蛋白磷酸酯酶2A通过MAPK信号通路抑制宫颈癌细胞侵袭

目的:探讨蛋白磷酸酯酶2A(PP2A)在宫颈癌细胞侵袭中的作用及其可能机制。方法:常规培养Hela细胞,通过药物或基因调节PP2A水平,药物实验中将Hela细胞分为3组,即正常对照组(DMSO处理,n=6)、DES组(10nM PP2A激动剂DES处理,n=6)和OA组(10nM PP2A抑制剂OA处理,n=6);质粒转染组中将Hela细胞分为对照组(DsRed组,n=6)、wtPP2A组(n=6)和siPP2A组(n=6),按分组要求转染相应质粒。采用小室穿孔实验观察其对Hela细胞侵袭的影响;蛋白免疫印迹实验(Western Blot)观察其对MAPK信号通路的作用。结果:研究发现上调PP2A可显著抑制Hela细胞侵袭,而下调PP2A则显著促进Hela细胞侵袭;激活PP2A可去磷酸化丝裂原激活的蛋白激酶家族(p-JNK, p-p38和p-ERK MAPK)及金属蛋白酶9(MMP-9)。结论:PP2A可能通过去磷酸化MAPK信号通路下调MMP-9起调节宫颈癌细胞侵袭的作用。     

Progesterone Inhibits Leptin-Induced Invasiveness of BeWo Cells

Background: This study investigated the roles of progesterone and leptin in placenta invasion, which is closely related to pregnancy prognosis. We examined the effects of leptin and progesterone on the invasion of BeWo cells, a human trophoblastic cell line, and the effect of concurrent treatment.Methods: Cells were treated with leptin (0, 5, 50, or 500 ng/mL) or progesterone (0, 2, 20, or 200 µM) and cultured in an invasion assay. Cells treated with 500 ng/mL leptin were also treated with progesterone (0, 2, 20, or 200 µM) in the invasion assay for 48 h. The number of cells that invaded the lower surface was counted in five randomly chosen fields using a light microscope with a 200× objective. The mRNA expression levels of MMP-9, TIMP1, TIMP2, and E-cadherin were detected by semi-quantitative PCR.Results: Invasion of BeWo cells was promoted by leptin and influenced by both leptin concentration and treatment duration. Invasion was most effective at 500 ng/mL leptin and 48 h culture. Leptin-induced invasiveness was suppressed by progesterone in a dose-dependent manner. Leptin significantly decreased the expression levels of TIMP1 and E-cadherin, whereas progesterone significantly decreased expression of MMP-9 and significantly increased levels of TIMP1, TIMP2, and E-cadherin.Conclusions: Leptin promotes invasion of BeWo cells, and progesterone suppresses leptin-induced invasion by regulating the expressions of MMP-9, TIMP1, TIMP2, and E-cadherin. The balance between leptin and progesterone may play an important role in human placenta formation during early pregnancy.     

Anti-Vibriocholerae IgY Antibody Inhibits Mortality in Suckling Mice Model

Background

Regarding to the importance of cholera in Iran and the potential advantages of egg yolk antibody (IgY) for immunotherapy, the aim of this study was to produce IgY antibody against V. cholerae Lipopolysaccharide (LPS) and determine its potential for V. cholerae treatment.

Glabridin from Chinese Herb Licorice Inhibits Fatigue in Mice

The inhibiting effect of glabridin from Chinese herb Licorice on fatigue was investigated in male BALB/c mice. Mice were divided into the following 4 experimental groups: control group (CG), low dose group (LG), middle dose group (MG) and high dose group (HG,). The control group was given 0.5% Tween 80 solution and the treatment groups (LG, MG, HG) were given various doses of glabridin (5, 10, 20 mg/kg) for 28 consecutive days. Body mass, blood lactic acid (BLA), serum blood urea nitrogen (BUN), liver glycogen and muscle glycogen concentrations in mice were determined. Results showed that glabridin significantly inhibited fatigue, which extended the exhaustive exercise time of mice, effectively delayed the elevation of blood lactic acid and increase in the storage of liver and muscle glycogen.     

Asiatic Acid Inhibits Lipopolysaccharide-Induced Acute Lung Injury in Mice

Asiatic acid (AA), a major triterpene isolated from Centella asiatica (L.) Urban, is known to exert various pharmacological activities, including anti-inflammatory and antioxidant effects. The aim of this study was to evaluate the anti-inflammatory effects of AA on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and clarify the underlying mechanisms. Lung pathological changes were assessed by H&E staining. The myeloperoxidase (MPO) activity was detected by MPO assay. The levels of inflammatory cytokines were measured by ELISA. TLR4 and NF-kB expression was detected by Western blot analysis. AA obviously inhibited LPS-induced lung histopathological changes, MPO activity, and inflammatory cell numbers in bronchoalveolar lavage fluid (BALF). Treatment of AA also inhibited LPS-induced TNF-α, IL-6, and IL-1β production. Furthermore, Western blot analysis showed that AA inhibited LPS-induced TLR4 expression and NF-kB activation. In conclusion, AA inhibited LPS-induced ALI in mice by inhibiting inflammatory cytokine production, which is mediated via blocking of the TLR4/NF-kB signaling pathway.     

重组人血管内皮抑制素对荷瘤裸小鼠的治疗作用

观察了重组人血管内皮抑制素 (YH - 16 )对人肝癌Be174 0 2 细胞系、人宫颈癌Hela细胞系和人胃癌SGC790 1细胞系在裸小鼠体内的抑制作用。试验采用每日尾静脉注射给药 ,YH - 16的剂量分别为 1.5、0 .75、0 .4mg/kg ,共给药 14次。结果表明YH - 16对上述三种人癌细胞系有明显抑制作用 ,其抑制率Be174 0 2 为 45 .6 7、43.0 8和 40 .0 0 % ;Hela为 40 .70、30 .15和2 4.12 % ;SGC790 1为 5 1.89、44 .34和 35 .85 %。YH— 16各给药组动物健康状况良好 ,无明显毒副反应     

改进的小鼠颈部心脏移植模型的制作

目的 探制小鼠颈部心脏移植模型手术技术的改进。方法 采用供心主动脉与受体颈总动脉侧壁长度相近的切口行端一侧连续吻合的方法进行４０次小鼠颈部心脏移植。结果 应用该方法使供心总缺血时间缩短至３０分钟；７天存活率提高到９７％。结论 新技术的采用降低了小鼠颈部心脏移植模型制作的难度，是一种简单实用的新方法。     

改进的小鼠颈部心脏移植模型的制作

目的探制小鼠颈部心脏移植模型手术技术的改进.方法采用供心主动脉与受体颈总动脉侧壁长度相近的切口行端-侧连续吻合的方法进行40次小鼠颈部心脏移植.结果应用该方法使供心总缺血时间缩短至30分钟;7天存活率提高到97%.结论新技术的采用降低了小鼠颈部心脏移植模型制作的难度,是一种简单实用的新方法.     

Specific IgM Enhances and IgG Inhibits the Induction of Immunological Memory in Mice

The effect of priming mice with IgM anti-SRBC (sheep erythrocytes) together with SRBC or IgG anti-SRBC together with SRBC on the development and expression of memory cells was studied. Mice primed with specific IgM and SRBC showed a much more efficient secondary plaque-forming cell and serum antibody response after challenge with SRBC in an adoptive transfer system than did controls primed with SRBC only. The expression of this enhanced memory of IgM-primed spleen cells was counteracted by the high levels of internal IgG anti-SRBC (also the result of priming with IgM) when the mice, instead of being tested in adoptive transfers, were challenged directly. The antigen-specific feedback suppression of the primary antibody response by specific IgG antibodies was also seen to inhibit partially the development of memory cells. The suppressive effect on priming could be demonstrated both in adoptive transfer systems and after direct boost of the same mice that received the primary immunization. Both the IgM enhancement and the IgG suppression of memory cell development were antigen-specific, since no effect on the antibody response to a non-cross-reacting antigen, horse erythrocytes, was seen. The effect of these up- or down-regulations of immunological memory could be demonstrated after secondary injections as long as 90-280 days after priming.     

有氧运动抑制高脂血症小鼠主动脉壁超微结构改变

目的　探讨有氧运动在动脉粥样硬化 (AS)形成过程中对血管壁的保护作用。方法　结合血脂水平的测定 ,对实验性高脂血症小鼠主动脉壁超微结构改变及有氧运动对其影响进行观察。结果　有氧运动(HE)组的甘油三酯 (TG) ,总胆固醇 (TC) ,低密度脂蛋白 (LDL)均低于高脂膳食致动脉粥样硬化 (AS)组 (P<0 .0 1) ;高密度脂蛋白 (HDL)高于AS组 (P <0 .0 1)。透射电镜显示AS组小鼠内皮细胞严重受损 ,甚至出现核固缩 ,受损的内皮细胞间有含脂滴的白细胞向内皮下穿入 ,平滑肌细胞内粗面内质网及高尔基体增多 ,弹力纤维变形。而HE组仅出现弹力纤维的轻度变形及脂质沉积。结论　有氧运动除可降低血脂外 ,同时对血管壁具有保护作用 ,但尚不能完全阻止动脉粥样硬化形成     

Baicalin Inhibits TLR2/4 Signaling Pathway in Rat Brain Following Permanent Cerebral Ischemia

Recent work from our laboratory demonstrated that baicalin attenuates inflammatory reaction and cerebral ischemia injury in rats. Toll-like receptor 2 and 4 (TLR2/4) and the downstream nuclear factor-kappa B (NF-κB) signaling pathway, which mediate the inflammatory reaction, are involved in the pathophysiological processes of cerebral ischemia. In this study, we investigated whether baicalin inhibits TLR2/4 signaling pathway in a rat model of permanent focal cerebral ischemia. Adult Sprague–Dawley rats underwent permanent middle cerebral artery occlusion (MCAO). Baicalin was administered by intraperitoneally injected twice at 2 and 12 h after the onset of ischemia. Cerebral infarct area and infarct volume were measured 24 h after MCAO. Expression of TLR2/4, NF-κB, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were determined by RT-PCR or western blot. NO and PGE2 production in rat brain were measured 24 h after MCAO. Serum content of tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) were detected by ELISA. Baicalin reduced cerebral infarct area and infarct volume. Baicalin reduced the expression of TLR2/4 and NF-κB, decreased the expression and activity of iNOS and COX-2 in rat brain. Baicalin also attenuated the serum content of TNF-α and IL-1β. Our results suggest that baicalin inhibits the TLR2/4 signaling pathway in cerebral ischemia, which may be a mechanism underlying the baicalin’s neuroprotection.     

Antigenic Stimulation and Blocking of Pregnancy in Mice by an Anti-Idiotypic Antibody to Progesterone

PROBLEM: The present study was carried out to see if the anti-idiotypic (anti-Id) antibody (Ab2) to progesterone could mimic the immunogenicity of the steroid hormone by giving rise to antiprogesterone antibodies (Ab3) and whether these antiprogesterone antibodies (Ab3) were biologically active. METHOD: Twenty virgin female Balb/C mice were actively immunized with the anti-Id antibody (Ab2). The antiprogesterone antibody (Ab3) titres in the serum were determined and the animals were used for fertility studies. In the passive immunization studies Balb/C female mice were injected i.p. with 100 μg of anti-Id antibody to see its effect on pregnancy. RESULTS: The actively immunized animals when mated showed 80% reduction in their fertility rate. The duration of infertility (20–121 days) in these animals could be directly correlated with the concentration of antiprogesterone antibody (Ab3). The anti-Id antibody (Ab2) blocked pregnancy in 80% of the passively immunized mice. CONCLUSION: The studies show that anti-Id antibody to progesterone could mimic the immunogenicity of progesterone and give rise to antiprogesterone antibodies (Ab3). The anti-Id antibody successfully blocked pregnancy in mice both after active and passive immunization.     

舒芬太尼诱导宫颈癌细胞自噬和凋亡并抑制癌细胞恶性增殖

目的 探讨舒芬太尼对宫颈癌细胞 SiHa 自噬、 凋亡及细胞增殖的影响。 方法 采用 CCK8 法检 测舒芬太尼梯度浓度 (3. 125、 6. 25、 12. 5、 25、 50、 100、 200、 400、 800 nmol / L) 作用下宫颈癌细胞 SiHa 细胞活力, 选取 4 个舒芬太尼作用浓度 (0、 25、 50、 100 nmol / L) 处理 SiHa 细胞 24 h, 采用流式细胞法、 克隆形成实验检测 SiHa 细胞凋亡及增殖情况, 免疫荧光法检测各组细胞 LC3 水平, 采用 Western 印迹检测 自噬相关蛋白 LC3Ⅱ/ LC3Ⅰ、 Beclin1、 ATG7 与增殖、 凋亡相关蛋白 P21、 Survivin 水平, 采用 RT-PCR 检测 细胞增殖相关基因 Ki67、 PCNA 表达水平。 结果 随着舒芬太尼处理浓度的增加, SiHa 细胞活力逐渐降低。 50、 100 nmol / L 舒芬太尼处理下 SiHa 细胞的 LC3Ⅱ/ LC3Ⅰ、 Beclin1、 ATG7 水平显著高于 0 nmol / L 舒芬太 尼处理, 免疫荧光检测结果显示 50、 100 nmol / L 舒芬太尼处理下 LC3 荧光信号高于 0 nmol / L 舒芬太尼处 理, 流式细胞仪检测结果显示 50、 100 nmol / L 舒芬太尼处理下 SiHa 细胞凋亡高于 0 nmol / L 舒芬太尼处理。 克隆形成实验结果显示 50、 100 nmol / L 舒芬太尼处理下 SiHa 细胞克隆形成率降低, Survivin 水平与 Ki67、 PCNA 表达水平降低, 而 P21 水平升高。 结论 舒芬太尼对宫颈癌细胞自噬、 凋亡有促进作用, 对细胞增 殖有抑制作用, 可能与调节细胞自噬、 凋亡、 增殖相关蛋白或基因水平有关。     

Adenovirus-Mediated Wnt5a Expression Inhibits the Telogen-To-Anagen Transition of Hair Follicles in Mice

The canonical Wnt/β-catenin pathway plays an important role in hair cycle induction. Wnt5a is a non-canonical Wnt family member that generally antagonizes canonical Wnt signaling in other systems. In hair follicles, Wnt5a and canonical Wnt are both expressed in cells in the telogen stage. Wnt5a has been shown to be critical for controlling hair cell fate. However, the role that Wnt5a plays in the transition from the telogen to anagen stage is unknown. In this study, using whole-mount in situ hybridization, we show that Wnt5a is produced by several other cell types, excluding dermal papilla cells, throughout the hair cycle. For example, Wnt5a is expressed in bulge and secondary hair germ cells in the telogen stage. Our studies focused on the depilated 8-week-old mouse as a synchronized model of hair growth. Interestingly, overexpression of adenovirus Wnt5a in the dorsal skin of mice led to the elongation of the telogen stage and inhibition of the initiation of the anagen stage. However, following an extended period of time, four pelage hair types grew from hairless skin that was induced by Wnt5a, and the structure of these new hair shafts was normal. Using microarray analysis and quantitative arrays, we showed that the expression of β-catenin and some target genes of canonical Wnt signaling decreased after Wnt5a treatment. These data demonstrate that Wnt5a may inhibit the telogen stage to maintain a quiescent state of the hair follicle.