健脾益气摄血颗粒对肠道出血斑马鱼内脏组织细胞因子影响研究

Study on Effect of Jianpi Yiqi Shexue Granules(健脾益气摄血颗粒)on Cytokines in Viscera of Zebrafish with Intestinal Hemorrhage

目的建立肠道出血斑马鱼模型,探讨健脾益气摄血颗粒对斑马鱼肠道组织免疫调控作用。方法首先建立肠道出血斑马鱼模型,并以健脾益气摄血颗粒低、中、高剂量组作为干预药物,实验结束后去除斑马鱼脾脏、心脏、肝脏组织,提取总RNA,并同逆转录,PCR法分析斑马鱼脾脏、肝脏、心脏组织内细胞因子浓度表达。结果①白介素-6(IL-6):与空白对照组比较,脾脏用药第2天、第7天各剂量组检测值明显升高,具有统计学意义;肝脏用药第2天各剂量组检测值明显升高,具有统计学意义,第7天167μg/mL、500μg/mL剂量组检测值明显升高,具有统计学意义;心脏用药第2天各剂量组检测值明显升高,具有统计学意义;用药第7天167μg/mL、500μg/mL剂量组检测值明显下降,具有统计学意义。②肿瘤坏死因子-α(TNF-α):与空白对照组比较,脾脏用药第2天167μg/mL、500μg/mL检测值明显下调,具有统计学意义;用药第7天167μg/mL、500μg/mL检测值明显下调,具有统计学意义;肝脏用药第2天各剂量组检测值明显下调,具有统计学意义,用药第7天各剂量组检测值上调;心脏用药第2天各剂量组检测值明显下调,具有统计学意义;用药第7天167μg/mL、500μg/mL剂量组检测值明显上调,具有统计学意义。③干扰素-γ(IFN-γ):与空白对照组比较,脾脏用药第2天各剂量组检测值上调,具有统计学意义;肝脏用药第2天、第7天各剂量组检测值明显下调,具有统计学意义;心脏用药第2天167μg/mL、500μg/mL剂量组检测值明显上调;用药第7天167μg/mL、500μg/mL剂量组检测值明显上调,具有统计学意义。④转化生长因子-β(TGF-β):与空白对照组比较,脾脏用药第2天、第7天各用药剂量组检测值下调,具有统计学意义。肝脏用药第2天各用药剂量组检测值下调,具有统计学意义,用药第7天各用药剂量组检测值明显上调,具有统计学意义;心脏用药第2天500μg/mL剂量组检测值上调,具有统计学意义,用药第7天167μg/mL、500μg/mL剂量组检测值下调,具有统计学意义。结论健脾益气摄血颗粒对肠道出血斑马鱼模型脾脏、肝脏、心脏组织细胞因子具有调节作用。

Objective We established a zebrafish model of intestinal hemorrhage to explore the immune regulation effect of Jianpi Yiqi Shexue Granules(健脾益气摄血颗粒)on the intestinal tissues of zebrafish,providing a basis for clinical promotion and application.Methods First,we established a zebrafish model of intestinal hemorrhage,and used the low,medium and high dose of Jianpi Yiqi Shexue Granules as the intervention drug.After the experiment,we removed spleen,heart and liver tissues,extracted total RNA,and analyzed the expression of cytokine concentration in spleen,liver and heart tissues by reverse transcription and PCR.Results①IL-6:Compared with that of the blank control group,the detection value of each dose group on the 2nd and 7th day of spleen medication was significantly increased.The detection value of each dose group on the 2nd day of liver medication was significantly increased.The detection value of the 167μg/mL and 500μg/mL dose groups on the 7th day was significantly increased.The detection value of each dose group on the 2nd day of cardiac medication was significantly increased.On the 7th day of medication,the detection values of 167μg/mL and 500μg/mL dose groups were decreased significantly,with statistical significance.②TNF-α:Compared with that of the blank control group,the detection values of 167μg/mL and 500μg/mL on the 2nd day of spleen medication were significantly lowered and had statistical significance.The detection values of 167μg/mL and 500μg/mL on the 7th day of medication were significantly lowered and statistically.The detection value of each dose group on the 2nd day of liver medication was significantly lowered and had statistical significance.The detection value of each dose group on the 7th day of medication was increased.The detection value of each dose group of the 2nd day of cardiac medication was significantly lowered and had statistical significance.On the 7th day of administration,the test values of the 167μg/mL and 500μg/mL dose groups were significantly increased.③IFN-γ:Compared with that of the blank control group,the detection value of each dose group on the 2nd day of spleen medication was increased,which was statistically significant.The detection value of each dose group of liver medication was significantly lowered on the second and 7th day of medication,with statistical significance.The test values of the 167μg/mL and 500μg/mL dose groups on the 2nd day of medication were significantly increased.The test values of the 167μg/mL and 500μg/mL dose groups on the 7th day of medication were significantly increased,which was statistically significant.④TGF-β:Compared with that of the blank control group,the test values of the spleen dosing groups on the 2nd and 7th day were lowered,which was statistically significant.The detection value of each dosage group on the 2nd day of liver medication was reduced,which was statistically significant.The detection value of each dosage group on the 7th day of medication was significantly increased,which was statistically significant.The detection value of the 500μg/mL dose group on the 2nd day of cardiac medication was increased.It had statistical significance.The detection value of the dose group of 167μg/mL and 500μg/mL on the 7th day of medication was lowered,which was statistically significant.Conclusion Jianpi Yiqi Shexue Granules can regulate the cytokines of spleen,liver and heart tissues of intestinal bleeding zebrafish model.