摘要
目的:探讨补肾助孕方对米非司酮诱导的黄体功能不全(LPD)模型大鼠卵巢内质网应激(ERS)的调控机制。方法:使用随机数表法将50只SD大鼠分为空白组,模型组,地屈孕酮(0.02 g·kg^(-1))组,补肾助孕方低(0.08 g·kg^(-1))、高剂量(0.24 g·kg^(-1))组。使用蛋白免疫印迹法(Western blot)及实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠卵巢免疫球蛋白重链结合蛋白(BIP),蛋白激酶R样内质网激酶(PERK),肌醇必需酶-1(IRE-1),活性转录因子6(ATF6),C/EBP同源蛋白(CHOP)蛋白和mRNA表达水平,酶联免疫吸附测定法(ELISA)检测各组大鼠血清孕酮(P)和雌二醇(E2)水平。结果:与空白组比较,模型组BIP,PERK,CHOP蛋白表达显著升高(P<0.01),PERK及CHOP mRNA表达明显下降(P<0.05);而IRE-1,ATF6蛋白表达,IRE-1,BIP,ATF6 mRNA表达,血清E2,P水平差异无统计学意义。与模型组比较,地屈孕酮组CHOP蛋白表达显著降低(P<0.01),而PERK,IRE-1,BIP,ATF6蛋白表达,PERK,IRE-1,BIP,ATF6,CHOP mRNA表达及血清E2,P水平差异无统计学意义;补肾助孕方低剂量组CHOP蛋白表达显著降低(P<0.01),CHOP mRNA表达水平明显升高(P<0.05),而PERK,IRE-1,BIP,ATF6蛋白和mRNA表达及血清E2,P水平差异无统计学意义;补肾助孕方高剂量组PERK,BIP,CHOP蛋白表达显著降低(P<0.01),CHOP mRNA表达水平显著升高(P<0.01),而IRE-1和ATF6蛋白表达,PERK,IRE1,BIP,ATF6 mRNA表达及血清E2,P水平差异无统计学意义。结论:补肾助孕方可以改善黄体功能不全,其机制可能与缓解内质网应激作用有关。
Objective: To investigate the regulatory mechanism of Bushen Zhuyun prescription(BSZYP) on endoplasmic reticulum stress(ERS) in rats with luteal phase defect(LPD) induced by mifepristone. Method: Fifty SD rats were randomly divided into a blank group,a model group,a positive control group(dydrogesterone,0.02 g·kg^(-1)),and low-(0.08 g·kg^(-1))and high-dose(0.24 g·kg^(-1))BSZYP groups.Western blot and Real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR)were used to detect the mRNA and protein expression levels of immunoglobulin binding protein(BIP),protein kinase R-like endoplasmic reticulum kinase(PERK),inositol-requiring enzyme 1(IRE-1),activating transcription factor 6(ATF6),and C/EBP homologous protein(CHOP). The enzyme-linked immunosorbent assay(ELISA)was used to detect the serum progesterone(P)and estradiol(E2)levels. Result: Compared with the blank group,the model group showed the elevated protein expression of BIP,PERK,and CHOP(P<0.01)and the dwindled mRNA expression of PERK and CHOP(P<0.05),while no significant difference was observed in the protein expression of IRE-1 and ATF6,mRNA expression of IRE-1,BIP,and ATF6,and serum E2 and P levels.Compared with the model group,the positive control group displayed diminished protein expression of CHOP(P<0.01),while no significant difference was observed in the protein expression of PERK,IRE-1,BIP,and ATF6,mRNA expression of PERK,IRE-1,BIP,ATF6,and CHOP,and serum levels of E2 and P. The protein expression of CHOP decreased(P<0.01)and the mRNA expression of CHOP increased(P<0.05)in the lowdose BSZYP group,while no significant difference was observed in the mRNA and protein expression of PERK,IRE-1,BIP,and ATF6,and serum E2 and P levels. In the high-dose BSZYP group,the protein expression of PERK,BIP,and CHOP was down-regulated(P<0.01),and the mRNA expression of CHOP was up-regulated(P<0.01),while no significant difference was observed in the protein expression of IRE-1 and ATF6,mRNA expression of PERK,IRE-1,BIP,and ATF6,and serum E2 and P levels. Conclusion: BSZYP can treat LPD by relieving ERS.
作者
史露露
周惠芳
SHI Lu-lu;ZHOU Hui-fang(The First Clinical College of Nanjing University of Chinese Medicine,Nanjing 210046,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Jiangsu Province Hospital of China Medicine,Nanjing 210029,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第16期84-89,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81973898,81774354)
江苏省中医药局重大项目(ZD201702)
江苏省中医药领军人才项目(SLJ0202)。