| With the extensive application of nuclear energy and the development of nuclear technology, people receive more and more artificial ionizing radiation. Acute radiation injury (ARI) abrupt onset, have rapid development, cause leukopenia and make immune function fail to restore, resulting in immune system disorders, which cause humans’s inflammatory response and severe bleeding,and inflammatory response and severe bleeding cause death in patients with ARI. So,it is very important to protect immune system especially increase WBC in ARI. Recent studies have found that TLR4 signaling pathway involves in inflammation reaction, and plays an important role in the body’s cellular and humoral immune process. Now, the effect of western medicine treatment is not satisfactory in ARI, and the effect of cytokines and other biological products is good, but expensive. Therefore, to develop an economical, efficient, and non-toxic radiation protective Chinese medicine, is an emergent necessary for people.ObjectiveWe want to use literature mining method to analyze the characteristics of Chinese medicine treatment preventing leukopenia caused by ARI, and summarize its regular pattern. we use Chinese medicine to prevent mice leukopenia caused by ARI in experimental research, and discover how Chinese medicine prevent mice from ARI and what is TLR4 signaling pathway’s function in ARI.MethodTheoretical study:Retrieving CNKI, Wanfang, VIP database of clinical research literature which is about Chinese herbs oral liquid treating leukopenia caused by IR. Then analyze frequency and characteristics of Chinese material medica. Experimental study:The animal models of ARI were replicated the male Balb/c mice.There are two groups including normal control group(NC), acute radiation injury group(IR). Mice in IR group were irradiated by 5 Gy 60Co γ-ray. In the 3 day after radiation,we detect indexes. The model is successfully copied, but leukopenia after IR is serious. In summary, we need to adjust the dose and sampling time, while adjust Chinese medicine prescription. Through literature review, with the theory of traditional Chinese medicine, we make two Chinese medicine prescription called Fang 1 and Fang 2 to experiment. There are five groups including NC group, IR group, berbamine group, Fang 1 group and Fang 2 group. Mice of NC and IR were treated with water, while the others were administered with the above medicines respectively for 14 days. In the 14th day by treated, mice except NC group were irradiated by 3.5 Gy 60Co γ-ray. In the 7 day after radiation, some relevant indicators would be detected, including WBC, LYMPH, the ratio of CD4+/CD8+ in blood; weight of mice, the mice Immune Organ Index and spleen biopsy; bone marrow biopsy, CD34+ cells in bone marrow, cell cycle and apoptosis of bone marrow. By the above experiment, we found that Fang 1 has a protective effect of the immune system, but there is still insufficient in the immune function of white blood cells protection. So, we use the theory of Chinese medicine to form a new prescription that is called Fang 3, while improving the pharmaceutical preparation process, adjust mouse sampling time. By the above method, we studied the protective effect of Fang 1 and Fang 3 on the immune function of WBC in irradiated mice and the function of TLR4 signaling pathway. The method is the same with above experiment, in the 4 day after radiation, some relevant indicators would be detected. The indicators are the same with the above, besides we added study on the mechanism, detecting mRNA level of TLR4, Myd88, NF-κB and TNF-α in spleen.Result1. Theoretical study:We included 54 articles,54 involving prescription, a total of 101 herbs, 582 frequency. We found that the Chinese medicine using more than 10 times treating radiation-induced leukopenia is Astragalus, Angelica,, etc.17 flavor; including 16 traditional Chinese medicines, such as tonic medicine, blood circulation drugs, diuresis Shenshi drugs, etc.; four qi of herbs before three is warm, flat, tepid; wu wei frequency before three is sweet, bitter, acrid; meridian frequency before three is spleen, kidney, liver.2. After irradiated with 5 Gy, the count of weight, WBC, LYMPH and spleen and thymus index in IR group decreased significantly. The ratio of CD4+/CD8+ in ARI increased significantly. Spleen atrophys with lymphocyte reduced, and the expression of TLR-4 and TNF-a in IR were significantly increased.3. In the 7 day after 3.5 Gy radiation, the pathology is a little slighter than part 1 IR mice. The percentage of CD34+ cells in bone marrow decreased, and bone marrow cell cycle arrest, with apoptosis rate increased. Berbamine can improve the mice WBC, LYMPH, reduce CD4+ /CD8+ ratio, and restore mice’s weight and thymus index. Berbamine can also protect lymphocytes in spleen and bone marrow, increase CD34+ cells,and reduce cell apoptosis in bone marrow. Fang 1 can reduce CD4+/CD8+ ratio, and restore mice’s weight. Fang 1 can also protect lymphocytes in spleen and bone marrow, and reduce cell apoptosis in bone marrow. Fang 2 can restore mice’s weight and spleen index, and protect lymphocytes in spleen. Fang 2 can also protect bone marrow, and reduce cell apoptosis in bone marrow. Finally, we choose Fang 1 to further study with synthetically consideration.4. In the 4 day after 3.5 Gy radiation, Fang 1 can reduce CD4+/CD8+ ratio, restore mice’s weight, and improve thymus index. Fang 1 can also protect lymphocytes in spleen and bone marrow. Lastly, Fang 1 can reduce the expression of TLR4, MyD88, NF-κB and TNF-α in spleen, and reduce peripheral TNF-α concentrations. Fang 3 can improve the mice WBC, LYMPH significantly, restore mice’s weight and protect lymphocytes in spleen. Fang 3 can also protect bone marrow, improve bone cell cycle arrest. Lastly, Fang 3 can reduce the expression of TLR4, MyD88, NF-κB and TNF-α in spleen, and reduce peripheral TNF-a concentrations.Conclusion1. Theoretical research:Through literature review, we found that the Chinese herbs treating leukopenia caused by radiation damage are sweet and warm tonics which meridian is spleen and kidney, aided by blood circulation drugs to achieve the purpose of regulating blood. Core herbs are Astragalus, Angelica, Millettia. Detecting from recipe, we find that TCM of leukopenia is mostly blood deficiency, spleen and kidney deficiency and blood stasis, and we should tonic Qi and blood, spleen and kidney, and stasis.2. Our experiment successfully replicated the experimental animal model of ARI, Fang 1 and Fang 3 can prevent the radiation injury of immune system in mice, and Fang 3 is better than Fang 1. Fang 3 can tonic Qi and spleen, be detoxification, tonic yin and kidney, tonic blood and promote blood circulation. Fang 3 consistenting with theoretical results further prove that using the theory of traditional Chinese medicine treatment of radiation-induced leukopenia is significant. |
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