大鼠嗅感觉上皮发育及嗅细胞凋亡的研究

目的探讨神经特异性烯醇酶（NSE）、嗅细胞标记蛋白（OMP）及细胞凋亡在不同胎龄大鼠嗅黏膜中的表达。方法免疫组化方法检测NSE及OMP在不同孕期大鼠嗅上皮中的表达及其规律。TUNEL方法检测不同孕期大鼠嗅上皮中凋亡细胞并计算细胞凋亡指数（AI）。结果 E13d鼻腔黏膜中即有NSE阳性表达,细胞数量多。E13d嗅上皮中未见OMP阳性表达细胞。在E14d,嗅上皮中出现嗅OMP阳性细胞,数量少,随胎龄增加阳性细胞数量增多,至17d达高峰并逐渐趋于稳定。E13E15d细胞凋亡数量较稳定,至E16d凋亡细胞数量明显增加,达到高峰,E18E21d凋亡细胞数量逐渐减少渐趋于稳定。E16dAI与其他d数差异有统计学意义。结论 E14d嗅黏膜中已有发育成熟的嗅细胞,数量少,胚胎发育后期大鼠嗅化学感受器发育已趋于成熟。在嗅上皮发育过程中存在细胞凋亡,E16d嗅细胞凋亡出现一高峰,细胞凋亡在嗅觉发育过程中起重要作用。

Berbamine induces apoptosis in human hepatoma cell line SMMC7721 by loss in mitochondrial transmembrane potential and caspase activation

Objective： To investigate the effect ofberbamine on human hepatoma cell line SMMC7721. Methods： The effects of 24 h and 48 h incubation with different concentrations （0-64 μg/ml） of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide （MTT） assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI （propidium iodide） staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential （△ψm）, the expression of activated caspase3 and caspase9 was analyzed by Western-blot. Results： The proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential （AVm） and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk. Conclusion： Berbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.

Effects of Bushenyiqihexue Formula on the Endometrial Gland Apoptosis in Mice with Blastocyst Implantation Dysfunction

Objective: To observe the effects of Bushenyiqihexue Formula (补肾益气和血方 Formula for Tonifying the Kidney, Replenishing qi and Harmonizing Blood, FTKRQHB) on the endometrial gland apoptosis in the mice with blastocyst implantation dysfunction. Methods: The mice with the first-day pregnancy were divided into the control, model and treatment groups, with 30 in each group, and blastocyst implantation dysfunction was induced by subcutaneous injection of mifepristone in the mice of the model and treatment groups. The pregnancy rate and implantation number of blastocysts were measured and the expressions of proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, and activated caspase-3 were detected in all the three groups. Results: The model group had significantly depressed pregnancy rate, implantation number of blastocysts and apoptosis index, and elevated proliferation index of endometrial gland as compared with the control group (P<0.05 or P<0.01). Administration of FTKRQHB (the treatment group) resulted in significant increases in pregnancy rate, implantation number of blastocysts and apoptosis index of the endometrial gland, and a significant decrease in the proliferation index of the endometrial gland as compared with the model group (P<0.05 or P<0.01). The differences in the four indexes between the treatment group and control group were not significant statistically. The Bax and activated caspase-3 expressions in endometrial gland in the model group became significantly lower than that of the control group (P<0.01), whereas those in the treatment group were significant higher than that of the model group (P<0.01). However, the Bax and activated caspase-3 expressions in endometrial gland were similar in both treatment and control groups. Conclusion: Promoting the increases in Bax and activated caspase-3 expressions in the endometrial gland and bringing into balance between apoptosis and proliferation of the glandular cells at the implantation window phase by FTKRQHB may contribute to the effects of promoting the establishment of endometrial receptivity and improving blastocyst implantation dysfunction.

Apoptosis within mouse eye induced by Toxoplasma gondii

Objective To investigate apoptosis induced by Toxoplasma gondii (T. gondii) in eyes of C57BL/6 (B6) mice. Methods Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) technique and pathological changes within eyes were analyzed at different time points after intraocular inoculation of either 50 or 500 of tachyzoites. Results In eyes that received 50 tachyzoites, a few apoptotic inflammatory cells in the anterior chamber and keratocytes in the cornea were seen at days 1 and 2, but no apoptosis was detected 4 days after inoculation. Significantly greater apoptosis of inflammatory cells was observed in the anterior chamber and in the vitreous of eyes injected with 500 parasites. Apoptosis of inflammatory cells in the anterior chamber and of keratocytes in the cornea was seen at day 1. The apoptotic stromal keratocytes strikingly increased at day 4. There were a number of apoptotic inflammatory cells in the vitreous at day 2, and a few apoptotic retinal cells along the internal limiting membrane and the nerve fiber layer of the retina 4 days after inoculation. Conclusion These results suggest that apoptosis of inflammatory cells infiltrated eye infected with this parasite may be a mechanism of eliminating the organism.