虎黄烧伤搽剂中白藜芦醇苷通过上调SIRT1对创伤后应激障碍模型大鼠血清miR-9表达及其ox-LDL诱导的巨噬细胞增殖和炎性因子的影响  

Effect of Resveratrol Glycosides in Tiger Yellow Burn Tincture(虎黄烧伤搽剂)on Serum miR-9 Expression and ox-LDL-induced Macrophage Proliferation and Inflammatory Factors by Up-Regulating SIRT

作　　者：李玉先[1] 汪开新 沙前坤 程先能[1] 湛奇 何小琴 殷国海 唐智权 LI Yuxian;WANG Kaixin;SHA Qiankun;CHENG Xianneng;ZHAN Qi;HE Xiaoqin;YIN Guohai;TANG Zhiquan(Chongqing Traditional Medicine Hospital,Chongqing 400021,China;Kaizhou Hospital of Traditional Chinese Medicine,Chongqing 405400,China;Chongqing Yangdu Biology Institute,Chongqing 408000,China;Chongqing Rongchang District Peoples Hospital,Chongqing 402460,China)

机构地区：[1]重庆市中医院,重庆400021 [2]重庆市开州区中医院,重庆405400 [3]重庆央都生物研究院药学部,重庆408000 [4]重庆市荣昌区人民医院,重庆402460

出　　处：《中华中医药学刊》2023年第11期49-52,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81873998);重庆市技术创新与应用发展专项面上项目。

摘　　要：目的 探讨虎黄烧伤搽剂中白藜芦醇苷通过上调沉默信息调节因子1(silent information regulator 1,SIRT1)对创伤后应激障碍模型大鼠血清微小RNA-9(microRNA-9,miR-9)表达及其氧化低密度脂蛋白(oxidized low density lipoprotein, ox-LDL)诱导的巨噬细胞增殖和炎性因子的影响。方法 选取100只大鼠,建立创伤后应激障碍大鼠模型,50只给予虎黄烧伤搽剂中白藜芦醇苷,作为虎黄烧伤搽剂中白藜芦醇苷组,另外50只不做任何处理作为模型组。培养大鼠巨噬细胞THP-1,对照组:加入普通培养基;ox-LDL组:80μmol/L ox-LDL处理细胞24 h;虎黄烧伤搽剂中白藜芦醇苷+ox-LDL组:100μmol/L虎黄烧伤搽剂中白藜芦醇苷预处理2 h后,再加入80μmol/L ox-LDL处理24 h。实时定量PCR(qRT-PCR)检测SIRT1 mRNA和miR-9表达水平;四甲基偶氮唑蓝(Tetramethylazolazole blue, MTT)检测细胞增殖情况;采用酶联免疫吸附实验(enzyme linked immunosorbent assay, ELISA)法检测肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白细胞介素-1β(Interleukin-1 β,IL-1β)、白细胞介素6(Interleukin 6,IL-6)炎性因子水平;蛋白质免疫印迹法(Western blot)检测细胞周期蛋白1(CyclinD1)、周期素依赖性激酶2(Cyclin dependent kinase 2,CDK2)、p21蛋白表达。结果 与模型组相比,虎黄烧伤搽剂中白藜芦醇苷组SIRT1 mRNA表达水平显著上升(P<0.05),miR-9表达水平显著降低(P<0.05)。与对照组相比,ox-LDL组细胞增殖率、CyclinD1、CDK2蛋白表达水平显著降低(P<0.05),而p21蛋白表达水平显著升高(P<0.05)。与ox-LDL组相比,虎黄烧伤搽剂中白藜芦醇苷+ox-LDL组细胞增殖率、CyclinD1、CDK2蛋白表达水平显著升高(P<0.05),而p21蛋白表达水平显著降低(P<0.05)。与对照组相比,ox-LDL组SOD含量显著降低(P<0.05),但MDA含量和ROS水平显著升高(P<0.05)。与ox-LDL组相比,虎黄烧伤搽剂中白藜芦醇苷+ox-LDL组SOD含量显著升高(P<0.05),但MDA含量和ROS水平显著降低(P<0.05)。与Objective To investigate the resveratrol glycosides in Tiger Yellow Burn Tincture(虎黄烧伤搽剂) up-regulating the expressions of microRNA-9(miR-9) and its oxidation in the serum of rats with post-traumatic stress disorder by up-regulating the silent information regulator 1(SIRT1),low-density lipoprotein(ox-LDL)-induced macrophage proliferation and the effects of inflammatory factors. Methods A total of 100 rats were selected to establish a rat model of post-traumatic stress disorder. Fifty rats were given resveratrol glycosides in Tiger Yellow Burn Tincture as the resveratrol group and another 50 rats with nothing intervention were as a model group. The macrophages THP-1 of rats were cultured. The control group was added common medium. The ox-LDL group was given ox-LDL treated cells(80 μmol/L) for 24 h. The resveratrol glycosides+ox-LDL group was given resveratrol glycosides in Tiger Yellow Burn Tincture(100 μmol/L) for 2 h and then added ox-LDL(80 μmol/L) for 24 h. Real-time quantitative PCR(qRT-PCR) was used to detect the expression levels of SIRT1 mRNA and miR-9. Tetramethyl azozolium blue(MTT) was used to detect cell proliferation and enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β) and interleukin 6(IL-6). Western blot detection was used for cyclin 1(CyclinD1),cyclin dependence Sex kinase 2(CDK2) and p21 protein expression. Results Compared with those of the model group, the expression level of SIRT1 mRNA in the resveratrol glycosides group significantly increased(P<0.05),and the expression level of miR-9 decreased significantly(P<0.05). Compared with those of the control group, the cell proliferation rate, CyclinD1 and CDK2 protein expression levels in the ox-LDL group were significantly reduced(P<0.05),while the p21 protein expression levels were significantly increased(P<0.05). Compared with those of the ox-LDL group, the cell proliferation rate, CyclinD1 and CDK2 protein expression levels of the resveratrol glycosi

关 键 词：虎黄烧伤搽剂 白藜芦醇苷 大鼠 MIR-9 OX-LDL诱导 巨噬细胞增殖 炎性因子 

分 类 号：R285.5[医药卫生—中药学] R282.71[医药卫生—中医学]