文献类型：期刊文献

中文题名：黄芪主要成分对TGF-β1损伤睾丸Leydig细胞保护效应的研究

英文题名：Research on Protective Effects of Main Components of Huangqi(Astragali Radix) on TGF-β1-damaged Testis Leydig Cells

作者：李彦荣[1];李能莲[1];杨丽红[1];高成英[1];张宝元[1];颜鲁林[1]

第一作者：李彦荣

机构：[1]甘肃中医药大学,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2021

卷号：39

期号：6

起止页码：200

中文期刊名：中华中医药学刊

外文期刊名：Chinese Archives of Traditional Chinese Medicine

收录：CSTPCD;;北大核心:【北大核心2020】；

基金：甘肃省高等学校科研项目(2018A-180);甘肃中医药大学定西校区项目(2017XJZD09)。

语种：中文

中文关键词：黄芪;Leydig细胞;保护效应

外文关键词：main components of Huangqi(Astragali Radix);Leydig cells;protective effect

摘要：目的通过研究黄芪总皂苷、黄芪总多糖、黄芪总黄酮对TGF-β1损伤睾丸Leydig细胞增殖和活性的影响,探究黄芪主要成分中对睾丸Leydig细胞保护具有先导性化合物地位的成分。方法体外培养睾丸Leydig细胞,将培养的细胞分为3组:正常对照组、模型组、实验组。正常对照组加入RPMI1640完全培养基2 mL;模型组加入含5 ng/mL TGF-β1及与中药等体积PBS的RPMI1640完全培养基2 mL;实验组分别加入黄芪总皂苷终浓度20、100μg/mL,黄芪总多糖终浓度10、50μg/mL,黄芪总黄酮终浓度10、50μg/mL的含有5 ng/mL TGF-β1的RPMI1640完全培养基2 mL。48 h后进行细胞爬片和ELISA法检测T、E2、P450aromd蛋白含量,显微镜观察细胞爬片,用SPSS 20.0、GraphPad Prism 6.0软件对数据进行统计分析。结果 TGF-β1能使Leydig细胞形态异常,数量减少,明显抑制Leydig细胞合成T、E2、P450aromd,与正常组比较差异有统计学意义(P<0.05)。黄芪总皂苷、黄芪总多糖、黄芪总黄酮都均能改善睾丸Leydig细胞病变,促进Leydig细胞的增殖,并且存在一定的量效关系。三成分都能提高TGF-β1损伤的睾丸Leydig细胞的活性,与模型组比较差异有统计学意义(P<0.05),单因素分析,终浓度10μg/mL的黄芪总黄酮缓解TGF-β1抑制睾丸Leydig细胞合成T的效果最显著;终浓度20μg/mL的黄芪总皂苷缓解TGF-β1抑制睾丸Leydig细胞合成E2的效果最显著;终浓度50μg/mL的黄芪总黄酮缓解TGF-β1抑制睾丸Leydig细胞合成P450arom的效果最显著。三者与模型组比较差异有统计学意义(P<0.05)。多因素分析,终浓度50μg/mL的黄芪总黄酮缓解缓解TGF-β1抑制Leydig细胞合成T、E2、P450aromd的效果最显著。结论黄芪总黄酮对睾丸Leydig细胞保护具有先导性化合物的地位。
Objective By studying the effects of main components of Huangqi(Astragali Radix) on proliferation and activity of TGF-β1-damaged testis leydig cells, the paper is designed to probe into the constituents of Huangqi(Astragali Radix) main chemical components that keep leading compounds working in Testis Leydig cells. Methods The leydig cells cultured in vitro were divided into three groups: control group, model group and experiment group. The normal control group was added with RPMI1640 full medium 2 mL. The model group added 2 mL of RPMI1640 full culture medium containing 5 ng/mL TGF β1 and PBS with the same volume of the Chinese medicine. The experimental group added 2 mL RPMI1640 with 5 ng/mL of TGF-β1,20 μg/mL and 100 μg/mL of total astragaloside final concentration, 10 μg/mL and 50 μg/mL of total astragalus polysaccharide, 10 μg/mL and 50 μg/mL of total flavonoids. After 48 hours, the contents of T, E2 and p450 aromd protein were detected by cell climbing tablets and ELISA. The cell climbing tablets were observed by microscope.The data was analyzed by SPSS 20.0 and GraphPad Prism 6.0 software. Results TGF-β1 can make Leydig cells abnormal in morphology and decrease in quantity, and significantly inhibit the synthesis of T, E2 and p450 aromd by Leydig cells. There was significant difference compared with the normal group(P<0.05). The results showed that the total saponins, polysaccharides and flavonoids of Huangqi(Astragali Radix) could improve the Leydig cell lesion and promote the proliferation of Leydig cells. The three components can improve the activity of Leydig cells in testis with TGF-β1 injury, which was statistically significant(P<0.05). Single factor analysis showed that the total flavone of Huangqi(Astragali Radix) with final concentration of 10 μg/mL can alleviate the effect of TGF-β1 on the synthesis of T of Leydig cells in testis. The final concentration was 20 μg/mL. The results showed that the total saponin of Huangqi(Astragali Radix) 20 μg/mL had the most significant effect on inhibiting the synthesis of E2 by Leydig cells in testis, and the total flavonoid of Huangqi(Astragali Radix) with the final concentration of 50 μg/mL was the most effective in inhibiting the synthesis of P450 arom by TGF-β1. The difference of the three groups and the model group was statistically significant(P<0.05). The results of multivariate analysis showed that the total flavonoid of Huangqi(Astragali Radix)with the final concentration of 50 μg/mL alleviated the inhibition of TGF-β1 on the synthesis of T, E2 and p450 aromd by Leydig cells. Conclusion Protective effects of the three extracts on Leydig cells vary and are most probably exerted by inhibiting TGF-β1, of which total astragalys flavonoids play a leading role on testis Leydig cells.