藻酸盐诱发的免疫反应对兔腮腺腺细胞的作用  

The effect of immune reaction induced by alginate on parotid acinar cells in vitro

作　　者：何巍[1] 吕继连[1] 李龙江[2] 

机构地区：[1]郑州大学第一附属医院口腔颌面外,450052 [2]四川大学口腔生物医学工程教育部重点实验室

出　　处：《实用口腔医学杂志》2009年第6期779-782,共4页Journal of Practical Stomatology

基　　金：国家教委博士点基金(编号:20040610079)

摘　　要：目的:探讨藻酸盐诱发的免疫反应对腮腺腺细胞的作用。方法:采用Alginate-BSA交联物免疫兔,ELISA法检测藻酸盐抗血清效价;实验分5组:A、空白对照组,B、牛血清白蛋白组,C、藻酸盐组,D、抗藻酸盐血清组和E抗藻酸盐血清+藻酸盐组,分别在1、6、12和24 h采用MTT法检测各组腮腺腺细胞增殖情况。倒置显微镜观察不同组腮腺腺细胞生长、形态和结构变化;扫描电镜观察E组腺细胞超微结构的改变。结果:Alginate-BSA交联物免疫兔约40 d,藻酸盐抗血清的效价达到1∶400;二者适宜反应浓度:藻酸盐为40μg/ml,抗藻酸盐血清的稀释度为1∶100;MTT检测结果A、B、C、D、E组在前3个时间点没有差异,而在24 h时,E组与A、B、C、D组差异显著(P<0.05),说明抗藻酸盐血清和藻酸盐的免疫反应对腺细胞的增殖有显著的抑制。倒置显微镜下在12 h和24 h可观察到E组个别腺细胞表面有破裂,胞质外溢,细胞形态不完整;其它组未见异常。扫描电镜观察E组在6 h即有个别腺细胞胞膜有破裂,呈圆孔形,细胞的轮廓仍清晰完整。12 h可见破裂细胞数增多,且细胞膜上破裂孔、裂也增多变大。24 h可见细胞的形态不完整,有较大范围胞膜破裂,细胞崩解。结论:抗藻酸盐血清和藻酸盐反应对腮腺腺细胞可造成免疫损伤,导致细胞死亡。Objective： To explore the effect of immune reaction induced by alginate on parotid acinar cells in vitro. Methods： Rabbits were immunized from the conjugated alginate-BSA （ 1.0 mg/kg） by 40-days routine immunity method. ELISA method was used to examine the titration （valence） of anti-alginate serum. Five groups （group A： contrast, group B： BSA, group C： alginate, group D： anti-alginate serum, group E： alginate ＋ anti-alginate serum） were examined by MTT method at four time points （ 1,6,12 and 24 h）. The growth and morphology of parotid acinar cells were observed under inverted phase contrast microscope and scanning electron microscope. Results： Antibody-serum was acquired by routine immunity method, and the titration （valence） of anti-alginate serum was 1： 400. M3T results showed that the proliferation of parotid acinar cells had been limited at 24 h（ P 〈 0.05 ）, the other three time points showed no difference. Under inverted phase contrast microscope, a few of acinar cells whose membranes were destroyed after 12 h, some cell contents leaked out. The holes in membrane could be seen early at 6h under scanning electron microscope. Most of the acinar cells were broken at 24 h. Conclusion： The antibody- serum to alginate and immunized rabbit was acquired by routine immunity method. The immune reaction induced by alginate can destroy parotid acinar cells in vitro.

关 键 词：免疫反应 藻酸盐 腮腺腺细胞 MTT法 兔 

分 类 号：R781.7[医药卫生—口腔医学]