| Hepatic cellular cancer (HCC) refers to the liver or intrahepatic bile duct cell cancer, the disease is insidious onset, rapid development, and a higher rate of death, known as the "King of cancers" said.Clinical treatment for HCC medical practitioners mainly use surgery or radiotherapy and chemotherapy methods, which has side effects and bad cure, therefore, there is an urgent need to find a point of early intervention for hepatocellular carcinoma, early intervention prevention.Chinese medicine believes that pricking blood therapy has the effect of reinforcing healthy qi and eliminating pathogen, dispelling stasis to promote regeneration, clearing heat and removing toxin and dissipating mass. Previous studies have confirmed that the effect of pricking blood therapy in prevention and treatment of liver disease is significant. Based on previous researches about pricking blood therapy study on t-cell subsets, as well as the significance of immune t cells against HCC, the paper put forward the academic hypothesis "pricking blood therapy could prevent HCC by regulating the immune t cells ". The study aims to establish a DEN induced rat HCC model, from the perspective of cellular immunology, with a focus on the use of flow cytometry (FCM) investigate the pricking blood effect on HCC rat T cell subsets blood, in order to explore pricking blood on the preventive effect of HCC carcinogenesis and related immune regulation mechanism.ObjectiveUsing diethylnitrosamine (DEN) established HCC model in rats induced by chemical. To explore the prevention of pricking blood therapy on HCC effect and the immunological mechanisms, further to provide some new ideas and methods for TCM in preventing and treating tumor.MethodsForty male Wistar rats were randomly divided into normal group with 10, and 15 rats in model group and blood-letting puncture prevention group. In addition to the normal group, the other two groups were injected DEN intraperitoneally to establish liver cancer model, pricking blood prevention group was pricked on or near Zusanli, Yanglingquan and Taichong acupoint,2 times per week, until the 13 week. Experiment rats observed under normal circumstances.13th week after the experiment, taking 5mL blood from the abdominal aorta and using chemical method for detection of serum liver function of alanine aminotransferase (ALT), alkaline phosphatase (ALP), transformation of glutamine peptide enzyme (GGT) levels; Enzyme-linked immunosorbent assay (ELESA) determination of serum levels of tumor markers AFP and SF; By Flow Cytometry(FCM) determination of peripheral blood t-lymphocyte subsets (CD3+T, and CD4+T, and CD8+T) level, Using SPSS 17.0 statistical analysis of the data.Results1. Rats in normal group were generally in good condition; but the model group were at listlessness, slower activity, yellow hair loss, diet reduce and sometimes bloody urine; Rats of blood-letting puncture prevention group in general were worse than normal, but better than the model group.2. Compared with normal group, the serum ALT, ALP, GGT in model group were significantly increased, the difference was statistically significant(.P<0.01). Compared with the model group, the serum ALT, ALP, GGT, in the blood-letting puncture prevention group were reduced (P<0.05),there was a statistically significant difference(P<0.05).3. Compared with the normal group,the model rat serum tumor markers AFP, SF content were significantly higher, the difference was statistically significant (P<0.01).Compared with the model group, serum AFP levels lower in rats with blood-letting puncture prevention group (T<0.05), and SF was significantly lower (P<0.01), the difference was statistically significant (P<0.05).4. Compared with the normal group, peripheral blood CD3+Tcell rate in the model group was statistically significant difference (P<0.05).CD4+Tcell was significantly reduced, and the difference was statistically significant (P<0.0l). CD8+Tcell had no significant difference in the two groups, no significant (P>0.05).Blood-letting puncture prevention group compared with the model group, CD3+Tcell, CD4+Tcell ratio in peripheral blood were increased, and the difference was statistically significant (P<0.05).CD8’Tcell had no significant difference in the two groups, no significant (P>0.05).Conclusion1. Pricking blood on or near Zusanli, Yanglingquan, Taichong superficial collateral can improve the HCC liver pathological changes in rats, lower serum liver function ALT, ALP and GGT, lower levels of tumor markers AFP and SF, to a certain extent, the treatment can inhibit the development of liver cancer.2. Pricking blood on or near Zusanli, Yanglingquan, Taichong superficial collateral Can effectively increase CD3+T, CD4+T levels, but no significant effect on CD8+T, description CD8+T has little significance on diagnosis of different immune status after pricking blood, pricking blood improve total HCC immune t-lymphocyte subsets in rats of blood (CD3+T) is mainly to improve the mechanism of the CD4+T subgroup, enhance immune function.3. Pricking blood therapy can improve immune function, to a certain degree of delay or prevent precancerous lesions of hepatocellular carcinoma, which may be related to promote cancer toxin and metabolites excreting, raise immune t cell subsets (CD3+Tcell, CD4+Tcell) level, enhance the body’s defense function and the ability to resist cancer. |
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