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

发布时间：2018-08-23 08:58

【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(P0.01);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(P0.01);中、高浓度组间,Bcl-2 mRNA相对表达量差异有统计学意义(P0.01)。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。
[Abstract]:Objective to investigate the effect of Wenyang Lishui prescription on apoptosis of podocyte in idiopathic membranous nephropathy (IMN) with deficiency of spleen and kidney yang. Methods CCK-8 was used to detect the effect of serum of IMN patients with different concentrations of spleen and kidney yang deficiency on the proliferation of immortalized podocytes in vitro. The model of podocyte injury with IMN serum of spleen and kidney yang deficiency type was established in vitro. Rats of grade SPF were perfused with water extract of different concentrations of Wenyangli water prescription, blood was taken from abdominal aorta, serum was separated and sterilized. Normal control group, model group, low, medium and high concentration serum group, Wenyang Lishui prescription drug serum and spleen and kidney yang deficiency type IMN serum injured podocytes were incubated for 48 h to detect the relative expression of apoptosis-related gene Caspase-3 and Bcl-2 mRNA by qRT-PCR. Results the model of podocyte injury was induced by 10% serum of IMN patients with deficiency of spleen and kidney yang. The relative expression of Caspase-3 mRNA was increased in the model group after 48 h incubation with different concentrations of WYLW serum. Compared with the normal control group, the expression of Caspase-3 was increased in the model group. However, there was no significant difference in the relative expression of Caspase-3 mRNA and Bcl-2 mRNA in the drug serum group. Compared with the model group, the relative expression of Bcl-2 mRNA was increased in the low, middle and high concentration groups, and there were significant differences between the two groups, and between the two groups, between the model group and the middle and middle groups, there was no significant difference in the relative expression of Bcl-2 mRNA between the two groups. The relative expression of Bcl-2mRNA in the middle and high concentration groups was significantly higher than that in the low concentration group (P0.01), and the difference was statistically significant (P0.01), the difference was significant (P0.01), and the relative expression of Bcl-2mRNA in the middle and high concentration groups was significantly higher than that in the low concentration group (P0.01). There was significant difference in the relative expression of Bcl-2 mRNA between high concentration groups (P0.01). Conclusion for 48 h, podocyte injury in serum of IMN patients with deficiency of spleen and kidney yang and co-incubation of serum containing Wenyang Lishui prescription for 48 h may play an anti-apoptotic effect by upregulating the level of Bcl-2.
【作者单位】：广州中医药大学第一附属医院肾病科;
【基金】：国家自然科学基金资助项目(编号:81373568)
【分类号】：R277.5

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