their LVEDs and LVEDd were reduced but suggested no statistically significant differences (F =2.258,F =0.3464,Pall >.05).At 4 weeks,both LVEF and LVFS were elevated in the YQHX prescription group (P =.008,.009) and the perindopril group (P =.279,.333),where differences in the later group indicated no statistical significance;the YQHX prescription group and the perindopril group were also featured by reduced LVEDs and LVEDd (Pall <.05).Morphology:Compared with the MI group at two time nodes,the YQHX prescription group and the perindopril group exhibited significant improvement in the pathological changes in myocardial cells and mitochondrial structure.Expression of AMPK signal pathway-related proteins and mRNA:The expression of both pLKB1 and pAMPK proteins followed a rising trend in the YQHX prescription group and the perindopril group at 1 week.The expression of LKB1mRNA and AMPKmRNA was elevated (Pall <.05).The increased expression of PGC-1α,NRF1 and mtTFA proteins demonstrated statistically significant differences (Pall <.05).The expression of mtDNA protein followed an increasing trend.At 4 weeks,the expression of both pLKB1 and pAMPK proteins was elevated (Pall <.05).Heightened expression was also reported in LKB1mRNA and AMPKmR-NA (Pall <.05).The increased expression of PGC-1α,NRF1,mtTFA and mtDNA proteins demonstrated statistically significant differences (Pall <.05).ATP synthesis:ATP synthesis was increased in the YQHX prescription group and the perindopril group at both 1 week and 4 weeks (Pall <.001).Conclusion:The possible mechanism of YQHX prescription for preventing and treating post-MI myocardial remodeling may function through strengthening the activation AMPK signal pathway by LKB1,thus further increasing the expression of downstream transcription factor proteins and initiating mitochondrial replication and transcription;as a result,the YQHX prescription can improve the post-MI damage in mitochondrial morphological structure in the tissue at heart marginal zone as well as enhance mitochondrial biosynthesis and ATP synthesis.

益气活血方对实验性大鼠子宫肌瘤的影响

目的观察益气活血方治疗实验性大鼠子宫肌瘤的作用．方法将三月龄未孕雌性大鼠随机分为空白对照组、模型对照组、阳性给药组0.465g／kg／d、益气活血方高剂量给药组40g／kg／d、低剂量给药组10g／kg／d，用苯甲酸雌二醇加黄体酮肌肉注射方法对模型对照组、阳性给药纽、益气活血方高、低剂量给药组进行子宫肌瘤模型的复制，一周后同时对给药组分别灌胃给予受试药益气活血方和阳性对照药桂枝茯苓胶囊，五周后测各组的血液流变学指标，解剖大鼠取出子宫．计算子宫系数，并对子宫进行病理组织形态学观察．结果益气活血方能抑制子宫肌瘤模型大鼠子宫的扩大及瘤样增生和改善血凝状态，结论益气活血方具有预防和治疗子宫肌瘤的作用．

益气活血中药对肺癌上皮细胞粘附分子表达及侵袭的影响

目的观察益气活血中药对肺癌细胞株体外生长抑制,上皮型钙粘素(E-CD)表达及侵袭能力的影响。方法采用细胞计数、S-P免疫组织化学、图像分析及侵袭小室等方法,观察人肺腺癌细胞株A549、小细胞肺癌细胞株NC-H446在益气活血中药、全反式维甲酸(ARTA)处理不同时间后细胞生长、E-CD表达及侵袭能力的影响。结果在益气活血中药干预下,NC-H446、A549两细胞株生长受到抑制,E-CD阳性表达率分别由原来的12%±8%、23%±11%提高到34%±9%、39%±13%,积分光密度分别由78.5±16.9、109.8±13.2增加到200.3±21.7、229.7±11,前后相比具有明显差异穴P<0.01雪鸦穿过基质胶的细胞数分别为126.5±43.61、106.9±55.48,与对照组相比差异显著穴P<0.01雪鸦上述情况与ARTA相比,无显著性差异(P>0.05)。结论益气活血中药能明显抑制NC-H446和A549生长熏能促进E-CD的正常表达熏减弱肿瘤细胞的侵袭转移能力。

益气活血中药联合西药治疗PCI术后的临床观察

[目的]观察益气活血中药联合西药治疗经皮腔内冠状动脉介入术（PCI）后的临床疗效。[方法]将40例PCI成功的冠心病患者，随机分为治疗组20例，对照组20例。对照组以西药常规治疗，治疗组以西药常规用药加益气活血中药治疗。疗程3个月。观察患者服药后的临床症状、心绞痛发生率及西药的停减情况。[结果]在改善临床症状、减少心绞痛发生率及西药停减方面，治疗组和对照组比较有显著差异（P〈0．05）。[结论]益气活血中药联合西药治疗能提高PCI术后临床疗效、防治PCI术后再狭窄。

益气活血颗粒对心肌梗死大鼠尿液代谢组学的影响

目的观察益气活血颗粒对心肌梗死后大鼠尿液中代谢物质的影响。方法采用结扎左冠状动脉前降支制备SD大鼠的心肌梗死(MI)模型,将造模成功的大鼠随机分为7 d组和28 d组。每组再次随机分为模型组、培哚普利组、芪参益气滴丸组和益气活血颗粒组,另设只穿刺不结扎的假手术组。术后第2天开始培哚普利组(0.4 mg/kg)、芪参益气滴丸组(30 mg/kg)、益气活血颗粒组(21 g/kg)每天给予相应药物灌胃,模型组和假手术组给予等量蒸馏水灌胃。分别于灌胃7 d和28 d后收集大鼠尿液。尿液样本中含有的代谢物质种类相同,但含量不同,通过核磁共振氢谱检测分别筛选出7 d组和28 d组中差异代谢物质,并在假手术组和模型组比较得到的共同差异代谢物基础上,比较益气活血颗粒组与模型组的差异。结果灌胃7 d后初步筛选出有差异的代谢物质16种(P<0.05)。模型组与假手术组相比,尿液中找到差异代谢物9种(P<0.05),其中模型组赖氨酸、β-羟丁酸、脯氨酸、谷氨酸、草酰乙酸酯含量较假手术组降低,而天冬酰胺、果糖、异亮氨酸、丝氨酸含量较假手术组升高。益气活血颗粒组与模型组比较,找到差异代谢物6种(P<0.05),益气活血颗粒组赖氨酸、β-羟丁酸、谷氨酸、草酰乙酸酯的含量较模型组升高,果糖、异亮氨酸含量较模型组降低。灌胃28 d后初步筛选出差异代谢物质25种(P<0.05)。模型组与假手术组相比,尿液中找到差异代谢物14种(P<0.05),其中模型组β-羟丁酸、谷氨酸、甲胺、半胱氨酸、甜菜碱、胆碱、乳酸、脯氨酸、苏氨酸、苹果酸、烟酸含量较假手术组降低,而鸟氨酸、γ-氨基丁酸、α-葡萄糖含量较假手术组升高。益气活血颗粒组与模型组比较找到差异代谢物12种(P<0.05),益气活血颗粒组谷氨酸、半胱氨酸、甜菜碱、胆碱、乳酸、脯氨酸、苏氨酸、苹果酸、烟酸含量较模型组升高,鸟氨酸、甲胺、α-葡萄糖的含量较模型组降低。结论 MI后大鼠出现多种代谢障碍,而益气活血颗粒可有效调节紊乱的代谢产物,促使其向正常范围转归。