山楂黄酮对小鼠睾丸间质细胞热损伤保护作用

Protective effect of hawthorn flavones on thermal damage of mice leydig cells

目的探讨山楂黄酮对离体小鼠睾丸间质细胞的热应激防护作用。方法选取对数生长期的小鼠睾丸间质细胞株(TM3),设对照组、热应激组(43℃1 h)、山楂黄酮高、中、低剂量组(800、200、50 mg/L)、阳性对照组(绒毛膜促性腺激素500 IU/L)。采用噻唑蓝法观察细胞活性,光学显微镜观察细胞形态学变化,检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性。结果与对照组比较,热应激组细胞增殖率[(77.74±5.10)%]明显下降,SOD和GSH-Px活性[分别为(19.27±1.02)、(246.03±13.53)U/mL]降低,MDA含量[(2.54±0.09)nmol/mL]明显升高(P<0.01),细胞数量明显下降,细胞收缩变形;与热应激组比较,山楂黄酮低剂量组细胞增值率[(91.33±5.50)%]、SOD和GSH-Px活性[分别为(28.31±1.30)、(270.91±10.34)U/mL]明显升高,MDA含量[(2.04±0.01)nmol/mL]明显下降(P<0.05),细胞数量虽明显减少,但细胞形状保持良好。结论山楂黄酮可增强细胞抗氧化能力,对睾丸间质细胞的热应激损伤具有一定保护作用。

Objective To examine protective effect of hawthorn flavones on thermal damage in mice leydig cells in vitro.Methods Mice leydig T cell lines （TM3）during logarithmic phase growth were selected and divided into different groups （control,thermal stress group[43℃ for 1 hour],thermal stress plus drug group and positive control group）.The mice in thermal stress plus drug group were then divided into high （flavones of 800 mg/L）,midium （200 mg/L）,and low dose groups（50 mg/L）.The mice in positive control group were treated with human chorionic gonadotropin（HCG）（500 IU/L）.The cell activity was observed with methyl thiazoletetrazolium （MTT）assay and cell proliferation rate was expressed by experimental groups/control ×100%.The morphological change was observed with optical microscopy.Molondialdehyde （MDA）,superoxide dismutase （SOD）,and glutathione peroxidase （GSH-Px）were determined.Results The cell activity of thermal stress model group（77.74±5.10%）reduced significantly（P〈0.01）compared with the control;the cell activity of high（84.28±3.47%）,midium（88.57±8.89%）and low dose drug group（91.33±5.50%）increased significantly（P〈0.01）compared with that of control group.The cell activity of HCG positive control group（93.52±0.75%）increased significantly （P〈0.01）.The leydig cells of control group grew well.The number of the cells in the thermal stress group decreased obviously and deformation was observed after heat treatment.The number of the cells was not obviously changed in HCG positive control group and so as the number of cells in drug groups but the cells were in good shape.The contents of SOD （19.27±1.02 U/ml）and GSH-Px（246.03±13.53 U/ml）of thermal stress group reduced significantly（P〈0.01）and MDA increased significantly（P〈0.01）compared with those of the control.The contents of SOD（28.12±1.02 U/ml）and GSH-Px （290.62±13.53 U/ml）of HCG positive control group increased significantly（P〈0.01）and MDA decreased significantly （P〈0.01）compared with thermal stress group.The contents of SOD and GSH-Px of drug groups increased significantly（P〈0.05）and MDA（1.94±0.08 nmol/ml）decreased significantly（P〈0.05）compared with those of thermal stress group.Conclusion Hawthorn flavones has antioxidant capacity and protective effect on thermal stress damage in mice leydig cells.