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

TU Hai-tao; LU Huan; LUO Yue-zhong; WU Xing-bo

doi:10.3969/j.issn.1001-9448.2017.18.001

Abstract：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 Ser-toli 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 10% 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)in the model group and the low,middle and high drug content serum groups. For comparison between the drug - containing serum groups,the relative expression values of bcl - 2 mRNA for the middle and high drug content serum groups were higher than that for the low drug content serum group,with statistical significance of the differences (P < 0. 01);and the difference of the relative expression values of bcl - 2 mRNA between the middle and high drug content serum group was of statistical significance (P < 0. 01). Conclusion Co - incubation of the spleen - kidney Yang insufficiency IMN serum - injured Sertoli cells and the serum containing the formulation warming Yang for diuresis for 48h may play anti - apoptosis effect by means of up - regulation of the expression level of bcl - 2.