【作者】 宋丽君；

【导师】 牛凤兰；

【作者基本信息】 吉林大学 ， 劳动卫生与环境卫生学， 2009， 硕士

【摘要】 本研究通过建立ICR小鼠Hep-A-22(以下称为H22)肝癌移植性实体肿瘤模型,对扶正肝康颗粒对H22荷瘤ICR小鼠肿瘤、抗肿瘤免疫功能、肝肾毒性进行了初步考察。结果表明,扶正肝康颗粒对H22荷瘤ICR小鼠有一定体内抑瘤作用,使H22实体瘤异型性有所改善;对H22荷瘤ICR小鼠抗肿瘤免疫功能无显著影响,胸腺、脾脏重量和脏器指数、血清TNF-α、IL-2无显著改变,血清IFN-γ显著降低;对H22荷瘤ICR小鼠肝脏无明显毒性,肝脏重量和脏器指数无显著改变,血清TP、ALB、GLO、ALT、ALP以及肝组织tSOD、GSH-PX无显著变化;中剂量表现为H22荷瘤ICR小鼠血清BUN显著下降。综上认为,扶正肝康颗粒对肝脏肿瘤未见有明显抑制作用,未发现有促进荷瘤小鼠免疫功能作用,未见对荷瘤小鼠肝功能有显著影响。本次试验结果表明扶正肝康颗粒在肝脏肿瘤研究方面结果不甚理想,但仍为该药物在肝病相关领域的应用提供了一部分基础性数据,对该药物在有效领域的研究提供一个对比参考。更多还原

【Abstract】 Objective:Fuzhenggankang Granule, an compound recipe, is determined as an assistant drug for the therapy of liver diseases. The preliminary study found that Fuzhenggankang Granule had an efficacy on liver cirrhotic ascites based on the rat model of that disease induced by CCL4 compositing gavage. In order to investigate whether this drug had some effects on the other types of common liver diseases besides the liver cirrhotic ascites, the ICR mouse was used in this study and also the mouse hepatoma cell line H22 to establish the H22-bearing ICR mouse model, and then we observed the effests of Fuzhenggankang Granule in the aspects of the tumor , the immune function and the function of other organs such as the liver and the kidney, looking forward to expanding the awareness and the therapeutic applications of the drug.Methods:The resuscitated H22 cell line was recovered in ICR mouse peritoneal cavity, with 1.0×108 cells per milliliter as the fina use concentration and then inoculated to ICR mouse by axillary subcutaneous transplantation with 0.2mL cell suspension of each model mouse to construct the hepatoma solid tumor model. The model mice were sent to positive control, negaive control and the drug assay groups randomly. The final concentration of the positive control drug CTX (powder) dissoved in the normal saline was 4 g·L-1, and its daily dose was 0.02 g for each kilogram body weight by intraperitoneal injection once a day, equal to the daily volume of 0.05 mL CTX solution for 10 g body weight. As the negative control, normal saline was given 0.2 mL for 10 g body weight once a day as the daily dose by gavage. The high, middle and low doses of Fuzhenggankang Granule are respectively 7.20 g, 3.60 g and 1.20 g for each kilogram body weight by gavage once a day as the daily doses, and the preparing concentrations were 0.36 g·mL-1, 0.12 g·mL-1 and 0.06 g·mL-1 equal to the daily volume of 0.2 mL for 10 g body weight respectively. Fuzhenggankang Granule, positive control and negative control drugs were began to given respectively at the following day after the H22 inoculation, lasting 10 days continually. After 24h of the latest drug administration, the model mice were executed and the specimens were collected. The drug control(middle) group also used the ICR mouse and were raised and administrated with the middle dosage at equal pace with others, but no H22 cell inoculation. During the 10 days of the drug administration, the body weight of each model mouse was measured as a reference to adjust the drug administration volume to it. The blood was collected by enucleation of eyeball ,and after separation the serum was stored at -80℃for the detection of TP, ALB, GLO, ALT, ALP, BUN, TNF-α, IFN-γand IL-2. The weights of fresh tissues including thymus, spleen, liver and solid tumor were measured and preserved in 4 % formaldehyde for paraffin section preparation and the further Hematoxylin-Eosin staining. Part of the fresh liver tissues was preserved at -80℃for the tissue homogenate preparation and the further detection of tSOD, GSH-PX in liver.Results:Compared with the negative control, the tumor inhibition rates of Fuzhenggankang Granule had a relatively better inhibition rate(24.5 %) in the middle dose(3.60 g·kg-1BW·d-1)of Fuzhenggankang Granule, but no statistical significance. Compared with the negative control, Hematoxylin-Eosin staining of the H22 solid tumor showed that both the high and the middle drug groups had remissions of atypia to some extent, revealed the clearer intercellular connections and cell contours, as well as the decrease staining of nuclei and the improvement in nucleo-cytoplasmic ratio. Compared with the negative control, the drug had no significant effects on the weight of thymus and the spleen as well as their organ indexes and histomorphology. Compared with the negative control, the drug had no significant effects on the levels of TNF-α, IL-2, but with a decrease of IFN-γin serum. Compared with the negative control, the drug had no significant effect on the ponderal growth. Compared with the negative control, the drug had no significant effect on the liver in terms of the organ weight, the organ index and the histomorphology. Compared with the negative control, there were no statistical changes in the aspects of TP, ALB, GLO and ALB/GLO, showing the protein synthesis function of liver. The ALT, ALP activities in the serm, the tSOD, GSH-PX activities in the liver had no significant changes as well. Compared with the negative control, the BUN in serum had significant decrease in the drug middle group.Conclusions:1. Fuzhenggankang Granule had a certain tumor inhibitory effect on the solid tumor in H22-bearing ICR mouse model and the atypia of the solid tumor eased at the same time. But the inhibitory rate had no statistical significance.2. Fuzhenggankang Granule had no significant effect on the anti-tumor immune function in H22-bearing ICR mouse model. The weight and the organ index of thymus and spleen had no significant changes, and also the levels of TNF-αand IL-2 in serum. But the IFN-γhad a significant decrease.3. Fuzhenggankang Granule had no significant toxic effect on the liver of H22-bearing ICR mouse model in the aspects of the organ weight and index, and the activities of the hepatic enzymes such as ALT, ALP in serum and tSOD, GSH-PX in tissue.4. Fuzhenggankang Granule showed out the significant decrease of BUN in serum in the middle drug dose.更多还原