益气解毒方对12 Gy^(60)Co γ射线辐射后Sertoli细胞分泌功能的防护作用及机制研究  

Protective effect and mechanism of Qi-Boosting Toxin-Resolving Decoction on secretory function of Sertoli cells after ^(60)Co γ-ray radiation at 12 Gy

作　　者：闫玥 叶一帆 王磊 姜荟茜 庄鑫丽 陈晓莹 何昶昊 张淑静 徐文慧 胡素敏 YAN Yue;YE Yifan;WANG Lei;JIANG Huiqian;ZHUANG Xinli;CHEN Xiaoying;HE Changhao;ZHANG Shujing;XU Wenhui;HU Sumin(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488;Beijing Institute of Traditional Chinese Medicine,Beijing University of Chinese Medicine)

机构地区：[1]北京中医药大学中医学院,北京100029 [2]北京中医药大学北京中医药研究院

出　　处：《现代中医临床》2022年第6期44-49,共6页Modern Chinese Clinical Medicine

基　　金：国家自然科学基金项目(No.12075035,No.11675027);北京中医药大学校级重点攻关项目(No.2020-JYB-ZDGG-008)。

摘　　要：目的 研究^(60)Coγ射线对睾丸支持细胞(Sertoli细胞)炎性因子分泌水平的影响,并探究益气解毒方治疗后Sertoli细胞分泌功能损伤的防护机制。方法 Sertoli细胞分为正常组(NC组)、模型组(IR组)、益气解毒高剂量组(YQJD-H组)、益气解毒中剂量组(YQJD组)、益气解毒低剂量组(YQJD-L组)、核因子激活的B细胞的k-轻链增强抑制剂组(Bay组)、中药+Bay组(Y+B组)。各组以相应条件培养24 h后,除NC组外,使用12 Gy^(60)Coγ射线进行一次性照射,建立Sertoli细胞辐射损伤模型。CCK8法检测细胞活力,Elisa法检测细胞上清液中白细胞介素-1β(Interleukin-1β, IL-1β)、白细胞介素-6(Interleukin-6, IL-6)、肿瘤坏死因子-α(Tumor Necrosis Factor-α, TNF-α)、转化生长因子-β(Transforming Growth Factor-β, TGF-β)的含量,WB法检测NF-κB/IκBα通路蛋白表达水平。结果 照射后24 h,与NC组比较,IR组Sertoli细胞增殖能力下降(P<0.01),IL-1β、IL-6、TNF-α、TGF-β的分泌量均明显升高(P<0.01);NF-κB、磷酸化NF-κB蛋白表达水平均明显升高(P<0.01),IκBα蛋白表达水平明显降低(P<0.01);与IR组比较,益气解毒方能够升高照射后Sertoli细胞的存活率(P<0.01),YQJD组、Bay组、Y+B组细胞分泌的IL-1β、IL-6、TNF-α、TGF-β的量均明显降低(P<0.05);YQJD组、Bay组、Y+B组细胞的NF-κB蛋白表达水平明显降低(P<0.05,P<0.01,P<0.01),YQJD组、Y+B组细胞的磷酸化NF-κB蛋白表达水平均明显降低(P<0.01),YQJD组、Bay组、Y+B组细胞的IκBα蛋白表达水平均明显升高(P<0.01)。结论 12 Gy^(60)Coγ射线降低Sertoli细胞增殖能力,使其分泌功能紊乱,而益气解毒方能通过调节NF-κB/IκBα通路,减轻Sertoli细胞受到的辐射损伤,在一定程度上促进睾丸炎性微环境恢复稳定。Objective To study the effect of ^(60)Co γ-ray on secretion of inflammatory factors by Sertoli cells, and to explore the protective mechanism of Yiqi Jiedu Decoction(Qi-Boosting Toxin-Resolving Decoction) on the secretory function of Sertoli cells injured by the radiation.Methods Sertoli cells were divided into Normal Control(NC) group, Ionizing Radiation(IR) group, Yiqi Jiedu Decoction high dosage(YQJD-H) group, Yiqi Jiedu Decoction middle dosage(YQJD) group, Yiqi Jiedu Decoction low dosage(YQJD-L) group, NF-κB inhibitor(Bay) group and Yiqi Jiedu + NF-κB inhibitor(Y+B) group. The groups were cultured under corresponding conditions for 24 hours. Then, radiation-damaged cell models were created by putting all groups except the NC group under^(60)Co γ-ray radiation at 12 Gy. The CCK8 method was used to detect the cell viability of the Sertoli cells, Elisa to detect the content of IL-1β, IL-6, TNF-α, TGF-β in cell supernatant, and Western Blot to detect the protein expression level of the NF-κB/IκB α pathway.Results 24 hours after irradiation, compared with the NC group, the proliferation of Sertoli cells in the IR group decreased(P < 0.01). The secretion of IL-1β, IL-6, TNF-α, TGF-β increased(P < 0.01). The protein expression level of NF-κB and phosphorylated NF-κB increased(P < 0.01). The protein expression level of IκB α decreased(P < 0.01). A comparison with the IR group reveals that Yiqi Jiedu decoction can increase the cell viability of Sertoli cells(P < 0.01). It can also significantly reduce the content of IL-1β, IL-6, TNF-α, TGF-β(P < 0.01, P < 0.01, P < 0.05, P < 0.01). The protein expression level of NF-κBin in the YQJD group, Bay group and Y+B group decreased(P < 0.05, P < 0.001, P < 0.01). The protein expression level of phosphorylated NF-κB in the YQJD group and Y+B group decreased(P < 0.01). The protein expression level of IκB α in the YQJD group, Bay group and Y+B group increased.Conclusion ^(60)Co γ-ray radiation at 12 Gy reduced the proliferation ability of Sertoli cells an

关 键 词：益气解毒方 电离辐射 SERTOLI细胞 炎性因子 NF-κB/IκBα通路 

分 类 号：R2-03[医药卫生—中医学]