大白鼠闭合性脑损伤后神经细胞的凋亡

目的：本实验在改良前脑局部闭合性脑硬膜外占位性颅脑损伤的创伤性模型基础上研究神经元细胞调亡。方法：５６ 只Ｗｉｓｔａｒ大白鼠,采用球囊注水在脑硬膜外造成占位及持续时间不同所形成的分级脑外伤,饲养２ｄ 后脑标本采用原位末端标记（ＴＵＮＥＬ）法进行ＰＣＤ阳性细胞的检测。结果：４０ 只存活鼠中,实验组损伤周围有ＴＵＮＥＬ阳性细胞的表达,在对侧及未损伤区几乎无表达。本实验造成脑损伤的机制中除直接受压区的坏死外受损周围区域有凋亡的参与。提示凋亡可能是脑创伤后脑损伤中神经元细胞死亡的方式之一。     

Studies on hepatocyte apoptosis,proliferation and oncogene c-fos expression in carbon tetrachloride-induced cirrhotic rat liver

Ａｐｏｐｔｏｓｉｓｈａｓｂｅｅｎｉｎｔｅｎｓｉｖｅｌｙｓｔｕｄｉｅｄｉｎｈｅｐａｔｏｂｉｌｉａｒｙｄｉｓｅａｓｅｓ．Ｈｏｗｅｖｅｒ,ｄａｔａａｂｏｕｔｔｈｅｒｅｌａｔｉｏｎｓｈｉｐｂｅｔｗｅｅｎｌｉｖｅｒｃｉｒｈｏ－ｓｉｓａｎｄａｐｏｐｔｏｓｉｓｉｓ...     

保尔佳对原发性肝癌患者sIL-2R、IL-6、TNFα的影响及促进肝癌细胞凋亡的初步研究

对２０例原发性肝癌患者（大剂量组１０例、小剂量组１０例）临床应用保尔佳１周，用ＥＬＩＳＡ法检测治疗组用药前后外周血及用药后门静脉血的ｓＩＬ－２Ｒ，ＩＬ－６，ＴＮＦα的水平变化及用末端转移酶标记法（ＴＵＮＥＬ法）检测治疗组肝癌细胞的凋亡情况，与未治疗组、正常对照组相比。结果表明，治疗组外周血ｓＩＬ－２Ｒ水平明显下降（大剂量组Ｐ＜０．０１、小剂量组Ｐ＜０．０５），ＩＬ－６水平明显上升（大剂量组Ｐ＜０．０１、小剂量组Ｐ＜０．０５），ＴＮＦα水平有所下降（大剂量组Ｐ＜０．０５、小剂量组Ｐ＞０．０５），治疗组门静脉血与外周血ｓＩＬ－２Ｒ、ＩＬ－６、ＴＮＦα两者间无明显差异（Ｐ＞０．０５）。ＴＵＮＥＬ法检测显示保尔佳能促进肝癌细胞的凋亡（Ｐ＜０．０１，大、小剂量组间Ｐ＞０．０５）。且保尔佳对肝癌患者的血常规、肝功能等无不良影响。保尔佳是一种具有增强机体免疫功能、促进肿瘤细胞凋亡且无明显毒副作用的新型抗癌药物。     

子宫内膜腺上皮细胞中增殖与凋亡相关基因表达初探

目的:探讨不同类型子宫内膜腺上皮细胞的增殖和凋亡情况。方法:采用免疫组化和TUNEL方法检测不同类型子宫内膜组织中增殖细胞核抗原(PCNA)与细胞凋亡表达。结果:(1)从正常子宫内膜直至子宫内膜样癌的PCNA表达逐渐增加。在子宫内膜样腺癌中随着癌细胞分化程度的降低、肌层浸润深度的增加和手术病理分期的升高,PCNA表达逐渐增加,且各组之间PCNA表达均有显著性差异(P<0.05)(。2)从正常子宫内膜、子宫内膜增殖症到子宫内膜样腺癌细胞凋亡的表达逐渐增加。子宫内膜样腺癌中不同分化程度的癌细胞、不同手术病理分期之间凋亡表达均有显著差异(皆P<0.01);而不同肌层浸润深度之间凋亡表达无显著差异(P>0.05)。(3)PCNA表达与细胞凋亡在不典型增生、子宫内膜样癌中均呈负相关(分别r=-0.943,P<0.01;r=-0.976,P<0.01)。结论:PCNA是内膜细胞癌变过程中一项重要的指标,其在内膜癌的发生和发展中起重要作用,可作为估计患者预后的一个重要参数。细胞凋亡情况在一定程度上可以反映肿瘤的生物学行为。内膜细胞增殖与凋亡相互作用共同促进内膜细胞过度增殖,从而导致肿瘤的形成和进展。     

Neurodegenerative changes and apoptosis induced by intrauterine and extrauterine exposure of radiofrequency radiation

Adverse health effects of radiofrequency radiation (RFR) on the ongoing developmental stages of children from conception to childhood are scientifically anticipated subject. This study was performed to identify the effects of global system for mobile communications (GSM) modulated mobile phone like RFR in 1800 MHz frequency on oxidative DNA damage and lipid peroxidation beside the apoptotic cell formation, using histopathological and immunohistochemical methods in the brain tissue of 1-month-old male and female New Zealand White rabbits that were exposed to these fields at their mother's womb and after the birth. Oxidative DNA damage and lipid peroxidation levels were investigated by measuring the 8-hydroxy-2′-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels, respectively. Histopathological changes were observed using by hematoxylin and eosin (HE) staining. Apoptotic cells were detected in the examined organs by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining.For both male and female infants; 8-OHdG levels increased in the group exposed to RFR in both intrauterine and extrauterine periods compared to the infants that were never exposed to RFR and the ones were exposed when they reached one month of age (p < 0.05). MDA results were different for male and female rabbits. There was no difference between all female infant groups (p > 0.05), while only intrauterine exposure significantly causes MDA level increase for the male infants. HE staining revealed mild lessions in neuronal necrobiosis in brain tissues of female rabbits that had only intaruterine exposure and male rabbits had only extrauterine exposure. Gliosis were mildly positive in brain tissues of rabbits that are exposed only intrauterine period, also the group exposed both intrauterine and extrauterine periods. However, there was no apoptotic change detected by TUNEL staining in the brain tissues of all groups.     

The antioxidants,vitamin A and E but not vitamin C and melatonin enhance the proapoptotic effects of irinotecan in cancer cells in vitro

Irinotecan is one of the camptothecin analog which has been shown to have a broad spectrum of antitumor activities against various malignancies. The aim of this study was to evaluate the effect of vitamin A, C, E and melatonin on proapoptotic activity of irinotecan in human cancer cells in vitro. We observed that irinotecan induced apoptosis in all types of analyzed cell lines when used as a single agent. Combination of selected antioxidants with various doses of irinotecan (7.5–60 μM) resulted in significant increase in apoptotic cell death in A549 and HT29 cancer cell lines. The highest killing efficiency was observed after co-incubation of the cells with irinotecan and vitamin A (10 μM), or vitamin E (25 μM), respectively. The addition of vitamin C and melatonin to irinotecan treatment did not promote increase in killing of cancer cells. Our results indicate that some antioxidants can enhance the proapoptoic activity (properties) of irinotecan in human cancer cells in vitro. These findings may be supportive for the optimization of therapeutic efficacy of irinotecan treatment.     

重组人P53融合蛋白抗肝癌细胞的实验研究

研究蛋白传导域(PTD)与人野生型P53(WtP53)融合表达的P53融合蛋白PTD-P53对HepG2的增殖抑制和凋亡的影响.用PTD-P53处理HepG2后,采用MTT法分析处理后细胞的生长增殖情况,流式细胞术(FCM)分析处理后细胞周期变化及脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测细胞凋亡情况.PTD-P53对:HepG2的生长有明显抑制作用.细胞周期也出现阻滞现象,细胞凋亡明显增加.PTD-P53有一定的抗肝癌细胞作用.     

葛根素诱导血管平滑肌细胞凋亡的实验研究

目的：观察葛根素促进人脐动脉血管平滑肌细胞凋亡的作用，探讨葛根素抑制平滑肌细胞增殖的机制。方法：分离培养人脐动脉平滑肌细胞，不同浓度葛根素与细胞孵育，TUNEL检测葛根素诱导细胞凋亡，rt-PCR实验检测促凋亡基因Bax和抗凋亡基因Bcl-XL。结果：随着葛根素浓度升高，细胞生长受抑制，TUNEL阳性细胞显著增加。rt-PCR实验发现促凋亡基因Bax随药物浓度和作用时间的增加而上升，而且Bax、Bcl-XL基因表达比例有一定升高。结论：葛根素通过调节经典的Bax、Bcl-XL通路对血管平滑肌细胞凋亡有一定诱导作用。     

G-CSF/SCF exert beneficial effects via anti-apoptosis in rabbits with steroid-associated osteonecrosis

Objectives

Osteonecrosis is also known as avascular necrosis, and two types of cell death are included in the pathogenesis of osteonecrosis: necrosis and apoptosis. Apoptosis in the osteonecrosis of femoral head is thought to be the key determinant of glucocorticoid-induced cortical bone loss. The present study was implemented to evaluate the anti-apoptotic effect of Granulocyte colony-stimulating factor and stem cell factor (G-CSF/SCF) in rabbits with steroid-induced osteonecrosis.

Methods

In the experiment, osteonecrosis was induced by low-dose lipopolysaccharide and subsequent pulsed high-dose methylprednisolone. Rabbits in preventive medicine group were treated with100 μg/kg/d G-CSF and 25 μg/kg/d SCF. Then hematological and histomorphometric methods were used to investigate the treatment effects of osteonecrosis. Apoptosis was assessed via quantitative TUNEL staining and activated caspase-3 immunostaining and immunoblotting.

Results

The results showed that G-CSF/SCF treatment could increase the secretion of serum osteocalcin, but inhibit the expression of serum tartrate-resistant acid phosphatase (TRAP5b). The incidence of osteonecrosis was significantly decreased in Preventive group when compared with Steroid group (42.1% vs. 88.2%). Histomorphometric analysis showed that G-CSF/SCF pre-disposal treatment was able to increase trabecular mineral appositional rate (MAR) and bone formation rate (BFR). Quantitative TUNEL and activated caspase-3 levels showed lower apoptosis in the Preventive group.

Conclusions

In conclusion, G-CSF/SCF treatment could inhibit caspase-3-dependent apoptosis in osteocytes to exert beneficial effects in preventing steroid-induced ON in rabbit models.     

Apurinic/apyrimidinic endonuclease immunoreactivity in germ cells of experimental varicocele-induced rat testes

Increased germ cell apoptosis is related to oxidative DNA damage; therefore, we investigated whether there was a significant change in apurinic/apyrimidinic endonuclease (APE) in varicoceles. Experimental varicoceles were created by partial ligation of the left renal vein of adult male Sprague-Dawley rats, which were sacrificed at 1, 3 and 6 weeks after varicocele creation. Testicular tissues were sampled for TUNEL, Western blotting and immunohistochemistry. There was a significant increase in apoptotic germ cells in the ipsilateral testes 6 weeks after varicocele creation. Increased activation of p53, Bax and cleaved caspase-3 in the left testes was also noted. APE increased activation until 3 weeks after varicocele creation, and then decreased at 6 weeks after varicocele surgery. The spermatocytes were immunostained for both 8-hydroxy-2′-deoxyguanosine and APE, but the spermatogonia revealed only APE immunopositivity in the defective tubules. These results suggest that repression of APE is an underlying mechanism of augmented p53-dependent apoptosis in varicocele-induced rat testes and that remaining APE in the spermatogonia plays a decisive role in regaining testicular spermatogenic function after varicocelectomy.