乌梅含漱液治疗尿毒症口干症的网络药理学研究及生物学验证

目的基于网络药理学并辅以初步生物学验证探讨乌梅含漱液治疗尿毒症口干症的分子机制。方法 通过网络药理学分析从乌梅含漱液中筛选出124 个活性成分,利用Genecards、OMIM、Disgenet、TTD、 DrugBank 等数据库检索尿毒症口干症的疾病靶点预测。采用Cytoscape 3.8.0 软件构建“药物活性成分-治疗尿 毒症口干症潜在靶点”网络,将活性成分与预测疾病靶点的二者进行分子对接验证,借助R 3.6.3 软件的 ggplot2 包及clusterProfiler 将关键靶点进行GO 和KEGG 富集分析。最后以硫酸阿托品诱导5/6 肾切除大鼠制备 尿毒症口干症大鼠模型为研究材料,采用低、高不同剂量乌梅含漱液干预,从大鼠唾液分泌、唾液腺病理组织 学以及关键靶标蛋白表达等多种指标,进行多个层面的验证性实验,验证前期计算机模拟结果的可靠性。 结果计算机模拟实验预测结果表明,124 个活性药效分子作用于46 个关键靶点,其中核心靶点为EGFR、 ESR1、TP53、MCM2、NTRK1、XPO1、FN1、CDK2、CUL3、APP,分子对接结果发现活性成分与ESR1、 CDK2 结合性能良好。靶标主要作用涉及炎症反应、氧化应激反应、受体活性及生物化学过程调控等多种生物 过程,参与炎症、氧化应激、细胞凋亡、信号转导及其他等20 条信号通路。生物学验证实验结果表明,经乌 梅含漱液的干预后,明显增加了尿毒症口干症大鼠的平均唾液流量,减轻了颌下腺组织受损程度,且增加了分 子对接筛选的靶标ESR1 蛋白的表达量。结论乌梅含漱液的药效分子可能作用于ESR1 核心靶标,通过调节 PI3K/Akt 等多条信号通路发挥药效,为进一步开展乌梅含漱液治疗尿毒症口干症作用机制的研究提供了新思路 和新方法。     

氯沙坦对肾间质纤维化的抑制作用

目的:肾小管间质纤维化(tubulointerstitial fibrosis,TIF)是慢性肾脏病进展为终末期肾病(end-stage renaldisease,ESRD)的重要共同通路。血管紧张素ⅡAT1受体阻断剂--氯沙坦对慢性肾脏病具有保护作用,但其能否抑制肾小管间质纤维化及其可能的机制尚不清楚。本实验通过在体内构建单侧输尿管梗阻(unilateral ureteral obstruction,UUO)大鼠模型,观察氯沙坦对肾小管间质纤维化的抑制作用及其可能机制。方法:50只SD大鼠分为3组:假手术组(n=10),UUO组(n=20),UUO氯沙坦治疗组(n=20)。术后氯沙坦治疗组予以氯沙坦灌胃,剂量20mg·kg-1·d-1。分别在术后第7天、第14天处死大鼠。冰冻组织行E-cadherin,Vimentin,α-SMA,β-catenin和ZEB1免疫荧光染色及Western blot分析。结果:UUO组大鼠第7天组织病理学改变为肾小管管腔扩大,小管萎缩,间质增宽及炎细胞浸润,部分小管间质纤维化;第14天间质纤维化表现更为严重。与对照组相比,UUO模型组大鼠第7天、第14天肾间质上皮标志物E-cadherin表达显著下降,而Vimentin,α-SMA,β-catenin和ZEB1表达增加。与UUO模型组比较,UUO氯沙坦治疗组大鼠肾小管间质纤维化改变明显减轻,肾组织E-cadherin表达增加,Vimentin,α-SMA,β-catenin和ZEB1的表达减少。结论:氯沙坦可抑制大鼠体内的肾小管间质纤维化,其机制可能与氯沙坦抑制小管上皮间充质转化相关因子β-catenin/ZEB1的表达有关。     

结合肠肾轴理论探讨中医药从结肠治疗慢性肾脏病的研究思路

慢性肾脏病（CKD）正日益成为全球性的社会问题,延缓CKD 的进展具有重要经济和社会意义。近年研究发现,结肠是产生尿毒症毒素的重要器官之一,肠源性尿毒素与CKD 的预后密切相关,因此国外学者提出了延缓CKD 进展的肠-肾轴理论（The Theory of Gut-kidney Axis）,成为研究的热点。中药结肠给药治疗CKD 在临床广泛应用,大量的临床报道提示其可延缓肾衰进展,但临床研究质量偏低,限制了其进一步推广应用。本研究在广东省中医院肾病科学术带头人——黄春林教授应用通腑泄浊法治疗CKD 思路的指导下,结合最新肠肾轴理论,开展了一系列探索性研究,并提出了进一步临床和实验研究的思路,为日后开展高质量的临床研究,获得高级别的临床证据提供数据和方法学基础。     

基于肠道菌群探讨补脾益肾方改善5/6肾切除大鼠肠道屏障延缓慢性肾脏病进展的作用机制

目的 探讨补脾益肾方延缓5/6肾切除大鼠慢性肾脏病进展的作用机制。方法 雄性SD大鼠随机分为4组：假手术组、5/6肾切除模型组、补脾益肾方低剂量组、补脾益肾方高剂量组。假手术组和5/6肾切除模型组予蒸馏水灌胃，补脾益肾方低剂量组和高剂量组分别给予低剂量和高剂量补脾益肾方治疗，灌胃为期4周。检测各组大鼠血肌酐、尿素氮、24 h尿蛋白定量、硫酸吲哚酚、二胺氧化酶；肾脏组织采用PAS和Masson染色；结肠组织采用HE染色；Western-blot检测ZO-1、Occludin、Claudin-1三种紧密连接蛋白及IL-22在结肠组织的表达水平；采用16S rDNA高通量测序并进行生物信息学分析。结果 与假手术组相比，5/6肾切除组大鼠的血肌酐、尿素氮、24 h尿蛋白定量、血清硫酸吲哚酚和二胺氧化酶水平明显升高；与5/6肾切除组相比，补脾益肾方高剂量组大鼠的血肌酐、尿素氮、24 h尿蛋白定量、血清硫酸吲哚酚和二胺氧化酶水平明显下降，肾脏病理损伤缓解；补脾益肾方亦可调整肠道菌群，提高益生菌Prevotella（普氏菌属）、Phascolarctobacterium（考拉杆菌属）和Megamonas（巨单胞菌属）的相对丰度，降低Clostridiaceae（梭菌科）、Haloplasmataceae（盐扁菌科）、Micrococcaceae（微球菌科）、Pseudomonadaceae（假单胞菌科）和Ruminococcus（瘤胃球菌）的相对丰度；促进紧密连接蛋白（ZO-1、Occludin和Claudin-1）及IL-22的表达，减轻结肠固有层和粘膜层水肿及炎症细胞浸润；减少毒素入血，血清中的硫酸吲哚酚水平下降，进而延缓CKD进程。结论 补脾益肾方延缓5/6肾切除大鼠的CKD进展可能是通过调节肠道菌群，修复肠道屏障功能，降低血清尿毒症毒素水平，以实现肾保护作用。     

补脾益肾法治疗慢性肾脏病的特色与优势

正随着经济发展,慢性病已成为我国国民的主要健康问题,慢性肾脏病(chronic kidney disease,CKD)是一类进展性疾病,在过去10年间,已成为世界主要的公共卫生健康问题而备受关注。调查结果显示,我国成年人群中慢性肾脏病的患病率为10.8%,据此估计     

尿毒清颗粒对体内及体外肾小管上皮细胞向间质细胞转化的抑制作用

Objective： To investigate the effect of the Uremic Clearance Granule （UCG, 尿毒清颗粒）, a Chinese patent medicine, on tubular epithelial-to-mesenchymal transition （EMT） in a unilateral ureteral obstruction （UUO） model in vivo and transforming growth factor （TGF）- 131 induced EMT of HK-2 cells in vitro. Methods： In vivo study, 50 Sprague Dawley rats were divided into three groups： a sham operation group （n=10）, a UUO group （n=20）, and a UUO with UCG treatment group （n=20）. The UCG was given at a dose of 4.5 g/kg body weight per day by gavage after surgery. In vitro study, HK-2 cells were cultured in 10% fetal bovine serum （FBS）, 10% healthy rat serum, 10% FBS and TGF-13 1 （10 ng/mL）, 10% healthy rat serum and TGF-13 1, or 10% rat serum containing the uremic clearance granule and TGF-13 1. The expression of the epithelial marker E-cadherin and the mesenchymal markers vimentin and e~-smooth muscle actin （ot-SMA） in kidney tissues and HK-2 cells were investigated by Western blot analysis and immunofluorescence staining. Results： The rats of the UUO group showed obvious tubulointerstitial fibrosis, compared with the sham operation group rats. Tubulointerstitial fibrosis score was reduced by 17.5%± 1.1% at day 7 and by 20.0%_＋ 1.2% at day 14 in the UCG-treated group, compared with the UUO group. The UCG could maintained expression of E-cadherin and suppressed expression of vimentin and α-SMA in kidney tissues of UUO rats at days 7 and 14, as determined by Western blot analysis and immunofluorescence staining. Rat serum containing the UCG partially inhibited TGF- β 1-induced fibroblast phenotype of HK-2 cells and maintained the epithelial morphology of HK-2 cells in vitro. This occurred partially through a reduction of vimentin expression and an increase of E-cadherin expression. Conclusion： These results suggest that the UCG prevents tubular EMT and may be a promising agent for treating tubulointerstitial fibrosis.     

Treatment of Fibrillary Glomerulonephritis by Corticosteroids and Tripterygium Glycoside Tablets: A Case Report

<正>Fibrillary glomerulonephritis(FGN)is a rare kidney disease characterized by the deposition of randomly arranged fibril deposits 15-30 nm in diameter,mainly in the mesangium and/or glomerular basement membranes(GBMs),and negative Congo red staining.~((1,2))The diagnosis of FGN is made in about 1%of native kidney biopsy specimens.~((3))     

Colon May Provide New Therapeutic Targets for Treatment of Chronic Kidney Disease with Chinese Medicine

Chronic kidney disease(CKD) has become a worldwide health and social problem.Retarding its progression to end-stage renal disease is beneficial both to the patients and the healthcare system.Plenty of clinical trials have indicated that enema with Chinese medicine could effectively prevent chronic renal failure,and was widely used in the clinical practice.However,studies on mechanism were still nearly blank,which may prevent further improvement of therapeutic efficacy.Recent studies had discovered that colon was an important organ where uremic toxins were generated.The uremic toxins involved could not only promote CKD progression,but also was closely correlated with CKD mortality.Reducing production and promoting excretion of toxins were confirmed to reduce renal tubule interstitial fibrosis and delay renal progression.On the basis of the theory of gut-kidney axis above,we had conducted pilot clinical researches to evaluate the effect of enema with Chinese medicine on the intestinal flora,gut barrier,enterogenous uremic toxins and renal protection.The preliminary results revealed that rheum enema through colon could accelerate intestinal dynamics,improve intestinal barrier function,regulate intestinal flora and reduce production and absorption of intestine-derived uremic toxins such as indoxyl sulfate,which may reduce renal fibrosis and delay renal progression.Further studies could provide more evidence for colon as a new therapeutic target for the treatment of CKD with Chinese medicine.     

荞麦壳药包加压包裹治疗肾性水肿31例

目的:观察荞麦壳药包加压包裹治疗肾性水肿的临床疗效。方法:采用荞麦壳药包加压包裹治疗肾性水肿31例,每天包裹时间为6 h,14 d为1个疗程,观察患者下肢水肿消失情况。结果:31例中,治愈10例,好转14例,未愈7例,总有效率达80%。结论:利用荞麦壳药包可以提高肾性水肿的治愈率,且不良反应少。     

黄春林教授从脾肾相关理论指导慢性肾脏病症状管理的经验

慢性肾脏病患者长期受到多种临床症状困扰,生活质量明显下降,需要科学有效的症状管理。本文介绍广东省名中医黄春林教授应用脾肾相关理论指导我科对慢性肾脏病患者进行症状管理的经验,为临床挖掘有效的干预策略提供思路。