Objective To investigate the impacts and mechanism of circular RNA polyribonucleotide nucleotidyltransferase 1(Circ-PNPT1)on glucose and lipid metabolism and offspring outcomes in rats with GDM through microRNA-345-3p(miR-345-3p)/lipid raft scaffold protein 2(FLOT2)axis.Methods 75 pregnant female rats were divided into normal control(NC)group,GDM group,si-NC group,si-Circ-PNPT1 group,and si-Circ-PNPT1+miR-345-3p-inhibitor group,with 15 rats in each group.Glucose and lipid metabolism indexes were detected in each group.FIns was detected by ELISA.The survival rate and body weight of fetal rats were compared in each group.The expression of Circ-PNPT1,miR-345-3p and FLOT2 mRNA in placenta was detected by qRT-PCR.The double luciferase reporter gene experiment verified the targeting relationship between miR-345-3p and Circ-PNPT1 and FLOT2.Western blot was used to detect the expression of FLOT2 protein in rat placenta.Results Compared with the NC group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expressions of Circ-PNPT1,FLOT2 in GDM group increased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p decreased(P<0.05).Compared with GDM group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expression of FLOT2 in si-Circ-PNPT1 group decreased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p increased(P<0.05);MiR-345-3p-inhibitor reversed the improvement of si-Circ-PNPT1 on GDM rats(P<0.05).Conclusions Knocking down Circ-PNPT1 can up-regulate miR-345-3p and down-regulate FLOT2 to improve glucose and lipid metabolism and offspring outcome in GDM rats.

Objective To investigate the impacts and mechanism of circular RNA polyribonucleotide nucleotidyltransferase 1(Circ-PNPT1)on glucose and lipid metabolism and offspring outcomes in rats with GDM through microRNA-345-3p(miR-345-3p)/lipid raft scaffold protein 2(FLOT2)axis.Methods 75 pregnant female rats were divided into normal control(NC)group,GDM group,si-NC group,si-Circ-PNPT1 group,and si-Circ-PNPT1+miR-345-3p-inhibitor group,with 15 rats in each group.Glucose and lipid metabolism indexes were detected in each group.FIns was detected by ELISA.The survival rate and body weight of fetal rats were compared in each group.The expression of Circ-PNPT1,miR-345-3p and FLOT2 mRNA in placenta was detected by qRT-PCR.The double luciferase reporter gene experiment verified the targeting relationship between miR-345-3p and Circ-PNPT1 and FLOT2.Western blot was used to detect the expression of FLOT2 protein in rat placenta.Results Compared with the NC group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expressions of Circ-PNPT1,FLOT2 in GDM group increased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p decreased(P<0.05).Compared with GDM group,the level of FPG,FIns,HOMA-IR,TC,TG,LDL-C,embryo weight,and the expression of FLOT2 in si-Circ-PNPT1 group decreased,the level of HDL-C,embryo survival rate and the expression of miR-345-3p increased(P<0.05);MiR-345-3p-inhibitor reversed the improvement of si-Circ-PNPT1 on GDM rats(P<0.05).Conclusions Knocking down Circ-PNPT1 can up-regulate miR-345-3p and down-regulate FLOT2 to improve glucose and lipid metabolism and offspring outcome in GDM rats.

Objective To study the influence of 4-aminopyridine(4-AP)on proliferation,production,and apoptosis through inhibiting voltage-gated K+channel(Kv)in ovarian luteinized granulosa cells.nethods Ovarian luteinized granulosa cells were recovered from 25 women with regular menses who underwent in vitro fertilization programme.The cultured granulosa cells were divided into 4 groups:blank group,4-AP treated group,human chorionic gonadotropin(hCG)-induced group and hCG+4-AP cotreated group.The final concentrations of hCG and 4-AP were 1250 U/L and 5 nmol/L respectively.The progesterone production WaS detected by the chemoluminescence method.The expression of Kv mRNA on human ovarian luteinized granulosa cell was detected by RT.PCR The influence on the early apoptosis of gTanulosa cells bv 4-AP was observed by flow eytometry.Cellular caSpage-3 activities were observed with colorimetric method and the inhibition of the cell proliferation was studied using methyl thiazolyl tetrazolium(MTT)method.Results(1)Kv mRNA wag expressed in granulosa cell.(2)The progesterone production64),(206±32),(1991±172)and(763±79)nmol/L,respectively after24 hours culture.Exposure of the(3)The flow cytometry analysis and the cellular caapase-3 A405 showed that 4-AP increased the percentage ofearly phase apoptosis(P<0.01):4-AP treated group VS blank group[(40±5)%and 0.049 ±0.009]VS[(17±4)% and 0.029±0.008],hCG+4-AP CO-treated group VS hCG-induced group[(25±4)%and0.039 ±0.0081 VS[(15±3)%and 0.022 ±0.007].(4)24 hours after treated with 4-AP and hCG,theinhibitory rate of cultured granulosa cells of 4-AP treated group was higher than the blank group(19.7%VS0).and that of hCG+4-AP co-treated group was obviously higher than hCG-induced group(34.6% VS O,P<0.01).Conclusions The voltage-gated K+ channels expressed by ovarian luteinized granulosa cellplay an important role in cell proliferation,production,and apoptosis.4-AP may inhibit differentiation ofprogesterone in granulosa ceHs through the inhibition of proliferation and induction of apoptosis.