淫羊藿含药血清对骨质疏松大鼠BMSCs的Sirt1及PPARγ mRNA表达的影响  

Experimental Study of Epimedium Medicated Serum on the Expression of Sirt1 mRNA and PPARγ mRNA in the BMSCs of Rats with Osteoporosis

作　　者：汪盛玉 陈超[2] 张文财[1] 刘海全[1] WANG Shengyu;CHEN Chao;ZHANG Wencai;LIU Haiquan(Affiliated Orthopedics Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510240 Guangdong, China;School of Chinese Medicine, Southern Medical University, Guangzhou 510515 Guangdong, China)

机构地区：[1]广州中医药大学附属骨伤科医院,广东广州510240 [2]南方医科大学中医药学院,广东广州510515

出　　处：《中药新药与临床药理》2019年第3期296-300,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(30901915);广东省自然科学基金项目(2015A030313367);广东省中医药局科研项目(20161164)

摘　　要：目的探索淫羊藿含药血清对骨质疏松(OP)大鼠骨髓间充质干细胞(BMSCs)沉默信息调节因子1(Sirt1)及过氧化物酶体增殖物激活受体γ(PPARγ)mRNA表达的影响。方法将20只雌性SD大鼠采用卵巢切除术制备OP模型,其中5只OP模型大鼠用于分离培养BMSCs,细胞分为5组,即空白组,慢病毒转染组,淫羊藿含药血清高、中、低剂量组。其余OP模型大鼠灌胃给予淫羊藿水提液(按生药量计算为12、6、3 g·kg-1)及等量蒸馏水,制备淫羊藿高、中、低剂量含药血清及空白血清。采用pLentiH1-Sirt1 shRNA慢病毒载体感染BMSCs后,以淫羊藿含药血清高、中、低剂量分别干预培养24、72 h。采用MTT法检测BMSCs细胞增殖情况;采用实时荧光定量PCR法检测Sirt1、PPARγmRNA表达。结果与空白组比较,慢病毒转染组24、72 h的细胞活力无明显变化(P> 0.05),BMSCs的Sirt1 mRNA表达明显下降(P <0.01,72 h),PPARγmRNA表达明显上升(P<0.01,24、72 h)。与慢病毒转染组比较,淫羊藿含药血清中剂量组24、72 h的细胞活力均有明显升高(P <0.05),淫羊藿含药血清高、中、低剂量组干预24、72 h后BMSCs的Sirt1 mRNA表达均明显升高(P<0.05,P<0.01),PPARγmRNA的表达均显著降低(P<0.05,P<0.01)。结论淫羊藿含药血清能促进OP大鼠BMSCs的Sirt1 mRNA表达,抑制PPARγmRNA表达,从而发挥抑制BMSCs成脂分化倾向,达到预防和治疗OP的目的。Objective To explore the effects of serum containing epimedium on the expression of silencing information regulator 1(Sirt1) and peroxisome proliferator-activated receptor γ(PPARγ) in bone marrow mesenchymal stem cells(BMSCs) of castrated rats. Methods Osteoporosis(OP) model was established by ovariectomy in 20 SD female rats. Five OP model rats were used to isolate and culture BMSCs. The cells were divided into five groups, including blank group, lentivirus transfection group and high, medium and low dose groups of epimedium containing serum. The other OP model rats were given epimedium water extract(calculated as12,6,3 g·kg^-1 according to the amount of crude drug) and distilled water in equal quantity by gavage to prepare high, medium and low doses of epimedium medicinal serum and blank serum. After BMSCs were infected with pLentiH1-Sirt1 shRNA lentivirus vector, the high, medium and low doses of epimedium serum were used to interfere with the culture for 24 and 72 hours. MTT assay was used to detect the proliferation of BMSCs cells,and Real-Time fluorescence quantitative PCR was used to detect the expression of Sirt1 and PPARγ. Results Compared with the blank group,the lentivirus transfection group did not change significantly after intervened for 24 h and 72 h(P > 0.05),the expression of Sirt1 in BMSCs decreased significantly(P < 0.01,72 h) and PPARγ increased significantly(P < 0.01,24 h,72 h). Compared with the lentivirus transfection group,after 24 and 72 hours of intervention,the cell viability of the medium dose group of epimedium medicinal serum increased significantly,the expression of Sirt1 in BMSCs of high,medium and low dose epimedium serum groups increased significantly(P<0.05,P<0.01) and PPARγ decreased significantly(P < 0.05,P < 0.01). Conclusion Epimedium medicated serum can promote expression of Sirt1 mRNA and inhibit expression of PPARγ mRNA,which can inhibit adipogenic differentiation of BMSCs to achieve the prevention and treatment of osteoporosis.

关 键 词：淫羊藿 骨质疏松 骨髓间充质干细胞 沉默信息调节因子1 过氧化物酶体增殖物激活受体Γ 

分 类 号：R285.5[医药卫生—中药学]