T型钙通道在感染性早产中的作用及机制研究

Preterm birth is the leading cause of infant morbidity and mortality, but at present there is no effective method to prevent preterm birth, the reason is that the mechanism of preterm delivery is not clear. Infection is the main cause of premature delivery. In our previous work, we injected lipopolysaccharide (LPS) into pregnant rats' abdominal cavity and successfully established infected preterm model. We found that LPS can upregulate the expression of T type calcium channel through NF- kappa B signaling pathway, LPS can also activate T type calcium channel to increase intracellular calcium concentration. Thus we speculate that T type calcium channel may be involved in the occurrence of preterm birth induced by LPS. This study intends to collect people's uterine smooth muscle tissue of preterm birth, through molecular biology methods and electrophysiological technique, such as cell culture, PCR, Western-blot, immunohistochemistry and patch clamp, will further prove the role of T- type calcium channel in preterm labor and clear the specific mechanism of T type calcium channel in preterm labor. Preventing and curing preterm labor is always a thorny problem in obstetrics. Calcium ion channel is the only and final way of calcium signal transduction of uterine contraction, starting from T- type calcium channel to study the mechanism of premature delivery, will provide a theoretical basis and a new target for finding effective drug to treat preterm birth.

早产是导致婴儿发病率及死亡率的首要原因，但目前尚无防治早产的有效手段，其原因在于早产的发生机制未明确。现已知感染是早产发生的主要原因。在我们前期工作中，向孕鼠腹腔注射脂多糖（LPS）成功建立了小鼠感染性早产模型，并发现LPS通过激活NF-κB信号通路上调T型钙通道的表达，同时LPS能够激活T型钙通道使细胞内钙浓度增加。由此我们推测T型钙通道可能参与了LPS诱导的感染性早产的发生。本研究拟收集人的早产子宫平滑肌组织，通过细胞培养、PCR、western-blot、免疫组化等分子生物学方法及膜片钳等电生理技术，进一步证实T-型钙通道在早产中的作用，并明确T型钙通道在早产发生中的具体机制。早产的治疗是多年来困扰产科的棘手问题。钙离子通道是子宫收缩的钙信号传导的的必经之路，从T-型钙离子通道入手，研究早产的发生机制，将为发现新的治疗早产的有效药物及靶点提供理论依据。

早产是导致婴儿发病率及死亡率的首要原因，但目前尚无防治早产的有效手段，其原因在于早产的发生机制未明确。现已知感染是早产发生的主要原因。钙离子通道是子宫收缩的钙信号传导的的必经之路，本课题从T-型钙离子通道入手，研究感染性早产的初步发生机制。本课题在国家自然基金青年基金的资助下，进展顺利，并根据专家评审建议，进行了适当调整及修改。. 首先收集人的早产子宫平滑肌组织，并进行了子宫平滑肌细胞的原代培养，通过IHC、Western-blot、PCR等方法比较了早产组及同孕周非早产组T型钙通道CAV3.2的表达变化，同时根据评审意见加入了L型钙通道的研究。其次通过张力实验、建立小鼠早产模型等方法比较了T型和L型钙通道拮抗剂对子宫平滑肌收缩的不同影响。最后利用ELLSA检测了T型及L型钙通道对NF-kB活性及下游产物表达的影响。在感染性早产中，T型钙通道cav3.2与L型钙通道cav1.2蛋白表达升高，且明确T型钙通道主要影响子宫平滑肌收缩的频率，L型钙通道主要影响其收缩的强度；同时阻断T-型与L-型钙通道可发挥协同抑制早产的作用，该抑制作用与抑制NF‑κB/Nrf-2信号通路的活性及炎症相关因子表达有关。T-型与L-型钙通道在感染性早产模型中具有不同的作用，同时应用两种钙通道拮抗剂可能将成为预防及治疗早产的新策略。.项目资助发表核心论文2篇，英文文章3篇。培养硕士生1名，博士2名。