温阳利水方含药血清对脾肾阳虚IMN患者血清致小鼠足细胞凋亡的作用  

Research of the effect of serum containing formulation warming Yang on apoptosis of mouse Sertoli cells induced by serum from patients with spleen-kidney Yang insufficiency IMN

作　　者：涂海涛[1] 鲁欢[1] 罗月中[1] 吴兴波[1] TU Hal - tao LU Hnan LUO Yue - zhong WU Xing - bo(The First Affiliated Hospital of Guangzhou University Of Chinese Medicin, Guangzhou 510405, Guangdong, China)

机构地区：[1]广州中医药大学第一附属医院肾病科,广东广州510405

出　　处：《广东医学》2017年第18期2745-2749,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81373568)

摘　　要：目的探讨温阳利水方含药血清对特发性膜性肾病(IMN)脾肾阳虚证足细胞凋亡的作用。方法CCK-8检测不同浓度脾肾阳虚型IMN患者血清对体外培养小鼠永生化足细胞增殖能力的影响,构建脾肾阳虚型IMN血清损伤足细胞体外模型。SPF级大鼠予不同浓度温阳利水方水提物灌胃,腹主动脉取血,分离血清并灭菌后备用。设正常对照组、模型组、低、中、高浓度含药血清组,温阳利水方含药血清与脾肾阳虚型IMN血清损伤足细胞共孵育48 h,qRT-PCR检测凋亡相关基因Caspase-3、Bcl-2 mRNA相对表达量。结果 10%脾肾阳虚型IMN患者血清造成足细胞损伤模型,加入不同浓度温阳利水方含药血清共孵育48 h后,Caspase-3 mRNA相对表达量,与正常对照组比较,模型组增多,但差异无统计学意义;含药血清组Caspase-3 mRNA相对表达量无明显增减。Bcl-2 mRNA相对表达量,与模型组比较,低、中、高浓度含药血清组Bcl-2 mRNA相对表达量均增加,组间比较,模型组与中、高浓度含药血清组差异均有统计学意义(P<0.01);含药血清组间比较,中、高浓度组Bcl-2mRNA相对表达量均较低浓度组增加,且差异均有统计学意义(P<0.01);中、高浓度组间,Bcl-2 mRNA相对表达量差异有统计学意义(P<0.01)。结论对脾肾阳虚型IMN患者血清损伤足细胞,温阳利水方含药血清共孵育48 h,可能通过上调Bcl-2的水平,发挥抗凋亡效应。Objective To investigate the effect of the serum containing the formulation warming Yang on apopto- sis of mouse Sertoli cells induced by serum from patients with spleen - kidney Yang insufficiency IMN （ Idiopathic mem- branous nephropathy）. Methods CCK - 8 （ Cell Counting Kit - 8） was used to detect the effect of the serum at various concentrations from patients with spleen - kidney Yang insufficiency IMN on the proliferative capacity of immortalized Set- toll cells from mice cultured in vitro, so as to construct an in - vitro model of Sertoli cells injured by the serum with spleen - kidney Yang insufficiency IMN. SPF （ specific pathogen free） grade rats was administered intragastrically with the water extract of the formulation warming Yang for diuresis formulation, and then blood samples were collected from the abdominal aorta to isolate the serum, and sterilized it for use. The normal control group, the model group, and the low, mediate and high drug content serum groups were set up. The serum containing the formulation warming Yang for diuresis and the spleen - kidney Yang insufficiency IMN serum - injured Sertoli cells were incubated together for 48h, the relative expres- sion of the apoptosis - related gene caspase - 3 and bcl - s mRNA was assessed with qRT - PCR. Results The I0% ser- um from spleen - kidney Yang insufficiency IMN caused the Sertoli＇s cell - injured model. After adding the serum samples containing the formulation warming Yang for diuresis at different concentrations to incubate together for 48h, the relative expression of caspase - 3 mRNA for the model group increased, compared to that for the control group, without statistical significance of the difference; and that of caspase - 3 mRNA for the drug - containing serum groups had no obvious chan- ges. As for the relative expression of bcl - 2 mRNA, it increased in the low, middle and high drug content serum groups compared to that in the model group. Comparison between groups showed statistical significance （P 〈0.01 ） i

关 键 词：温阳利水方含药 脾肾阳虚证 足细胞 特发性膜性肾病 

分 类 号：R285.5[医药卫生—中药学]