Nanoparticle-facilitated delivery of BAFF-R siRNA for B cell intervention and rheumatoid arthritis therapy

The present study was designed to explore the effects of B-cell activating factor receptor (BAFF-R) siRNA encapsulated nanoparticles on collagen-induced arthritis (CIA). BAFF-R siRNA encapsulated nanoparticles (NP-_siBAFF-R) were constructed using a double emulsion method and was characterized by dynamic light scattering and transmission electron microscopy. Cellular uptake of nanoparticles was determined using flow cytometry. The CIA mouse model was established and the mice were intravenously injected with nanoparticles. NP-_siBAFF-R effectively decreased the expression of BAFF-R in B cells and facilitated the delivery of siRNA into B cells. Treatment of NP_siBAFF-R ameliorated rheumatoid arthritis (RA) symptoms in the CIA mouse model via decreasing the arthritis score, mean ankle diameter, the levels of anti-collagen IgG in serum and increasing the expression of collagen type II and osteocalcin in dissected joint tissues. Additionally, treatment of NP_siBAFF-R decreased the percentage and number of B cells and inhibited the production of pro-inflammatory cytokines in RA mice. These results demonstrate that NP-_siBAFF-R may provide an effective strategy for RA treatment.     

藿朴夏苓汤治疗早期新型冠状病毒肺炎（COVID-19）湿邪郁肺证物质基础及功效的网络分析

目的筛选藿朴夏苓汤治疗早期新型冠状病毒肺炎(COVID-19)湿邪郁肺证的物质基础,预测其作用机制。方法查阅文献及临床报道分析藿朴夏苓汤与早期COVID-19湿邪郁肺证的方证关系。运用TCMSP数据库筛选藿朴夏苓汤中潜在活性成分,将活性成分与新型冠状病毒(SARS-CoV-2)3CL水解酶、血管紧张素转化酶Ⅱ(ACE2)进行分子对接,根据结合能筛选与两者均有较好结合作用的核心成分。借助Cytoscape软件构建关键成分靶点蛋白互作网络,筛选出核心靶点;通过STRING数据库进行核心靶点的GO分析,Cytoscape软件Clue GO插件进行Pathway、KEGG富集分析。结果方证关系分析藿朴夏苓汤用于治疗早期COVID-19湿邪郁肺证,筛选出藿朴夏苓汤中潜在作用成分12个,核心靶点67个。其中通草中的通脱木皂苷元Ⅰ,茯苓、猪苓中的过氧麦角甾醇,半夏中的黄芩苷与SARS-CoV-2 3CL水解酶、ACE2均具有较好的结合活性。GO、Pathway、KEGG富集分析结果显示藿朴夏苓汤中12个潜在作用成分参与调节刺激反应、信号转导、细胞死亡等生物过程以及白介素信号通路、癌症EGFR信号通路、酪氨酸激酶信号转导途径、编程性细胞死亡途径、MAPK信号通路等。结论藿朴夏苓汤以化湿解毒、宣肺透邪治疗早期COVID-19湿邪郁肺证患者,菲酮、黄芩苷、酸枣仁皂苷、啤酒甾醇、常春藤皂苷元、过氧麦角甾醇、柠檬二烯醇、麦角甾-7,22-二烯-3-酮、通脱木皂苷元Ⅰ、泽泻醇B-23-醋酸酯、泽泻醇B、新橙皮苷可能为其主要的物质基础,通过阻断SARS-CoV-2病毒蛋白合成,阻止病毒进入宿主细胞,通过调控白介素信号通路、MAPK信号通路、PI3K-Akt信号通路、T细胞受体信号通路、C型凝集素受体信号通路,抑制相关炎症因子的表达发挥作用。     

Pharmacological evaluation and mechanistic study of compound Xishu Granule in hepatocellular carcinoma

ObjectiveIn this study, we used HepG2 human hepatocellular carcinoma cells to study the effects of Compound Xishu Granule (CXG) on cell proliferation, apoptosis, and the cell cycle in vitro. We also used a xenograft tumor model to study the anti-tumor effects of CXG and related mechanisms in vivo.MethodsThe effect of CXG on cell viability was measured using Cell Counting Kit-8 and a colony formation assay. The effect of CXG on apoptosis and the cell cycle was analyzed using flow cytometry. The in vivo anti-tumor effect of CXG was assessed by measuring the volume change in xenograft tumors after drug administration. The CXG anti-tumor mechanism was studied using western blotting assay to detect cell cycle and apoptotic associated proteins.ResultsCXG suppressed HepG2 cell proliferation in a time- and dose-dependent manner in vitro. Colony formation experiments showed that CXG administration for 24 h significantly reduced HepG2 cell formations (P < .01). Flow cytometric analysis showed that CXG treatment for 48 h promoted apoptosis and blocked HepG2 cells in the G2/M phase. Western blotting results showed that Bax was significantly up-regulated and Bcl-2 was down-regulated in graft tumor tissues and HepG2 cells after CXG administration, which increased the Bax/Bcl-2 ratio. PLK1, CDC25C, CDK1, and Cyclin B1 expression were up-regulated. CXG had a good inhibitory effect on graft tumor growth in vivo.ConclusionCXG has good anti-tumor effects in vitro and in vivo. In vitro, CXG promoted HepG2 cell apoptosis and induced G2/M phase arrest. In vivo, CXG significantly inhibited graft tumor growth. The CXG mechanism in treating hepatocellular carcinoma may be that CXG can induce abnormal apoptotic and cell cycle associated protein expression, leading to mitotic catastrophe and apoptosis.     

黄芪甲苷调节PI3K/Akt/FoxO1通路抑制糖尿病大鼠肝糖异生

目的:探讨黄芪甲苷对2型糖尿病(T2DM)大鼠肝损伤保护潜在机制及肝组织磷酯酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/叉头框转录因子1(FoxO1),磷酸烯醇式丙酮酸羧基酶(PEPCK)和葡萄糖6-磷酸酶(G6Pase)蛋白表达的影响。方法:6周高糖高脂饮食后,链脲佐菌素(STZ)一次性腹腔注射(0.035 g·kg^-1)建立T2DM大鼠模型,随机分为正常组、模型组、黄芪甲苷低、中、高剂量组及二甲双胍组。黄芪甲苷低、中、高剂量组灌胃黄芪甲苷0.02,0.04,0.08 g·kg^-1·d^-1,二甲双胍组灌胃二甲双胍0.2 g·kg^-1·d^-1;给药8周后,于末次灌胃24 h禁食不禁水后处死,测血清肝生化指标,肝脏指数等;采用苏木素-伊红(HE)染色观察肝脏组织病理形态学变化;马松(Masson)染色观察纤维化程度;过碘酸希夫(PAS)染色反应观察细胞内糖原等变化;免疫组化及蛋白免疫印迹法(Western blot)检测各组肝中PI3K/Akt/FoxO1信号蛋白及PEPCK,G6Pase蛋白表达水平。结果:与正常组比较,模型组肝脏指数显著升高(P<0.01),肝功能指标丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST),总胆固醇(TC),甘油三酯(TG)的含量显著升高(P<0.01),高密度脂蛋白胆固醇(HDL-C)显著降低(P<0.01),大鼠体质量显著减轻(P<0.01),PI3K/Akt/Fox O1信号减弱(P<0.01),PEPCK,G6Pase蛋白表达水平显著增强(P<0.01);与模型组比较,黄芪甲苷中、高剂量组及二甲双胍组肝功能指标ALT,AST,TC,TG的含量明显降低(P<0.05,P<0.01),大鼠体质量明显增加(P<0.05,P<0.01),肝组织Fox O1,PEPCK及G6Pase蛋白水平明显降低(P<0.05,P<0.01),Fox O1的磷酸化水平明显增强(P<0.05,P<0.01)。结论:黄芪甲苷可能通过调节PI3K/Akt/Fox O1信号通路抑制高脂高糖加小剂量STZ诱导T2DM肝糖异生,从而起到延缓T2DM大鼠肝脏损伤作用。     

CSTB、A-FABP在膀胱癌诊断与预后的价值

目的 分析CSTB、A-FABP在膀胱癌诊断和预后的意义。方法 选取2014年1月至2016年7月在本院进行手术、病理证实的膀胱癌患者45例、良性膀胱病变患者88例，检测膀胱癌样本癌组织、癌旁组织、良性膀胱病变组织中CSTB、A-FABP表达水平。评价CSTB、A-FABP在鉴别诊断膀胱癌的效用。对比膀胱癌进展对象（2年内转移、复发、肿瘤死亡）以及未进展对象组织中CSTB、A-FABP表达水平。结果 癌组织、癌旁组织、良性病变对象组织中CSTB、A-FABP表达水平差异有统计学意义（P<0．05）。CSTB、A-FABP作为诊断标准（任意一项为阳性），鉴别诊断膀胱癌与良性病变的灵敏度为84．4%、特异度为90．9%、阳性预测值82．6%、阴性预测值92．0%、符合率88．7%。2年内转移、复发、肿瘤死亡的患者共30例，无转移、复发、死亡的患者共15例，膀胱癌进展对象CSTB、A FABP表达量高于未进展对象，差异有统计学意义（P<0．05）。结论 CSTB、A-FABP在膀胱癌诊断、预后预测中均有一定的临床价值。     

养阴清热化瘀方对急性放射性肺炎大鼠TLR-4/NF-κB信号通路的影响

目的:探讨养阴清热化瘀方对大鼠急性放射性肺炎的防护作用及可能机制。方法:采用随机数字表法将48只雌性Wistar大鼠随机分为空白对照组(NC)、模型组(MD)、养阴清热化瘀方提前干预组(YHF-EI)及养阴清热化瘀方组(YHF-ST),分别于照射后4周时采集血清及取出右肺。观察大鼠的一般情况,应用HE染色观察肺组织病理改变,ELISA法检测血清中含量, RT-PCR和Western-blotting法分别检测肺组织内TLR4、NF-κB p65 mRNA和蛋白的水平。结果:养阴清热化瘀方能显著减轻大鼠放射性肺炎的炎症表现; 与空白对照组比较,血清中TNF-α、IL-6、TGF-β的含量升高,TLR4和NF-κB p65 mRNA转录活性增强,肺组织中TLR4和NF-κB p65的蛋白表达水平升高; 与模型组比较,两组养阴清热化瘀方组血清中TNF-α、IL-6、TGF-β的含量降低,TLR4和NF-κB p65 mRNA转录活性下降,肺组织中TLR4和NF-κB p65的蛋白表达水平降低。结论:养阴清热化瘀方可能通过抑制“TLR4/NF-κB”信号通路,从而抑制TNF-α、IL-6、TGF-β的产生,起到抵抗放射性肺炎的发生,从而发挥放射防护作用。     

MPMBP down-regulates Toll-like receptor (TLR) 2 ligand-induced proinflammatory cytokine production by inhibiting NF-κB but not AP-1 activation

MPMBP is a novel non-nitrogen-containing bisphosphonate (non-NBP) which possesses anti-bone resorptive activity and an antioxidant side chain. This study aimed to assess the effects of MPMBP on the production of proinflammatory cytokines and chemokines by the macrophage-like cell line, J774.1, in the presence of Toll-like receptor (TLR) agonists. J774.1 cells were pretreated with or without MPMBP for 5 min, and then incubated with or without Pam₃Cys-Ser-(Lys)₄ (Pam₃CSK₄, a TLR2 agonist) or lipid A (a TLR4 agonist) for 24 h. MPMBP down-regulated TLR2 ligand-induced production of IL-6, MCP-1, MIP-1α, and TNF-α, but not TLR4 ligand-induced proinflammatory cytokine production, and was not cytotoxic in J774.1 cells. Cu-CPT22, a TLR2 antagonist, down-regulated Pam₃CSK₄-induced production of IL-6, MCP-1, and MIP-1α, but not TNF-α. MPMBP inhibited the translocation of NF-κB p65, but not p50, RelB, or p52, and inhibited the activation of JNK, but not p38 MAPK or ERK, in J774.1 cells stimulated with Pam₃CSK_4. Moreover, MPMBP did not down-regulate AP-1 activation in J774.1 cells stimulated with Pam₃CSK₄ or lipid A. Our findings suggest that MPMBP inhibits proinflammatory cytokine production in J774.1 cells by suppressing NF-κB p65 activation in the TLR2, but not TLR4, pathway.