| Primary liver cancer is one of the common malignant tumors in clinical medicine. It has high mortality and poor treatment effect, and there are no effective cure measures. Surgical resection is still one of the most effective means of treatment. Although researches for postoperative treatment programs in recent years have made great progress, but the overall the effects are still not satisfactory. Thus, for us country with high Morbidity of primary liver cancer, researches of postoperative rehabilitation to reduce the recurrence rate of liver cancer patients and improve the quality of life has great social significance and economic value.Modern medicine has considered that immune function in patients had a high degree of correlation with recurrence. It has been preliminarily confirmed that perioperative immune status and postoperative cellular immune function are related of their disease progression, recurrence and metastasis. The Way to improve the immune status of patients with liver cancer after surgery is one of the current research directions in the field of cancer rehabilitation.Since ancient times, TCM promoted sports. For cancer patients, the importance of sports was embodied in "both diseases anti-change" and "more defense complex".Tai chi, Ba Duan Jin and other sports therapy were applied to the crowd. Modern medicine respected exercise therapy as one of the main treatments for cancer rehabilitation. Tai chi and other movement clearly demonstrated therapy of cancer patients can improve the function of the immune system.Although the movement has an important position and plays an irreplaceable role in cancer rehabilitation. However, including the latest edition of "Cancer Survivor Exercise Guide" by the US, there is no set of clear and standardized exercise program for patients of liver cancer to guide rehabilitation after surgery. Also there is a lack of theoretical foundation about the relationship of exercise and the immune status.In conclusion, this study established orthotopic liver xenograft mouse model for exploring suitable time to do exercising intervention. Using T cell in the spleen as the main object of study, we discuss how exercise intensity regulates the function of T cell in mice and observe the effects of different exercise intensities on metabolites for mice. Using the method of westerning blot, we discuss how exercise intensity regulates the mechanism of the effect of exercise on the lymphocyte function of mice. Meanwhile, we used metabolomics to investigate how aerobic exercise effected the metabolites of orthotopic xenograft mouse models and initially revealed positive impacts of reasonable physical prescriptions on postoperative rehabilitation for liver cancer in mice to verify the scientific significance of physical prescriptions in postoperative rehabilitation programs.Part One The establishment of the exercise model which is post-operative of orthotopic transplantation tumor of hepatocellular carcinoma in miceObjective:To explore the postoperative state of orthotopic transplantation tumor resection, and observe the effect of exercise on the postoperative state in mice.Methods:The establishment and grouping of the exercise model which is post-operative of orthotopic transplantation tumor of hepatocellular carcinoma in mice should be made. Mice were evaluated from the weight rehabilitation, independent of the activity, bldood biochemistry, liver function and other basic indicators.Results:Weight difference between before and after exercise groups was no statistical significance. Starting time of the influence of autonomic activity in mice with a difference, p<0.05 statistically significant. Blood biochemical and liver function indicators, such as visible in the late postoperative movement of mice with postoperative late model of moderate intensity exercise mice propagated in serum was32.00±2.53g/l. Late after high intensity exercise model mice propagated in serum was32.12±2.53 g/1. Both were higher than in the late postoperative serum content of normal motion model mice:26.44±4.10 g/1 and p<0.05. The difference is statistically significant. Postoperative late model of moderate intensity exercise mice serum TP was57.83±2.71 g/1. Late postoperative and high intensity exercise model mice serum TP was 52.62±4.37 g/1. Both were higher than in the late postoperative serum content of normal motion model mice:48.77±4.41 g/1 and p<0.05. The difference is statistically significant.Conclusion:It is best to restore the overall state of the mice when breaking 7 days after the surgery. The model is successfully established. Exercise intensity and liver function were inversely proportional to the trend.Part Two Experimental study of how aerobic exercise effects the immune function in mice after orthotopic transplantation tumor of hepatocellular carcinomaObjective:To observe the influence on immune function in mice in different sports involvement and different exercise intensity.Methods:1. Using concanavalin A to stimulate spleen lymphocytes of model mice. Use MTT method to detect the influence of different exercise intervention methods on T lymphocyte proliferation in model mice.2. Suspension model mice spleen lymphocytes, magnetic bead separation T lymphocytes, flow cytometry instrument to detect the expression of CD69+,25+rates.3. Suspension preparation model mice spleen lymphocytes, magnetic bead separation T lymphocytes, ELISA method to detect PMA and Ion induction of T cell secretion of IL-10, the TNF-a secretion, etc.Results:1. The influence of time and strength on lymphocyte proliferation in mice:For insitu liver transplantation model mice after tumor resection, early postoperative moderate intensity exercise mice lymphocyte proliferation ability for (227.42±0.62)%. Early postoperative high intense exercise mice lymphocyte proliferation ability for (188.38±0.23)%. Both lower than normal postoperative exercise mice, namely (348.03±0.89)%. Late postoperative moderate intensity exercise mice lymphocyte proliferation ability for (233.77 ±0.30)%, and late postoperative normal movement of the mouse lymphocyte proliferation ability (235.03±0.89)%, compared to both numerical approach, and there was no statistically significant difference between the two. Late postoperative high intensity exercise mice lymphocyte proliferation ability for (329.12±0.43)%, postoperative late is better than that of the normal movement of the mouse lymphocyte proliferation ability (235.03±0.89)%, but there was no significantly statistical difference between the two. Interaction analysis shows that the start time and motion strength combined resection of liver transplantation in situ tumor model mice spleen lymphocyte proliferation ability of p<0.05, statistically significant.2. Intervention time and strength of the model mice factor CD69+and CD25+T cells surface:For liver transplantation in situ early motion model of mice after tumor resection, moderate intensity treadmill exercise intervention model surface expression of CD69+T lymphocytes in mice rate was (36.54±0.46)%, high intensity treadmill exercise intervention model surface expression of CD69+T lymphocytes in mice rate was (38.72± 0.20)%, both the surface expression of CD69+T lymphocytes were below the normal movement of mice (53.38±0.25)%.In the late postoperative exercise mice, the high intensity exercise intervention model under the surface of T lymphocytes in mice CD69+ expression rate was (40.31±0.33)%, moderate intensity exercise intervention model under the surface of T lymphocytes in mice CD69+expression rate was (45.21±0.36)%, were greater than the normal movement of the surface of the model T lymphocytes in mice after CD69+expression rate, namely (37.15±0.18)%, and p<0.05, the difference is statistically significant. For liver transplantation in situ early model mice after tumor resection, moderate intensity treadmill exercise intervention model surface expression of CD25+T cells in mice to (37.82±0.26)%, high intensity treadmill exercise intervention model surface expression of CD25+T cells in mice to (41.63±0.37)%, both the surface expression of CD25+T cells were below the normal movement of mice (45.65±0.22)%.In the late postoperative exercise mice, of the moderate intensity exercise intervention model the surface expression of CD25+T cells in mice to (42.72±0.29)%, of the high intensity exercise intervention model the surface expression of CD25+T cells in mice to (46.40± 0.43)%, were greater than normal after motion model surface expression of CD25+T cells in mice, namely (41.63±0.37)%, and p<0.05, the difference is statistically significant.3. In time and strength of the model T lymphocyte secretion of cytokines in mice:For in situ liver transplantation model in mice after tumor resection, early postoperative exercise mice, the intensity of the T cells to secrete the influence of TNF-a no statistical difference. Late postoperative exercise mice, the moderate intensity exercise group with high intensity exercise group with normal movement has a bigger difference, and p<0.05, the difference is statistically significant. Each specific T lymphocytes in the TNF-a respectively:late postoperative moderate intensity treadmill motion model of mice was 188.74±96.10 pg/ml, postoperative late for high intensity treadmill exercise model mice was 205.93±110.04 pg/ml, normal movement for late postoperative model in mice was 450.30±116.37 pg/ml.IL-10 on the strength of the difference is not statistically significant, but the interaction analysis found after the late postoperative exercise mice, IL-10 production is lower than the early postoperative exercise mice. IL-10/TNF-a in the late postoperative exercise mice, moderate intensity exercise 10.68, high intensity exercise 9.21, were greater than normal movement in mice:5.72.IL-2 in the late postoperative exercise mice, moderate intensity exercise group lymphocyte of IL-2 was 748.22±354.11 pg/ml, less than normal mice campaign:1257.49±279.85 pg/ml.IL-6, IFN-y there was no statistically significant difference between the groups of mice.Conclusion:1. Late after surgery or in high intensity exercise has promoting effect to mice spleen lymphocyte proliferation ability, especially the effect of high intensity exercise.2. Rest period after surgery or in high intensity exercise on model surface CD69+T lymphocytes in mice, expression of CD25+all have positive role, intensity is proportional to the expression rate.3. From the lymphocytes of TNF-a, IL-10 secretion of cytokines, such as the difference is visible in the late postoperative liver or high intensity exercise may be the process of postoperative recurrence of orthotopic transplantation tumor mice.Part Three Mechanism of the effect of exercise on the lymphocyte of model mice.Objective:This study focuses on different intervention time and intensity of exercise on postoperative function of lymphocytes in mice liver transplantation in situ tumor effects of mechanism.Methods:Suspension preparation model mice spleen lymphocytes, protein after adopting Wersten blot method detection, JNK, PARP, Caspase3, Bax, Bcl-2 protein, etc.Results:Western blot method detected, compared with normal postoperative exercise mice phosphorylation JNK expression, early postoperative moderate intensity exercise reduce spleen lymphocytes in mice phosphorylation JNK expression; Compared with postoperative period normal movement in mice the expression of PARP, early postoperative moderate intensity exercise increases the expression of lymphocyte PARP in mice, the late high intensity exercise also has promoting effect to the rise of PARP. Compared with normal postoperative exercise mice capase3 expression, the expression of caspase3 in early postoperative intensive movement rose.Conclusion:Movement may be mediated by inhibiting the mitochondrial apoptosis pathway affect lymphocyte apoptosis in mice, that movement has certain antiapoptotic effect.Part Four The Effects of Exercise on Metabolism of Post-operative of Orthotopic Transplantation Tumor of Hepatocellular Carcinoma In MiceObjective:This part of the study focuses on the stability of the model from the perspective of metabolomics. And make a useful exploration for the objective study of liver cancer patients to guide the exercise rehabilitation.Methods:Use the exercise model which is post-operative of orthotopic transplantation tumor of hepatocellular carcinoma in mice and high performance liquid chromatography and mass spectrometry (UP LC-QT o F-/MS) research platform. Compare the metabolites spectral differences between postoperative multiply different amount of exercise groups and the control group by using principal component analysis and partial least squares discriminant analysis. Through variable important indicators and t-test, screen potential metabolic different matters, and then use the database to retrieve the potential biomarkers for analysis and identification.Results:By multivariate statistical analysis, compared with the control mice, only a small difference in substance was elected in low-intensity exercise group and high-intensity exercise group. Metabolites are generally no significantly different.Conclusion:Differences metabolomic analysis based on the characteristic metabolites may related. |
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