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目的:利用日本血吸虫童虫与宿主细胞的共培养模型,研究不同日本血吸虫童虫密度对共培养宿主细胞乳酸脱氢酶(LDH)活性的影响。方法:将机械断尾后的日本血吸虫童虫以0条(对照)、200条、400条、600条及800条/ml的虫体密度与接种于小盖玻片上、密度为2.0×105个/ml、预培养24h的静脉内皮细胞(ED-25)共培养,72h后对宿主细胞ED-25进行LDH细胞化学染色,Olympus-BH2显微镜下观察并拍照,采用HIPAS-2000图像分析仪测定其含量,并作统计分析。结果:随着日本血吸虫童虫密度的增加,共培养宿主细胞的LDH着色逐渐加深;童虫密度为400条/ml时,共培养宿主细胞的LDH着色最深;随后随着童虫密度的继续增加,宿主细胞的LDH着色不再加深,反而变浅。定量分析结果显示,虫体密度为400条/ml时,与其共培养的宿主细胞ED-25LDH的平均吸光值(A值)最大(0.1676±0.0378);虫体密度为800条/ml时的ED-25LDH的A值最小(0.1471±0.0216)。统计分析发现,虫体密度为200条/ml时宿主细胞LDH的A值(0.1566±0.0156)与对照的(0.1558±0.0323)差异不明显(P>0.05),虫体密度为400条/ml时宿主细胞LDH的A值显著大于对照组(P<0.01),各虫体密度间A值差异均有统计学意义(P<0.01或P<0.05)。结论:以400条/ml日本血吸虫童虫与2.0×105个/ml宿主细胞ED-25共培养,可明显增强宿主细胞LDH活性,促进其新陈代谢。
OBJECTIVE: To study the effects of different densities of Schistosoma japonicum on the lactate dehydrogenase (LDH) activity of co-cultured host cells using the co-culture model of Schistosoma japonicum and host cells. Methods: Schistosoma japonicum Schistosoma japonicum after mechanical tails was inoculated into small coverslips with density of 2.0 × 105 (control), 200, 400, 600 and 800 / ml. / ml and pre-cultured for 24 hours. ED-25 cells were stained with LDH after 72 hours, observed under Olympus-BH2 microscope and photographed. The HIPAS-2000 image analyzer Content, and make statistical analysis. Results: With the increase of the density of Schistosoma japonicum, LDH coloration of co-cultured host cells gradually deepened. When the density of sperm was 400 per ml, LDH coloration of co-cultured host cells was the deepest. , LDH coloration of host cells no longer deepened, but lighter. Quantitative analysis showed that the average absorbance value (A value) of host cells ED-25LDH co-cultured with the parasitoid was the highest (0.1676 ± 0.0378) when the parasites density was 400 / ml; ED A value of -25LDH is the smallest (0.1471 ± 0.0216). Statistical analysis showed that there was no significant difference (P> 0.05) between the A value of host cell LDH (0.1566 ± 0.0156) and the control (0.1558 ± 0.0323) at the density of 200 cells / ml The A value of LDH in host cells was significantly higher than that in control group (P <0.01), and there was significant difference in A value between host densities (P <0.01 or P <0.05). CONCLUSION: Co-culture of Schistosoma japonicum with ED-25 of 2.0 × 105 / ml host cells at 400 / ml could significantly enhance the LDH activity of host cells and promote their metabolism.