瘦素、神经肽Y对离体人卵巢颗粒细胞分泌功能的影响

目的　探讨瘦素、神经肽Y(NPY)对离体人卵巢颗粒细胞雌、孕激素生成的影响。　方法　将来自体外受精 胚胎移植的 2 5例离体人卵巢颗粒细胞纯化后 ,在不同浓度瘦素 (1、10、10 0 μg/L)、NPY(1× 10 -8、1× 10 -7、1× 10 -6mol/L)单独或与人绒毛膜促性腺激素 (hCG ,0、2 0IU/L)联合作用下体外培养 ,采用放射免疫方法测定培养液中雌二醇 (E2 )、孕酮(P)的水平。　结果　 (1)无论单独或与hCG联合不同浓度瘦素组E2 、P水平均无显著性差异 (P >0 .0 5 ) ;(2 )NPY浓度 1× 10 -7mol/L时 ,E2 水平显著低于空白对照组 (P <0 .0 5 ) ,但与hCG联合组显著高于单独组 (P <0 .0 5 ) ;(3)NPY浓度 1× 10 -7mol/L、瘦素浓度 10 μg/L组 ,E2 水平明显高于相应浓度NPY单独作用组 (P <0 .0 5 )。　结论　 (1)瘦素不直接影响体外培养颗粒细胞E2 、P的分泌 ,可能通过影响其它因子的作用而间接调节卵巢功能 ;(2 )NPY单独作用时 ,对颗粒细胞E2 的分泌有一定抑制作用 ,但当hCG存在时作用消失 ,说明在正常月经周期中NPY对黄体的生成和维持可能无明显不利影响 ;(3)瘦素可能阻断NPY对颗粒细胞分泌E2 的抑制作用 ,表明二者可能对卵巢功能有相互协调作用。     

4-羟基他莫昔芬对前列腺基质细胞增殖与凋亡的作用

目的:研究4-羟基他莫昔芬(OHT)对原代培养的前列腺基质细胞增殖与凋亡的影响。方法:以10-8~10-5mol/L的雌二醇(E2)、己烯雌酚(DES)、OHT以及10-8~10-6mol/L的E2与10-7mol/L的OHT混合物分别作用于原代培养的前列腺基质细胞,采用MTT法和TUNEL法分别检测细胞增殖和凋亡。结果:OHT对前列腺基质细胞增殖和凋亡的作用与E2和DES比较均有显著差异(P均<0.05),在10-7~10-5mol/L浓度下表现出与浓度相关(r=-0.383,P=0.005)的抑制增殖作用(P<0.05),10-7mol/L的OHT可以抑制相同甚至更高浓度(10-6mol/L)E2的促增殖作用(P<0.05);OHT在10-8~10-5mol/L浓度下显示与浓度相关(r=0.349,P=0.012)的促凋亡作用(P<0.05),且10-7mol/L OHT的促凋亡作用不能被相同甚至更高浓度(10-6mol/L)E2所逆转(P>0.05)。结论:OHT在一定浓度范围内对原代培养的前列腺基质细胞具有明显的抑制增殖和促凋亡作用,该作用可能不完全是通过竞争性抑制雌激素受体而实现的。     

17β-雌二醇对豚鼠胆囊平滑肌收缩的影响

目的 观察17p-雌二醇对豚鼠胆囊平滑肌的基因组和非基因组效应,初步探究其机制。方法 构建去卵巢豚鼠动物模型,皮下注射17β-雌二醇,检测各组豚鼠血清雌二醇和八肽胆囊收缩素(CCK-8)含量;采用张力换能器观察17p-雌二醇对胆囊离体肌条收缩的影响。观察17β-雌二醇对豚鼠胆囊平滑肌肌条的急性效应及可能机制。结果 与假手术组相比,去卵巢组豚鼠血清雌二醇和CCK-8含量降低(P＜0.05),离体胆囊肌条对CCK-8以及乙酰胆碱的敏感性升高(P＜0.05)。随着皮下注射17β-雌二醇时间（1、3和7 d）的延长,二者含量升高(P＜0.05),离体胆囊肌条对CCK-8和乙酰胆碱的敏感性下降(P＜0.05)。10-9～10-7 mol/L的17β-雌二醇对正常豚鼠离体胆囊肌条收缩无明显影响(P＞0.05),10 6～10 5mol/L的17β-雌二醇可以抑制胆囊肌条收缩(P＜0.05),这种抑制效应可以被尼莫地平、阿托品、地伐西匹、ICI 182,780和Y-27632等阻断剂阻断。结论 17β-雌二醇与胆囊动力相关,可以通过基因组和非基因组机制影响胆囊平滑肌的收缩。     

c-ras与大鼠颗粒细胞和黄体细胞生成孕酮的关系

应用离体细胞培养 ,研究了反义 c-ras寡脱氧核苷酸 (反义 c-ras ODN)对人绒毛膜促性腺激素 (h CG)诱导的大鼠颗粒细胞和黄体细胞生成孕酮 (P)的影响及其与外源性 c AMP和 Ca2 +的关系。结果发现 ,2 0 μmol/ L 反义 c-ras ODN能显著抑制 h CG诱导颗粒细胞的 P生成量 (P<0 .0 5 ) ,而对黄体细胞的 P生成无明显影响 ;反义 c-ras ODN对h CG诱导的颗粒细胞生成 P的抑制作用能被加入 1 0 - 4mol/ L 二丁酰 c AMP所逆转 ,钙通道阻断剂维拉帕米对抑制作用具有协同效应。结果提示 ,c-ras癌基因参与 h CG诱导的颗粒细胞生成 P的调控 ,而对 h CG诱导的黄体细胞 P生成关系不大     

山楂叶总黄酮对离体大鼠胸主动脉环功能的影响

目的 观察山楂叶总黄酮(HLF)对大鼠胸主动脉血管环的作用,并探讨其作用机制.方法 采用大鼠胸主动脉环张力测定法,观察HLF对高浓度KCl(6×10-2mol/L)和去甲肾上腺素(NE,1×10-6mol/L)预收缩的离体大鼠胸主动脉血管环作用;并观察左旋硝基精氨酸甲酯(L-NAME,1×10-5mol/L)和吲哚美辛(1×10-4mol/L)对其作用的影响.结果 HLF(5×10～50×10-3g/L)对高浓度氯化钾(KCl,6×10-2mol/L)预收缩的内皮完整或去内皮的血管环均无明显作用(P＞0.05);对去甲肾上腺素(NE,1×10-6mol/L)预收缩的血管环产生内皮依赖性的舒张作用(P＜0.01);HLF对NE预收缩血管的舒张作用能被L-NAME(1×10-5 mol/L)显著阻断(P＜0.01),而不能被吲哚美辛(1×10-4mol/L)阻断(P＞0.05).结论 HLF对胸主动脉血管有舒张作用,其机制可能是通过抑制受体操纵的钙通道(ROC)的激活而直接抑制细胞外钙内流降低细胞内Ca2+浓度而起作用的;HLF的内皮依赖性舒张作用可能与血管内皮一氧化氮(NO)合成有关.

Abstract：

Objective To investigate the effects and mechanisms of hawthorn leaf flavonoids (HLF) on isolated thoracic aorta in rats. Methods The tension of rat thoracic aorta tings was measured.The effects of HLF on the thoracic aorta preconstricted by potassium chloride ( KCl, 6 × 10-2 mol/L) and norepinephrine (NE, 1 × 10-6 mol/L), as well as the effects of L-nitro arginine methyl ester (L-NAME,1 × 10-5 mol/L) and indomethacin ( 1 × 10-4 mol/L) on the relaxation response of HLF were observed.Results HLF completely relaxed the constriction induced by NE ( 1 × 10 -6 mol/L) in endothelium-intact thoracic aorta ( P ＜ 0. 01 ), but had no effect on those preconstricted aorta rings by a high concentration of KCl (6 × 10-2 mol/L) in endothelium-intact and endothelium-denuded rat aorta (P ＞ 0. 05). The relaxation response of HLF was significantly inhibited by L-NAME (P ＜ 0. 01 ), but not by indomethacin (P ＞0. 05). Conclusion The vasorelaxation induced by HLF in rat aorta rings may involve the reduction of Ca2+ influx through the calcium channels operated by receptor (ROC). The endothelium-dependent relaxation of HLF may be related to the generation of nitric oxide (NO).     

Decreased gallbladder response in leptin-deficient obese mice

Obesity is a major risk factor for gallstone formation, but the pathogenesis of this phenomenon remains unclear. Human data on gallbladder emptying are conflicting, and no animal data exist on the effect of obesity on gallbladder motility. Leptin, a hormone produced by adipocytes, is known to have central effects on neuropeptide Y and cholecystokinin, but the influence of leptin on the biliary effects of these hormones is unknown. Therefore we tested the hypothesis that leptin-deficient C57BL/6J-lep^ob obese mice would have decreased gallbladder responses to excitatory stimuli. Twelve-week-old lean control (C57BL/6J) (n = 22) and C57BL/6J-lep^ob obese (n = 20) female mice were fed a nonlithogenic diet. The mice were fasted overnight and underwent cholecystectomy. Whole gallbladders were placed in 3 ml muscle baths. After optimal length was determined with acetylcholine (10^-5 mol/L, responses to increasing doses of neuropeptide Y (10^-8 to 10^-6 mol/L) and cholecystokinin-8 (10^-10 to 10^-7 mol/L) were measured. Student’s t test and two-way analysis of variance were used where appropriate. Results were expressed as Newtons per cross-sectional area. The lean control mice had significantly greater excitatory responses to acetylcholine than the obese mice (0.37 ± 0.05 vs. 0.16 ± 0.02, P < 0.01). The gallbladder responses were also greater when mice were treated with neuropeptide Y (10^-8 mol/L: 0.00 ± 0.00 vs. 0.00 ± 0.00, NS; 10^-7 mol/L: 0.12 ± 0.02 vs. 0.05 ± 0.01, P < 0.01; 10^-6 mol/L: 0.26 ± 0.08 vs. 0.06 ± 0.01, P < 0.01) and cholecystokinin (10^-10 mol/L: 0.27 ± 0.04 vs. 0.13 ± 0.02, P < 0.01; 10^-9 mol/L: 0.59 ± 0.08 vs. 0.27 ± 0.04, P < 0.01; 10^-8 mol/L: 0.80 ± 0.11 vs. 0.37 ± 0.05, P < 0.01; 10^-7 mol/L: 0.86 ± 0.11 vs. 0.44 ± 0.06, P < 0.01). These data suggest that genetically obese, leptin-deficient mice have decreased responses to acetylcholine, neuropeptide Y, and cholecystokinin. We conclude that decreased gallbladder motility contributes to the increased incidence of gallstones associated with obesity. Presented at the Forty-Second Annual Meeting of The Society for Surgery of the Alimentary Tract, Atlanta, Georgia, May 20–23, 2001 (oral presentation). Supported by grant RO1-DK442 79–07 from the National Institutes of Health.     

The expression of TGF-βmRNA and its biological function in rat osteoblasts with exposure to raloxifene

Objective:To investigate the effects of raloxifene and estradiol on the proliferation,differentiation and the expression of transforming growth factor-β(TGF-β)of osteoblasts in vitro.Methods:Different doses of raloxifene and estradiol were added into the medium of the second generation osteoblasts obtained from the skull of newborn SD rats.The following parameters including cell proliferation,activity of alkaline phosphatase(ALP),the levels of type Ⅰ collagen(Col-I)mRNA and TGF-β1 mRNA in different groups were measured and analyzed.Results:Raloxifene and 17-βestradiol(17-β E2)showed no significant effect on stimulating the proliferation of osteoblasts(P>0.05 vs the control).However,raloxifene could significantly improve ALP activity and Col-I mRNA expression in high consistency group(P<0.01)in dose-dependent manner.Raloxifene group in 10-8 mol/L increased the expression of TGF-β1 mRNA(vs the control,P<0.05).Estradiol significantly increased ALP activity,Col-I mRNA expression and TGF-β1 mRNA expression(P<0.05 or P<0.01 vs the control).Conclusions:Both of raloxifene and estradiol could stimulate differentiation of osteoblasts and expression of bone matrix,but showed no effect on proliferation of cultured osteoblasts.The expressions of TGF-β1 mRNA were different,which might imply their different mechanisms by means of estrogen receptor.     

姜黄素拮抗瘦素损伤小鼠足细胞的实验研究

目的：探讨瘦素对小鼠足细胞的损伤效应,及姜黄素拮抗这一效应的可能.方法：小鼠足细胞分为四组,分别为空白对照组,瘦素刺激组,姜黄素加瘦素组,及姜黄素对照组.mRNA检测采用实时定量PCR方法,蛋白检测采用免疫印迹法.结果：瘦素（15 ng/ml）能显著抑制足细胞nephrin,podocin,podoplanin,podocalyxin表达,与对照组比较,mRNA表达的抑制率分别为31％、28％、33％及29％;蛋白质表达的抑制率分别为26％、30％、24％及32％,P均＜0.05.加入姜黄素（5μmol/L）后,上述足细胞标志蛋白均被上调,与瘦素组比较,mRNA表达分别上调1.25、1.15、1.34及1.20倍;蛋白质表达分别上调1.15、1.31、1.14及1.32,P均＜0.05.瘦素（15 ng/ml）能显著活化Wnt/β-catenin信号通路,与对照组比较,Wnt1、Wnt2b和Wnt6及其下游蛋白β-catenin的mRNA表达分别上调1.54、1.29、1.52及1.25倍,P均＜0.05.β-catenin的蛋白磷酸化水平抑制率为17％,P＜0.05.加入姜黄素（5μmol/L）后,上述信号通路分子均被抑制,与瘦素组比较,Wnt1、Wnt2b和Wnt6及β-catenin的mRNA表达抑制率分别为12％、12％、22％及10％,P＜均0.05.磷酸化的β-catenin上调1.13倍.结论：瘦素能通过Wnt/β-catenin信号通路损伤足细胞,而姜黄素能逆转这一反应.     

山莨菪碱对离体大鼠腹主动脉舒缩张力的作用及其机制

目的 观察山莨菪碱对离体大鼠腹主动脉舒缩张力的作用,探讨其作用机制.方法 采用Wistar大鼠离体腹主动脉环(长4～5 mm的血管)灌流技术,观察山莨菪碱累积浓度(每5 min加入山莨菪碱,使营养液中山莨菪碱浓度分别达3×10-6,10-5,3×10-5,10-4,3×10-4 mol/L)对单剂量苯肾上腺素(PE)(10-5 mol/L)预处理的血管舒张作用的影响,及不同浓度山莨菪碱(10-6,10-5,10-4 mol/L)预孵对累积浓度PE(每5 min加入PE,使营养液中PE浓度分别达10-8,3×10-8,10-7,3×10-7,10-6,3×10-6 mol/L)收缩血管作用的影响,分别运用一氧化氮合酶抑制剂左旋硝基精氨酸甲酯(L-NAME)(100 μmol/L)(对照组加入等容积的蒸馏水)和KATP通道阻断剂格列苯脲(Gly)(100 μmol/L)(两对照组分别加入等容积的蒸馏水及溶剂二甲基亚砜DMSO)处理血管环,观察对山莨菪碱舒张血管的抑制作用.结果 山莨菪碱累积浓度对单剂量PE预收缩内皮完整及去内皮血管环均有舒张作用,最大舒张分别为(78.6±6.9)%和(65.76±11.39)%.程度相似,差异无统计学意义(P＞0.05).不同浓度山莨菪碱预孵对累积浓度PE收缩血管环作用呈浓度依赖性抑制,浓度越大,抑制作用越明显.用阻断剂L-NAME及Gly处理后,L-NAME可阻断山莨菪碱的舒张血管(内皮完整)作用(P＜0.05),而Gly对去内皮血管具阻断作用,但只在2 min时与对照组差异有统计学意义(P＜0.05).结论 山莨菪碱具有内皮及平滑肌依赖性舒张血管作用,即通过内皮上一氧化氮-环鸟苷酸(NO-cGMP)及平滑肌上α受体发挥作用.山莨菪碱抑制血管收缩主要通过ATP依赖性K+通道发挥作用.     

雌激素对基质成纤维细胞和乳腺癌细胞系SDF-1水平的影响

目的：探讨雌激素对基质成纤维细胞和乳腺癌细胞SDF-1的影响,从而揭示雌激素是否通过SDF- 1/CXCR4信号通路影响乳腺癌的生物学特性。方法 选取基质成纤维细胞MRC5和乳腺癌细胞MCF-7为研究对象,分成对照组、雌激素组和雌激素+雌激素受体阻断组,分别加入不同生理浓度的17-β雌二醇作用一定时间以及同一浓度17-...     

胰岛素对脂肪干细胞生长因子分泌功能的影响

Objective To study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells (ADSCs). Methods ADSCs were isolated from human ab-dominal adipose tissue and cultured. The immunophenotype and adipose induced-differenciation were identi-fied, and the third generation cells were collected. The collected cells were assigned to 1 × 10-8, 1 × 10-7, 1 × 10-6 mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs' nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry. Results The secretion amounts of VEGF and HGF of ADSCs in 1 × 10-8 and 1 × 10-7 mol/L insulin groups [ (471±41, 762±66 ng/L), (643±64, 930±67 ng/L) , respectively ] were significantly higher as compared with those in control group (286±47, 577±84 ng/L) ( P <0.05 orP <0.01). No change occurred in the secretion amount of VEGF and HGF of ADSCs in 1±10-6 mol/L insulin group ( P >0.05 ). The supernatant fluid of ADSCs' nutrient medium of 1 ± 10-8 ,1 ± 10-7 mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1 ± 10-7 mol/L group ( P < 0.05 or P < 0. 01 ). Conclusions Insulin in the concentrations of 1 ± 10-8 and 1 ± 10-7 mol/L can notably promote ADSCs' function of secreting VEGF and HGF.     

Effect of Bisphenol A on steroid production in human granulosa cells in vitro

Objective:To study the effects of environmental estrogen-like chemical bisphenol A(BPA)on ovarian function with the model of the cultured human granulosa cells in vitro.Methods:The granulosa cells from healthy women were collected and cultured in DMEM with 10% heat-inacti-vated fetal bovine serum.BPA,final concentration 10-7 to 10-4 mol/L,was added to the cell-culture medium and treat cells for 48 hours.The levels of estrogen and progesterone in the supernatant of the cultured cells were meas-ured by DELFIA.Total RNA was extracted from the cultured cells.The expression levels of P450 scc and P450 arom mRNA were measured by RT-PCR.Results:The cultured human granulosa cells could express high levels of P450scc and P450arom.The levels of estrogen and progesterone increased after BPA treatment,expression of P450 arom mRNA was reduced significantly at 10-5 to 10-4mol/L of BPA,but the expression of P450 scc mRNA was increased at 10-6 to 10-5 mol/L of BPA.The levels of estrogen and progesterone in the supernatant of the cultured cells were not affected significantly by BPA at the same concentrations incubated for 48 hours.Conclusion:BPA could affect steroid hormone synthesis and transformation in granulosa cells,such as the ex-pression of P450 scc and P450 atom,so it can affect ovarian function.Although we did not find a significant effect of BPA on the final estrogen and progesterone levels in this studying model,noxious effects of BPA on ovarian function may be exist in the human granulose cells.     

降钙素基因相关肽混合去甲肾上腺素对大鼠心肌细胞L-型钙电流的影响

目的 探讨降钙素基因相关肽(CGRP)混合去甲肾上腺素(NE)对大鼠心肌细胞L-型钙电流(ICa-L)的影响.方法 SD大鼠,雌雄不拘,体重260～280 g,急性分离其心肌细胞,用全细胞膜片钳方法记录ICa-L.采用随机数字表法,将心肌细胞随机分为3组(n=6):CGRP组用含CGRP 1×10-7mol/L的台氏液灌流;NE组用含NE 1×10-6mol/L的台氏液灌流;CGRP+NE组(CN组)用含NE 1×10-6mol/L+CGRP 1×10-7mol/L的台氏液灌流.各组均灌流1 min,流速2 ml/min;然后用台氏液洗脱1 min.于灌流前1 min、灌流1 min和洗脱1 min时,记录ICa-L峰值,并绘制CGRP或NE灌流后ICa-L的电流-电压曲线.结果 CGRP可抑制心肌细胞ICa-L(P<0.05).NE可促进心肌细胞ICa-L(P<0.05).与CN组比较,灌流1 min时CGRP组ICa-L降低,NE组ICa-L升高(P<0.05).CGRP使心肌细胞ICa-L的电流-电压曲线明显上移;NE使心肌细胞ICa-L的电流一电压曲线明显下移.结论 CGRP可削弱NE对大鼠心肌细胞ICa-L的促进作用.

Abstract：

Objective To investigate the effects of calcitonin gene-related peptide (CGRP) combined with norepinephrine (NE) on L-type calcium current (LCa-l) in rat ventricular myocytes. Methods Ventricular myocytes were isolated from SD rats (weighing 260-280 g) by retrograde perfusion of the heart via the aorta with an enzyme-containing solution as previously described. Whole-cell patch-clamp recording was made using Axopatch 200B amplifier. The cells were perfused for 1 min with Tyrode solution containing CGRP 1 × 10-7 mol/L (group CGRP) , NE 1 × 10-6 mol/L (group NE), or CGRP 1 × 10-7 mol/L + NE 1 × 10-6 mol/L (group CN) and again washed with Tyrode solution. ICa-L was recorded 1 min before and 1 min after the cells were perfused and 1 min after the cells were washed. I-V curve of ICa-L was made after the cells were perfused with solution containing CGRP or NE alone. Results CGRP significantly inhibited the peak of ICa-L, while NE significantly promoted the peak of ICa-L(P < 0.05) . The peak of ICa-L was significantly decreased 1 min after the cells were perfused in group CGRP,while increased 1 min after the cells were perfused in group NE compared with group CN ( P < 0.05). CGRP made the I-V curve of ICa-L move up-ward, while NE made the I-V curve of ICa-L move down-ward. Conclusion CGRP can weaken the promotion of ICa-L induced by NE in rat ventricular myocytes.     

一氧化氮抑制ghrelin诱导的生长激素腺瘤GH3细胞生长激素分泌和细胞增殖及其机制

目的 检测一氧化氮(NO)对ghrelin诱导的大鼠GH3细胞的生长激素(GH)分泌和细胞增殖的影响,探讨NO的作用机制.方法 首先应用ghrelin在不同浓度分别作用2 h;应用ghrelin在工作浓度1×10-7mol/L,分别作用不同时间,检测对GH3细胞GH分泌的影响;然后检测NO的供体(SNAP,1×10-5 mol/L)和NO合成酶的抑制剂(NAME,1×10-5mol/L)对ghrelin诱导的GH分泌和细胞增殖的影响;用酶联免疫吸附试验(ELISA)方法检测GH水平,噻唑蓝(MTT)比色法检测细胞增殖,Western免疫印迹法检测细胞内信号通路蛋白的活性变化.结果 ghrelin刺激GH3细胞分泌GH呈时间和浓度依赖性(P＜0.01),ghrelin明显刺激GH3细胞的增殖(P＜0.05);SNAP可抑制基础的和ghrelin刺激的GH分泌(P＜0.01),对细胞增殖也有明显影响(P＜0.01),而NAME对GH的分泌和细胞增殖无影响;ghrelin可诱导细胞外信号调节激酶(ERK)的活化,SNAP可以抑制这种效应.结论 NO抑制ghrelin诱导GH3细胞GH分泌和细胞增殖,其机制可能阻断了ghrelin激活的ERK信号通路.     

中脑胶质细胞神经营养因子前多肽对大鼠神经元的保护作用

目的 观察中脑胶质细胞神经营养因子(MANF)前多肽对由谷氨酸所致大鼠下丘脑神经元损伤的保护作用及量效关系.方法 将培养Wistar大鼠下丘脑神经元分为正常对照组(CG)、谷氨酸组(GG)、MANF前多肽+谷氨酸组(MGG1～4,MANF前多肽浓度分别为1×10-7、1×10-8、1×10-9、1×10-10 mol/L).各组神经元培养24 h后,分别检测培养细胞死亡率、神经元轴突长度、最长树突长度和树突数量.结果 GG组神经元死亡率为(0.342±0.055)%,显著高于CG组的(0.065±0.017)%(P＜0.01);MGG1组细胞死亡率最低(0.157±0.038)%,并随MANF浓度降低依次增高,MGG4的细胞死亡率最高,达(0.285±0.035)%,MGG1组神经元轴突长度、最长树突长度和树突数量均为最高,分别为(133.5±21.6)μm、(128.4±18.6)μm、(1.84±0.27)个/细胞,并随MANF浓度降低依次减少,MANF组细胞死亡率和神经元形态均较GG组明显改善(P＜D.05),但仍较CG组较差(P＜0.05),MANF前多肽浓度越高者,神经元情况改善越好.结论 MANF前多肽对谷氨酸所致神经元损伤有显著的保护作用,且在1×10-7～1×10-10 mol/L浓度范围内随浓度增高,保护效果增强.

Abstract：

Objective To observe the protective effects of pre-mesencephalic astrocyte-derived neurotrophic factor ( MANF) polypeptide on glutamate-induced rat hypathalamus neuron injury and the dose-effect relationship. Methods Cultured Wistar rats hypothalamic neurons were divided into normal control group (CG) , glutamate group (GG), pre-MANF polypeptide + glutamate groups (MGGM, preMANF polypeptide concentrations were 1 × 10 -7 , 1 × 10 -8, 1 × 10-9 and 1 ×10 -l0 mol/L) randomly. After culture for 24 h, mortality of neurons was tested, and length of neuron axons, the maximum length and number of dendrites were measured. Results The neuronal death rate in GG group was (0. 342 ± 0. 055 ) % , significantly higher than that in CG group (P ＜ 0. 01) . The cell mortality, neuron axon length,the maximum length and number of dendrites in MGG1 groups were (0. 157 ±0.038)% , (133. 5 ±21.6) μm, (128. 4 ± 18. 6) μm, 1. 84 ±0. 27 per cell respectively, and significantly improved as compared with GG group (P ＜ 0. 05) , but were still worse than in CG group (P ＜0. 05). Higher the pre-MANF polypeptide concentration was, better the neuron figure showed. Conclusion Pre-MANF polypeptide has a significant protective effect on neuron injury caused by glutamate, and the protective effect is stronger as the concentration higher within 1 × 10-7-1×10 -10 mol/L.     

c-myc对大鼠黄体细胞人绒毛膜促性腺激素诱导的孕酮和雌二醇生成的影响

应用离体细胞体外孵育法研究了反义ｃ ｍｙｃ寡脱氧核苷酸（反义ｃ ｍｙｃＯＤＮ）对大鼠黄体细胞人绒毛膜促性腺激素（ｈＣＧ）诱导的孕酮（Ｐ）和雌二醇（Ｅ２）生成的影响及其与外源性ｃＡＭＰ和Ｃａ２＋的关系。结果发现，反义ｃ ｍｙｃＯＤＮ能呈剂量相关方式抑制黄体细胞ｈＣＧ诱导的Ｐ和Ｅ２的生成。在浓度分别为１０和５μｍｏｌ／Ｌ时的抑制作用即具有显著意义（Ｐ＜０．０５）；而无义ｔａｔ寡脱氧核苷酸则无此作用。反义ｃ ｍｙｃＯＤＮ对黄体细胞ｈＣＧ诱导的Ｐ和Ｅ２产生的抑制作用能被加入１０－４ｍｏｌ／Ｌ二丁酰ｃＡＭＰ逆转，钙离子通道阻断剂维拉帕米对此种抑制作用具有协同效应。结果提示，ｃ ｍｙｃ癌基因参与黄体细胞ｈＣＧ诱导的Ｐ和Ｅ２生成的调控。     

内皮素对大鼠睾丸间质细胞睾酮生成的影响

本研究采用大鼠睾丸间质细胞体外培养的技术 ,观察了内皮素 1对离体间质细胞睾酮分泌的影响。研究发现 ,10 -9mol/L的ET 1可显著抑制间质细胞睾酮的基础分泌 (P <0 .0 5 ) ,并且ET 1对人绒毛膜促性腺激素 (hCG)刺激睾丸间质细胞睾酮分泌也有抑制作用 ,其有效抑制浓度为 10 -10 mol/L(P <0 .0 5 )。本实验结果提示 ,ET 1呈剂量依赖性抑制睾丸间质细胞睾酮的基础分泌和hCG诱导的分泌 ,ET 1可能为睾丸内的一种局部调节多肽     

Interaction between porcine granulosa and thecal cells in steroidogenesis in an amnion dual chamber culture system

IntroductionInordertoinvestigatetheovariansteroidogenicfunction,scientistsusedtocul-turegranulosacellsorthecalcellsinvit-roLl~5].Thereisagrowingawarenessthatinvivogranulosaandthecalcellsintheovaryareseparatedjustbyabasementmetnbrane.Bothtypesofcellsareimportantduringthefollicledevelopmentandovummaturation,interact-ingoneachother.The"twocells"theoryhasbeenknownformorethantwentyyears['].Thesinglecellculturewitheithergranulosacellsorthecalcellswasnotaperfectmodelforinvitrostudy,Itisnecessarytoe…     

姜黄素保护大鼠肾小管上皮细胞对抗氧化应激引起的损伤作用

目的 探讨姜黄素对氧化应激诱导大鼠近端肾小管上皮细胞 (NRK-52E) 损伤作用的影响。 方法 用不同浓度的H2O2处理NRK-52E细胞,建立氧化应激损伤NRK-52E的实验模型。应用Hoechst 33258染色法观察凋亡细胞的形态学改变;PI染色流式细胞仪检测细胞凋亡率;Western印迹法检测Bcl-2蛋白的表达。 结果 H2O2在100~500 μmol/L浓度范围内处理NRK-52E细胞24 h,呈浓度依赖性地增加细胞的凋亡率;500 μmol/L H2O2能显著抑制NRK-52E细胞Bcl-2的表达（P < 0.05）。20 μmol/L和40 μmol/L姜黄素能显著阻断H2O2对NRK-52E细胞的致凋亡作用[（32.9±8.1）%、（22.23±9.3）%比（72.7±10.5）%,均P < 0.05],并能显著拮抗H2O2对Bcl-2蛋白表达的下调作用（P < 0.05）。40 μmol/L姜黄素本身也能上调Bcl-2蛋白的表达（P < 0.05）。 结论 姜黄素能保护NRK-52E细胞对抗氧化应激引起的细胞凋亡,此肾小管上皮细胞保护作用可能与其抑制H2O2对Bcl-2蛋白表达的下调作用有关。     

载银珊瑚羟基磷灰石的抑菌活性与细胞毒性研究

目的 评价负载不同浓度银离子后的珊瑚羟基磷灰石(CHA)对大肠杆菌和金黄色葡萄球菌的抑菌效果与细胞毒性.方法 采用真空冷冻干燥法制备1×10-2～1×10-5 mol/L Ag+浓度的载银CHA(Ag-CHA),然后采用琼脂平板抑菌法评价Ag-CHA对大肠杆菌和金黄色葡萄球菌的抑菌效果,并测量24 h后的抑菌圈直径.采用甲基噻唑基四唑(MTT)法评价8×10-5mol/L的Ag-CHA在2、4、7d对L929小鼠成纤维细胞的毒性作用. 结果 载银浓度为1×10-2～8×10-5mol/L的Ag-CHA对大肠杆菌和金黄色葡萄球菌的抑菌圈直径均大于12 mm,有明显的抑菌作用;而载银浓度低于8×10-5mol/L的Ag-CHA无抑菌作用.载银浓度低于8×10-5mol/L的Ag-CHA对L929细胞无毒性作用;8×10-5mol/LAg-CHA浸提液作用细胞2、4、7 d后,细胞生长状态良好,相对增殖率均大于90%,毒性等级为0或1级.结论 最佳载银浓度为8×10-5mol/L,该条件下的Ag-CHA有良好的抑菌作用,且无细胞毒性.