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

doi:10.11843/j.issn.0366-6964.2019.07.020

摘要/Abstract

摘要： 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。

Abstract: 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.

中图分类号:

S853.74

王忠清, 林春发, 钟文杰, 罗艺晨, 朱兆荣, 刘娟. 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[J]. 畜牧兽医学报, 2019, 50(7): 1500-1508.

WANG Zhongqing, LIN Chunfa, ZHONG Wenjie, LUO Yichen, ZHU Zhaorong, LIU Juan. Effect of Zhu Qin Extractive Fluid on the Proliferation and Transcription of Inflammatory Factors in LPS-injured IPEC-J2 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1500-1508.

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