膜细胞对化疗损伤性颗粒细胞的影响及左归丸含药血清干预作用  

Effect of theca cells on granulosa cells injured by chemotherapy and intervention with serum containing Zuogui pill components

作　　者：孙晓峰 黄欣怡 曾贵荣 龚力民[3] 乔江[4] 阳松威 周琳[5] SUN Xiaofeng;HUANG Xinyi;ZENG Guirong;GONG Limin;QIAO Jiang;YANG Songwei;ZHOU Lin(School of Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Center of Drug Safety Evaluation and Research of Drugs&Hunan Key Laboratory of Pharmacodynamics and Safety Evaluation of New Drugs,Changsha 410331;College of Medicine,Hunan University of Traditional Chinese Medicine,Changsha 410208;Department of Obstetrics and Gynecology,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410007;Changsha Medical College,Changsha 410219.)

机构地区：[1]湖南中医药大学医学院,长沙410208 [2]湖南省药物安全评价研究中心&新药药效与安全性评价湖南省重点实验室,长沙410331 [3]湖南中医药大学药学院,长沙410208 [4]湖南中医药大学附属第一医院妇产科,长沙410007 [5]长沙医学院,长沙410219

出　　处：《中国比较医学杂志》2022年第2期16-21,共6页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(81774362,81303004);湖南省自然科学基金(2018JJ2290)。

摘　　要：目的探讨膜细胞对化疗损伤性颗粒细胞的影响以及左归丸含药血清的干预作用。方法制备左归丸含药血清,培养大鼠卵巢颗粒细胞和膜细胞,分8组,分别是(1)空白对照组;(2)共培养组;(3)ZGW血清组;(4)ZGW血清+共培养组;(5)模型组;(6)模型+共培养组;(7)模型+ZGW血清组;(8)模型+ZGW血清+共培养组。实时荧光定量PCR法(qRT-PCR)及蛋白免疫印迹法(Western blot)分别检测Bax、Caspase-3、Beclin-1、p62、LC3B在转录水平和翻译水平上的表达。结果模型组的Beclin-1、LC3B、Bax及Caspase-3基因在转录水平及翻译水平上均有高表达,加入10%左归丸含药血清处理或与膜细胞共培养可下调上述基因的表达;p62基因的表达较对照组降低,加入10%左归丸含药血清处理或与膜细胞共培养可上调模型组p62基因表达。当实验组与膜细胞共培养时加入10%左归丸含药血清后,5种基因表达的变化程度更明显。结论膜细胞能缓解体外颗粒细胞的化疗性损伤,且与左归丸含药血清有协同作用,与抑制颗粒细胞自噬和减少其凋亡有关。Objective To investigate the effect of theca cells on granulosa cells injured by chemotherapy and the intervention effect of serum containing Zuogui pill(ZGW).Methods Serum containing Zuogui pill components was prepared.Granulosa and theca cells were cultured and divided into 8 groups,including blank control group,coculture group,ZGW serum group,ZGW serum+coculture group,model group,model+coculture group,model+ZGW serum group and model+ZGW serum+coculture group.The expression levels of Bax,Caspase-3,Beclin 1,p62 and LC3B were detected by qRT-PCR and Western blot.Results Beclin-1,LC3B,Bax and Caspase-3 were highly expressed in the model group at the transcriptional and translational levels,which were downregulated after adding 10%serum containing Zuogui pill components or co-culturing with theca cells.The gene expression of p62 was lower in the model group than that in the control group,and was upregulated by adding 10%serum containing Zuogui pill components or co-culturing with theca cells.When the experimental group was co-cultured with theca cells,the expression of these genes changed more substantially after the addition of 10%serum containing Zuogui pill components.Conclusions Theca cells alleviated the chemotherapy-induced injury of granulosa cells in vitro and demonstrated a synergistic effect with serum containing Zuogui pill components.These effects were related to the inhibition of autophagy and reduction in apoptosis of granulosa cells.

关 键 词：左归丸 化疗损伤 颗粒细胞 膜细胞 自噬 凋亡 

分 类 号：R-33[医药卫生]