哮喘嗜酸性粒细胞凋亡与抗凋亡

哮喘的气道炎症以嗜酸性粒细胞（ＥＯＳ）持续聚集为特征，当ＥＯＳ被激活后，可诱发哮喘时组织的损害。ＥＯＳ性炎症的消退与临床哮喘的改善有关，但气道ＥＯＳ清除的机制仍不清楚。细胞凋亡———一种程序化细胞死亡（ＰＣＤ）的形式，在促进炎细胞清除和炎症的消退方面...     

Overexpression of TRB3 gene in adipose tissue of rats with high fructose-induced metabolic syndrome

Insulin resistance is the physiopathologic foundation of metabolic syndrome. TRB3 has been revealed to be involved in insulin resistance in the liver by interacting directly with Akt and blocking its activation. Our investigation aims at exploring the relationship between metabolic syndrome and TRB3 mRNA expression in adipose tissue of rats. Two groups were studied as follows: the control group (Control, n = 12) was fed a standard rodent chow, and the experimental group (Fructose n = 9) was fed a high-fructose diet. Body weight and systolic blood pressure were measured per 4 weeks. At the end of 38 weeks, levels of tribbles mRNAs in adipose tissue were determined by quantitative real-time polymerase chain reaction (PCR), and Akt/phospho-Akt expression was assessed by Western blot. Results show that levels of TRB1-3 mRNAs were expressed in adipose tissue of rats of both groups, and tribbles mRNAs were TRB1 (Control: 0.00515, Fructose: 0.00497), TRB2 (Control: 0.02104, Fructose: 0.01988), and TRB3 (Control: 0.00457, Fructose: 0.00822), respectively. Of the three, TRB3 mRNA alone significantly increased by 94% in adipose tissue of fructose-fed rats compared with those in adipose tissue of the controls (P<0.05), and there was significant positive correlation between TRB3 mRNA levels and HOMA-R in fructose group (r = 0.68, P<0.05). Western blot analysis showed that phospho-Akt (Ser-473) expression was significantly decreased in adipose tissue of fructose-fed rats compared with controls (P<0.001). The present study suggests that TRB3 may be involved in metabolic syndrome by inhibiting activation of Akt in adipose tissue.     

3.0T MR IDEAL序列成像应用于腰骶神经根受压的初步临床研究

目的探讨IDEAL技术在腰椎间盘突出症中显示腰骶神经受压的临床应用价值。方法将明显腰腿痛并腰椎间盘突出的患者40例作为观察组,正常志愿者10例作为对照组,行腰骶神经根IDEAL序列冠状位薄层扫描和常规扫描,所有图像均经最大信号强度投影(MIP)和曲面重建(CPR)后处理,观察腰骶神经根的受压特点及其与突出椎间盘与邻近组织的关系。结果观察组40例中有92组腰椎间盘不同程度的疝出,而由此引起的神经根受压共81组,其中神经根侧隐窝段受压的概率显著大于椎管内段,但是任何部位的神经根受压好发生于同侧还是双侧受压并没有统计学意义(P=0.338)。另有4组神经根受压由增厚的黄韧带或骨质增生引起;有6组由神经根鞘囊肿引起。受压神经根的影像学表现为:(1)神经根局部缺损样压迹;(2)走行的改变;(3)形态的改变,包括局部肿胀,甚至截断样改变;(4)与邻近组织局部粘连,边界不清,边缘模糊;(5)局部脑脊髓间隙变窄或消失。结论 IDEAL序列能直观显示腰骶神经根与邻近结构的关系及受压特点,能够对神经根受压作出准确的定位和受压程度评估,对判断腰骶神经受压、损伤等具有一定的临床诊断价值,也能为临床早期选择正确的治疗方案及疗效观察提供可靠的参考依据。     

两种脓毒症模型小鼠血清白细胞介素-6与其早期生存率的关系

目的探讨盲肠结扎穿孔( CLP)、腹腔持续置管引流( CASP)脓毒症模型小鼠血清白细胞介素-6(IL-6)与其早期生存率的关系。方法将60只雄性BALB/c小鼠编号,采用数字表法随机分为3组,即假手术组( Sham组)、CLP组、CASP组,各20只。 CLP组小鼠麻醉后,于中下腹正中线做纵行切口,充分暴露并结扎盲肠,用18 G注射器针头在结扎部位远端穿孔并来回贯穿2次,将少量肠内容物挤出,回纳盲肠,缝合腹壁切口。 CASP组术前准备、开腹方式同CLP模型,暴露小鼠升结肠系膜,在其对侧处插入静脉导管,退出针芯,荷包缝合并固定、剪断静脉导管,将游离端放置于腹腔内。用棉签挤压肠管,使少量肠内容物从导管口溢出,以确认导管通畅。 Sham组术前准备、开腹、术后处理同CLP组,不行盲肠结扎、穿孔。观察各组小鼠术后12、24、36、48、60、72 h的生存率。术后6 h采集各组小鼠尾静脉血,采用 ELISA 检测血清 IL-6,比较此时间点3组存活小鼠间 IL-6水平以及CLP组、CASP组的存活和死亡小鼠间IL-6水平。结果 Sham组、CLP组和CASP组术后72 h生存率分别是100.0%(20/20)、45.0%(9/20)和10.0%(2/20),3组间总体比较差异有统计学意义(字2=32.970,P<0.01),两两比较3组间差异均有统计学意义(P值均<0.05);3组小鼠生存时间曲线经Log-rank检验,差异有统计学意义(字2=34.030,P<0.05)。术后6 h,Sham组、CLP组、CASP组存活小鼠血清 IL-6水平分别是(36.62依10.30) ng/L、(2443.47依970.50) ng/L、(4057.93依827.41)ng/L,差异均有统计学意义(Hc=29.270,P<0.01);CLP组和CASP组各组内存活小鼠血清IL-6水平平均分别为(1348.80依276.25) ng/L、2100.00 ng/L,明显低于死亡小鼠的(3157.29依330.94)ng/L、(4275.48依512.71)ng/L,CLP组差异有统计学意义(t=13.071,P<0.01)。结论两种脓毒症模型小鼠的转归都有一定的病死率,较好地模拟了脓毒症的病理生理过程,CLP模型侧重于模拟局限性腹腔脓肿,CASP模型则侧重于模拟弥漫性腹膜炎。 CLP组模型术后6 h血清IL-6水平较低的小鼠,术后72 h生存率较高。     

瘦素促进肺成纤维细胞向肌纤维母细胞的转分化

目的探讨瘦素对肺成纤维细胞向肌纤维母细胞转分化的影响及其作用机制。方法体外培养HFL-1人胚肺成纤维细胞,用(0~200)ng/m L重组人瘦素(r HL)干预或联合转化生长因子β1(TGF-β1)处理HFL-1细胞,Western blot法测定α平滑肌肌动蛋白(α-SMA)、蛋白激酶B(AKT)、磷酸化的AKT(p-AKT)的表达。CCK-8法测定细胞增殖,ELISA测定细胞上清液1型胶原蛋白含量。结果 (50、100、200)ng/m L的r HL处理HFL-1细胞48 h,α-SMA的蛋白表达水平均较对照组明显上调;与200 ng/m L r HL或5 ng/m L TGF-β1单独处理的细胞相比,200 ng/m L r HL和5 ng/m L TGF-β1联合处理HFL-1细胞48 h,HFL-1细胞α-SMA蛋白表达水平显著上调。r HL能够上调HFL-1细胞p-AKT的表达,LY294002可抑制r HL对HFL-1细胞α-SMA表达的诱导作用。与对照组相比,(12.5、25、50、100、200)ng/m L作用72 h,细胞存活率无显著性差异。(50~200)ng/m L r HL作用48 h,细胞上清液中1型胶原蛋白含量明显升高。结论瘦素能够促进肺成纤维细胞向肌纤维母细胞转分化,其作用机制与激活PI3K/AKT信号通路有关。     

Herpes Simplex Virus 1 Suppresses the Function of Lung Dendritic Cells via Caveolin-1

Caveolin-1 (Cav-1), the principal structural protein of caveolae, has been implicated as a regulator of virus-host interactions. Several viruses exploit caveolae to facilitate viral infections. However, the roles of Cav-1 in herpes simplex virus 1 (HSV-1) infection have not fully been elucidated. Here, we report that Cav-1 downregulates the expression of inducible nitric oxide synthase (iNOS) and the production of nitric oxide (NO) in dendritic cells (DCs) during HSV-1 infection. As a result, Cav-1 deficiency led to an accelerated elimination of virus and less lung pathological change following HSV-1 infection. This protection was dependent on iNOS and NO production in DCs. Adoptive transfer of DCs with Cav-1 knockdown was sufficient to confer the protection to wild-type (WT) mice. In addition, Cav-1 knockout (KO) (Cav-1^−/−) mice treated with an iNOS inhibitor exhibited significantly reduced survival compared to that of the nontreated controls. We found that Cav-1 colocalized with iNOS and HSV-1 in caveolae in HSV-1-infected DCs, suggesting their interaction. Taken together, our results identified Cav-1 as a novel regulator utilized by HSV-1 to evade the host antiviral response mediated by NO production. Therefore, Cav-1 might be a valuable target for therapeutic approaches against herpesvirus infections.     

基于Ras/Raf-1/ERK信号转导通路调控VEGF的表达探讨活血解毒方对糖尿病大鼠视网膜的影响

目的：观察活血解毒方对糖尿病大鼠视网膜血管形态的影响并探讨其可能的作用机制。方法：一次性腹腔注射链脲佐菌素(Streptozotocin,STZ,65 mg/kg)诱导糖尿病大鼠模型,造模后随机分为模型组和活血解毒方组。另设正常对照组。药物干预12周后,应用视网膜消化铺片法观察视网膜血管形态;应用Western blot法检测视网膜VEGF蛋白的表达,应用Real-Time PCR法检测视网膜VEGF、Ras、Raf-1与ERK mRNA的表达。结果：模型组视网膜毛细血管面积密度及内皮细胞/周细胞比例与较正常组升高(P<0.001),VEGF蛋白及VEGF、Ras、ERKmRNA表达升高(P<0.05),Raf-1mRNA表达升高(P>0.05)。与模型组比较,活血解毒方组视网膜毛细血管面积密度和内皮细胞/周细胞比例降低(P<0.01和P<0.001),VEGF蛋白与VEGF、Ras、Raf-1、ERK mRNA表达下降(P<0.05)。结论：活血解毒方能明显抑制视网膜毛细血管和内皮细胞的增生,其机制可能是通过干预Ras/Raf-1/ERK信号转导通路,下调VEGF在视网膜中的表达,抑制血管新生,从而改善DR。     

HPLC法同时测定苁蓉总苷胶囊中松果菊苷、毛蕊花糖苷、异毛蕊花糖苷的含量

目的：建立同时测定苁蓉总苷胶囊中松果菊苷、毛蕊花糖苷、异毛蕊花糖苷三种成分含量的HPLC法。方法：Waters C18色谱柱（4.6 mm×150 mm,5 μm）,流动相：甲醇-0.1%甲酸溶液梯度洗脱;流速：1.0 mL·min-1;检测波长：330 nm;柱温30 ℃。结果：松果菊苷、毛蕊花糖苷、异毛蕊花糖苷分别在27.792-277.92 μg·mL-1 （r=0.999 6,n=6）,2.418 4-24.184 μg·mL-1 （r=0.999 6,n=6）,5.106-51.06 μg·mL-1 （r=0.999 8,n=6）范围内呈良好的线性关系,加样回收率均符合含量测定要求。结论：该方法简便、准确,可以用于苁蓉总苷胶囊中松果菊苷、毛蕊花糖苷、异毛蕊花糖苷三个成分的含量测定。     

HPLC同时测定栀子药材中两类活性成分

目的：本文建立了同时测定栀子药材中有机酸类（新绿原酸、绿原酸、隐绿原酸等）、环烯醚萜苷类（京尼平苷酸、去乙酰车叶草酸甲酯、京尼平龙胆双糖苷、栀子苷等）2类有效成分的高效液相色谱含量测定方法,用于栀子药材的质量控制。方法：采用Kromasil C₁₈（250 × 4.6 mm,5 μm）;流动相：乙腈（A）-0.1%磷酸溶液（B）,梯度洗脱,0-10 min,5% → 10%（A）;10-20 min,10% → 18%（A）;20-30 min,18% →20%（A）;流速1 mL·min^-1,进样量10 μL,柱温35℃,检测波长为324 nm 和238 nm。结果：20 min 内绿原酸、绿原酸、隐绿原酸、京尼平苷酸、去乙酰车叶草酸甲酯、京尼平龙胆双糖苷、栀子苷等被测组分均出峰且均达到基线分离;在一定浓度范围内具有良好的线性关系,新绿原酸：Y=24.62X-0.462（r=0.999 9）;绿原酸：Y=27.76X-3.045（r=0.999 9）;隐绿原酸：Y=24.07X-0.569（r=0.999 8）;京尼平苷酸：Y=12.95X-1.088（r=1.000 0）;去乙酰车叶草酸甲酯：Y=14.57X-0.693（r=0.999 8）;京尼平龙胆双糖苷：Y=7.261X-0.117（r=0.999 9）;栀子苷：Y=13.60X-13.59（r=0.999 9）,平均回收率均在95%-105%。结论：该检测方法快速、准确、灵敏,可为栀子药材多成分质量控制提供了一定的参考。