益气解毒方对12 Gy60Co 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 60Co γ-ray radiation at 12 Gy
- 现代中医临床 2022年29卷第6期 页码：44-49
- 作者机构：
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
- 作者简介：
闫玥，女，在读硕士生
#胡素敏，女，博士，教授，博士生导师，E-mail：husm@bucm.edu.cn
- 基金信息：国家自然科学基金项目(12075035;11675027);北京中医药大学校级重点攻关项目(2020－JYB-ZDGG－008)
DOI：10.3969/j.issn.2095-6606.2022.06.010
中图分类号： R2-03纸质出版日期：2022-11-30，
收稿日期：2022-02-26，
- 稿件说明：
移动端阅览
闫玥, 叶一帆, 王磊, 等. 益气解毒方对12 Gy60Co γ射线辐射后Sertoli细胞分泌功能的防护作用及机制研究[J]. 现代中医临床, 2022,29(6):44-49.
YAN Yue, YE Yifan, WANG Lei, et al. Protective effect and mechanism of Qi-Boosting Toxin-Resolving Decoction on secretory function of Sertoli cells after 60Co γ-ray radiation at 12 Gy[J]. Modern Chinese Clinical Medicine, 2022,29(6):44-49.
闫玥, 叶一帆, 王磊, 等. 益气解毒方对12 Gy60Co γ射线辐射后Sertoli细胞分泌功能的防护作用及机制研究[J]. 现代中医临床, 2022,29(6):44-49. DOI： 10.3969/j.issn.2095-6606.2022.06.010.
YAN Yue, YE Yifan, WANG Lei, et al. Protective effect and mechanism of Qi-Boosting Toxin-Resolving Decoction on secretory function of Sertoli cells after 60Co γ-ray radiation at 12 Gy[J]. Modern Chinese Clinical Medicine, 2022,29(6):44-49. DOI： 10.3969/j.issn.2095-6606.2022.06.010.
摘要
目的
2
研究
60
Co γ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
2
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
Co γ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/IκB α通路蛋白表达水平。
结果
2
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
2
12 Gy
60
Co γ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/IκB α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。
Abstract
Objective
2
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
2
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
2
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
2
60
Co γ-ray radiation at 12 Gy reduced the proliferation ability of Sertoli cells and disordered their secretory function. Yiqi Jiedu Decoction can significantly reduce the radiation damage to Sertoli cells by regulating NF-κB/IκB α pathway
and promote the restoration and stability of testicular microenvironment to a certain extent.
关键词
益气解毒方电离辐射Sertoli细胞炎性因子NF－κB/IκB α通路
Keywords
Yiqi Jiedu DecoctionirradiationSertoli cellsinflammatory factorNF-κB/IκB α pathway
references
徐郁琴，丁艳平，刘淑宁，等.黄芪甲苷通过toll样受体4/核因子κB通路介导辐射所致肾损伤的防护作用[J].解剖学报，2021,52(4):621-627.
LI Y H，HE Q，CHEN Y Z, et al. p-Coumaric acid ameliorates ionizing radiation-induced intestinal injury through modulation of oxidative stress and pyroptosis[J]. Life Sciences, 2021, 278(10):119546.
王晓玲，黄冬榕，郑倩倩，等.“芪归药对”干预NF－κB通路调控骨髓造血干细胞辐射旁效应损伤的机制研究[J].时珍国医国药，2021,32(5):1051-1054.
尤耀东，张玲，俞旭君，等.基于“生精微环境”探讨强精片对不育小鼠Sertoli细胞炎性因子IL－1β、TNF－ɑ分泌的影响[J].中华男科学杂志，2020,26(8):740-744.
翁碧霞，娄江涛，余亮亮，等.红景天苷减轻手机电磁辐射对大鼠睾丸支持细胞损伤的实验研究[J].浙江中医药大学学报，2018,42(11):951-956.
潘斌，席晔斌，陈广洁，等.锌指蛋白265基因对睾丸Sertoli细胞UU感染后IL－1α、IL－6及TGF－β1表达的影响[J].现代免疫学，2013,33(3):219-224.
杨文军. 姜黄素对电离辐射致雄性小鼠生殖系统损伤的保护作用及相关机制研究[D].上海：中国人民解放军海军军医大学，2018.
匡振坤，杨云霜，张蓉，等.补肾解毒方对核辐射所致生殖系统损伤的防护作用[J].中医杂志，2017,58(24):2128-2131.
赵雪. 补肾解毒方对电离辐射致小鼠生殖系统损伤的防治作用研究[D].锦州：锦州医科大学，2018.
陈晓莹，何昶昊，胡素敏，等.益气解毒方对2Gy 60Coγ射线诱导的雄性小鼠生精功能损伤的防护作用[J].湖南中医药大学学报，2020,40(12):1456-1461.
卢曦，李盼飞，胡素敏，等.益气解毒方对2.0 Gy～(60)Coγ射线致小鼠睾丸急性损伤的防护效应[J].北京中医药大学学报，2020,43(9):746-753.
DUNPHY G, FLANNERY SM, ALMINE JF, et al. Non-canonical activation of the dna sensing adaptor STING by ATM and IFI16 mediates NF-kappaB signaling after nuclear DNA damage [J]. Mol Cell, 2018, 71(5):745-760.
ZHAO J, ZHANG L, LU A, et al. ATM is a key driver of NF-kappaB-dependent DNA-damage-induced senescence, stem cell dysfunction and aging [J]. Aging (Albany NY), 2020, 12(6):4688-4710.
SWETA S，DAMODARr G . Mannan oligosaccharide requires functional ETC and TLR for biological radiation protection to normal cells[J]. BMC Cell Biology, 2018, 19:9.
MULERO M C, HUXFORD T, GHOSH G. NF-kappaB, IkappaB, and IKK: Integral Components of Immune System Signaling [J]. Adv Exp Med Biol, 2019, 1172:207-226.
ZHANG Q, LENARDO MJ, BALTIMORE D. 30 Years of NF-kappaB: A Blossoming of Relevance to Human Pathobiology [J]. Cell, 2017, 168(1-2):37-57.
KANAREK N, BEN-NERIAH Y. Regulation of NF-kappaB by ubiquitination and degradation of the IkappaBs [J]. Immunol Rev, 2012, 246(1): 77-94.
CECILIA GARLANDA，CHARLES A. DINARELLO, ALBERTO MANTOVANI. The Interleukin－1 family: back to the future[J]. Immunity, 2013, 39(6): 1003-1018.
DINARELLO CHARLES A. Overview of the IL－1 family in innate inflammation and acquired immunity.[J]. Immunological reviews, 2018, 281(1) : 8-27.
杨晓炜，刘凤海，刁慧杰，等.抗肿瘤坏死因子－α对大鼠脑缺血再灌注损伤的干预作用[J].中国老年学杂志，2021,41(21):4780-4784.
陈宣维，贾连顺，林建华，等.大鼠脊髓损伤后白细胞介素－1β、肿瘤坏死因子－α基因表达的变化[J].骨与关节损伤杂志，2003,18(11):764-766.
D MONSIVAIS, MATZUK MM, PANGAS SA. The TGF－β Family in the Reproductive Tract[J]. Cold Spring Harbor Perspectives in Biology, 2017:a022251.
RIHAM SOLIMAN SAID，HEBA A. MOHAMED, DINA HAMADA KASSEM. Alpha-lipoic acid effectively attenuates ionizing radiation-mediated testicular dysfunction in rats: Crosstalk of NF－B, TGF－β，and PPAR-pathways[J]. Toxicology, 2020, 442(prepublish) : 152536-152536.
郝钰，万红娇，邝枣园.医学免疫学与病原生物学[M].北京：科学出版社，2017:40-41.
王也，张力木，齐帅，等.IL－6、IL－33、IL－10在牙周炎和类风湿关节炎中的表达和相关性分析[J].上海口腔医学，2021,30(5):498-503.
陈纪烨，周国锋，王永成，等.桂枝汤桂枝－白芍不同比例配伍通过调节TGF－β_1/Smads信号通路及慢性炎症改善盐敏感高血压大鼠心肌纤维化[J].中国实验方剂学杂志，2020,26(1):50-58.
JIANG K, YAO G, HU L, et al. MOB2 suppresses GBM cell migration and invasion via regulation of FAK/Akt and cAMP/PKA signaling [J]. Cell Death Dis, 2020, 11(4): 230.
王磊，王安，胡素敏.试论急性辐射损伤的中医学病因——电离毒[J].北京中医药大学学报，2017,40(1):27-30.
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