碱性核蛋白及增殖性核抗原Ki-67在基底细胞癌中的表达

碱性核蛋白(basonuclin)是一种锌指样蛋白,在具有分化能力的细胞核中表达,Ki-67是一种增殖性核抗原,是早期评估肿瘤细胞增殖的方法之一,在增殖细胞的核内表达.基底细胞癌(BCC)是临床上较常见恶性程度较低的皮肤肿瘤,其细胞来源仍有争议,预后有很大的差异.我们研究碱性核蛋白和Ki-67在BCC中的表达类型及其意义.

氧化苦参碱对银屑病模型雌激素周期小鼠阴道上皮细胞增殖和凋亡的影响

目的:探讨氧化苦参碱对银屑病模型雌激素周期小鼠阴道上皮细胞增殖和凋亡的影响。方法:采用注射雌激素刺激小鼠阴道上皮组织增生的方法复制银屑病小鼠动物模型,同时给予不同剂量的氧化苦参碱干预,实验10 d后,应用免疫组化法和末端转移酶介导的缺口末端标记(TUNEL)技术分别检测各组小鼠阴道上皮细胞细胞增殖性核抗原(PCNA)表达水平和凋亡情况。结果:与正常组相比,模型组PCNA表达显著增加(P<0.01),细胞凋亡指数明显增加(P<0.01),氧化苦参碱80、40和20mg/kg组PCNA表达显著减少(P<0.01),凋亡指数显著增加(P<0.01)。结论:氧化苦参碱对实验性银屑病具有保护作用,该作用可能与其降低表皮细胞PCNA表达水平,促进表皮细胞凋亡有关。

血小板源性生长因子对体外培养子宫肌瘤细胞增殖的影响

目的: 研究血小板源性生长因子 (Platete-DerivedGrowthFactor, PDGF) 对体外培养的子宫肌瘤细胞和子宫平滑肌细胞增殖性核抗原 (PCNA) 和细胞内前胶原α1多肽的表达的影响。方法: 用基因重组人PDGF-AB (10ng/ml) 作用体外培养的子宫肌瘤细胞, 以邻近正常子宫肌层细胞作对照, 用RT-PCR法从mRNA水平检测在有无PDGF-AB作用下, PCNA、前胶原α1多肽 (Ⅰ、Ⅲ) 表达的差异; 用免疫组化SABC法从蛋白水平显示PDGF对PCNA表达的影响。结果: 两种细胞受PDGF-AB刺激后, PCNA、collagenα1 (Ⅰ) 表达均有增加, 但两者在肌瘤细胞中表达的增加显著高于子宫平滑肌细胞 (P<0. 01); 而collagenα1 (Ⅲ) 在子宫肌瘤细胞和子宫平滑肌细胞中的变化无显著性差异。结论: PDGF在体外可同时促进子宫肌瘤细胞增殖和细胞外基质分泌的增加, 从而使肌瘤组织不断增长。

当归多糖对豚鼠银屑病样皮损中PCNA表达的影响

目的研究当归多糖对豚鼠耳部银屑病样皮损中细胞增殖性核抗原(PCNA)表达水平的影响。方法50g/L普萘洛尔乳剂外涂豚鼠耳背皮肤,制备银屑病样模型,然后随机分成模型对照组(I)、低剂量组(II)、中剂量组(III)、高剂量组(IV),依次给予生理盐水、100mg/kg,200mg/kg,400mg/kg当归多糖腹腔注射。应用免疫组化法分别检测各组治疗前后皮损中PCNA表达水平。结果造模后豚鼠耳部上皮PCNA阳性表达率与涂药前相比明显增加(P<0.05)。II,III,IV组治疗后皮损中PCNA阳性表达率较治疗前均明显降低(P<0.05)。结论当归多糖对普萘洛尔诱发的豚鼠耳背银屑病样模型有治疗作用,可能与其降低皮损中PCNA表达水平有关。

细胞凋亡在脂质肾损害中的作用

目的 :研究细胞凋亡在脂质肾损害中的作用。方法 :高脂组大鼠饲以 4%胆固醇及 1%胆酸钠的鼠料 ,正常组大鼠饲以普通鼠料 ,分别于 4周及 8周时处死大鼠。采用TUNEL法检测肾组织中凋亡细胞 ,增殖性核抗原及层粘连蛋白在肾组织中的表达用免疫组化法检测 ,血脂及尿蛋白检测用生化法。结果 :与正常对照组相比 ,4周时高脂组增殖性核抗原表达增加 ,8周时高脂组凋亡细胞明显增多 (P <0 0 5 )。 8周时高脂组大鼠肾组织中层粘连蛋白质表达增加 ,2 4h尿蛋白定量明显增多 (P <0 0 5 ) ,肾小球、肾小管及其肾小管间质细胞凋亡指数与 2 4h尿蛋白定量呈正相关 (r =0 978,P <0 0 0 1;r =0 986 ,P <0 0 0 1;r =0 975 ,P <0 0 0 1)。结论 :细胞凋亡在脂质肾损害中起着重要作用 。

细胞周期蛋白依赖性激酶抑制剂p27kip1在翼状胬肉中的表达

目的 探讨细胞周期蛋白依赖性激酶抑制剂p2 7kip1在翼状胬肉中的表达及其在翼状胬肉发病机制中的作用。方法 采用免疫组化法对手术切除的翼状胬肉标本和正常结膜组织进行增殖性核抗原及细胞周期蛋白依赖性激酶抑制剂 p2 7kip1蛋白检测。结果 正常结膜组织中和翼状胬肉中增殖性核杭原表达分别为 0 .2 6 8± 0 .0 2 1和 0 .4 75± 0 0 4 6 (P<0 .0 5 ) ,p2 7kip1分别为 0 .4 2 3± 0 .0 31和 0 .2 4 2± 0 0 17(P <0 .0 5 )。结论 结膜中细胞周期蛋白依赖性激酶抑制剂p2 7kip1的低表达可能与翼状胬肉的发生、发展有关。

食源性L-精氨酸对高胆固醇血症大鼠肾损害的影响

目的 探讨L-精氨酸在脂质肾损害中的作用。方法 26只雄性Wistar大鼠随机分为3组，正常对照组饲以普通饲料。高脂组饲以含4％胆固醇和1％胆酸钠的鼠料，L-精氨酸组除饲以高胆固醇鼠料外饮用含1％精氨酸的自来水。8周后生化法测24小时尿蛋白定量、肌酐清除率、血脂及血丙二醛、肾皮质丙二醛，免疫组化法测肾组织中层粘连蛋白、增殖性核抗原表达及巨噬细胞浸润情况。结果 8周时高胆固醇血症组及L一精氨酸组血清总胆固醇、低密度脂蛋白胆固醇浓度较正常组高(P<0．01)，前二者尿蛋白定量较正常组增高(P<0．05)。高胆固醇血症组肌酐清除率较正常组明显下降(P<0．05)，L-精氨酸组较正常组明显上升(P<0．05)。L-精氨酸组层粘连蛋白表达较正常组及高胆固醇血症组表达增强，增殖性核抗原阳性细胞较正常组及高胆固醇血症组增多。L-精氨酸组及高胆固醇血症组肾组织中浸润的巨噬细胞数较正常组增多(P<0．05)，前二者无显著差异。结论 食源性L-精氨酸可引起高胆固醇血症大鼠肾组织增生，不能改善高胆固醇血症大鼠早期肾损害．

丙酸氯倍他索对咪喹莫特诱导银屑病小鼠中PCNA表达的影响

目的通过丙酸氯倍他索处理咪喹莫特诱导的银屑病小鼠模型,研究其对PCNA过度表达的影响。方法将30只雌性BALB/c小鼠随机分成3组:正常组、模型组(咪喹莫特组)、丙酸氯倍他索处理组,每组10只。所有小鼠实验前48 h背部剃毛。正常组不作处理;模型组每天上午涂抹咪喹莫特65 mg/只(背部涂药55 mg,双耳10 mg);丙酸氯倍他索处理组每天上午给咪喹莫特65 mg/只(背部55 mg,双耳10 mg),4 h后涂抹丙酸氯倍他索55 mg/只(背部45 mg,双耳10 mg),连续给药9 d。第10天取所有小鼠背部皮肤组织采用HE染色病理切片并观察,免疫组化法检测小鼠皮肤中PCNA表达情况。结果模型组小鼠皮肤皮损严重,银屑病症状明显;丙酸氯倍他索治疗后银屑病症状减轻,表皮厚度明显薄于模型组,皮肤PCNA的表达较模型组减少。结论丙酸氯倍他索的抗银屑病作用机制可能与抑制PCNA在皮肤组织中过度表达有关。

Role of TGF-β1/Smads pathway in carotid artery remodeling in renovascular hypertensive rats and prevention by Enalapril and Amlodipine

Objective To investigate the role of transforming growth factor-β1 （TGF-β1）, Smad2/3 and Smad7 expressions in carotid artery remodeling in renovascular hypertensive rats, and also the therapeutic effect of Enalapril and Amlodipine. Methods The renovascular hypertensive rat （RHR） models with ＂two-kidney and one-clip＂ were established, including model group （n = 6）, sham-operated group （n = 6）, Enalapril group （10 mg/kg per day, n = 6）, Amlodipine group （5 mg/kg per day, n = 6） and combination group （Amlodipine 2.5 mg/kg per day ＋ Enalapril 5mg/kg per day, n = 6）. The medication were continuous administrated for six weeks. Carotid artery morphological and structural changes in the media were observed by HE staining, Masson staining and immuno histochemical staining. Media thickness （MT）, MT and lumen diameter ratio （MT/LD）, and the expression levels of media a-smooth muscle actin （α-actin）, proliferating cell nuclear antigen （PCNA）, TGF-β1, phosphorylated Smad2/3 （p-Smad2/3） and Smad7 in carotid arteries were measured. Results The media of carotid arteries in RHR model group was significantly thickened, the volume of smooth muscle cell was increased, and the array was in disorder; MT, MT/LD, the proliferation index of smooth muscle cell and collagen fiber area percentage of carotid arteries in the model group were significantly higher than those in the sham-operated group （P 〈 0.01）. Compared to sham-operated group, the model group had significantly higher expressions of TGF-β1 and p-Smad2/3 （P 〈 0.05） and lower Smad7 expression. Both Enalapril and Amlodipine improved smooth muscle hypertrophy and collagen deposition, reduced RHR carotid MT, MT/LD, proliferation index of smooth muscle cell, collagen fiber area percentage and the expressions of TGF-β1 and p-Smad2/3 （P 〈 0.05）, increased Smad7 expression （P 〈 0.05）. Moreover, the combination treatment of Enalapril and Amlodipine had significantly better effects than single Amlodipine group （P 〈 0.05）, but not single Enalapril group. Conclusions TGF-β1/Smads pathway may participate in the mechanism of carotid artery remodeling in RHR; the role of Amlodipine and Enalapril in inversing carotid artery remodeling may be related to the change of TGF-β1/Smads pathway, the combination treatment of Amlodipine and Enalapril had better effects than single administration of Amlodipine.

Proliferation in rat gastric mucosal cells induced by chronic ethanol feeding through the ROS/BMK1 pathway

Objective: To investigate the correlation between the gastric mucosal cell proliferation and low-concentration alcohol intake in a chronic drinking rat model, and to investigate the possible role of ROS/BMK1 pathway in this process. Methods: SD rats were randomly divided into 4 groups: control group, administered with tap water; ethanol group, with 6% ethanol in the drinking water; quercetin group, with quercetin (100 mg/kg) by intragastric gavage twice a day; ethanol+quercetin group, administered with quercetin combined with 6% ethanol. The cell proliferation in rat gastric mucosa was analyzed by flow cytometery and proliferating cell nuclear antigen (PCNA) immunohistochemical staining. Activation of ERKs and BMK1 was evaluated by the expression and phosphorylation of these kinases using Western Blot analysis. Results: Compared to the controls, the cell proliferation in gastric mucosa of rats exposed to the ethanol for 7 d was enhanced, and the activation of BMK1 was also increased in this period. Otherwise quercetin, as a free radical scavenger, attenuated increased cell proliferation and activation of BMK1 in rat stomach treated with ethanol. However, no changes of ERKs expression and phosphorylation occurred in the rats in all groups. Conclusion: These results suggested that the ROS and BMK1 activation may be a central mechanism, which underlies cell proliferation in rat gastric mucosa stimulus with the chronic low-concentration ethanol.

How to use progestin in hormone replacement therapy: an animal experiment

Objective To determine whether continuous or cyclic hormone replacement therapy (estrogen and progestogen) is better.Methods One hundred and forty Sprague-Dawley rats were randomly divided into seven groups. The 1st and 2nd groups were normal estrous and ovariectomy (OVX) controls. Treatment of the other groups imitated the clinical regimen (continuous and cyclic) with estradiol valerate (E2V) and medroxy progesterone (MPA) in different ratios of combination. The rats were sacrificed and sections of uterus were stained with HE and histochemical metheds to detect mitosis and proliferating cell nuclear antigen (PCNA), respectively. The mitotic index (MI) and PCNA index were calculated.Results The MI and PCNA index were similar in luminal and glandular cells. Both markers were low in the two control groups. When E2V was given for 1 to 6 days, both the MI and PCNA index increased with duration of treatment. When MPA was added, both markers were reduced to a very low level. In the continuous regimen, both markers decreased as the MPA dosage increased. The ratio of E2V∶MPA=1∶0.5 was enough to suppress markers to a low level similar to that of normal estrous rats. A further increase in the ratio to 1∶1.0 showed no further decrease in PCNA index. In the cyclic regimen, MPA was added for the last 5 days. The mitotic index reached a significantly low level near 0 in all ratios, but the PCNA index in each subgroup was still as high as the positive control, even though the dosage of MPA was increased several times to 1∶8.0. When MPA was added for the last 10 days, the PCNA index at a ratio of 1∶4.0 could be reduced to a low level.Conclusion The results of this study suggest that the continuous regimen was better than the cyclic regimen in postmenopausal hormone replacement therapy (HRT). Progestin should be given for at least 10 days in the cyclic regimen.