三棱总黄酮对大鼠子宫平滑肌收缩的调节作用研究  

Regulatory effect of common burreed rhizome flavone on uterine smooth muscle contraction in rats

作　　者：刘素军 贺克[1,2] 刘姣[3] 宋慧英[2] 艾冰 葛军[2] LIU Sujun;HE Ke;LIU Jiao;SONG Huiying;AI Bing;GE Jun(Department of Pharmacy,the Fourth Hospital of Shijiazhuang,Shijiazhuang,Hebei 050021;Clinical Medical Research Center,the Fourth Hospital of Shijiazhuang,Shijiazhuang,Hebei 050021;Department of Pharmacology of Traditional Chinese Medicine,College of Pharmacy,Hebei University of Chinese Medicine,Shijiazhuang,Hebei 050200)

机构地区：[1]河北省石家庄市第四医院药剂科,河北石家庄050021 [2]河北省石家庄市第四医院临床医学研究中心,河北石家庄050021 [3]河北中医学院中药药理教研室,河北石家庄050200

出　　处：《河北中医》2023年第5期821-825,830,共6页Hebei Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(编号:81803822);河北省卫健委医学科研项目(编号:20200150);河北省高等学校科学技术研究项目(编号:QN2017112);河北省自然科学基金面上项目(编号:H2020423222)。

摘　　要：目的观察三棱总黄酮对大鼠子宫平滑肌收缩的调控作用及其机制。方法取雌性SD大鼠20只,分离子宫平滑肌段,置于装有洛氏液的恒温浴槽中,施加1.0 g负荷,观察终浓度为0.5、5、50、500、5000μmol/L的三棱总黄酮对子宫平滑肌自主收缩以及诱导剂[前列腺素F 2α(PGF 2α)、乙酰胆碱(Ach)、BayK8644和缩宫素(OT)]引起的痉挛收缩幅度和频率的影响。原代分离大鼠子宫平滑肌细胞,观察三棱总黄酮对OT引起的胞浆中钙离子浓度变化和钙离子衰退时间的影响,以及对细胞中钙/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)和肌球蛋白轻链20(MLC20)的蛋白磷酸化水平的影响。结果终浓度为5、50、500和5000μmol/L的三棱总黄酮均能够显著降低子宫平滑肌自主收缩幅度(P<0.05),且随着浓度的增大,收缩幅度缩小,但对收缩频率没有明显的影响(P>0.05);终浓度为50μmol/L的三棱总黄酮对PGF 2α、Ach、BayK8644和OT引起的子宫平滑肌痉挛收缩有抑制作用,可使子宫平滑肌收缩幅度明显减小(P<0.05),收缩频率降低(P<0.05);0.5和5μmol/L的三棱总黄酮对OT引起的子宫平滑肌细胞内钙荧光强度的变化率有显著的抑制作用(P<0.05),可使细胞内钙离子衰退的时间延长(P<0.05),并使CaMKⅡ和MLC20的蛋白磷酸化水平降低(P<0.05)。结论三棱总黄酮能够抑制子宫平滑肌的痉挛性收缩,其作用机制与阻断钙通道,抑制并减慢胞浆中钙离子浓度变化,进而下调钙调控相关蛋白磷酸化的水平有关。Objective To investigate the regulatory effect of common burreed rhizome flavone(CBRF)on the contraction of uterine smooth muscle in rats and the underlying mechanism.Methods Uterine smooth muscle segments were separated fromfemaleSD rats and placed in a constant temperature bath containing Rockwell's solution.The effects of 0.5,5,50,500,5000μmol/L CBRF on spontaneous contraction of uterine smooth muscle and spastic contraction amplitude and frequency induced by inducers(Prostaglandin F 2α[PGF 2α],Acetylcholine[Ach],BayK8644 and Oxytocin[OT])were observed under 1.0 g load.Primary uterine smooth muscle cells were isolated from rats to observe the effects of CBRF on OT induced changes in calcium ion concentration in cytoplasma and the duration of calcium ion decay.Meanwhile,protein phosphorylation levels of calcium ion/calmodulin-dependent kinaseⅡ(CaMKⅡ)and myosin light chain 20(MLC20)in cells were detected by western blot(WB).Results Final concentration of 5,50,500 and 5000μmol/L CBRF could significantly reduce the amplitude of spontaneous contraction of uterine smooth muscle(P<0.05).With the increase of concentration,the amplitude of contraction decreased.However,no significant effect was observed on the frequency of contraction(P>0.05).50μmol/L of CBRF had inhibitory effects onthe spastic contraction of uterine smooth muscle induced by PGF 2α,Ach,BayK8644 and OT,which significantly reduced the contraction amplitude and frequency of uterine smooth muscle(P<0.05).0.5 and 5μmol/L CBRF significantly inhibited the change rate of intracellular calcium fluorescence intensity in uterine smooth muscle cells induced by OT(P<0.05),and prolonged the decay time of intracellular calcium ions(P<0.05).Moreover,phosphorylation levels of CaMKⅡand MLC20 decreased remarkably(P<0.05).Conclusion CBRF can inhibit spasmodic contraction of uterine smooth muscle in rats.The possible mechanism is related to blocking calcium channels,suppressing and slowing down changesin calcium ion concentration in the cytoplasm,thereby down

关 键 词：三棱 大鼠 子宫 肌 平滑 肌细胞 子宫收缩 

分 类 号：R285[医药卫生—中药学]