含药血清对HGSMCs中MLC磷酸化水平的调节机制研究.pdfVIP

运脾开胃方含药血清对HGSMCs中MLC磷酸化水平的调节

机制研究

白宇;赵智强

【摘要】目的观察运脾开胃方含药血清对人胃窦平滑肌细胞(HGSMCs)内Ca2+、

MLC、p-MLC的干预作用以及与Ghrelin受体(GHSR)的关系,研究运脾开胃方治

疗厌食症的机制.方法培养HGSMCs,制备低、中、高浓度的运脾开胃方含药血清;

采用激光共聚焦、Westernblot法检测运脾开胃方含药血清对HGSMCs内

Ca2+、MLC、p-MLC的影响;阻断HGSMCs表面GHSR并干扰细胞内外Ca2+

的正常调控,研究运脾开胃方含药血清对HGSMCs内p-MLC的干预机制.结果低、

中、高剂量组运脾开胃方含药血清能在一定时间范围内上调HGSMCs内Ca2+的

相对荧光强度(P0.05~0.01);低、中、高剂量组运脾开胃方含药血清能在10min

时显著上调HGSMCs内p-MLC水平(P0.05~0.01);(D-Lys3)-GHRP-6、

Thapsigargin、D-HBSS液能显著抑制高剂量运脾开胃方含药血清在10min时对

p-MLC的上调作用(P0.01).结论运脾开胃方含药血清可能通过激活GHSR提高

细胞内Ca2+水平来促进MLC磷酸化.%OBJECTIVEToobservetheeffectof

spleenappetizerformula(SAF)containingserum(SAF-CS)onCa2+,MLCand

thep-MLCinhumangastricsmoothmusclecells(HGSMCs)andthe

relationshipwithGhrelinreceptor(GHSR),andtoexplorethemechanismof

SAFfortreatinganorexia.METHODSHGSMCswerecultured,whilethe

low,mediumandhighconcentrationsofSAF-CSwereprepared;Confocal

laserdetectionmethodandWesternblotwereusedtotesttheeffectsof

SAF-CSonCa2+,MLCandp-MLC;TheGHSRexistedonthesurfaceof

HGSMCswasblockedandthenormalrgulationofintracellularand

extracellularCa2+wereabolishedtostudythemechanismofSAF-CSeffect

onp-MLC.RESULTSLow,me-diumandhighdosegroupofSAF-CSup-

regulatedtherelativefluorescenceintensityofCa2+inacertaintime

range(P0.01,P0.05)inHGSMGs;low,mediumandhighdosegroupof

SAF-CSmarkedlyincreasedthelevelofp-MLCat10minin

HGSMCs(P0.01,P0.05);(D-Lys3)-GHRP-6,Thapsigargin,D-HBSSsolution

significantlyinhibitedtheup-regula-tionofp-MLCbythehighdoseof

SAF-CSat10min(P0.01).CONCLUSIONTheSAF-CSmayincrease

intracellularCa2+levelsandpromotephosphorylationofMLCvia

activatingGHSR.

【期刊名称】《南京中医药大学学