Effects of platelet-derived growth factor and interleukin-10 on Fas/Fas-ligand and Bcl-2/Bax mRNA expression in rat hepatic stellate cells in vitro

AIM: To investigate the effects of platelet-derived growth factor(PDGF) and interleukin-10 (IL-10) on Fas/Fas-ligand and Bcl-2/Bax mRNA expressions in rat hepatic stellate cells.METHODS: Rat hepatic stellate cells (HSCs) were isolated and purified from rat liver by in situ digestion of collagenase and pronase and single-step density Nycodenz gradient.After activated by culture in vitro, HSCs were divided into 4 groups and treated with nothing (group N), PDGF (group P),IL-10 (group I) and PDGF in combinalJon with IL-10 (group C),respectively. Semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis was employed to compare the mRNA expression levels of Fas/FasL and Bcl-2/Bax in HSCs of each group.RESULTS: The expression levels of Fas between the 4 groups had no significant differences (P>0.05). FasL mRNA level in normal culture-activated HSCs (group N) was very low.It increased obviously after HSCs were treated with IL-10(group I) (0.091±0.007 vs 0.385±0.051, P<0.01), but remained the low level after treated with PDGF alone (group P)or PDGF in combination with IL-10 (group C). Contrast to the control group, after treated with PDGF and IL-10, either alone or in combination, Bcl-2 mRNA expression was downregulated and Bax mRNA expression was up-regulated, both following the turn from group P, group I to group C.Expression of Bcl-2 mRNA in group C was significantly lower than that in group P (0.126±0.008 vs0.210±0.024, P<0.01).But no significant difference was found between group C and group I, as well as between group I and group P (P>0.05).Similarly, the expression of Bax in group C was higher than that in group P (0.513±0.016 vs0.400±0.022, P<0.01).No significant difference was found between group I and group P (P>0.05). But compared with group C, Bax expressions in group I tended to decrease (0.449±0.028 vs 0.513±0.016,P<0.05).CONCLUSION: PDGF may promote proliferation of HSCs but is neutral with respect to HSC apoptosis. IL-10 may promote the apoptosis of HSCs by up-regulating the expressions of FasL and Bax and down-regulating the expression of Bcl-2, which may be involved in its antifibrosis mechanism.

Nimodipine Modulates Bcl-2 and Bax mRNA Expression after Cerebral Ischemia

In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA. The increased ratio of Bcl-2 and Bax mRNA may contribute to the anti-apoptic effect of nimodipine. The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new strategy to manage neuronal damage.

Influence of Gossypol on Ultrastructure and mRNA Expression of Bax and Bcl-2 in Mice Testis

The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups： control group, L-group （30 mg-kgt. d）, M-group （60 mg·kg-1 ·d） and H-group （120 mg·kg-1· d） and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose （SCC） or SCC （control group） for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased （p〈0.05） in L-group and then significantly decreased （p〈0.05, p〈0.01） in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased （p〈0.05, p〈0.01） than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease （p〈0.05） compared with that in M-group. On the other hand, mRNA expression of Bax significant increased （p〈0.05,p〈0.01） in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01） in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.

Effect of basic fibroblast growth factor and danshen on bcl-2 and p53 mRNA expression in the brain of rats exposed to repeated, high, positive acceleration (+Gz)

BACKGROUND： Both animal experiments and clinical studies have shown that basic fibroblast growth factor （bFGF） and danshen （Salvia miltiorrhiza） can exhibit protective effects on ischemia-reperfusion cerebral injury. OBJECTIVE： To test whether bFGF and danshen can protect cerebral injury induced by exposure to repeated, high, positive acceleration （＋Gz） in an animal model and to analyze the possible mechanisms. DESIGN, TIME AND SETTING： Randomized controlled animal study. The experiment was performed at the Research Center for Molecular Biology, Air-force General Hospital of Chinese PLA from April to August 2000. MATERIALS： A total of 20 clean grade, healthy, Sprague Dawley rats of both genders, weighing （200 ± 15） g, were provided by our experimental animal center. Rats were randomly divided into 5 groups： the control group, ＋Gz exposure group, bFGF group, danshen group, and saline group, with 4 animals per group. bFGF （Beijing Bailuyuan Biotechnology Co. Ltd.） and danshen solution （Shanghai Zhongxi Pharmaceutical Co. Ltd.） were used. METHODS： All rats were fixed on a rotary arm of a centrifugal apparatus （2 m in radius） with their heads oriented towards the center of the apparatus. Except for rats in the control group, the value of ＋Gz exposure was ＋14 Gz with an acceleration rate of 1.5 G/s. The peak force lasted for 45 seconds. ＋Gz exposure was performed three times with intervals of 30 minutes. Rats in the control group received the same ＋Gz procedure, but the G value was ＋1 Gz. Rats in bFGF group and danshen group were intraperitoneally injected with 100 μg/kg bFGF or 15 g/kg danshen solution, respectively, at 30 minutes prior to centrifugation and immediately after centrifugation. Rats in saline group were injected with the same volume of saline. Six hours after exposure, rats were decapitated. One hemisphere was preserved in liquid nitrogen for RNA extraction and the other was processed for apoptosis detection. MAIN OUTCOME MEASURES： mRNA levels of bcl-2 and p53 were measured by semi-quantitative reverse-transcription polymerase chain reaction. Apoptotic cell death was detected by terminal deoxynuleotidyl transferase-mediated dUTP nick end labeling. RESULTS： Changes in mRNA expression of bcl-2 and p53 and apoptotic cells were observed in rat brain six hours after repeated ＋Gz exposures, bFGF and danshen were able block the changes of bcl-2 and p53 expression and inhibit apoptotic cell death. CONCLUSION： The data suggest that apoptosis and changes in bcl-2 and p53 expression in the rat brain can be induced by repeated ＋Gz exposures. Apoptosis is, therefore, one of the molecular mechanisms of brain damage induced by repeated ＋Gz exposures, bFGF and danshen were of the equal potency in preventing brain injury induced by repeated ＋Gz exposures.

子宫内膜癌组织中IGF2BP1mRNA,PEG10mRNA表达及与增殖基因表达的相关性和预后研究

目的研究子宫内膜癌(endometrial carcinoma,EC)组织中胰岛素样生长因子2 mRNA结合蛋白1(insulinlike growth factor 2 mRNA binding protein 1,IGF2BP1)mRNA,父系表达遗传印记基因10(patrilineal expression of genetic imprinting gene 10,PEG10)mRNA表达及与增殖基因表达的相关性及预后。方法选取2017年1月~2019年1月湖北理工学院附属妇幼保健院诊治的100例EC患者。实时荧光定量PCR检测EC癌组织和癌旁组织中IGF2BP1 mRNA,PEG10 mRNA及增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)mRNA,细胞周期素D1(cyclin D1)mRNA,细胞周期蛋白依赖激酶4(cyclin dependent kinase 4,CDK4)mRNA表达。免疫组织化学检测IGF2BP1,PEG10蛋白表达。相关性采用Pearson相关分析。Kaplan-Meier曲线分析不同IGF2BP1,PEG10表达组EC患者的预后差异。COX回归分析EC患者的预后影响因素。结果EC癌组织中IGF2BP1 mRNA(1.84±0.33),PEG10 mRNA(2.12±0.40),PCNA mRNA(3.14±0.42),cyclinD1 mRNA(2.81±0.36),CDK4 mRNA(2.37±0.34)高于癌旁组织(0.78±0.21,0.91±0.25,0.74±0.13,0.67±0.21,0.59±0.18),差异具有统计学意义(t=25.652~54.588,均P<0.05)。癌组织中IGF2BP1(70.00%),PEG10(72.00%)蛋白阳性率高于癌旁组织(100%,9.00%),差异具有统计学意义(χ^(2)=75.000,82.363,均P<0.05)。EC中IGF2BP1 mRNA,PEG10 mRNA表达与PCNA mRNA,cyclinD1 mRNA,CDK4 mRNA表达呈正相关(r=0.562~0.625,均P<0.05)。EC中IGF2BP1 mRNA与PEG10 mRNA表达呈显著正相关(r=0.663,P<0.05)。FIGO分期Ⅲ期、并发淋巴结转移EC癌组织中IGF2BP1(86.49%,87.50%),PEG10(89.19%,90.63%)阳性率高于FIGO分期Ⅰ~Ⅱ期(60.32%,61.90%)、无淋巴结转移(61.77%,63.24%),差异具有统计学意义(χ^(2)=6.863~8.608,均P<0.05)。IGF2BP1阳性组患者三年总体生存率70.00%(49/70)低于阴性组的90.00%(27/30);PEG10阳性组患者三年总体生存率为69.44%(50/72),低于阴性组的92.86%(26/28),差异具有统计学意义(Log-rankχ^(2)=4.133,5.491,P=0.042,0.019)。FIGO分期Ⅲ期(OR=1.449,95%CI:1.148~1.830)、并发淋巴结转移(OR=1.442,95%CI:1.124~1.850),IGF2BP1阳性(OR=1.637,95%CI:1.239~2.163)及PEG10阳性(OR=1.576,95%CI:1.136~1.187)是影响EC患者生存预后的独立危险因素(均P<0.05)。结论EC中IGF2BP1,PEG10表达升高,两者与增殖基因表达呈正相关,是EC预后评估的肿瘤标志物。

Overexpression of sterol carrier protein-2 mRNA in patients with cholesterol gallstones

BACKGROUND: Hypersecretion of biliary cholesterol is believed to be one of the important causes of lithogenic bile. Sterol carrier protein-2 ( SCP2 ) participates in choles- terol trafficking and metabolism and may play a key role in cholesterol gallstone formation. This study was undertaken to investigate the expression of liver SCP2 mRNA in pa- tients with cholesterol gallstone and those patients with non-cholesterol gallstone. METHODS: The expression of liver SCP2 mRNA was studi- ed in 36 patients with cholesterol gallstone and 30 patients with non-cholesterol gallstone by reverse transcription-poly- merase chain reaction (RT-PCR). RESULT: The expression of SCP2 mRNA was increased more significantly in patients with cholesterol gallstone than in patients with non-cholesterol gallstone. CONCLUSION: The SCP2 gene was overexpressed in pa- tients with cholesterol gallstone, indicating that SCP2 may be one of the important causes of cholesterol gallstone.

Effects of L-Tetrahydropalmatine on the Expressions of bcl-2 and bax in Rat after Acute Global Cerebral Ischemia and Reperfusion

To investigate the effects of L-Tetrahydropalmatine (L-THP) on the expressions of bcl-2, bax and neuronal apoptosis after cerebral ischemia and reperfusion, 60 Wistars rats were randomly divided into 3 groups: sham-operation group (group S, n = 20), ischemic-reperfusion group treated with saline (group I, n=20) and ischemia-reperfusion group treated with L-THP (group T, n=20) .The rat model of global cerebral ischemia and reperfusion was induced by Pulsinelli's four-vessel occlusion method. The expression of bcl-2 and bax mRNA was detected by in situ hybridization and reverse transcriptional polymerase chain reaction (RT-PCR). The number of apoptotic neurons was examined by terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) method. Compared with group S, the expression of bcl-2 and bax mRNA in group I was increased significantly (P<0.01), and the number of apoptotic neurons increased either (P< 0.01). After L-THP treatment, the expression of bcl-2 mRNA was up-regulated (P<0.01) and that of bax mRNA was down-regulated (P<0.01); the number of apoptotic neurons was decreased (P<0.01). Our results indicated that bcl-2 may suppress apoptosis and bax promote apoptosis after cerebral ischemia and reperfusion. L-THP could ameliorate cerebral ischemia and reperfusion damage by reducing the apoptosis through regulating bcl-2 and bax.

ER、bcl-2和p53在鸡与鹌鹑属间杂交种早期胚胎中的mRNA表达

通过人工授精获得鸡（♂10只）与鹌鹑（♀100只）属间杂交种蛋并同机入孵，采用Wpkci引物和多重PCR鉴定66～120h的鸡（♂）与鹌鹑（♀）属间杂交种活胚的性别后，选取不同时间点雌、雄胚胎共300枚，以β-actin为内标，通过RT-PCR分别测定雌激素受体（职）和细胞凋亡因子（bcl-2、p53）的mRNA相对丰度；探讨ER、bcl-2和p53对杂交种早期胚胎发育及性别分化的影响。结果表明：（1）杂交种胚胎ERmRNA表达在66～84h期间雌性极显著高于雄性（P〈0．01=，由此推测杂交种的性分化时间大致在胚胎发育的66～84h范围内；（2）bcl-2和p53的mRNA表达在杂交胚胎发育过程中具有明显的时序性，说明bcl-2和p53基因对早期杂交胚的发育具有重要的影响。

夹脊电针对大鼠脊髓损伤后bcl-2mRNA表达的影响

为了探讨夹脊电针治疗脊髓损伤的机制,选用Wistar健康雄性大鼠,随机分为模型组、电针密波治疗组、地塞米松组及假手术组.采用改良的Allen's捶击法致大鼠T11-12脊髓损伤,应用原位杂交化学方法观察脊髓损伤早期bcl-2mRNA的表达.研究结果表明:伤后6h内进行治疗较伤后12h进行治疗,能更好地促进损伤大鼠的运动功能恢复,并且脊髓损伤早期,地塞米松治疗疗效优于密波组;脊髓损伤中晚期,密波组疗效好于药物组.结论表明:夹脊电针可上调bcl-2mRNA表达,从而减少或抑制细胞凋亡的发生.

密蒙花总黄酮对去势导致干眼症雄鼠泪腺Bax mRNA、Bcl-2 mRNA表达的影响

目的观察密蒙花总黄酮对雄激素水平下降所致干眼症雄鼠的泪腺上皮细胞中Bax mRNA、Bcl-2mRNA表达的影响。方法将健康1月龄、体质量150～180gWistar雄性大鼠150只随机分为A1、B1、C1、D1、E1、A3、B3、C3、D3、E3、A5、B5、C5、D5、E5共15组,每组10只。采用去势的方法建立雄激素水平下降所致干眼症动物模型,分别观察各组SchimerI试验、泪膜破裂时间及角膜荧光素染色情况,以确定干眼症模型是否建立成功。其中,A代表正常组;B代表假手术组;C代表模型对照组;D代表雄激素对照治疗组(造模成功后,丙酸睾酮1mg.kg-1大腿肌肉注射,每3d1次);E代表密蒙花总黄酮治疗组(造模成功后,密蒙花总黄酮0.045g.kg-1灌胃,每3d1次);1代表饲养1个月;3代表饲养5个月;5代表饲养7个月。用RT-PCR法对各组泪腺组织中Bax mRNA、Bcl-2mRNA的表达进行检测,并比较。结果SchimerI试验、泪膜破裂时间及角膜荧光素染色结果显示,大鼠均在造模3个月后形成干眼症。D3组、D5组泪腺组织中的BaxmRNA相对含量较C3组、C5组降低,差异均有统计学意义(均为P<0.01);E3组、E5组较C3组、C5组亦降低,差异亦均有统计学意义(均为P<0.01);且E3组(3.787±0.165)与E5组(5.707±0.411)BaxmRNA相对含量差异有统计学意义(P<0.01);D5组(4.610±0.481)与E5组间比较差异亦有统计学意义(P<0.01)。D3组、D5组Bcl-2mRNA的相对含量较C3组、C5组增加,差异均有统计学意义(均为P<0.01);E3组、E5组较C3组、C5组亦增加,差异均有统计学意义(均为P<0.01);D5组(5.713±0.339)与E5组(3.147±0.057)间比较Bcl-2mRNA表达差异有统计学意义(P<0.01);且E3组(8.427±0.055)与E5组比较差异亦有统计学意义(P<0.01)。结论密蒙花总黄酮可使大鼠泪腺中Bcl-2mRNA表达增加,降低BaxmRNA的表达,但随病程的延长其作用弱于雄激素。密蒙花总黄酮治疗雄激素水平下降所致干眼症的机制可能与其产生拟雄激素效应后,对凋亡相关基因Bcl-2mRNA表达的上调和Bax mRNA表达的下调有关。

氨茶碱对哮喘豚鼠嗜酸细胞凋亡及bcl-2 mRNA表达的影响

目的观察氨茶碱对体外不同密度嗜酸细胞 ( Eos)凋亡及 bcl- 2 m RNA表达的影响 ,探讨其促进哮喘 Eos凋亡的机制。方法卵蛋白激发哮喘豚鼠动物模型 4 8h后行支气管肺泡灌洗 ,分离不同密度 Eos。氨茶碱 ( AM) ( 10 - 6 ～ 10 - 3m ol/L)干预 2 4 h,原位杂交检测 Eos的 bcl- 2 m RNA表达 ,TU NEL 法检测细胞凋亡。结果 AM干预 2 4 h,可见 Eos凋亡增加 ,以低密度 Eos( HEos)为明显 ,bcl- 2 m RNA表达降低 ,呈剂量依赖性。bcl- 2 m RNA的表达与 Eos凋亡呈负相关。结论 AM可抑制 bcl- 2 m RNA表达 ,促进 Eos凋亡。 bcl- 2参与了 AM对 Eos凋亡的调节。

酸枣仁汤对失眠大鼠中脑中缝背核Bcl-2及脑源性神经营养因子mRNA的影响

目的观察对氯苯丙氨酸(PCPA)失眠大鼠中脑中缝背核Bcl-2及脑源性神经营养因子mRNA(BDNF mRNA)表达的改变,以及酸枣仁汤(SZRD)对PCPA所致失眠大鼠中脑中缝背核Bcl-2及BDNF mRNA表达改变的影响,探讨酸枣仁汤可能的作用环节。方法 SD大鼠42只,随机分为空白对照组、模型组、SZRD大剂量组(15 g.kg-1)、SZRD中剂量组(7.5 g.kg-1)、SZRD小剂量组(3.75 g.kg-1)、西药治疗组(艾司唑仑2 mg.kg-1)。采用腹腔注射PCPA的方法建立失眠大鼠模型,各组治疗7d后,取脑组织,采用免疫组织化学染色法检测Bcl-2蛋白表达变化,采用原位杂交法检测BDNF mRNA表达的变化。结果 PCPA失眠模型组大鼠中脑中缝背核Bcl-2及BDNF mRNA表达增强,与空白对照组相比,差异具有统计学意义(P<0.01);酸枣仁汤高剂量组中缝背核内Bcl-2及BDNF mRNA表达增加,与模型组相比具有显著性差异(P<0.01),艾司唑仑组中缝背核BDNF mRNA表达增加,与模型组相比具有显著性差异(P<0.001),艾司唑仑组中缝背核Bcl-2表达与模型组相比无显著性差异。结论 PCPA所致失眠能导致中脑中缝背核Bcl-2及BDNF mR-NA表达增强,酸枣仁汤能调节中脑中缝背核Bcl-2及BDNF mRNA表达量,从而发挥对失眠的干预作用。

传染性法氏囊病毒对SPF雏鸡免疫器官细胞凋亡及bcl-2mRNA表达的影响

为证实传染性法氏囊病毒(IBDV)感染SPF雏鸡后,其免疫器官细胞凋亡与bcl-2mRNA动态表达的关系,应用TUNEL和实时荧光定量PCR方法分别对雏鸡免疫器官细胞凋亡数量和bcl-2mRNA表达的动态变化进行了检测,结果发现,IBDV感染雏鸡后3～7d,法氏囊、胸腺和脾脏中凋亡细胞数量显著或极显著高于相应对照雏鸡(P<0.01或P<0.05);同时在上述3种免疫器官中bcl-2mRNA的表达均不同程度增加,并于病毒感染后3～7d差异显著或极显著(P<0.01或P<0.05)。表明IBDV可增加感染雏鸡免疫器官的细胞凋亡数量和bcl-2mR-NA的表达。

密蒙花总黄酮含药血浆对干眼症细胞模型Bax mRNA及Bcl-2 mRNA表达的影响

目的:观察密蒙花总黄酮含药血浆对干眼症细胞模型中泪腺上皮细胞Bax mRNA及Bcl-2 mRNA表达的影响。方法:将培养生长状态良好的泪腺上皮细胞,随机分为无雄激素培养黄酮治疗组(以下简称密蒙花总黄酮组),含雄激素培养对照组(以下简称雄激素组),无雄激素培养空白组(以下简称空白组)。分别加入含密蒙花总黄酮血浆、丙酸睾酮、含空白血浆开始进行干预。建立干眼症细胞模型。用Trizol试剂提取细胞总RNA。RT-PCR法检测泪腺上皮细胞中Bax mRNA及Bcl-2 mRNA的表达。结果:密蒙花组、雄激素组能够抑制泪腺上皮细胞中BaxmRNA的表达,与空白组比较差异有显著性(P<0.01);密蒙花组Bax mRNA的表达低于雄激素组,比较差异有显著性(P<0.01)。密蒙花组、雄激素组Bcl-2 mRNA的表达均高于空白组,比较差异有显著性(P<0.01);密蒙花组Bcl-2 mRNA的表达高于雄激素组,比较差异有显著性(P<0.01)。结论:密蒙花总黄酮含药血浆作用于雄激素水平下降所致干眼症的细胞模型可抑制Bax mRNA的表达,使Bax mRNA的表达量降低,同时促进Bcl-2 mRNA的表达,使Bcl-2 mRNA的表达量增加。密蒙花总黄酮含药血浆很可能是通过抑制泪腺上皮细胞中Bax mRNA及促进Bcl-2 mRNA表达的途径而达到抑制细胞凋亡的作用。

芪黄煎剂对缺血-再灌注大鼠肠黏膜上皮细胞Bcl-2、Bax及Caspase-3、9 mRNA表达的影响

目的观察中药芪黄煎剂对缺血-再灌注(ischemia-reperfusion,I/R)损伤大鼠肠黏膜上皮细胞凋亡基因Bcl-2、Bax mRNA表达和信号转导分子Caspase-3、9 mRNA表达的影响。方法 40只Wistar大鼠随机分为假手术组、I/R损伤组、谷氨酰胺(Gln)组和芪黄煎剂组,每组10只。假手术组管饲生理盐水3天后,仅腹壁切开而不行I/R手术;I/R损伤组与假手术组同样方法干预后,行I/R损伤手术;Gln组和QHD组分别管饲Gln和芪黄煎剂3天后,行I/R损伤手术。RT-PCR方法检测肠上皮细胞凋亡基因Bcl-2、Bax mRNA表达和凋亡信号转导分子Caspase-3、9 mRNA表达。结果与假手术组比较,I/R损伤组大鼠肠黏膜细胞内Bcl-2、Bax、Caspase-3、9 mRNA均明显增多,差异有统计学意义(P<0.05)。与I/R损伤组比较,Gln组、芪黄煎剂组Bcl-2 mRNA表达增多,Bax、Caspase-3、9 mRNA表达减少,差异均有统计学意义(P<0.05)。与Gln组比较,芪黄煎剂组Bax mRNA表达更低(0.281±0.087vs0.350±0.053),Bcl-2/Bax值更高(1.648vs1.374),差异无统计学意义(P>0.05)。结论芪黄煎剂通过抑制细胞凋亡减轻I/R对大鼠肠黏膜上皮造成的损伤,其作用机制可能与增加Bcl-2 mRNA表达和降低Bax、Caspase-3、9 mRNA表达有关。

金雀异黄素对多囊卵巢综合征大鼠卵巢组织中Bcl-2及Bax mRNA表达的影响

本试验旨在探讨金雀异黄素(genistein,GEN)对多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠卵巢组织中抗凋亡基因Bcl-2及促凋亡基因Bax mRNA表达的影响。将90只雌性Wistar大鼠随机分为6组:对照组,模型组,GEN低、中、高剂量组及雌激素组。按照胰岛素联合人体绒毛膜促性腺激素造模法建立PCOS试验动物模型,造模成功后,每组选择10只大鼠进行试验。剂量组分别给予5、10、20 mg/kg GEN,雌激素组给予0.5 mg/kg己烯雌酚,受试物灌胃给予,持续15 d,观察动情周期10 d。采用原位杂交法检测各组大鼠卵巢组织中Bcl-2和Bax mRNA表达量。同时测定大鼠卵巢组织中超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶活性及丙二醛含量。结果表明:与模型组相比,GEN能提高大鼠卵巢组织中Bcl-2 mRNA表达量,降低Bax mRNA表达量,并且可调节卵巢组织中抗氧化酶活性,其作用效果与雌激素相似。GEN可以增强大鼠卵巢组织中Bcl-2 mRNA表达量,拮抗Bax mRNA表达量,推测这是GEN改善PCOS卵巢功能的作用途径之一。

大豆异黄酮对青年雌性大鼠卵巢、子宫组织中Bcl-2 mRNA和Bax mRNA表达的影响

目的:通过动物实验研究,观察Bcl-2 mRNA和Bax mRNA在青年雌性大鼠卵巢和子宫组织中的表达情况,从细胞凋亡角度研究大豆异黄酮对青年雌性大鼠的抗衰老作用。方法:选用2月龄青年雌性大鼠50只,按体质量分成5组,每组10只,分别为对照组(基础饲料)、低剂量组(大豆异黄酮100mg/(kgbw·d))、中剂量组(大豆异黄酮200mg/(kgbw·d))、高剂量组(大豆异黄酮300mg/(kgbw·d))、雌激素组(己烯雌酚0.5mg/kg饲料),实验周期7周。采用原位杂交法检测各组大鼠卵巢和子宫组织中Bcl-2 mRNA和Bax mRNA的表达水平。结果:大豆异黄酮能够增强大鼠卵巢和子宫组织中Bcl-2 mRNA的表达水平;而各剂量组大鼠卵巢和子宫组织中Bax mRNA水平呈下降趋势。结论:大豆异黄酮可以增强抗凋亡基因Bcl-2mRNA,拮抗促凋亡基因Bax mRNA的水平,推测这是大豆异黄酮对青年雌性大鼠发挥抗衰老作用的途径之一。

碘对大鼠甲状腺细胞凋亡及bax、bcl-2基因mRNA表达的影响

目的探讨碘对甲状腺细胞凋亡及凋亡相关基因表达的影响。方法Wistar大鼠分为6组,即低碘组(LI)、正常碘组(NI)、5倍碘组(5HI)、10倍碘组(10HI)、50倍碘组(50HI)、100倍碘组(100HI),用含不同碘的自来水喂养,6个月后取甲状腺。末端标记法(TUNEL)进行原位细胞凋亡检测,反转录-聚合酶链反应(RT-PCR)法对甲状腺细胞凋亡相关基因bax、bcl-2进行半定量测定。结果原位细胞凋亡检测结果表明,LI组细胞凋亡明显,而NI组与其他组均未见凋亡细胞。baxmRNA表达水平以bax/β-actin光密度比表示,LI组为1.096±0.089,NI组为0.647±0.062,5HI组为0.638±0.191,10HI组为0.676±0.081,50HI组为0.644±0.092,100HI组为0.751±0.152;同样方法表示,bcl-2mRNA表达水平分别为0.273±0.185、0.172±0.115、0.236±0.107、0.235±0.071、0.267±0.065、0.313±0.062。结果表明,在LI组大鼠甲状腺bax、bcl-2mRNA表达水平均明显增加(P<0.05);在其他组,bcl-2mRNA表达水平随给碘量增加而增加,而bax无明显变化。但bax/bcl-2比值随摄入碘量增加而呈降低趋势。结论碘缺乏易诱发大鼠甲状腺细胞凋亡;而碘过量不易引起细胞凋亡,甲状腺细胞对碘过量有一定耐受能力。bcl-2家族参与甲状腺细胞凋亡过程。

电针调节脑缺血再灌注大鼠大脑皮层Bcl-2和Bax mRNA的表达

目的:探讨电针是否通过干预Bax/Bcl-2 mRNA的表达,来调控缺血再灌注大鼠大脑皮层细胞凋亡,发挥电针对脑神经元保护作用。方法:将清洁级健康雄性SD大鼠19只,按随机数字表随机分成假手术组(n=5)和用于造模动物14只,采用改良Longa线栓大脑中动脉法制作局灶性脑缺血再灌注模型。在成功制作出脑缺血再灌注模型并作神经缺损综合评分为2分以上后随机分为模型组、电针组,每组各7只。取电针组大鼠"百会"及"大椎"穴,采用疏密波刺激30min,电流强度1～3mA,频率20Hz/80Hz,疏、密波交替时间各为1.5s。用实时荧光定量聚合酶链反应(RT-PCR)技术分别检测大鼠大脑皮层组织Bcl-2和Bax mRNA的表达。结果:与假手术组相比,模型组Bcl-2、Bax mRNA表达均显著升高,而Bcl-2/Bax比值显著下降;与模型组相比,电针组Bax mRNA表达降低,Bcl-2 mRNA表达虽无统计学差异,但Bcl-2/Bax比值显著上升,形成以Bcl-2占优势的Bcl-2/Bax二聚体。结论:电针可以通过调控内源性凋亡途径,抑制Bax的促凋亡作用,降低缺血再灌注区的细胞凋亡,促进细胞的存活,这可能是电针拮抗局灶性脑缺血再灌注损伤后的神经细胞凋亡,保护脑神经元的分子生物学机制之一。

抗痫灵对戊四氮诱导癫痫大鼠海马区Bcl-2mRNA及其蛋白表达的影响

目的:观察中药复方抗痫灵对戊四氮致痫大鼠海马Bcl-2mRNA及其蛋白表达的影响。方法:观察大鼠痫性发作行为;采用免疫组化方法检测大鼠脑组织中海马Bcl-2蛋白的表达;采用实时荧光定量(Real timePCR)方法检测大鼠海马组织Bcl-2mRNA的相对表达。结果:经过中药复方抗痫灵治疗6周后,大鼠痫性发作行为明显减少;海马区抑制凋亡基因Bcl-2mRNA及其蛋白表达均升高。结论:抗痫灵能减少癫痫发作次数,上调海马区抑凋亡基因Bcl-2mRNA及其蛋白表达。