桃红四物汤通过GLUL/NOTCH通路促进BMMSCs成骨分化及核糖体生成研究
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Taohong Siwu Decoction promoting osteogenic differentiation and ribosome biogenesis of BMMSCs through the CLUL/NOTCH pathway
CHEN Pei’， JIAN Gonghui²， YANG Lei'， CHEN Zhenhua’， ZHANG Tianze³， YE Zhengxian³， QI Xinyu²， XIONG Hui²， LU Xiaolongl* 1.TheFirstHospitalof Hunan UniversityofChineseMedicine，Changsha，Hunan41Oo7，China; 2.HunanUniversityof
Chinese Medicine， Changsha， Hunan 41O2O8， China; 3. The Second Hospital of Hunan University oj Chinese Medicine，Changsha， Hunan 41ooo5，China
[Abstract]ObjectiveToexplorethe specific mechanisms of Taohong Siwu Decoction （THSWD）inpromoting osteogenic diferentiationofbonemarrow mesenchymalstemcells（BMMSCs）andafectingfracture healing.Methods BMMSCsfromrats were cultured in vitro.WellgrowingBMMSCswereselectedanddividedintothefollowinggroups：thecontrol（CONTROL）group，the THSWD intervention （THSWD）group，the THSWD intervention+glutaminesynthetase（GLUL） knockdownoroverexpresionblank control （THSWD+sh-NCor THSWD+OE-NC） group，andthe THSWD intervention+GLULknockdown oroverexpression（THSWD+shGLUL or THSWD+OE-GLUL） group. Celsin the CONTROL group werecultured in complete BMSC medium.The THSWD group was treated with10O ng/mL THSWD-containing serum.The THSWD+sh-NCand THSWD+OE-NC groups were incubated with100 ng/mL THSWD-containing serumandthen transfected with sh-NCandOE-NC lentiviruses，respectively.The THSWD+sh-GLUL andTHSWD+OE-GLUL groupswere alsotreated with10O ng/mLTHSWD-containingserum followed bytransduction with sh-GLUL andOE-GLULlentivirusesrespectively.Theproteinexpresionlevelsofalkalinephosphatase （ALP），Osterix（Osx），rutelated transcriptionfactor2（Runx2），GLULNOTCHintracelulardomain （NCD），neurogeniclocusnotchhomolgprotein1（OH1）， andhairyandenhancerofsplit-1（HES1）weredetectedby Westernblot.Mineralizationwasassessdusing AlizarinRedstaining. Theexpresionof5SrRNA，5.8SrRNA，18SrRNA，and28SrRNAwasmeasuredviaqRT-PCR.Networkpharmacologyanalysis wasemployedtopredictandidentifyGLULasakeytargetthroughwhichTHSWDregulatesribosomebiogenesistoaffect BMMSCs.Cellviabilitywasevaluatedusing theCCK-8assayandcellmigrationdistancewasquantifiedusingthe Transwelassay. Results ComparedwiththeCONTROLgroup，theexpressionlevelsofALP，Osx，andRunx2 significantlyincreasedintheTHSWD group （ P
[Keywords]fracture healing; Taohong Siwu Decoction;bone marow mesenchymal stemcells;ribosome biogenesis；NOTCH signaling pathway
骨折作为临床实践中极为常见的一种病症，广泛存在于各类年龄段的人群中，可导致机体出现局部肿胀、疼痛、畸形、活动受限等症状。（剩余12912字）
