Protein kinase C isoforms in the porcine corpus luteum: Temporal and spatial expression patterns

Porcine corpora lutea (CL) fail to show a luteolytic response to prostaglandin-F-2α (PGF-2α) (ie, luteolytic sensitivity, or LS) until ∼day 13 of the estrous cycle. In view of the importance of protein kinase C (PRKC) in PGF-2α signal transduction, it was hypothesized that limiting levels of 1 or more PRKC isoforms may explain the lack of LS before day 13. This hypothesis was tested by examining expression of mRNA and protein, and the cellular localization patterns of the 11 PRKC isoforms throughout the porcine estrous cycle, to determine whether PRKC expression correlates with and thus may be associated with the control of the acquisition of LS in the pig. The expression patterns show that for most PRKC isoforms (ie, PRKC alpha, beta 1, beta 2, delta, epsilon, theta, iota, and zeta), mRNA was maximally expressed on day 7 or day 10 (protein kinase D1 only) of the cycle, whereas PRKCs gamma, eta, and lambda were unchanged. At the protein level, only PRKC epsilon (PRKCE) significantly changed during the estrous cycle and was elevated on day 13 (versus days 4, 7, and 15; P < 0.05). By immunofluoresence, most PRKC isoforms, including PRKCE, were localized to steroidogenic large luteal cells (LLC) and small (nonendothelial cell) luteal cell subtypes (SLC). In conclusion, since the increase in PRKCE protein expression (day 13) occurred coincidentally with the onset of LS (≥day 12), these results support a potential role for PRKCE in control of the acquisition of LS in the pig.     

低分子质量壳寡糖对蛋鸡生产性能、蛋品质、血清生化指标、盲肠微生物数量及脾脏白细胞介素-2、肿瘤坏死因子-α基因表达的影响

本试验旨在研究饲粮中添加不同水平的低分子质量(1 000 u)壳寡糖对蛋鸡生产性能、蛋品质、血清生化指标、盲肠微生物数量以及脾脏白细胞介素-2(IL-2)和肿瘤坏死因子-α(TNF-α)基因表达的影响。选取体重和产蛋率相近的58周龄海兰褐壳蛋鸡600只,随机分为4组,每组5个重复,每个重复30只。对照组饲喂基础饲粮,试验组分别饲喂在基础饲粮中添加300、600、900 mg/kg壳寡糖的试验饲粮。预试期为7 d,正试期为42 d。结果表明:1)添加300、600和900 mg/kg壳寡糖组的产蛋率分别比对照组提高了4.52%(P0.05)、2.99%(P0.05)和4.08%(P0.05)。2)试验第3、6周末,添加600和900 mg/kg壳寡糖组的鸡蛋哈夫单位分别比对照组提高了6.87%、6.69%和6.47%、6.60%(P0.05)。3)与对照组相比,饲粮添加600、900 mg/kg壳寡糖显著降低了血清葡萄糖、胆固醇含量和谷草转氨酶活性(P0.05)。4)与对照组相比,饲粮添加600、900 mg/kg壳寡糖显著提高了盲肠双歧杆菌和乳酸杆菌的数量(P0.05),显著降低了盲肠金黄色葡萄球菌的数量(P0.05)。5)与对照组相比,饲粮添加300、600 mg/kg壳寡糖显著提高了脾脏IL-2 m RNA表达水平(P0.05),饲粮添加600 mg/kg壳寡糖显著提高了脾脏TNF-αm RNA表达水平(P0.05)。由此可见,饲粮中添加不同水平的壳寡糖,提高了蛋鸡的产蛋率和哈夫单位,调节了肠道微生物菌群,增强了蛋鸡的免疫力,适宜壳寡糖添加水平为600 mg/kg。     

α-酮戊二酸和大蒜素对生长猪生长发育及养分表观消化率的影响

本试验旨在研究α-酮戊二酸(AKG)和大蒜素对生长猪生长性能、养分表观消化率、血清生化指标以及肠道形态结构的影响。选用40头初始平均体重为(30.64±1.35)kg的健康三元(杜×长×大)杂交生长猪,按体重相近、性别比例相同原则随机分为5组,每组8个重复,每个重复1头猪。5组试验猪分别饲喂无抗基础饲粮(对照组)、无抗基础饲粮+25 mg/kg抗生素(Ⅰ组)、无抗基础饲粮+1%AKG(Ⅱ组)、无抗基础饲粮+0.5%大蒜素(Ⅲ组)、无抗基础饲粮+1%AKG+0.5%大蒜素(Ⅳ组)。预试期7d,正试期21d。结果表明:1)与对照组相比,Ⅰ组和Ⅳ组生长猪的平均日增重分别提高了10.80%和9.91%,但差异不显著(P0.05)。2)与对照组相比,Ⅱ组钙的表观消化率显著提高(P0.05),Ⅳ组粗蛋白质、钙和磷的表观消化率显著提高(P0.05),且Ⅳ组较Ⅰ组显著提高了磷的表观消化率(P0.05)。3)与对照组相比,Ⅱ组血清免疫球蛋白G(IgG)含量显著提高(P0.05);Ⅲ组血清甘油三酯(TG)含量显著降低(P0.05);Ⅳ组血清总胆固醇(TCHO)和TG含量显著降低(P0.05),血清碱性磷酸酶(AKP)、谷草转氨酶(AST)和谷丙转氨酶(ALT)活性及血清免疫球蛋白A(IgA)、IgG含量显著提高(P0.05)。与Ⅰ组相比,Ⅳ组血清TCHO和TG含量有降低趋势(P0.05),血清AST和ALT活性显著提高(P0.05)。4)与对照组相比,Ⅳ组回肠绒毛高度及空肠和回肠的绒毛高度/隐窝深度值显著提高(P0.05)。5)与对照组相比,Ⅳ组回肠中杯状细胞和淋巴细胞数目显著增加(P0.05)。综上,无抗饲粮中联合添加1%AKG和0.5%大蒜素可通过改善肠道形态结构,提高对粗蛋白质、钙和磷等养分的消化吸收能力,促进肠道健康,进而改善生长猪的生长性能。     

59份野生桑桑叶中的DNJ含量及粗提物对α-糖苷酶的抑制活性

对来自云南省不同地区的59份野生桑桑叶中的1-脱氧野尻霉素(DNJ)含量,以及桑叶粗提物对淀粉酶、蔗糖酶和麦芽糖酶的体外抑制活性进行了测定,同时采用液-质联用(LC-MS)仪分离纯化桑叶粗提物中的DNJ,并对其抑制蔗糖酶的机制进行了初步研究,期望筛选出适用于治疗糖尿病药物开发的药用桑资源,以及为解析桑叶粗提物和纯化的桑叶DNJ抑制α-糖苷酶活性的机制提供线索。研究结果表明:不同来源的野生桑桑叶中的DNJ含量存在明显差异,其中来自开远地区的岩桑桑叶中DNJ的质量分数高达0.791 1%,是具有药用开发价值的野生桑资源;不同来源的野生桑桑叶粗提物对淀粉酶、蔗糖酶和麦芽糖酶活力的体外抑制率差异显著,但抑制活性的大小与样品中的DNJ含量不成正比,推测桑叶中除含有DNJ外还含有具有明显的α-糖苷酶抑制活性的多种DNJ类似物;从野生桑桑叶粗提物中分离纯化的DNJ对淀粉酶无明显抑制活性,但对蔗糖酶和麦芽糖酶有显著的抑制活性,对蔗糖酶表现为非竞争性抑制作用。     

甘蔗不同品种(种)个体发育过程中叶片淀粉酶活性的变化

甘蔗品种(种)叶片α-淀粉酶活性变化的基因型效应较大,12个供试材料间的差异明显,种间的差异大于品种间的差异,在个体发育过程中变化幅度较小。β-淀粉酶的活性变化与生长发育的关系较为密切,其变化呈典型的单峰曲线,活性的最高峰出现在甘蔗的旺长时期。几乎所有供试材料酶活性变化趋势是一致的,但变化幅度有差异。分蘖末期至伸长初期,同一材料的β-淀粉酶活性通常高于α-淀粉酶。     

CYP450 epoxygenase/EET pathway attenuates adipose inflammation of obese mice through HIF-1α

AIM To investigate the effects of cytochrome P450 （CYP450） epoxygenase/epoxyeicosatrienoic acid （EET） pathway on insulin resistance in obese mice, and to explore the possible mechanisms. METHODS High-fat diet-induced obesity model was established in C57BL/6Cnc mice, and the obese mice were randomly divided into 3 groups, including obesity group （treated with saline; n=10）, EET group （treated with 11,12-EET; n=10） and EET inhibitor 14,15-epoxyeicosa-5（Z）-enoic acid （EEZE） group （n=10）. Normal C57BL/6Cnc mice （n=10） treated with saline served as control. Protein expression of CYP2J2 （one of CYP450 epoxygenases） and hypoxia-inducible factor-1α （HIF-1α） was measured by Western blot. Vessel-like structure was detected by immunofluorescence staining. The serum levels of insulin, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β, IL-6 and monocyte chemoattractant protein-1 （MCP-1） were measured by ELISA. RESULTS In obese mice, homeostasis model assessment of insulin resistance （HOMA-IR） values were increased, the protein level of CYP2J2 was reduced, and the protein level of HIF-1α was increased in adipose tissues as compared with the controls （P<0.05）. The serum levels of MCP-1, IL-1β, IL-6 and TNF-α were also significantly increased in obese mice （P<0.05）. After treatment with 11, 12-EET, the HOMA-IR values were decreased compared with vehicle-treated obese mice, HIF-1α expression levels were decreased in the adipose tissue, and the serum levels of MCP-1, IL-1β, IL-6 and TNF-α were reduced （P<0.05）. Immunohistochemical results of adipose tissue from vehicle-treated obese mice showed a marked decrease in vessel-like structures （CD31-positive） compared with normal control mice （P<0.05）. EET treatment significantly increased the newly formed vessel-like structures in the visceral adipose tissues of obese mice as compared with vehicle-treated obese mice （P<0.05）. CONCLUSION High-fat diet-induced obesity and insulin resistance are closely related to the CYP450 pathway. Exogenous EETs effectively decrease obesity-induced insulin resistance possibly through pro-angiogenesis and attenuation of hypoxia and inflammation.     

Three new water-soluble alkaloids from the leaves of Suregada glomerulata (Blume) Baill

Three new water-soluble alkaloids (1-3) together with twelve known compounds (4-15) have been isolated from the water extract of leaves of Suregada glomerulata. Their structures were determined by spectroscopic analysis and chemical method. Compounds 1-3 were evaluated for their in vitro inhibitory activity against α-glucosidase and HIV-1 replication. However, no significant activities were found.     

Antinociceptive and antiplasmodial activities of cassane furanoditerpenes from Caesalpinia volkensii H. root bark

The chloroform and ethyl acetate extract (100 mg/kg) of Caesalpinia volkensii H. exhibited significant (P ≤ 0.05) antinociceptive activities using hot plate and writhing tests in mice while the later showed antiplasmodial activity (IC₅₀ 0.23 ± 0.07 and 4.39 ± 2.49 μg/ml) against chloroquine sensitive (D6) and chloroquine-resistant (W2), respectively. Two new furanoditerpenes [rel. 1β,5α-dihydroxyvoucapane (1) and rel. 1β,6β-dihydroxyvoucapane; 19β-methyl ester (2)] together with seven known compounds [voucapane (3), voucapan-5-ol (4), deoxycaesaldekarin C (5), caesaldekarin C (6), 5-hydroxyvinhaticoic acid (7), triacontanyl-(E)-ferulate (8), triacontanyl-(E)-caffaete (9) and 30′-hydroxytriacontanyl-(E)-ferulate (10)] were isolated from the two extracts. The administration of 3, 4, 5 and 6 (100 mg/kg i.p) caused a significant (P ≤ 0.05) reduction in the number of writhing episodes induced by acetic acid and (P ≤ 0.01) increased pain latency threshold in hot-plate test compared to control. However, the pure compounds indicated relatively (P ≤ 0.05) low antiplasmodial activity. The phytochemical constituents from the root bark of C. volkensii had better analgesic properties than antimalarial properties, justifying the use of the plant root bark as a remedy for pain.     

Effects of the exopolysaccharide fraction (EPSF) from a cultivated Cordyceps sinensis on immunocytes of H22 tumor bearing mice

In order to explore the effects of exopolysaccharide fraction (EPSF) from one of the anamorph strains of Cordyceps sinensis on immunocyte activity of H22 tumor bearing mice, ICR mice were treated with EPSF for 7 days by intraperitoneal injection at doses of 15 mg/kg (low-dose), 30 mg/kg (mid-dose) and 60 mg/kg (high-dose) after H22 tumor cells were implanted. At the end of the experiments, the tumor weight of each mouse was measured. Phagocytosis of mouse peritoneal macrophages was tested by neutral red uptake. The TNF-alpha expression of macrophages was assayed by ELISA. Proliferation ability and cytotoxicity of spleen lymphocytes were determined by MTT methods. The mRNA levels of cytokine TNF-alpha and IFN-gamma mRNA of spleen lymphocytes were detected by RT-PCR. The results indicated that EPSF not only significantly inhibited the H22 tumor growth, but also significantly elevated immunocytes' activity. It significantly enhanced the phagocytosis capacity of peritoneal macrophages and proliferation ability of spleen lymphocytes at all the three doses; it significantly promoted macrophages' TNF-alpha expression and spleen lymphocytes' cytotoxicity. EPSF also significantly elevated TNF-alpha and IFN-gamma mRNA expression of splenic lymphocytes. This experimental finding suggests that EPSF could elevate the immunocytes' activity in H22 tumor bearing mice.     

团花树α扩展蛋白基因的克隆及表达分析

采用扩增保守区域、基因组步移及3'RACE技术,在团花树中克隆到15个α-expansin基因的cDNA序列,命名为AcEXPA2-16(GenBank注册号JF922686-JF922700),其相对应的基因组序列GenBank注册号为JF922701-JF922715.包括已知的AcEXPA1,这16个EXPA蛋白的大小及序列高度保守,包括N端的信号肽、3个外显子和2个内含子.通过氨基酸序列比对和系统进化树分析,AcEXPA1-16分为4个亚族,分别为A、B、C和D亚族.定量PCR分析显示AcEXPA1-16基因在同一组织中的表达存在冗余现象,而单个基因在不同的组织中又存在特异表达,尤其AcEXPA8在形成层中的表达水平极高.这表明AcEXPA8在木质部发生的过程中可能起到重要的作用.研究结果为在团花树中研究α-expansin基因与木质部的生长和材质的关系提供信息,并最终为林木分子育种提供潜在的候选基因.