The Effect Of Jianpiliqi Chinese Herbal On Rat Liver Micro-environment Of Regulatory T Cells

Objective:To observe the Jianpiliqi Chinese herbal medicines' effect on the liver micro-environment of regulatory T cells (regulatory T, Treg) and explore the effect of Jianpiliqi herb on liver immune regulation of Treg cellular mechanism of micro-environment.Methods:1. Group modeling:(1) select 60 clean grade Wistar male rats with 6-week-old, weighing 140g±10g, divide randomly into blank group, control group, Jianpiliqi group, each of Section 20. Each five rats in the same group were vested in one cage, marking the cages.(2)The blank group rats were fed the normal stuffs and water; control group and Jianpiliqi group rats were fed 80ppm diethylnitrosamine (DEN) aqueous solution all the time, and start to accept the treatment after 16 weeks. The Jianpiliqi group rats fed with stuffs containing the Chinese Medicines (consisting of Codonopsis 10g, Poria 20g, Atractylodes 20g, Chinese yam 20g, tangerine peel 10g, bupleurum chinense 10g, white peony root 10g, citrus aurantium 10g and licorice 10g nine Chinese herbal medicines, being processed into 1g crude drug per ml with the standard water decoction and alchol sedimentation technics, Reserved in 4℃refrigerator), intake dose is about 10 times to human in terms of body kg/weight. The control group were fed quondamly.2. Specimen collecting and producting:Weighing the rats and killed them by neck off when the experiment expire to 20 weeks. cutting open the abdominal cavity, clipping the whole liver after the rats being killed, recording general situation and registering the number and size of cancerous nodes, weighing the liver. Prepared respectively the specimens of the normal group rats'hepatic tissue, the control group and Jianpiliqi group rats'hepatic celluar carcinoma tissue (taking the largest one while multiple carcinoma nodes), carcinoma adjacent tissues (within 3mm away cancerous side) and the distal carcinoma tissues (lcm away from cancer organizations) under axenic conditions, then fixed with 10% formalin, paraffin-embedded routinely and cut by 4um thick slices continuously, hematoxylin staining lining, hot Aqua Blue-based, Seal the slices with resin after drying. Detecting the Treg cells and the CD4+ T, CD8+ T cells of the slices by immunohistochemical method. Taking pictures and preserving the preferable samples.Results:1. To the Jianpiliqi group rats, in weight lost (P=0<0.01) and liver weight increased (P=0.021<0.05) are less than the control group rats significantly, and the number of hepatic carcinoma nodes are less than the control group rats (P=0.011<0.05), the tumor are smaller than the control group (P=0.026<0.05)2. By immunohistochemistry, the number of Jianpiliqi group rats'Treg cells in the micro-environment of the hepatic celluar carcinoma are less than the control group rats (P=0.0072<0.01), while CD4+ T cells in the micro-environment of the hepatic celluar carcinoma are increased more than the control group rats (P=0.000<0.01), but the Jianpiliqi Chinese Medcine had no effect on the CD8+ T cells in the micro-environment of the hepatic celluar carcinoma (P=0.527>0.05).Conclusion:Jianpiliqi Chinese herbal medicines to slow tumor growth and proliferation may be one mechanism of micro-environment by reducing the number of Treg cells enhance CD4+ T cells and improve the micro-environment of immunosuppression to achieve.