The Variation Of PI3K-AKT-mTOR And AMPK-mTOR Signaling Pathway In Human Uterine Smooth Muscle During Labor

IntroductionDelivery is a complicated and long process.Uterine contraction is one of the key points of delivery.Abnormal uterine contraction is one of the important reasons leading to dystocia and postpartum hemorrhage.Studies on the initiation,maintenance and regulation of uterine contraction are unclear.Our previous studies have found autophagy activation in uterine smooth muscle cell during delivery.Rhythmic contractions during labor cause paroxysmal and transient hypoxia of uterine smooth muscle tissue,autophagy plays an important role in the process of ischemia and hypoxia.Autophagy is a "double-edged sword",which can promote cell metabolism and maintain cell stability.Excessive autophagy can also cause cell death.Is PI3K-AKT-mTOR and AMPK-mTOR signaling pathways involved in autophagy-activated uterine smooth muscle cells during labor? What is the change in the signal path of uterine smooth muscle cell autophagy during abnormal delivery?Part one Changes of Autophagy in Human Uterine Myometrium in non-labor/labor/long labor groupsObjective:In this part,the preliminary work of the experiment was checked and the changes of autophagy in the long labor group were further understood.Materials and Methods: The Ethics Number.2018041701.Human uterine myometrium strips were selected.The upper edge of the incision of lower uterine segment was cut off when doing the cesarean section.The size of muscle strips wasabout 5 cm×1 cm×1 cm.There are three groups: non-labor group,labor group and long labor group.Part of the uterine smooth muscle was made into electron microscopic specimens and autophagty was observed under electron microscope.The remaining tissues were preserved in two parts at-80? for subsequent experiments.Results: The number of autophagosomes in uterine myometrium cell increased in labor group,and the number of autophagosomes in uterine myometrium cell decreased in long labor group compared with labor group.Conclusion: In human uterine fibroids,the autophagy is activated during labor.Part two Changes of PI3K-AKT-mTOR and AMPK-mTOR signaling pathways in human uterine myometrium during laborObjective: the previous studies of our research group found that the autophagy of uterine muscle was upregulated during delivery,but the pathway of autophagy regulation is still unclear.In this section,Real Time-PCR and Western Blot will be used to detect the expression of signals associated with autophagy-related pathways in non-labor and in labor groups.Materials and Methods: The material is the same as the previous part.The uterine smooth muscle strips are separeated into 2 parts.One part of the tissue RNA,was extracted.Another part of tissue protein,was extracted.Real Time-PCR and Western Blot are used to detect the expression of the regulatory factors associated with autophagy,mTOR,PI3K,AKT and AMPK in uterine myometrium.Result:1.The expression of mTOR in uterine myometrium of in labor group was lower than that of non-labor group,the expression of AMPK was significantly higher than that of non-labor group,the difference among the two groups was significant(P < 0.05).2.Compared with the non-labor group,there was no significant difference in the expression of PI3K and AKT in uterine myometrium tissue between the in labor group and the non-labor group,P > 0.05.Part three Changes of PI3K-AKT-mTOR and AMPK-mTOR signaling pathways in human uterine smooth muscle during long labor groupObjective: The core three factors during childbirth are productivity,fetus and birth canal.The most important factor affecting the process of labor is uterine contraction.With the prolongation of delivery time and the stagnation of labor,the main clinical cause is uterine atony.Is the signal pathway associated with autophagy changes when uterine atony occurs? This section will study the expression of autophagy-related pathway-related factors in uterine atony in long labor group.Materials and Methods: Same as part II.Result:1.The expression of mTOR in the long labor group was higher than that in the in labor group,the expression of AMPK was lower in the long labor group than that inthe in labor group.There are significant differences between the three groups(P <0.05).2.The mRNA expression of PI3K decreased in the long labor group,there are significant differences between the three groups(P < 0.05).The expression of AKT in the long labor group did not change with those in the in labor and non-labor groups and there was no significant difference among the three groups(P > 0.05).Full text conclusions1.The activation of autophagy-related AMPK-mTOR signaling pathway may mediate the activation of autophagy in uterine myometrium cells during delivery.2.PI3K-AKT-mTOR pathway related to autophagy of uterine myometrium cells was not activated during labor.3.The signal pathway of AMPK-mTOR related to autophagy in uterine smooth muscle cells was inhibited and the degree of autophagy was inhibited in long labor group.4.The PI3K pathway associated with autophagy and glucose transport and metabolism may be inhibited in uterine myometrium cells during long labor group.