散结镇痛胶囊对PGF_(2α)诱发小鼠子宫平滑肌细胞收缩的抑制作用探讨

Inhibitory effects of Sanjie Zhentong capsule on primary mouse myometrial cells contraction induced by prostaglandin F_(2α)

目的研究散结镇痛胶囊对前列腺素F_(2α)致原代小鼠子宫平滑肌细胞收缩变化的影响,初步探讨其对痛经的治疗作用。原代培养小鼠子宫平滑肌细胞并进行纯度鉴定,给药后监测动态钙离子变化,染色法观察细胞收缩的情况,免疫荧光法测定细胞钙调蛋白(calmodulin,CaM)、肌球蛋白轻链激酶(myosin light chain kinase,MLCK)、磷酸化的肌球蛋白轻链2(phosphorylation of myosin light chain 2,p-MLC_(20))的蛋白表达水平。结果平滑肌肌动蛋白免疫荧光染色呈强阳性。与PGF_(2α)组比较,散结镇痛胶囊组的钙离子荧光值明显降低(P≤0.05);细胞面积明显增加(P≤0.01);CaM的蛋白表达水平明显降低(P≤0.05);MLCK的蛋白水平有下调趋势;p-MLC_(20)的蛋白表达水平明显降低(P≤0.01)。结论散结镇痛胶囊能治疗痛经,可能与其能抑制平滑肌细胞内钙离子内流,缓解细胞的收缩,降低CaM,p-MLC_(20)的蛋白表达水平有关。

Aim To investigate the effects of Sanjie Zhentong capsule on cultured mouse myometrial cell contraction induced by prostaglandin F2α( PGF2α) , and to elucidate the mechanism of Sanjie Zhentong capsule in treating dysmenorrheal. Methods Primary mouse myometrial cells were cultured and identified. Intracel-lular calcium ( [ Ca2+] i ) was monitored under a Flex-Station 3 Benchtop Multi-Modo Microplate Reader u-sing Calcium 6-QF. Myometrial cells were labeled to observe the changes of contraction. The expressions of calmodulin ( CaM ) , myosin light chain kinase ( ML-CK) , myosin light chain phosphorylation( p-MLC20 ) in mouse uterine smooth muscle cells ( USMCs ) were determined by immunofluorescence. Results Sanjie Zhentong capsule suppressed the intracellular[Ca2 + ]i inflow and reduced the areas of myometrial cells induced by PGF2α. Then it significantly decreased CaM and p-MLC20 levels. Conclusion Our results indicate that Sanjie Zhentong capsule has inhibitory effect on dysmenorrheal induced by PGF2α. Furthermore,its major mechanism may be related with the regulation of intracellular[ Ca2 + ]i inflow and the levels of CaM and p- MLC20.