目的探讨采用两种超排卵剂量下的鼠胚实验对辅助生殖实验室进行质控的可行性。方法对SPF级昆明雌性小白鼠行腹腔注射孕马血清促性腺激素(PMSG)和绒毛膜促性腺激素(HCG)超排卵,在采用8 IU PMSG+8 IU HCG和16 IU PMSG+16 IU HCG两种不同注射剂量下进行体外受精和体内受精,观察不同剂量、不同受精方式下鼠胚发育的优胚率和囊胚形成率。结果分别实施8 IU PMSG+8 IU HCG超排卵剂量与16 IU PMSG+16 IU HCG超排卵剂量后,雌性小鼠体外受精,鼠胚优胚率分别为90.965%和81.877%,囊胚形成率分别为82.341%和75.081%,差异有统计学意义(P<0.05);雌性小鼠体内受精,鼠胚优胚率分别为93.096%和85.889%,囊胚形成率分别为83.432%和76.994%,差异有统计学意义(P<0.05)。结论采用两种超排卵剂量下的鼠胚实验对辅助生殖实验室进行质控具有可行性,恰当的超排卵剂量,能获得较高的优胚率和囊胚形成率。
Objective:This study aimed to evaluate the effects of acupuncture on embryo implantation and maternal–fetal immune tolerance in rats with ovarian hyperstimulation,identify the mechanisms of acupuncture treatment,and promote application of acupuncture in the field of assisted reproduction.Methods:Eighty female Wistar rats were randomized into normal(N),model(M),acupuncture treatment(A),and progesterone treatment(P)groups,with 20 rats in each group.An ovarian hyperstimulation model was established in groups M,A,and P by the peritoneal injection of pregnant mare serum gonadotropin(PMSG)combined with human chorionic gonadotropin(HCG).Bilateral“Sanyinjiao(SP6)”,“Zusanli(ST36)”and“Taichong(LR3)”as acupoints were needled in rats in group A.Rats in group P received an intramuscular injection of progesterone(4 mg/kg).All pregnant rats were treated for seven days.On the 8th day,the pregnant rats were sacrificed for testing.The expression levels of estrogen(E2)and progesterone(P4)in serum and their receptors in the endometrium,Th1 and Th2 cytokines in serum and endometrium,RT-BM1(the counterpart of HLA-G in rats)mRNA,Ly49 activating receptors(Ly49s3,Ly49s5)and inhibitory receptors(Ly49i3,Ly49i4,Ly49i5)on immunocytes in the maternal–fetal interface were detected using ELISA,Western blot,and real-time polymerase chain reaction(PCR).Results:Compared with group N,the pregnancy rate and the number of implanted embryos were significantly decreased in group M(P<0.05);E2 and P4 levels in serum,and the protein levels of ER-αand Progesterone Receptor(PGR)in the implantation site of endometrial tissue were significantly reduced in group M(P<0.05);the expression of INF-γand IL-2 in serum and endometrium were significantly increased,while the expression of IL-4,IL-6,and IL-10 in serum and endometrium were significantly decreased in group M(P<0.05).The mRNA levels of RT-BM1,Ly49i3,and Ly49i4 receptors in group M were significantly higher than those in group N,whereas the mRNA levels of Ly49i5 and Ly49s5 receptors in gro
