膜肾1号方对大鼠肾脏病理及PI3K/Akt/mTOR信号通路表达的影响

[目的] 观察膜肾1号方对膜性肾病大鼠肾脏病理的改善作用及其对自噬通路磷脂肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/雷帕霉素靶蛋白（mTOR）相关蛋白表达的影响。[方法] 将大鼠随机分为对照组、模型组、膜肾1号方高剂量组、中剂量组、低剂量组,盐酸贝那普利组。采用大鼠尾静脉注射阳离子化牛血清白蛋白（C-BSA）的方法建立MN大鼠模型,灌胃、取材。苏木精-伊红（HE ）染色法观察大鼠肾脏组织病理改变;免疫球蛋白G（IgG）免疫荧光染色观察大鼠IgG沉积;蛋白免疫印迹法（Western Blot）检测PI3K/Akt/mTOR 信号通路相关蛋白及自噬相关蛋白轻链3（LC3）表达。[结果] 药物干预后,膜肾1号方组大鼠24 h尿蛋白、三酰甘油、总胆固醇、低密度脂蛋白下降且低于模型组,并具有统计学差异（P＜0.05）。光镜下观察,HE染色示正常组肾组织整体结构基本正常,膜性肾病（MN）模型组肾小球毛细血管丛充血,系膜增生,基底膜出现增厚,部分肾小管细胞空泡变、组织内可见炎症细胞浸润,可见嗜复红蛋白及IgG沉积。经膜肾1号方和盐酸贝那普利干预后,大鼠肾脏病理学改变均有所减轻。各组大鼠IgG沉积显示,与对照组比较模型组IgG沉积明显,IgG荧光表达升高,差异具有统计学意义（P＜0.05）,盐酸贝那普利组和膜肾1号方组IgG荧光表达下降且低于模型组,具有统计学差异（P＜0.05）。Western Blot检测显示,药物干预后,盐酸贝那普利组和膜肾1号方组大鼠PI3K-Akt信号通路相关蛋白表达下降,LC3 表达增加,并具有统计学差异（P＜0.05）。[结论] 膜肾1号方可改善大鼠肾脏病理损伤,膜肾1号方干预后PI3K-Akt信号通路相关蛋白磷酸化磷酸肌醇3激酶（p-PI3K）,磷酸化Akt蛋白（p-Akt）,磷酸化雷帕霉素靶蛋白（p-mTOR）表达明显降低,自噬相关蛋白LC3表达升高,其分子机制与自噬信号通路的调控有关。     

四君子汤非多糖功效组分的体外人源肠、肝微粒体代谢研究

目的 对四君子汤非多糖组分的主要活性成分进行人源肠、肝微粒体中的代谢过程研究,明确其在肠、肝中的代谢轮廓及代谢差异,为其临床应用提供科学证据。方法 应用Acquity UPLC I-class/VION IMS QTOF技术分析四君子汤非多糖组分中主要活性成分（异甘草苷、甘草素、甘草酸、甘草次酸、人参皂苷Rb₁、人参皂苷CK、白术内酯III）在人源肠、肝微粒体孵育体系下的代谢产物以及代谢转化途径。结果 甘草酸、甘草次酸在人源肠、肝微粒体中均以I相代谢为主;而甘草素、异甘草苷、人参皂苷Rb₁、人参皂苷CK则以肠II相代谢为主。白术内酯III在肠、肝微粒体中代谢均不显著,而四君子汤非多糖功效组分中其他化合物在肠微粒体中代谢速度均比在肝微粒体中更快。此外,甘草皂苷、人参皂苷主要进行脱糖基反应;异甘草苷以脱糖、羟基化等反应为主;甘草素则以葡萄糖醛酸结合为主;白术内酯III则主要发生水解、羟基化反应。结论 阐明了四君子汤非多糖组分中主要活性成分在人体肠道及肝脏中的代谢轮廓及代谢差异,为后续进行四君子汤非多糖功效组分的药效研究提供了数据支持,也为中药复方的功效组分研究提供了思路和策略。     

清咳平喘颗粒治疗痰热郁肺型咳嗽的临床疗效观察

目的 观察清咳平喘颗粒治疗痰热郁肺型咳嗽的临床有效性。方法 选取2020年1月—2021年6月上海中医药大学附属龙华医院呼吸科门诊就诊的痰热郁肺型咳嗽患者100例,按随机数字表法分为对照组和治疗组各50例,对照组为西医常规治疗,治疗组在西医常规治疗基础上联合清咳平喘颗粒治疗。两组均连续治疗2周。观察两组患者治疗前后的中医证候、咳嗽症状、气道反应（血清免疫球蛋白E、外周血嗜酸性粒细胞）、肺功能［最大呼气流量、第1秒用力呼气容积（forced expiratory volume for 1 second,FEV1）与用力肺活量（forced vital capacity,FVC）的比值］、生活质量及不良反应情况。结果 治疗后,对照组和治疗组的总有效率分别是90%、94%,两组比较差异无统计学意义（P>0.05）。治疗后,两组患者中医证候评分、日间及夜间咳嗽积分较治疗前降低,且治疗组明显低于对照组（P<0.05）。治疗后,两组血清免疫球蛋白E、外周血嗜酸性粒细胞较治疗前明显下降,且治疗组气道反应指标改善好于对照组（P<0.05）。治疗后,两组最大呼气流量、FEV1/FVC较治疗前明显上升（P<0.05）,但两组比较差异无统计学意义（P>0.05）。随访时两组患者的生活质量评分较治疗前均显著升高,治疗组评分改善优于对照组,两组比较差异具有统计学意义（P<0.05）。治疗期间两组均未发生不良反应。结论 清咳平喘颗粒在治疗痰热郁肺型咳嗽安全性良好,有助于改善咳嗽症状,改善肺通气,提高患者生活质量。     

落花生根内生真菌Fusarium oxysporum LHS-P1-3中恩镰孢菌素类化合物及生物活性研究

目的 研究落花生根内生真菌Fusarium oxysporum LHS-P1-3的次级代谢产物及其生物活性。方法 使用硅胶色谱柱、高效液相色谱等对代谢粗提物进行分离纯化,采用质谱、核磁共振等方法测定化合物的结构,采用CCK-8法测试化合物的抗肿瘤细胞活性,使用表面等离子共振实验（surface plasmon resonance,SPR）研究化合物与胆固醇转运蛋白1（niemann-pick c1-like 1,NPC1L1）相互作用关系。结果 从落花生根内生真菌Fusarium oxysporum LHS-P1-3分离得到6个化合物,包括1个新化合物（1）与5个已知化合物：白僵菌素（2）、恩镰孢菌素I（3）、恩镰孢菌素MK1688（4）、恩镰孢菌素H（5）及恩镰孢菌素B（6）。化合物1～4对HepG2细胞和HeLa细胞的IC₅₀均小于10 μmol/L,化合物2～4则对HCT116细胞的IC₅₀小于10 μmol/L,SPR研究表明化合物2、4在20 μmol/L浓度下,可与NPC1L1蛋白相互作用。结论 化合物1为新的恩镰孢菌素类化合物,命名为恩镰孢菌素W。化合物1～5具有抗肿瘤活性,化合物2、4与NPC1L1蛋白存在相互作用,可能是NPC1L1蛋白的潜在抑制剂。     

卡氏乳香中1个新的降二萜

目的 研究卡氏乳香Boswellia carterii中的化学成分及其抗骨质疏松活性。方法 运用硅胶柱色谱、ODS柱色谱和制备高效液相色谱等方法进行化学成分分离,通过波谱数据及文献对比对其进行结构解析。结果 卡氏乳香的95%乙醇提取物中分离得到11个化合物,分别鉴定为 (1R,2E,8S,9R)-1,9-环氧-8-乙酰氧基-4,5-二氧代降西松烷型-2-烯（1）、rel(1S,3R,7E,11S,12R)-1,12-epoxy-4-methylenecembr-7-ene-3,11-diol（2）、boscartin F（3）、boscartene K（4）、fidmansumbin-13(17)-en-3,16-diene（5）、3α-acetoxyl-7-oxo-tirucalla-8,24-dien-21-oic acid（6）、boscartene N（7）、3β-hydroxy-tirucallic acid（8）、α-boswellic acid（9）、boscarterol O（10）、boscarterol F（11）。结论 化合物1是新化合物,命名为卡氏乳香素,化合物5为首次从乳香中分离得到,化合物2和10首次从该属植物中分离得到。对11个化合物进行抗骨质疏松活性筛选,发现化合物1、3、6和10具有良好的抑制破骨细胞活性。     

人参皂苷Rg1对心肌细胞氧化应激损伤的抑制作用

目的 通过构建H₂O₂诱导的H9c2细胞氧化应激模型,观察人参皂苷Rg₁对氧化应激的抑制作用,并探讨mi R-499c是否参与人参皂苷Rg₁抑制氧化应激的作用机制。方法 采用600μmol/L的H₂O₂诱导H9c2细胞氧化应激模型,给予人参皂苷Rg₁预处理24 h,检测乳酸脱氢酶(lactate dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)活性及丙二醛(malondialdehyde,MDA)、活性氧(reactive oxygen species,ROS)水平和线粒体膜电位(mitochondrial membrane potential,MMP)变化;采用Western blotting检测凋亡相关蛋白表达。采用Lipofiter 3.0将miR-499c转染至H9c2细胞再制备H₂O₂诱导的氧化应激模型,给予人参皂苷Rg     

Rare multiple congenital anomalies-hypotonia-seizures syndrome type 1 (MCAHS1) – the clinical and molecular summary

Multiple congenital anomalies-hypotonia-seizures syndrome type 1 (MCAHS1) is a rare autosomal recessive genetic disease belonging to glycosylphosphatidylinositols biosynthesis defects (GPIBD), a group of recessive disorders characterized by intellectual disability, hypotonia, and seizures. Glycosylphosphatidylinositols (GPIs) are glycolipids that anchor and remodel cell proteins. These processes are highly conserved and fundamental in the metabolism of all eukaryotes, including humans. Here, we have reported a male patient presenting with hypotonia, intellectual disability, and epilepsy, who underwent whole exome sequencing (WES). The analysis revealed the presence of two deleterious variants in PIGN that encodes GPI ethanolamine phosphate transferase-1 – one novel (c.1247_1251delAAGTG; p.Glu416Glyfs*22), and one that has been previously reported in the medical literature (c.1434+5G>A) resulting in MCAHS1. The detailed clinical assessment followed by the medical literature review also pointed out transient macrosomia and unreported in MCAHS1 advanced bone age and postnatal tall stature. These symptoms suggest that MCAHS1 shares a phenotypic overlap with disorders associated with overgrowth. To conclude, our case report and summary of the medical literature may be helpful for clinicians and geneticists who diagnose patients presenting with hypotonia accompanied by tall stature, advanced bone age, and transient macrosomia.     

Protective effect of pinocembrin from Penthorum chinense Pursh on hepatic ischemia reperfusion injury via regulating HMGB1/TLR4 signal pathway

Hepatic ischemia–reperfusion injury (HIRI) is of common occurrence during liver surgery and transplantation. Pinocembrin (PIN) is a kind of flavonoid monomer extracted from the local traditional Chinese medicine Penthorum chinense Pursh (P. chinense). However, the effect of PIN on HIRI has not determined. We investigated the protective effect and potential mechanism of PIN against HIRI. Model mice were subjected to partial liver ischemia for 60 min, experimental mice were pretreated with PIN orally for 7 days, and H₂O₂-induced oxidative damage model in AML12 hepatic cells was established in vitro. Histopathologic analysis and serum biochemical levels revealed that PIN had hepatoprotective activities against HIRI. The variation of GSH, SOD, MDA, and ROS levels indicated that PIN treatments attenuated oxidative stress in tissue. PIN pretreatment obviously ameliorated apoptosis, and restrained the expression of HMGB1 and TLR4 in vivo. In vitro, compared with H₂O₂ group, the contents of ROS, mitochondrial membrane potential, apoptotic cells, and Bcl-2 protein were decreased, while the Bax protein expression was increased. Moreover, HMGB-1 small interfering RNA test and western blotting showed that PIN pretreatment reduced HMGB1 and TLR4 protein levels. In conclusion, PIN pretreatment effectively protected hepatocytes from HIRI and inhibited the HMGB1/TLR4 signaling pathway.     

A randomized,double-blind placebo-controlled add-on trial to assess the efficacy,safety, and anti-atherogenic effect of spirulina platensis in patients with inadequately controlled type 2 diabetes mellitus

The efficacy of spirulina platensis (S. platensis) as an add-on therapy to metformin and its effect on atherogenic keys in patients with uncontrolled Type 2 Diabetes Mellitus (T2DM) was evaluated. Sixty patients were randomly assigned to S. platensis (2 g/day) or placebo group for three months while continuing metformin as their usual treatment. The efficacy of S. platensis was determined using the pre- and post-intervention HbA1c levels (primary outcome) as well as tracking FBS and lipid profiles levels (TC, LDL-C, TG, and HDL-C) as secondary outcomes at the different treatment time points (0,30,60,90 days). During the three–month intervention period, supplementation with S. platensis resulted in a significant lowering of HbA1c (↓1.43, p < 0.001) and FBS (↓ 24.94 mg/dL, p < 001) levels. Mean TG in the intervention group was found to be significantly lower in the intervention group than in controls (p < 0.001). Total cholesterol (TC) and its fraction, LDL-C, exhibited a fall (↓41.36 mg/dL and ↓38.4 mg/dL, respectively; p < 0.001) coupled with a marginal increase in the level of HDL-C (↑3 mg/dL; p < 0.001). Add-on therapy with S. platensis was superior to metformin regarding long-term glucose regulation and controlling blood glucose levels of subjects with T2DM. Also, as a functional supplement, S. platensis has a beneficial effect on atherogenic keys (TG and HDL-C) with no adverse events.