川芎嗪对脂肪干细胞体外增殖及小鼠颗粒脂肪移植存活的影响  

Effects of ligustrazine on proliferation of adipose stem cells in vitro and survival of granular fat grafts in nude mice

作　　者：周景和[1] 徐慧芳[2] 祝飞 贾明 李金晟 ZHOU Jinghe;XU Huifang;ZHU Fei;JIA Ming;LI Jinsheng(不详;Department of Medical Cosmetology,Hangzhou First People's Hospital Affiliated to Zhejiang University School of Medi-cine,Hangzhou 310006,China)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院医疗美容科,310006 [2]浙江省中医院耳鼻咽喉科

出　　处：《浙江医学》2022年第13期1356-1361,I0003,共7页Zhejiang Medical Journal

基　　金：浙江省中医药科技计划项目(2015ZA125)。

摘　　要：目的探讨川芎嗪对脂肪干细胞(ADSC)体外增殖,以及添加川芎嗪对小鼠颗粒脂肪移植物存活的影响。方法将ADSC分组干预,分为ADSC组、ADSC-透明质酸(HA)组(0.2 ml ADSC+0.2 ml HA)、ADSC-HA+川芎嗪组(0.2 ml ADSC+0.2 ml HA+0.1 mg/ml川芎嗪),分别于培养的第1、3、5、7天,采用CCK-8法检测各组细胞增殖能力。将12只裸鼠按随机数字表法分为颗粒脂肪-HA+PBS组(200μl PBS+颗粒脂肪+200μl HA)、颗粒脂肪-HA+川芎嗪注射组(200μl PBS+颗粒脂肪+200μl HA+术后川芎嗪注射)、颗粒脂肪-HA+川芎嗪浸泡组(200μl 100 mg/L的川芎嗪+颗粒脂肪+200μl HA)、颗粒脂肪-HA+川芎嗪注射+浸泡组(200μl 100 mg/L的川芎嗪+颗粒脂肪+200μl HA+术后川芎嗪注射),各3只。使用1 ml注射器将各组混合物注射到裸鼠左右背部皮下,每点注射0.4 ml。术后川芎嗪注射是在皮下注射颗粒脂肪后,以2 mg/d的剂量注射川芎嗪,持续注射7 d。术后8周处死裸鼠解剖刮取出移植组织。组织学HE染色、油红O染色观察各组裸鼠颗粒脂肪移植组织切片。荧光定量PCR法检测各组脂肪形成相关转录因子过氧化物酶体增殖物激活受体γ(PPAR-γ)和CCAAT增强结合蛋白α(C/EBP-α)mRNA表达水平;透射电子显微镜下观察组织HA残留情况。结果培养第7天时,ADSC-HA+川芎嗪组细胞增殖能力较ADSC组、ADSC-HA组增强(均P<0.05),而ADSC组、ADSC-HA组细胞增殖能力比较差异无统计学意义(P>0.05)。与颗粒脂肪-HA+PBS组相比,颗粒脂肪-HA+川芎嗪注射组和颗粒脂肪-HA+川芎嗪浸泡组裸鼠皮下颗粒脂肪移植物重量较重(均P<0.05)。颗粒脂肪-HA+川芎嗪注射+浸泡组裸鼠皮下颗粒脂肪移植物重量较颗粒脂肪-HA+川芎嗪注射组、颗粒脂肪-HA+川芎嗪浸泡组更重(均P<0.05),而颗粒脂肪-HA+川芎嗪注射组与颗粒脂肪-HA+川芎嗪浸泡组比较差异无统计学意义(P>0.05)。组织学染色显示,颗粒脂肪-HA+川芎嗪注射组与颗粒脂肪-HA+川芎嗪�Objective To explore the effect of ligustrazine on the proliferation of adipose-derived stem cells(ADSCs)in vitro,and survival of mouse granular fat grafts.Methods Cultured ADSCs were treated with 0.2 ml hyaluronic acid(HA group)or 0.2 ml HA and 0.1 mg/ml ligustrazine(HA+ligustrazine group);while the untreated ADSCs served as control group.Proliferation of ADSC in each group was evaluated using CCK-8 assay at d 1,3,5 and 7.Twelve BALB/c nude mice were randomly divided into 4 groups(n=3 in each group)and subcutaneously injected with 200μl PBS+fat granules+200μl HA(granular fat-HA group),200μl PBS+fat granules+200μl HA+ligustrazine in vivo treatment(granular fat-HA+ligust-razine soaking group);200μl 100 mg/L ligustrazine+fat granules+200μl HA(granular fat-HA+ligustrazine injection group);and 200μl 100 mg/L ligustrazine+fat granules+200μl HA+in vivo ligustrazine treatment(granular fat-HA+ligus-trazine injection+ligustrazine soaking group),respectively.After 8 weeks,the mice were sacrificed to collect the transplant-ed tissue.Hematoxylin-eosin staining and oil red O staining were used to observe the transplanted granular fat tissue sec-tions of nude mice in each group.RT-quantitative PCR was used to detect genes expression of peroxisome proliferator-ac-tivated receptor-γ(PPAR-γ)and CCAAT enhancer binding protein-α(C/EBP-α).HA residue in transplanted tissue was de-tected by transmission electron microscopy.Results Proliferation ability of ADSCs in ADSC-HA+ligustrazine group was higher than that of ADSC control group and ADSC-hyaluronic acid(HA)group at d 7(all P<0.05);while there were no sig-nificant differences in proliferation ability of ADSCs between ADSC control group and ADSC-hyaluronic acid(HA)group(P>0.05).Compared with granular fat-HA+PBS group,weight of subcutaneous fat grafts in nude mice in granular fat-HA+ligust-razine injection group and granular fat-HA+ligustrazine soaking group significantly increased(all P<0.05).While weight of subcutaneous fat grafts in nude mice in granular fat-HA+ligustr

关 键 词：脂肪干细胞 颗粒脂肪 脂肪移植体 川芎嗪 脂肪形成 

分 类 号：R285.5[医药卫生—中药学]