Functional Coupling of Slack Channels and P2X3 Receptors Contributes to Neuropathic Pain Processing

The sodium-activated potassium channel Slack (K_Na1.1, Slo2.2, or Kcnt1) is highly expressed in populations of sensory neurons, where it mediates the sodium-activated potassium current (I_KNa) and modulates neuronal activity. Previous studies suggest that Slack is involved in the processing of neuropathic pain. However, mechanisms underlying the regulation of Slack activity in this context are poorly understood. Using whole-cell patch-clamp recordings we found that Slack-mediated I_KNa in sensory neurons of mice is reduced after peripheral nerve injury, thereby contributing to neuropathic pain hypersensitivity. Interestingly, Slack is closely associated with ATP-sensitive P2X3 receptors in a population of sensory neurons. In vitro experiments revealed that Slack-mediated I_KNa may be bidirectionally modulated in response to P2X3 activation. Moreover, mice lacking Slack show altered nocifensive responses to P2X3 stimulation. Our study identifies P2X3/Slack signaling as a mechanism contributing to hypersensitivity after peripheral nerve injury and proposes a potential novel strategy for treatment of neuropathic pain.     

What Can We Learn from FGF-2 Isoform-Specific Mouse Mutants? Differential Insights into FGF-2 Physiology In Vivo

Fibroblast growth factor 2 (FGF-2), ubiquitously expressed in humans and mice, is functionally involved in cell growth, migration and maturation in vitro and in vivo. Based on the same mRNA, an 18-kilo Dalton (kDa) FGF-2 isoform named FGF-2 low molecular weight (FGF-2LMW) isoform is translated in humans and rodents. Additionally, two larger isoforms weighing 21 and 22 kDa also exist, summarized as the FGF-2 high molecular weight (FGF-2HMW) isoform. Meanwhile, the human FGF-2HMW comprises a 22, 23, 24 and 34 kDa protein. Independent studies verified a specific intracellular localization, mode of action and tissue-specific spatiotemporal expression of the FGF-2 isoforms, increasing the complexity of their physiological and pathophysiological roles. In order to analyze their spectrum of effects, FGF-2LMW knock out (ko) and FGF-2HMWko mice have been generated, as well as mice specifically overexpressing either FGF-2LMW or FGF-2HMW. So far, the development and functionality of the cardiovascular system, bone formation and regeneration as well as their impact on the central nervous system including disease models of neurodegeneration, have been examined. This review provides a summary of the studies characterizing the in vivo effects modulated by the FGF-2 isoforms and, thus, offers a comprehensive overview of its actions in the aforementioned organ systems.     

Vitamin D Compounds PRI-2191 and PRI-2205 Enhance Anastrozole Activity in Human Breast Cancer Models

1,25-Dihydroxycholecalciferol, the hormonally active vitamin D₃ metabolite, is known to exhibit therapeutic effects against breast cancer, mainly by lowering the expression of estrogen receptors and aromatase activity. Previously, the safety of the vitamin D active metabolite (24R)-1,24-dihydroxycholecalciferol (PRI-2191) and 1,25(OH)₂D₃ analog PRI-2205 was tested, and the in vitro activity of these analogs against different cancer cell lines was studied. We determined the effect of the two vitamin D compounds on anastrozole (An) activity against breast cancer based on antiproliferative activity, ELISA, flow cytometry, enzyme inhibition potency, PCR, and xenograft study. Both the vitamin D active metabolite and synthetic analog regulated the growth of not only estrogen receptor-positive cells (T47D and MCF-7, in vitro and in vivo), but also hormone-independent cancer cells such as SKBR-3 (HER-2-positive) and MDA-MB-231 (triple-negative), despite their relatively low VDR expression. Combined with An, PRI-2191 and PRI-2205 significantly inhibited the tumor growth of MCF-7 cells. Potentiation of the antitumor activity in combined treatment of MCF-7 tumor-bearing mice is related to the reduced activity of aromatase by both An (enzyme inhibition) and vitamin D compounds (switched off/decreased aromatase gene expression, decreased expression of other genes related to estrogen signaling) and by regulation of the expression of the estrogen receptor ERα and VDR.     

抗抑郁样及焦虑样行为乳酸菌的筛选

越来越多的证据表明益生菌可以通过微生物-肠-脑轴来调节大脑功能和行为。本试验利用行为模型从益生菌库中初筛选出7株具有抗抑郁样/焦虑样行为潜能的乳酸菌菌株。进一步采用明暗箱和高架十字迷宫实验研究乳酸菌对健康小鼠焦虑样行为的影响;利用悬尾实验和强迫游泳实验研究乳酸菌对健康小鼠抑郁样行为的影响。研究显示补充马乳酒样乳杆菌ZW3,植物乳杆菌MA2和植物乳杆菌299(10~8CFU)4周后,与空白对照小鼠相比,可以增加小鼠在明暗箱中亮箱的停留时间和高架十字迷宫实验中开臂的停留时间,改善小鼠的焦虑样行为;此外这3株菌还可以降低小鼠在悬尾实验和强迫游泳实验中的不动时间,改善小鼠的抑郁样行为;并且它们可以降低小鼠脑区生物标记物5-羟色胺含量以及血清皮质酮水平。这些结果表明马乳酒样乳杆菌ZW3,植物乳杆菌MA2和植物乳杆菌299作为益生菌,具有改善焦虑样和抑郁样行为的潜能。     

短链脂肪酸改善2型糖尿病小鼠胰岛素抵抗和胰腺损伤

以链脲佐菌素(Streptozotocin,STZ)诱导的糖尿病小鼠为研究对象,研究短链脂肪酸对糖尿病小鼠血糖代谢的改善作用。通过腹腔注射200 mg/kg STZ建立糖尿病小鼠模型,研究乙酸钠、丙酸钠和丁酸钠对糖尿病小鼠进食量和体重变化、口服葡萄糖耐量(OGTT)、血糖水平、血清胰岛素水平、胰岛素抵抗(HOMA-IR)、胰岛β细胞功能(HOMA-β)指数及胰腺组织结构的影响。结果显示:与模型组相比,乙酸钠和丙酸钠可显著降低累计进食量(p0.05),分别降低了10.09%和8.90%;乙酸钠和丁酸钠,对2型糖尿病小鼠其他指标包括体重、空腹血糖、葡萄糖耐量、胰岛素抵抗和胰腺组织损伤修复都没有显著改善作用(p0.05)。丙酸钠可以显著降低血糖水平(20.65%)和胰岛素抵抗(11.19%),增强胰岛β细胞功能(64.50%),提高葡萄糖耐量,对胰腺组织损伤起到改善作用。     

樟芝多糖通过降低NLRP3 Caspase1炎性小体表达改善帕金森小鼠神经行为学的机制研究

目的:研究樟芝多糖通过降低NLRP3-Caspase1炎性小体表达改善6-OHDA构建的帕金森小鼠模型的行为学机制。方法:利用6-OHDA脑内注射构建帕金森小鼠模型，通过酪氨酸羟化酶（TH）免疫组化染色和行为学判定小鼠模型的构建成功。利用樟芝多糖进行干预，分别在干预前、干预后的第1、3、7天4个时间点进行神经行为学实验，分别采用转棒实验、爬杆实验检测小鼠自主行为能力以及协调能力，4个时间点取小鼠尾静脉外周血采用ELISA法检测外周血中Caspase1和IL-1β的表达，樟芝多糖干预第7天时待进行完行为学实验后小鼠断颈处死，取小鼠脑组织-纹状体，Western blot法检测纹状体中Caspase1、proCaspase1、NLRP3的表达，高效液相色谱检测纹状体中单胺类神经递质的表达，RT-QPCR检测Caspase1、NLRP3、IL-1β、IL-4、IL-6的mRNA表达。NISSl染色检测小鼠脑组织神经细胞凋亡情况。 结果:6-OHDA脑内注射可以造成小鼠帕金森样病变，且TH蛋白表达显著下调，樟芝多糖干预后小鼠的行为学得到显著改善（P<0.05），纹状体中Caspase1、proCaspase1、NLRP3的表达显著下调，与模型小鼠相比具有统计学差异（P<0.05），且相关炎症因子Caspase1、NLRP3、IL-1β、IL-4、IL-6的mRNA表达下调（P<0.05），纹状体中单胺类神经递质表达上升（P<0.05）。结论:樟芝多糖可以通过下调NLRP3-Caspase1炎性小体表达来改善6-OHDA构建的帕金森小鼠模型行为改善，这可能是樟芝多糖治疗帕金森的机制之一。     

Induction of adaptive response <i>in utero </i>by ionizing radiation: A radiation quality dependent phenomenon

Investigation on possible induction of adaptive response (AR) by high-liner energy transfer (LET) particle radiation for protection against low-LET photon radiation-induced detrimental effects has not yet been performed in utero. This study verified if an AR could be induced by high-LET particle radiation from accelerated heavy ions against low-LET X-ray radiation-induced detrimental effects on fetal mice. Total body irradiation of pregnant C57BL/6J mice were performed by delivering a priming dose ranging from 10 mGy to 320 mGy of particle radiation on gestation day 11 followed one day later by a challenge dose at 3500 mGy from X-ray radiation. The monoenergetic beams of carbon, silicon and iron with the LET values of about 15, 55, and 200 KeV/μm, respectively, were examined. Significant suppression by the priming radiation of the detrimental effects (fetal death, malformation, or low body weight) was used as the endpoints for judgment of a successful AR induction on gestation day 18. Existence of AR was not observed. On the other hand, the priming dose of high-LET particle radiation, in some cases, even increased the detrimental effects induced by the challenge dose from low-LET X-ray radiation. Although existence of AR induced by high-LET radiation in cultured mammalian cells in vitro and in certain tissues of laboratory mice in vivo was demonstrated, the present study did not suggest that low dose of high-LET particle radiation could induce an AR in fetal mice in utero under the setup of our experimental system.     

复方牛磺酸维生素饮料缓解小鼠体力疲劳作用研究

目的研究复方牛磺酸维生素饮料缓解小鼠体力疲劳的作用。方法将200只健康雄性ICR小鼠分成4组,第1组负重游泳,第2组测肝糖原和肝脏丙二醛(hepatic malondialdehyde,MDA),第3组测高尿素模型小鼠血清尿素氮(urea nitrogen,UREA)、乳酸脱氢酶(lactate dehydrogenase,LDH)、肝脏MDA,第4组测血乳酸(blood lactic acid,BLA),每组50只,组内按体重分成去离子水阴性对照组、糖水对照组、3个复方牛磺酸维生素饮料剂量组[3.33、6.67、20.00 mL/(kg·d) 6.25倍浓缩液]。各组连续灌胃30 d后,分别检测小鼠负重游泳时间、肝糖原、肝脏MDA、血清UREA、LDH、BLA等疲劳相关的指标。结果复方牛磺酸维生素饮料对各组小鼠体重无影响(P 0.05),在中剂量能显著延长小鼠负重游泳时间(P 0.05),显著增加小鼠肝糖原含量(P0.05),显著降低高尿素模型小鼠血清LDH(P0.01),在高剂量能显著降低高尿素模型小鼠血清中UREA(P0.01)、血清LDH(P0.05)、肝脏MDA含量(P0.05),对BLA曲线下面积无影响(P0.05)。结论此配方的复方牛磺酸维生素饮料对ICR小鼠具有缓解体力疲劳的功能。     

Male-Dependent Promotion of Colitis in 129 Rag2−/− Mice Co-Infected with Helicobacter pylori and Helicobacter hepaticus

The prevalence of gastric Helicobacter pylori (Hp) infection is ~50% of the world population. However, how Hp infection influences inflammatory bowel disease in humans is not fully defined. In this study, we examined whether co-infection with Hp influenced Helicobacter hepaticus (Hh)–induced intestinal pathology in Rag2^−/− mice. Rag2^−/− mice of both sexes were infected with Hh, of which a subgroup was followed by infection with Hp two weeks later. Co-infected males, but not females, had significantly higher total colitis index scores in the colon at both 10 and 21 weeks post-Hh infection (WPI) and developed more severe dysplasia at 21 WPI compared with mono-Hh males. There were no significant differences in colonization levels of gastric Hp and colonic Hh between sexes or time-points. In addition, mRNA levels of colonic Il-1β, Ifnγ, Tnfα, Il-17A, Il-17F, Il-18, and Il-23, which play important roles in the development and function of proinflammatory innate lymphoid cell groups 1 and 3, were significantly up-regulated in the dually infected males compared with mono-Hh males at 21 WPI. These data suggest that concomitant Hp infection enhances the inflammatory responses in the colon of-Hh-infected Rag2^−/− males, which results in more severe colitis and dysplasia.