Effects of Bisphenol A on expression of genes related to amino acid transporters,insulin- like growth factor,aquaporin and amino acid release by porcine trophectoderm cells

The peri-implantation period of pregnancy is critical for conceptus development, implantation, and signaling for establishment of pregnancy. This study evaluated the effects of bisphenol A (BPA) on proliferation, adhesion, and migration of porcine trophectoderm (pTr2) cells, expression of transporters of arginine and synthesis of amino acids. All concentrations of BPA decreased proliferation and adhesion of pTr2 cells after 96 h compared to the control group. Lower concentrations of BPA (1 × 10⁻⁹, 1 × 10^-8, 10^-7M) increased (P < 0.05), but higher concentrations of BPA (1 × 10^-5, 1 × 10^-4 M) decreased migration of pTr2 cells. BPA increased expression of SLC7A1 mRNA at lower concentrations (1 × 10⁻⁹ to 1 × 10^-6M) and SL7A6, another cationic acid transporter, at higher concentrations (1 × 10^-5, 1 × 10^-4 M). BPA also down-regulated the expression of IGF1 and IGF1 receptor at concentrations of 1 × 10^-7 to 1 × 10^-4 M compared to the control group. The expression of mRNAs for aquaporins (AQP) 3 and 4 were reduced at all concentrations of BPA, but at lower concentrations of BPA, (1 × 10⁻⁹ to 1 × 10^-8M) expression of AQP9 mRNA increased and the expression of AQP11 was not affected by BPA (P > 0.05). There was an inhibitory effect of BPA on the release of synthesis of asparagine, threonine, taurine, tryptophan, and ornithine into the culture medium by pTr2 cells. Collectively, BPA adversely affected the expression of transporters for cationic amino acids like arginine, as well as AQPs, IGF1, and IGF1R associated with proliferation, migration, and adhesion of pTr2 cells. Those adverse effects would likely increase pregnancy losses during the peri-implantation period of pregnancy.     

Emodin attenuates acute lung injury in Cecal-ligation and puncture rats

Acute lung injury (ALI) is a major cause of sepsis-induced acute respiratory failure. Emodin has been considered to play a protective role for acute lung edema in cecal ligation and puncture (CLP)-induced sepsis model. In this study we aimed to investigate whether emodin could improve CLP-induced lung sepsis via regulating aquaporin (AQP) and tight junction (TJ), inflammatory factors, and pulmonary apoptosis. The results showed that sepsis-induced pulmonary pathological changes were significantly improved after emodin treatment. Emodin was found to upregulate AQP and TJ expression in the CLP model. Meanwhile, inflammatory cytokine release and pulmonary apoptosis was remarkably reduced after emodin treatment in lung sepsis. Our data demonstrated that emodin could suppresse inflammation, restore pulmonary epithelial barrier and reduce mortality in CLP-induced ALI, suggesting the potential therapeutic application of emodin in sepsis.     

Evidence for a glycerol pathway through aquaporin 1 (CHIP28) channels

Permeabilities to glycerol and small non-electrolytes of three Aquaporin 1 CHIP (AQP1) water channels were measured in AQP1 cRNA-injected Xenopus laevis oocytes and in human AQP1 channels reconstituted in proteoliposomes. By an osmotic swelling assay, significant increases of ethylene glycol, glycerol and 1,3-propanediol apparent permeability coefficients (P_solutes) were found in oocytes expressing human, rat and frog AQP1. p-Chloromercuribenzene sulphonate (PCMBS) and CuSO₄ inhibited, by 95% and 58% respectively, apparent glycerol permeability (P _gly) in oocytes expressing human AQP1. pCMBS inhibition was reversed by -mercaptoethanol and CuSO₄ inhibition was partly reversed by the Cu²⁺-binding peptide Gly-Gly-His. Tritiated glycerol uptakes confirmed the augmented P _gly value of AQP1 cRNA-injected oocytes. In contrast, no increases of urea, meso-erythritol, D- or L-threitol, xylitol and mannitol uptakes were detected. Stopped-flow light scattering experiments performed with human AQP1 proteoliposomes also revealed a much greater increase of P _gly than did those with protein-free liposomes; the initial rate of proteoliposomes also swelling was inhibited by 96.2% with HgCl₂ and by 72.5% with CuSO₄. In AQP1 cRNA-injected oocytes and in proteoliposomes, the value of the glycerol reflection coefficient was 0.74–0.80, indicating that water and glycerol share the same pathway. All these results provide strong evidence that water and certain small solutes permeate the AQP1 channels expressed at the surface of X. laevis oocytes or reconstituted in proteoliposomes. The urea exclusion suggests that the selectivity of the AQP1 channels not only depends on the size of the solutes but probably also on their flexibility and their ability to form H-bonds.     

水通道蛋白3在人胎盘和胎膜中的表达及分布

目的 研究正常妊娠晚期人胎盘和胎膜水通道蛋白3(AQP3)的表达及分布.方法 收集5例正常足月妊娠剖宫分娩的胎盘和胎膜样本,用RT-PCR测定AQP3 mRNA在胎盘和胎膜组织中的表达;用Western印迹和免疫组织化学方法检测AQP3蛋白质水平在胎盘和胎膜的表达.结果 RT-PCR显示AQP3 mRNA在胎盘和胎膜组织均有表达.Western印迹结果显示胎盘组织在29 000左右有一特异性条带.免疫组织化学结果显示AQP3表达于合体滋养细胞,而在羊膜上皮细胞未见表达.结论 AQP3在胎盘母儿液体平衡中可能发挥重要作用.     

Bronchiolar expression of aquaporin-3 (AQP3) in rat lung and its dynamics in pulmonary oedema

Aquaporins (AQPs) are water channel proteins that permit osmotically driven water movement. To determine their dynamics in pulmonary oedema, we examined the expression of mRNA and protein for AQP1, AQP3, AQP4, and AQP5 in the lungs of normal and thiourea-treated rats. In the thiourea group, lung water content increased significantly (vs. controls) with the peak at around 4 h. Semi-quantitative RT-PCR showed that AQP3 mRNA in the thiourea group rose significantly, peaking at around 4–8 h. The expression of AQP1, AQP4, AQP5, ENaC and CFTR mRNA each decreased significantly some time after the peak in lung water content. Immunoblot analysis showed that glycosylated AQP3 protein was increased 4–10 h after treatment. Expression of the other AQP proteins was not significantly altered, except for that of AQP4. Immunohistochemical examination revealed that AQP1 was expressed in endothelia, AQP3 in the basal cells of the large airways and in cuboidal cells in the bronchioles, AQP4 in the basolateral membrane of airway cells and AQP5 in type-I pneumocytes. Our results suggest that AQP3 is expressed not only in large airways, but also in bronchioles, and is related to water movement in pulmonary oedema.     

健脾祛湿方对脾虚湿盛型高脂血症大鼠血脂、胃肠功能、水液代谢的影响及作用机制

目的：观察健脾祛湿方对脾虚湿盛型高脂血症大鼠血脂、胃肠功能、水液代谢的影响及作用机制。方法：将56只SD大鼠随机分为空白组（n=8）和造模组（n=48）,采用“劳倦过度+饮食不节+高脂饲料喂养”复制脾虚湿盛型高脂血症大鼠模型。造模4周后,根据总胆固醇（TC）水平将造模组随机分成6组：即模型组,血脂康组,参苓白术颗粒组,健脾祛湿方低、中、高剂量组,每组8只。分组后开始给药,灌胃剂量为1 mL/100 g,空白组、模型组给予生理盐水,其余组给予相应的受试物,连续给药6周。测定血脂四项TC、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-Ch）和高密度脂蛋白胆固醇（HDL-Ch）,胃肠激素,即胃动素（MTL）、促胃液素（GAS）和血管活性肠肽（VIP）,水液调节激素醛固酮（ALD）、抗利尿激素（ADH）和心房利尿钠肽（ANP）,以及白蛋白（ALB）和总蛋白（TP）,采用苏木精-伊红（HE）染色法观察胃、结肠组织形态,免疫荧光法检测结肠水孔蛋白(AQP)3、胃AQP4的表达位置及水平,采用蛋白质印迹法（Western Blotting）检测结肠、胃组织中闭合蛋白（Occludin）的表达量。结果：与正常组比较,模型组大鼠血清TC、TG、LDL-Ch水平升高（P<0.001）,HDL-Ch水平降低（P<0.05）,血清MTL、GAS水平降低（P<0.001,P<0.01）,VIP升高（P<0.001）,ALD、ADH升高（P<0.05,P<0.001）,ANP水平显著降低（P<0.001）,TP、ALB水平降低（P<0.001,P<0.05）。结肠绒毛、胃黏膜出现大量脱落情况,结肠AQP3荧光强度降低,胃AQP4荧光表达增强,结肠、胃组织Occludin表达水平降低。与模型组比较,血脂康组、参苓白术颗粒组、健脾祛湿方低、中、高剂量组大鼠血清TC、TG、LDL-Ch水平呈下降趋势,HDL-C水平升高,MTL、GAS显著升高,VIP水平显著降低,ALD、ADH水平降低,ANP显著升高,结肠、胃组织形态结构有所改善。结肠AQP3荧光表达增强,胃AQP4荧光强度减弱,结肠、胃组织Occludin表达水平增强。结论：健脾祛湿方可降低脾虚湿盛型高脂血症大鼠血脂,有健脾祛湿之功,其作用机制可能是通过调节Occludin、AQP3、AQP4的表达,保护紧密连接结构的完整性达到促进胃肠消化吸收功能,改善水液代谢障碍的作用。     

低氧环境对椎间盘自发性吸收的影响及其作用机制

目的探讨低氧环境对椎间盘自发性吸收的影响及其作用机制。方法取SPF级成年日本大耳兔9只,雌雄不限,平均体质量2 kg。将兔处死后取脊柱髓核组织,经消化、分离、培养后获得传代髓核细胞,将生长良好的髓核细胞制成细胞悬液。根据不同时效的低氧环境将细胞分为5组对照组(常氧浓度下培养6 h)、低氧6 h组(2%O2浓度下培养6 h)、低氧12 h组(2%O2浓度下培养12 h)、低氧24 h组(2%O2浓度下培养24 h)和低氧48 h组(2%O2浓度下培养48 h)。采用实时聚合酶链反应法检测缺氧诱导因子(HIF)-1α、3型酸敏感离子通道(ASIC3)及水通道蛋白3(AQP3)mRNA表达水平,采用流式细胞仪检测各组髓核细胞凋亡情况。采用SPSS 24.0软件对数据进行分析。结果与对照组比较,低氧各组细胞凋亡率均明显升高,差异均有统计学意义(均P<0.01);与低氧12、24和48 h组比较,低氧6 h组细胞凋亡率最高,差异均有统计学意义(均P<0.01)。与对照组比较,低氧各组细胞HIF-1α和ASIC3 mRNA表达水平均明显上升,AQP3 mRNA表达水平均明显下降,差异均有统计学意义(均P<0.01);与低氧12、24和48 h组比较,低氧6 h组HIF-1α和ASIC3 mRNA表达水平最高,差异均有统计学意义(均P<0.01)。结论短时间低氧环境可以促进髓核细胞凋亡,从而加速椎间盘突出组织自发性吸收进程,其机制可能与HIF-1α和ASIC3的表达增加及AQP3的表达下降有关。     

鼻腔应用丙酸氟替卡松对鼻息肉组织中水通道蛋白-2表达的影响

目的 :探讨鼻腔局部应用丙酸氟替卡松 (FP)喷雾剂对鼻息肉组织中水通道蛋白 2 (AQP 2 )表达的影响及类固醇激素治疗鼻息肉的作用机制。方法 :将 2 6例慢性鼻窦炎鼻息肉患者分为Ⅱ型组 (3期Ⅱ型 14例 )和Ⅲ型组 (Ⅲ型 12例 ) ;Ⅱ型组又分为FP组 (7例 )和对照组 (7例 ) ,Ⅲ型组亦分为FP组 (6例 )和对照组 (6例 )。Ⅱ型组和Ⅲ型组中的FP组术前应用FP喷雾剂喷鼻 7d ,对照组术前鼻腔不予任何处理。术中取 2 6例患者的鼻息肉组织 ,采用SP免疫组化技术检测鼻息肉组织中AQP 2的表达。结果 :Ⅱ型组和Ⅲ型组中的FP组鼻息肉体积明显缩小 ,鼻息肉组织中AQP 2表达的阳性细胞数和面密度值均明显低于对照组 (均P <0 .0 1)。结论 :在FP作用下鼻息肉水肿减轻、体积缩小 ,可能与AQP 2表达减少有关 ,这为类固醇激素治疗鼻息肉作用机制的阐明提供了新的方向     

豚鼠耳蜗和内淋巴囊水通道蛋白的表达

目的 研究豚鼠耳蜗和内淋巴囊组织中水通道蛋白(aquaporin，AQP)不同亚型的定位表达及其意义。方法 用兔抗大鼠AQP1、AQP2、AQP3、AQP4的多克隆抗体，采用免疫组化SP法分别检测相应AQP蛋白亚型在豚鼠耳蜗和内淋巴囊组织中的表达模式。结果 在耳蜗组织中，AQP1、4广泛分布于耳蜗的各个区域，如血管纹、螺旋韧带、Corti器、螺旋缘、螺旋神经节等，AQP3除了在血管纹表达呈弱阳性外，其余区域的表达与AQP1和AQP4相似，AQP2则仅表达于Reissner膜。在内淋巴囊组织中，AQP1、AQP3和AQP4在内淋巴囊上皮细胞和上皮下纤维组织均呈强阳性表达，只是AQP3的表达强度稍弱于AQP1和AQP4，而AQP2在内淋巴囊上皮细胞和上皮下组织均呈阴性表达。结论 水通道蛋白1、3、4以相似的方式广泛分布于豚鼠耳蜗和内淋巴囊组织中，而AQP2则仅表达于Reissner膜，提示不同亚型的AQP可能在不同区域协同作用参与内淋巴的调节，从而保持内耳内环境的稳定。