肾康灵对系膜细胞炎症因子的影响  

Effects of Shenkangling on Inflammatory Factors Secreted by Mesangial Cells

作　　者：艾斯[1,2,3] 郑健[1,2,3] 林青[2,3] 邱彩霞[2] 林雄[2] 宋艳芳[2,3] 

机构地区：[1]福建中医药大学,福建福州350122 [2]福建中医药大学附属人民医院,福建福州350004 [3]福建省中西医结合肾脏病重点实验室,福建福州350004

出　　处：《中药新药与临床药理》2014年第4期431-437,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金青年基金项目(81202835);福建省自然科学基金面上项目(2011J01199);福建省卫生厅青年基金项目(2012-1-30)

摘　　要：目的观察氧化低密度脂蛋白(Oxidized Low-Density Lipoprotein,ox-LDL)对系膜细胞(Mesangial Cells,MCs)分泌炎症介质功能的影响,并从细胞分子生物学水平阐明肾康灵的作用机理。方法采用肾康灵含药血清干预增殖系膜细胞的方法,按照随机数字表法分为7组,即正常对照组:DMEM-F12培养液;ox-LDL组:DMEM-F12培养液+ox-LDL(100滋g·mL-1);ox-LDL+趋化因子受体(CXCR6)组:DMEM-F12培养液+ox-LDL(100滋g·mL-1)+CXCR6;肾康灵低浓度组:DMEM-F12培养液+ox-LDL(100滋g·mL-1)+肾康灵低浓度含药血清;虞肾康灵高浓度组:DMEM-F12培养液+ox-LDL(100滋g·mL-1)+肾康灵高浓度含药血清;阿托伐他汀低浓度组:DMEM-F12培养液+ox-LDL(100滋g·mL-1)+阿托伐他汀(50滋mol·L-1);阿托伐他汀高浓度组:DMEM-F12培养液+ox-LDL(100滋g·mL-1))+阿托伐他汀(100滋mol·L-1)。以上各组标本培养24 h后收集各组细胞及上清液。每组实验细胞重复培养6次。分别利用ELISA法、RT-RCR法和Western blot法检测趋化因子配体16(CXCL16)、清道夫受体-B(CD36)、干扰素-酌(IFN-酌)、白细胞介素-6(IL-6)、肿瘤坏死因子-琢(TNF-琢)基因水平和蛋白含量。结果利用ox-LDL(100滋g·mL-1))诱导大鼠系膜细胞增殖并加入CXCR6受体后,CXCL16、CD36、IFN-酌、IL6、TNF-琢的基因表达均显著升高(P<0.01),肾康灵含药血清呈浓度依赖性降低其表达水平,肾康灵高浓度组降低最显著(P<0.01)。结论 ox-LDL诱导系膜细胞增殖时通过CXCR6介导,可促进CXCL16、CD36、IFN-酌、IL6、TNF-琢等炎症介质的释放,肾康灵可通过抑制炎症介质的释放,保护系膜细胞的功能。Objective To observe the effect of oxidized low density lipoprotein （ox-LDL） on the secretion of inflammatory cytokines in mesangial cells （MCs） and to clarify the therapeutic mechanism of Shenkangling at the level of cellular molecular biology. Methods Shenkangling-containing serum （SCS） was obtained from Sprague-Dawley rats which were administered Shenkangling. DMEM-FI2 culture fluid was used as the basic medium for the culture of MCs （normal control group） , and then according to the grouping, 100 μg/mL of ox-LDL, 100 μg/mL of ox-LDL＋ CXCR6, 100 μg/mL of ox-LDL＋low-dose SCS, 100 μg/mL of ox-LDL＋high-dose SCS, 100 μg/mL of ox-LDL＋atorvastatin 50 μmol/L, 100 μg/mL of ox-LDL＋atorvastatin 100 μmol/L were added separately, namely ox-LDL group, CXCR6 group, low-dose SCS group, high-dose SCS group, low-dose atorvastatin group, and high-dose atorvastatin group. After culturing for 24h, the MCs and MCs supernatant were collected. The cell culture repeated for 6 times. Expression levels of CXCL16, CD36, IFN- ν,/ , IL-6, TNF- α mRNA and protein in MCs were detected by RT-PCR and Western blotting, respectively. Results CXCL16, CD36, IFN-ν , IL-6 and TNF-α mRNA expression levels were increased obviously in CXCR6 group, and SCS could counteract the expression levels in a dose-dependent manner （P 〈 0.05）. Conclusion ox-LDL may promote the release of inflammatory mediators of CXCL16, CD36, IFN-ν, IL-6, and TNF-α in mesangial cells mediated by CXCR6, and Shenkangling has protective effect for the function of MCs through inhibiting the release of inflammatory cytokines.

关 键 词：肾康灵 系膜细胞 氧化低密度脂蛋白 CXCL16 CD36 

分 类 号：R285.5[医药卫生—中药学]