基于miR-370-3p与JAK2/STAT3通路相关性探讨活血荣络方促缺血性脑卒中后血管新生的机制

目的基于miR-370-3p与JAK2/STAT3通路相关性探讨活血荣络方促缺血性脑卒中后血管新生的机制。方法将大鼠随机分为6组,MCAO/R法造模,灌胃给药7 d后免疫荧光染色观察脑组织CD31、vWF及血管内皮生长因子(vascular endothelial growth factor,VEGF)表达;Western blot法检测脑组织JAK2、p-JAK2、STAT3、p-STAT3蛋白的表达;Real-time PCR(RT-PCR)法检测脑组织JAK2、STAT3 mRNA及miR-370-3p的表达;Pearson相关性分析脑组织miR-370-3p与JAK2/STAT3通路的相关性;培养大鼠脑血管平滑肌细胞,实时荧光定量PCR(RT-qPCR)检测LncRNA-H19和miR-370-3p表达;荧光素酶报告实验检测LncRNA-H19和miR-370-3p的靶向关系。结果活血荣络方能增加缺血区微血管密度及VEGF平均荧光强度,上调JAK2、STAT3 mRNA,下调miR-370-3p表达,促进JAK2、p-JAK2、STAT3、p-STAT3表达,且miR-370-3p分别与JAK2、STAT3 mRNA呈高度负相关,此过程能被STAT3 SH2结构域抑制剂Stattic逆转。结论活血荣络方可能通过下调miR-370-3p的表达、激活JAK2/STAT3通路、促进下游VEGF的表达而刺激缺血性脑卒中后血管新生,从而改善神经功能缺损症状。     

LncRNA Neat1调控miR-128-3p表达对创伤性脑损伤小鼠认知功能和神经炎症的影响

目的 探讨长链非编码RNA(LncRNA)Neat1靶向调节微小RNA(miR)-128-3p表达对创伤性脑损伤(TBI)小鼠认知功能和神经炎症的影响。方法 通过自由落体法建立TBI小鼠模型。将建模后的小鼠随机分为模型组、腺病毒空载体组(Ad-NC)组、Neat1过表达腺病毒(Ad-Neat1)组、miR-128-3p抑制物阴性对照(inhibitor NC)组、miR-128-3p抑制物(miR-128-3p inhibitor)组、Ad-Neat1+miR-128-3p模拟物(Ad-Neat1+miR-128-3p mimic)组、Ad-Neat1+miR-128-3p模拟物阴性对照(Ad-Neat1+mimic NC)组，10只/组，另取10只假手术组小鼠。水迷宫实验检测小鼠认知能力；苏木精-伊红(HE)染色观察小鼠伤灶周围脑组织病理学变化；实时荧光定量聚合酶链式反应(qRT-PCR)检测Neat1、miR-128-3p表达水平；酶联免疫吸附(ELISA)法测定血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平；双荧光素酶实验验证Neat1、miR-128-3p...     

LncRNA CBR3-AS1调节miR-409-3p/hnRNPA2B1轴对肝细胞癌细胞恶性生物学行为的影响

目的 探讨长链RNA CBR3-AS1(LncRNA CBR3-AS1)靶向微小RNA-409-3p(miR-409-3p)/核不均一核糖核蛋白A2B1(hnRNPA2B1)轴对肝细胞癌(HCC)细胞恶性生物学行为的影响。方法 qRT-PCR法分析肝癌组织中LncRNA CBR3-AS1和miR-409-3p表达水平。双荧光素酶检测LncRNA CBR3-AS1和miR-409-3p及hnRNPA2B1和miR-409-3p的靶向关系。将Hep G2细胞分为si-NC组、si-CBR3-AS1组、si-CBR3-AS1+anti-miR-NC组、si-CBR3-AS1+anti-miR-409-3p组、miR-NC组、miR-409-3p mimics组、miR-409-3p mimics+pcDNA组、miR-409-3p mimics+hnRNPA2B1组。平板克隆检测细胞增殖；Annexin V-FITC/PI法检测细胞凋亡；划痕实验检测细胞迁移；Transwell实验检测细胞侵袭；Western blot分析法检测细胞核增殖抗原标记物(Ki67)、细胞周期负调控因子(P21)、...     

miR-141-3p通过调节Keap1-NRF2/ARE信号通路对急性呼吸窘迫综合征大鼠肺纤维化的影响

目的 探讨miR-141-3p对急性呼吸窘迫综合征（ARDS）大鼠肺纤维化的影响及作用机制。方法 将大鼠按随机数字表法分为对照组、模型组、agomir-NC组、miR-141-3p agomir组，每组10只；除对照组外，其余大鼠采用脂多糖（LPS）滴注法构建ARDS模型；将大鼠肺泡Ⅱ型上皮细胞RLE-6TN细胞分为NC组、LPS组、miR-NC组、miR-141-3p mimics组、miR-141-3p mimics+pcDNA组、miR-141-3p mimics+NRF2与Kelch样环相关蛋白1(Keap1)组，除NC组外，其余各组建立LPS细胞模型；qPCR检测肺组织和细胞中miR-141-3p、Keap1 mRNA表达；Western blot检测肺组织和细胞上皮型钙黏附素（E-cadherin）、神经型钙黏附素（N-cadherin）、微管相关蛋白轻链3B(LC3B)、自噬相关基因Beclin-1、α平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原（Col-Ⅰ）、Keap1、核因子E2相关因子2(NRF2)、血红素氧合酶1(HO-1)表达；HE和Masson染色观察肺组织病理变化...     

木犀草素通过调节细胞凋亡和炎症发挥对内毒素性所致急性肾损伤的保护作用

目的 探讨木犀草素对急性肾损伤的保护作用。方法 通过脂多糖（LPS构建小鼠内毒素性所致的急性肾损伤模型，使用木犀草素对肾损伤小鼠进行治疗；给小鼠尾静脉注射miR-30c-5p antagomir以敲低miR-30c-5p的表达。动物分组如下：正常对照组、LPS组、木犀草素治疗组（LPS+Lu组）、木犀草素治疗的阴性对照组（LPS+DMSO组）、miR-30c-5p表达敲低组（antagomir 30c-5p组）、miR-30c-5p表达敲低对照组（antagomir NC组）。采用自动化学分析仪测定小鼠血清肌酐和尿素氮水平，通过HE染色观察肾组织病理损伤。采用ELISA试剂盒检测小鼠血清胱抑素C(Cys-C)、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-4水平，通过TUNEL染色和Western blot检测细胞凋亡。结果 与正常对照组相比，LPS组小鼠急性肾损伤病理情况严重，血清肌酐、尿素氮和Cys-C、促炎因子IL-6、TNF-α、促凋亡因子B淋巴细胞瘤-2基因（Bcl-2）相关X蛋白（Bax）的表达水平显著升高（P <0.05），而抗炎因子IL-4、抗凋亡因子...     

红车轴草总黄酮通过调控miR-99a-3p/CD36表达对高糖诱导的大鼠胰岛β细胞的保护作用北大核心CSCD

目的研究红车轴草总黄酮对高糖诱导的大鼠胰岛β细胞损伤的影响,及其潜在的分子机制。方法本实验分为对照组,模型组,低、中、高剂量实验组,miR-NC组,miR-99a-3p组,anti-miR-NC,anti-miR-99a-3p组,模型+miR-NC组,模型+miR-99a-3p组,模型+si-NC组,模型+si-CD36组,高剂量实验+anti-miR-NC组和高剂量实验+anti-miR-99a-3p组。对照组给予5.5 mmol·L^(-1)葡萄糖处置;模型组给予25 mmol·L^(-1)葡萄糖处置;低、中、高剂量实验组分别给予12.5,25.0和50.0 mg·L^(-1)红车轴草总黄酮和25 mmol·L^(-1)葡萄糖处置;miR-NC组、miR-99a-3p组、anti-miR-NC组、anti-miR-99a-3p组分别转染miR-NC、miR-99a-3p、anti-miR-NC、anti-miR-99a-3p;模型+miR-NC组、模型+miR-99a-3p组、模型+si-NC组、模型+si-CD36组均给予25 mmol·L^(-1)葡萄糖处理,并分别转染miR-NC、miR-99a-3p、si-NC、si-CD36;高剂量实验+anti-miR-NC组、高剂量实验+anti-miR-99a-3p组均给予50.0 mg·L^(-1)红车轴草总黄酮+25 mmol·L^(-1)葡萄糖,并分别转染anti-miR-NC或anti-miR-99a-3p。用流式细胞术测定细胞凋亡率,用定量实时聚合酶链反应检测细胞中miR-99a-3p和CD36 mRNA的表达水平。结果模型组INS-1细胞中CD36 mRNA(2.74±0.27 vs 1.01±0.08)和细胞凋亡率[(27.63±2.71)%vs(5.69±0.58)%]均较对照组显著升高,miR-99a-3p(0.38±0.03 vs 1.00±0.09)较对照组显著降低,差异均有统计学意义(均P<0.05);与模型组比较,低、中、高剂量实验组的上述结果相反。模型+miR-99a-3p组的细胞凋亡率[(12.48±1.19)%vs(26.84±2.41)%]较模型+miR-NC组显著降低,miR-99a-3p表达量(0.79±0.07 vs 0.34±0.03)较模型+miR-NC组显著升高,差异均有统计学意义(均P<0.05)。模型+si-CD36组INS-1细胞中CD36 mRNA(1.35±0.12 vs 2.81±0.26)及细胞凋亡率[(13.54±1.32)%vs(25.87±2.52)%]均较模型+si-NC组显著降低,差异均有统计学意义(均P<0.05)。高剂量实验+anti-miR-99a-3p组INS-1细胞中miR-99a-3p(0.43±0.14 vs 0.88±0.06)较高剂量实验+anti-miR-NC组显著降低,CD36 mRNA(2.51±0.22 vs 1.41±0.13)及细胞凋亡率[(20.45±2.03)%vs(10.56±1.02)%]均较高剂量实验+anti-miR-NC组显著升高,差异均有统计学意义(均P<0.05)。结论红车轴草总黄酮可能通过调控miR-99a-3p/CD36以减轻高糖对大鼠胰岛β细胞INS-1的损伤,进而抑制细胞的凋亡。     

miR-149-3p在先天性心脏病胎鼠心脏组织中的表达及其对P19细胞分化的影响

目的 探讨miR-149-3p在先天性心脏病（CHD）胎鼠心脏组织中的表达及其对小鼠畸胎瘤P19细胞分化的影响。方法 建立CHD室间隔缺损胎鼠模型,于妊娠第19天,HE染色观察心脏组织病理学变化,实时荧光定量PCR(qPCR)检测心脏组织中miR-149-3p和热休克蛋白蛋白家族B成员6(HSPB6)mRNA表达水平。二甲基亚砜（DMSO）诱导P19细胞向心肌细胞分化,并收集分化第0、5、10天的细胞,qPCR检测诱导分化过程中心肌分化标志物GATA结合蛋白4(GATA4)、心肌肌钙蛋白T(cTnT)、心房利钠尿多肽（ANP）的mRNA表达水平以及HSPB6 mRNA和miR-149-3p表达水平。miR-149-3p过表达慢病毒和空载慢病毒感染P19细胞,DMSO诱导分化第10天,免疫荧光染色检测细胞中cTnI蛋白表达情况,qPCR检测心肌分化相关标志物mRNA表达水平。双荧光素酶报告基因实验验证miR-149-3p与HSPB6之间的靶向关系。结果 与正常组比较,CHD组胎鼠出现心脏室间隔缺损,心脏组织中miR-149-3p高表达,而HSPB6低表达（P<0.01）。P19细胞...     

miR-579-3p过表达抑制宫颈癌HeLa细胞增殖、迁移和侵袭的研究

目的 探讨miR-579-3p在宫颈癌HeLa细胞中的表达及对宫颈癌细胞增殖、迁移和侵袭的影响，及其潜在机制。方法 将细胞分为H8组[正常培养正常宫颈上皮永生化细胞(H8细胞)]、HeLa组[正常培养宫颈癌细胞(HeLa细胞)]、HeLa+mimic-NC(HeLa细胞转染mimic-NC)和HeLa+mimic-miR-579组(HeLa细胞转染mimic-miR-579-3p)。用细胞计数试剂盒检测各组细胞的增殖情况，用实时荧光定量聚合酶链反应检测miR-579-3p和上皮细胞-间充质转化(EMT)相关基因mRNA的表达水平，用迁移实验和侵袭小室法检测细胞的迁移和侵袭情况。结果 H8组中miR-579-3p mRNA相对表达水平(1.00±0.04)显著高于HeLa组(0.12±0.01),差异有统计学意义(P<0.001)。HeLa+mimic-NC组和HeLa+mimic-miR-579组的miR-579-3p mRNA相对表达水平分别为0.13±0.01和0.83±0.10,第2天的增殖能力分别为165.91±9.46和111.35±9.66,侵袭细胞数分别为(124....     

Let-7e-5p调控Fas/FasL影响小鼠变应性鼻炎发病的机制研究

目的 探讨let-7e-5p对小鼠变应性鼻炎的影响及机制。方法 将18只BALB/c小鼠按随机数字表法均分为对照（Control）组、变应性鼻炎（AR）组、let-7e-5p激动剂阴性对照（AR+agomir-NC）组、let-7e-5p激动剂（AR+agomir）组、let-7e-5p抑制剂阴性对照（AR+antagomir-NC）组和let-7e-5p抑制剂（AR+antagomir）组。除Control组外，其余组应用卵白蛋白（OVA）致敏建立AR模型，AR+agomir组和AR+antagomir组同时给予let-7e-5p agomir或let-7e-5p antagomir进行干预，末次激发致敏后对小鼠变应性鼻炎进行评分，HE染色观察鼻黏膜组织，统计嗜酸性粒细胞数量，real-time PCR检测let-7e-5p、Fas、FasL m RNA表达，Western blot检测Fas、FasL蛋白表达，酶联免疫吸附试验检测血清免疫球蛋白E(IgE)、特异性IgE(sIgE)、干扰素γ(IFN-γ)、白细胞介素（IL）-12、IL-4、IL-13水平，双荧光素酶实验分析let...     

荆芥挥发油抗内毒素中毒小鼠NLRP3炎症小体通路的机制研究

目的以NLRP3炎症小体通路为切入点,探究荆芥挥发油抗炎效应的分子机制。方法荆芥挥发油低、高剂量(0.226、0.452 g·kg~(-1))连续灌胃给药5 d,末次给药后30 min,小鼠腹腔注射LPS (0.015 g·kg~(-1),10 mL·kg~(-1))构建内毒素中毒小鼠模型,造模后12 h取材,测定相关指标。Griess法测定肺组织NO含量;qPCR法检测肺组织中NLRP3、ASC、caspase-1、IL-1β、iNOS、p65 mRNA的表达;免疫组化法检测肺组织中P2X7R、Cathepsin B的表达;Western blot法测定肺组织中NLRP3、caspase-1(p20)、pro-IL-1β、COP1蛋白表达。结果 0.226 g·kg~(-1)荆芥挥发油明显降低模型小鼠肺组织Cathepsin B、NLRP3蛋白表达;0.452 g·kg~(-1)荆芥挥发油明显降低模型小鼠肺组织中NO水平,明显下调小鼠肺组织中NLRP3、iNOS、p65、IL-1βmRNA的表达及P2X7R蛋白表达;荆芥挥发油(0.226、0.452 g·kg~(-1))均能明显下调caspase-1(p20)蛋白水平,升高COP1蛋白水平。结论荆芥挥发油对内毒素中毒小鼠的保护作用与其抗炎效应密切相关,抗炎机制与抑制NLRP3炎症小体的激活有关。     

^3H—天麻素和^3H—天麻甙元的制备

本文介绍了一种催化氚法制备^3H-天麻素和^3H-天麻甙元的方法，所得产物放射性比活度为510。6GBq(13.8Ci)/mmol。放化纯度大于95%。     

3-二甲胺基-3-苯基丙醇的制备

苯甲醛和丙二酸经Knoevenagel缩合、NaBH4-I2体系还原得3-氨基-3-苯基丙醇，再经Eschweiler-Clark反应制得3-二甲胺基-3-苯基丙醇，总收率29％。     

Cyanidin-3-rutinoside increases glucose uptake by activating the PI3K/Akt pathway in 3T3-L1 adipocytes

In this study, the effect of cyanidin-3-rutinoside (C3R) on glucose uptake by 3T3-L1 adipocytes was studied. C3R significantly increased glucose uptake, which was associated with enhanced plasma membrane glucose transporter type 4 (PM-GLUT4) expression in 3T3-L1 adipocytes. The potentiating effect of C3R on glucose uptake and PM-GLUT4 expression was related to enhanced phosphorylation of insulin receptor substrate 1 (IRS-1) and Akt, as well as augmented activation of phosphatidylinositol-3-kinase (PI3K) in the insulin signaling pathway. C3R induced glucose uptake was inhibited only by the PI3K inhibitor, but not by an AMPK inhibitor in 3T3-L1 adipocytes. Therefore, C3R likely up-regulates glucose uptake and PM-GLUT4 expression in 3T3-L1 adipocytes by activating the PI3K/Akt pathways.     

蚯蚓蛋白对NIH3T3细胞的促进增殖作用研究

目的探讨蚯蚓蛋白(EFP)对NIH3T3细胞的促进增殖作用。方法 MTT法测定EFP对大鼠成纤维细胞(NIH3T3)的促进增殖作用;细胞划痕法测定EFP促进NIH3T3细胞划痕创面愈合作用;测定NIH3T3细胞培养液中胶原降解产物羟脯氨酸的含量。结果 EFP具有促进NIH3T3细胞的增殖作用,促进划痕创面愈合作用,增加了NIH3T3细胞培养液中羟脯氨酸的含量。结论 EFP具有促进NIH3T3细胞增殖和划痕创面愈合作用。     

Synthesis and Activity of the Glutathione Analogue γ-(L-γ-Oxaglutamyl)-L-cysteinyl-glycine

An efficient synthesis of the backbone modified glutathione analogue γ-(L-γ-oxaglutamyl)-L-cysteinyl-glycine ( 7 ), characterized by the presence of an urethane O-CO-NH linkage replacing the γ-glutamylic CH₂CO-NH fragment is described. The new analogue has been fully characterized by ¹H- and ¹³C-NMR, and FAB-MS. Compound 7 was tested for inhibition of γ-glutamyl-transferase activity and was found to be a non-competitive inhibitor of hog kidney γ-glutamyltransferase (EC 2.3.2.2).     

3-（3-羟基-1-甲基丙氧基）-1-丁醇和3-（3-羟基丁氧基）-1-丁醇的合成及其工艺改进

目的合成3-（3-羟基-1-甲基丙氧基）-1．丁醇（Ⅰ）和3-（3-羟基丁氧基）-1-丁醇（Ⅰ），并改进工艺提高产品纯度。方法以1，3-丁二醇为起始原料，通过硫酸催化双分子缩合、三苯甲基选择性保护、分离提纯、脱保护基4步反应得到目标化合物。结果与结论化合物Ⅰ的结构经。H．NMR13C-NMR谱确证，其含量经C-C测定，大于99．5％。化合物Ⅰ的纯度为90．0％，与文献相比纯度都有相应提高。     

Synthesis and activity of the glutathione analogue γ-(L-γ-azaglutamyl)-L-cysteinyl-glycine

The backbone-modified glutathione analogue γ-(L-γ-azaglutamyl)-L-cysteinyl-glycine 7, characterized by the presence of a NHCONH urea linkage deriving from the replacement of the native Glu γ-CH₂ with the aza (NH) group, was synthesized and fully characterized by FAB-MS, ¹H- and ¹³C-NMR. Potential of 7 and its oxidized form 6 as γ-glutamyltransferase inhibitors was investigated. Both compounds 7 and 6 were found to be competitive inhibitors of hog kidney y-glutamyltransferase (EC 2.3.2.2.) by binding at the donor site: the reduced analogue is a more efficient inhibitor than glutathione of the γ-glutamyl transfer reaction. Inhibition at the acceptor site, which is also present, appears to be more complex. In particular, un-competitive inhibition is observed for compound 7. The results indicate that γ-azapeptides of type 7 may represent interesting targets in the search for stable inhibitors of γ-glutamyltransferases. © Munksgaard 1995.     

Arctigenin promotes bone formation involving PI3K/Akt/PPARγ signaling pathway

This study investigated the mechanisms through which arctigenin promotes osteogenesis. Bone marrow mesenchymal stem cells (BMSCs) from ovariectomized (OVX) rats were differentiated into osteoblasts, and osteogenesis was evaluated via Alizarin Red S (ARS) staining and alkaline phosphatase (ALP) measurements in cultured BMSCs. The levels of phosphorylated AKT serine/threonine kinase 1 (p‐Akt), and peroxisome proliferator‐activated receptor gamma (PPARγ) expression were quantified by Western blot analysis. The levels of urine calcium (U‐Ca), urine phosphorus (U‐P), serum ALP, and bone mineral density (BMD) of OVX rats were assessed in vivo. The results showed that treatment with arctigenin in rat BMSCs enhanced mineralization, increased ALP activity, increased the expression of Akt and p‐Akt, and decreased PPARγ expression, consistent with its ability to promote osteoblast differentiation. Furthermore, arctigenin prevented OVX‐induced osteoporosis in rats by increasing BMD and ALP activity and inhibiting the loss of Ca and P. In contrast, treatment with LY294002, a selective inhibitor of the phosphatidylinositol 3‐kinase (PI3K), produced the opposite phenotype. These data suggest that the protective effects of arctigenin on BMSCs and OVX rat models result from the induction of osteogenesis involving the PI3K/Akt/PPARγ axis.     

PI3K-Akt-mTOR信号通路对Foxp3基因表达的影响

目的研究PI3K-Akt-mTOR信号通路对Foxp3基因表达的影响。方法将SPF级昆明系小鼠60只随机分为对照组(A)和实验组(B、C、D),B、C、D三组分别灌胃雷帕霉素0.2、0.4、0.6 mg.d-1,A组每天予以无菌水灌胃,共3周。3周后,无菌条件下心脏采血,EDTA抗凝,分离脾脏,制备单细胞悬液,采用流式细胞仪检测小鼠外周血和脾脏中CD4+CD25+调节性T细胞水平(CD4+CD25+Treg细胞占CD4+T细胞的百分比),RT-PCR检测小鼠脾细胞Foxp3mRNA的表达,免疫组织化学SP法染色观察小鼠脾脏中p-mTOR蛋白表达的变化,并利用计算机图像分析技术测量各实验组及对照组中p-mTOR蛋白表达的平均光密度。结果 Foxp3 mRNA在实验组(B、C、D)小鼠脾脏中表达的程度分别为(0.4853±0.0574、0.7886±0.1085、0.9639±0.2015),明显高于对照组A(0.1345±0.0271)(P<0.05);实验组(B、C、D)小鼠脾脏中p-mTOR蛋白表达的平均光密度分别为(0.2326±0.0431、0.1156±0.0223、0.0556±0.0041),明显低于对照组A(0.4223±0.0534)(P<0.05);两指标经pearson相关分析,小鼠脾脏中Foxp3mRNA的表达与p-mTOR蛋白表达呈负相关。结论抑制PI3K-Akt-mTOR信号通路会促使Foxp3的表达增强;PI3K-Akt-mTOR信号通路的激活程度与Foxp3的表达程度呈负相关。     

SYNTHESIS OF γ-L-GLUTAMYL-TAURINE*

Glu(Tau), a bioactive substance previously isolated from the protein free aqueous extract of bovine parathyroid powder, has been synthesized. The intermediate derivative Z-Glu(Tau)-OBzl was prepared in three different ways from Z-Glu-OBzl and (1) cystamine by using the mixed anhydride method followed by oxidation, (2) Tau by the active ester procedure via Z-Glu(ONp)-OBzl, (3) Tau applying mixed anhydride coupling. The protecting groups were removed by hydrogenolysis.