术芩提取液对脂多糖损伤的IPEC-J2细胞增殖及炎症因子转录的影响

Effect of Zhu Qin Extractive Fluid on the Proliferation and Transcription of Inflammatory Factors in LPS-injured IPEC-J2 Cells

为探究术芩提取液(Zhu Qin extractive fluid,ZQEF)对脂多糖(LPS)损伤的IPEC-J2细胞增殖及炎症因子转录的影响,采用薄层色谱法对ZQEF中主要活性成分进行定性鉴定,高效液相法测定黄芩苷含量。将IPEC-J2细胞随机分为空白组、LPS模型组、药物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ组(各组ZQEF浓度分别为10^1、10^2、10^3、10^4、10^5g·mL^-1),每组5孔,104个细胞·孔^-1。MTT法测定细胞增殖率,qRT-PCR测定TFF3、TNF-α和IL-8的mRNA转录变化。结果显示,ZQEF含有黄芩苷、黄芪甲苷和白术内酯Ⅰ,其中黄芩苷含量为1.469 8%;LPS模型组细胞增殖率降低,TFF3、TNF-α和IL-8的mRNA转录量显著增高,与空白组比较差异均极显著(P<0.01);与LPS模型组比较,药物Ⅱ组(10^-2g·mL^-1)和药物Ⅲ组(10^-3 g·mL^-1)细胞增殖率均增高,差异分别达到极显著和显著(P<0.01和P<0.05)水平,药物Ⅱ组TFF3和TNF-α的mRNA转录量降低,差异极显著(P<0.01),IL-8的mRNA转录量降低,差异不显著(P>0.05)。结果提示,ZQEF能促进LPS损伤的IPEC-J2细胞增殖,抑制炎症因子转录,推测与修复肠黏膜作用有关。

The objective of this study was to investigate the effect of Zhu Qin Extractive Fluid(ZQEF) on the proliferation and transcription of inflammatory factors in IPEC-J2 cells injured by lipopolysaccharide(LPS). Principal active component were identified by TLC and the content of baicalin was determined by HPLC in ZQEF. IPEC-J2 cells were randomly divided into control group, LPS model group and drug group Ⅰ,Ⅱ,Ⅲ,Ⅳ, and Ⅴ(concentration of ZQEF in each group was 10^-1, 10^2, 10^3, 10^4, 10^5 g·mL^-1, respectively), each group set up 5 holes, 104 cells·hole^-1. The proliferative rate of cells were determined by MTT method, and the mRNA transcription changes of TFF3, TNF-α and IL-8 were measured by qRT-PCR. The results showed: the main therapeutic ingredients include baicalin, astragaloside and atractylenolide-Ⅰ, and the content of baicalin was 1.469 8% in ZQEF. Compared with the control group, the proliferative rate of cells highly decreased, and the mRNA transcription of TFF3, TNF-α and IL-8 extremely increased in LPS model group(P<0.01). Compared with the LPS model group, the proliferative rate of cells highly enhanced in drug group Ⅱ(10^-2g·mL^-1) and Ⅲ(10^-3 g·mL^-1)(P<0.01 and P<0.05 respectively), the mRNA transcription of TFF3 and TNF-α significantly down-regulated(P<0.01), and the mRNA transcription of IL-8 down-regulated in drug group Ⅱ, non-significantly(P>0.05). The results suggest that ZQEF could facilitate the proliferation of IPEC-J2 cells injured by LPS, inhibit the transcription of inflammatory factors, which may be connected with effect of repairing the intestinal mucosa.