女贞子多糖在脂多糖诱导大鼠睾丸支持细胞炎性损伤中保护作用的研究  

Protective effect of Ligustri Lucidi Ait Polysaccharide against lipopolysaccharide-induced inflammatory injury of Sertoli cells in rats

作　　者：余亮亮[1] 娄江涛[1] 魏任雄[1] 陈建伟 YU Liang-liang;LOU Jiang-tao;WEI Ren-xiong;CHEN Jian-wei(Laboratory for Male Diseases,Ningbo Hospital of Traditional Chinese Medicine,Zhejiang University of Chinese Medicine,Ningbo,Zhejiang 315012,China;Ningbo Institute of Drug Control,Ningbo,Zhejiang 315000,China)

机构地区：[1]浙江中医药大学附属宁波市中医院男科实验室,浙江宁波315012 [2]宁波市药品检验所,浙江宁波315000

出　　处：《中华男科学杂志》2018年第10期871-877,共7页National Journal of Andrology

基　　金：宁波市自然科学基金(2016A610200)~~

摘　　要：目的:探究女贞子多糖(LLP)对脂多糖(LPS)诱导的大鼠睾丸支持细胞炎性损伤的影响。方法:体外分离和培养大鼠支持细胞,分对照组、LPS组、LPS+低剂量LLP组、LPS+中剂量LLP组和LPS+高剂量LLP组,处理细胞48 h。CCK-8检测细胞增殖,流式细胞术检测细胞凋亡,紫外分光光度法检测细胞培养液上清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化酶(GSH-Px)活性和丙二醛(MDA)浓度; ELISA检测各组细胞培养液上清中IL-1α,IL-6和TGF-β含量后,去除LPS继续培养48 h,再次检测各组细胞培养液上清中IL-1α,IL-6和TGF-β含量。结果:各组细胞增殖组间差异有统计学意义(F=153. 93,P <0. 01),LPS组与对照组相比细胞增殖率明显降低(P <0. 01),而中剂量和高剂量LLP+LPS组与LPS组相比细胞增殖率明显增高(P <0. 01);各组细胞凋亡率组间差异有统计学意义(F=64. 06,P <0. 01); LPS组与对照组相比细胞凋亡率明显增高(P <0. 05),而与LPS组相比,中剂量和高剂量LLP+LPS组细胞凋亡率有不同程度的下降(P <0. 01)。各组细胞SOD、CAT、GSH-Px和MDA组间差异均有统计学意义(F值分别为56. 07、41. 57、238. 46、285. 31,P <0. 01),LPS组与对照组相比SOD、CAT和GSH-Px活性均明显降低(P <0. 01),MDA浓度明显增加(P <0. 01);中剂量和高剂量LLP+LPS组与LPS组相比,SOD和CAT活性均有不同程度升高,而3种剂量LLP+LPS组与LPS组相比,GSH-Px活性逐渐升高,MDA浓度逐渐下降(P <0. 01)。去除LPS前,LPS组与对照组相比IL-1α、IL-6和TGF-β浓度明显增加(P <0. 01),中剂量和高剂量LLP+LPS组与LPS组相比IL-1α浓度逐渐增加(P <0. 05或<0. 01),而3种剂量LLP+LPS组与LPS组相比,IL-6和TGF-β浓度变化不明显(P> 0. 05)。去除LPS后LPS组IL-1α、IL-6明显降低,而TGF-β明显增加(t值分别为25. 26、61. 43和-18. 16,P <0. 01); LLP+LPS组IL-1α和IL-6较LPS组下降更明显,TGF-β浓度增加更显著(P <0. 01)。结论:LLP在LPS诱导大鼠睾丸支持细胞Objective： To explore the effect of Ligustri Lucidi Ait Polysaccharide （LLP） on lipopolysaccharide （LPS）-induced inflammatory injury of Sertoli cells. Methods ： Rat Sertoli cells were isolated and cultured in vitro and then divided into five groups, blank control, LPS, LPS ＋ low-dose LLP, LPS ＋ medium-dose LLP, and LPS ＋ high-dose LLP. After 48 hours of treatment, the proliferation of the cells was detected by CCK-8, their apoptosis determined by FMC, and the levels of superoxide dismutase （SOD）, catalase （CAT）, glutathione peroxidase （GSH-Px） activity and malondialdehyde （MDA） in the supematant of the cell culture medium measured by ultraviolet spectrophotometry. The contents of IL-lc~, IL-6 and TGF-[3 in the culture medium were detected by ELISA be- fore and after removal of LPS. Results ： The proliferation of the cells showed statistically significant differences among different groups （ F = 153.93, P 〈 0.01 ）, markedly reduced in the LPS group as compared with the blank control （ P 〈 0.01 ）, but remarkably in- creased in the high- and medium-dose LLP groups in comparison with the LPS group （ both P 〈 0.01 ） , and so did the apoptosis of the cells （F = 64.06, P 〈 0.01 ）, significantly increased in the LPS group as compared with the blank control （P 〈 0.05）, but markedly decreased in the high- and medium-dose LLP groups in comparison with the LPS group （ both P 〈 0.01 ）. Statistically significant differences were also observed among different groups in the levels of SOD （ F = 56.07, P 〈 0.01 ）, CAT （ F = 41.57, P 〈 0.01）, GSH-Px activity （F = 238.46, P 〈 0.01）, and MDA （F = 285.31, P 〈 0.01）, with decreased SOD, CAT and GSH-Px activity （ P 〈 0.01 ） and increased MDA （ P 〈 0.01 ） in the LPS group as compared with the control, but elevated SOD and CAT in the high- and medium-dose LLP groups and increased GSH-Px activity and decreased MDA concentration in all the three LLP groups in comparison with the LPS g

关 键 词：支持细胞 炎性损伤 脂多糖 免疫因子 女贞子多糖 大鼠 

分 类 号：R364.5[医药卫生—病理学] R321.1[医药卫生—基础医学]