| Hepatocellular carcinoma,a dangerous type of cancer,it`s incidence rate of malignant tumors ranked fourth and mortality ranked third in 2015,China.The best treatment is still early surgery.According to relevant statistics,the recurrence rate of hepatocellular carcinoma after 5 years after resection is still higher than 60%,small hepatocellular carcinoma after complete resection,there are more than 40% of the recurrence rate of [1].Studies have shown that after resection of hepatocellular carcinoma recurrence and immune function in patients with has a high correlation,in the peri operation period,the stress reaction caused by surgical trauma can inhibit the immune function of patients,the immune function of the body can not effectively suppress proliferation and diffusion of residual cancer cells,resulting in increased the risk of postoperative metastasis.Compared with other cancer sites and patients with 5 year survival rate is low,life quality difference [2].Therefore,how to avoid the recurrence and metastasis of liver cancer has become the focus of the treatment of liver cancer,improve the postoperative immune function of patients with liver cancer is important to improve the long-term survival rate of patients with hepatocellular carcinoma.Modern medical advice exercise therapy as a tumor postoperative rehabilitation treatment of moderate and regular long-term exercise on the body immune function,especially in improving the activity of immune cells and immune factors.And animal experiments showed that moderate intensity of long-term exercise to alleviate and inhibit the proliferation of tumor cells to help[3].Chinese medicine as one of the main means of complementary and alternative medicine,greatly enriched the content of tumor rehabilitation.As early as the spring and autumn and the Warring States period,the guidance of the operation,as a combination of respiratory movement and physical exercise,medical and sports methods.Daoyin was first seen in "Zhuang Zi-deliberately",in Synopsis of prescriptions of the Golden Chamber appeared guided surgery in the treatment of diseases related to entry for.In the Mawangdui Han tomb unearthed in the "guide map" in the detailed records of the 44 pieces of small body guide figure.To the Sui Dynasty,guiding operation formally as a means of medical by too hospital issued,recorded in the Chao Yuanfang general treatise,and puts forward the guiding methods in the treatment of liver diseases.But the tumor occurrence and development mechanism is very complex,at presentabout movement of after hepatic resection for hepatocellular carcinoma patients is still a lack of relevant data,there is not a clear and standardized exercise program for hepatocellular carcinoma patients after rehabilitation,but also the lack of the stages of exercise and immunity related objective theoretical basis.In summary,this study constructed after operation in a mouse model of orthotopic transplantation tumor of liver resection,and the mouse model of postoperative spontaneous activity,liver function and lymphocyte cell function were studied,exploration model mice in the postoperative general status and immune function recovery.In the mouse model of postoperative state analysis,after different time of mice of different intensity treadmill exercise in observed molecular mechanism of the effect of exercise on lymphocyte proliferation in mice and further study of exercise effect on lymphocyte proliferation in mice,lay the foundation for the subsequent experiments.Part One: the effect of surgery on the general condition and immune function of mice with orthotopic liver tumor resectionTarget:To construct orthotopic liver resection after tumor mouse model to observe the effect of surgery on the general condition and the immune function of miceMethods:1,Building model in mice after liver transplantation in situ tumor resection,using the spontaneous activity instrument observed spontaneous activity level of model mice after different time node(on the first day after operation,the third day,the fifth day,the seventh day,the nineth day).2,Collect the peripheral blood of modle mice,detect the serum albumin(albumin,propagated),bilirubin(total bilirubin,TB),ALT(alanine aminotransferase,ALT),aspertate aminotransferase(aspartate transaminase,AST),lactate dehydrogenase,lactate dehydrogenase,LDH)levels.3,Preparing spleen lymphocytes suspension of model mice,using enzyme-linked immunosorbent experiment(enzyme-linked immunosorbent assay,ELISA)method to detect the IL-2,IL-4,IL-6,IL-10,TNF-?,IFN-gamma cytokine secretion level.4,Using MTT colorimetry to detect each model mice spleen lymphocyteproliferation ability.5,Using surface flow cytometry to detect model mice spleen lymphocytes CD4+/CD8+ ratio.Results:1,The study results show that spontaneous activity in mice after one,three,five days,a total distance respectively for(782.54±142.95)cm,(932.22±46.90)cm,(1040.57±208.83)cm,compared with the distance in normal mice(1291.06±7.996)cm,the difference has statistical significance(P<0.05).Seventh day model of spontaneous activity in mice(1212.07±85.50)cm and ninth day model of spontaneous activity in mice(1144.11±59.79)cm and normal mice,the difference was not statistically significant.2,Detecting the serum of mice found that the first day of model mice after operation is most severe,except ALB,TB were(2.4±0.16)umol/L,alanine aminotransferase(ALT)for(364.00±46.5)u/L,AST for(1453.60±295.29)u/L,LDH is(2853.60±249.73)u/L are far higher than normal mice,and the difference is statistically significant.Among them,ALT,AST,LDH trends are consistent,both in the rapid rise,the first day that reached the peak,followed by a gradual decline.TB increased late in aminotransferase levels.The results showed that,TB in the postoperative first day rise(2.4±0.16)umol/L,the third day reached a peak(2.42 ± 0.23)umol/L,then dropped gradually.ALB there was no significant difference between the groups.3,the level of cytokine secretion was much higher than that of normal mice,and the difference was statistically significant(P<0.05).In addition to TGF-beta,the various cytokines were rapidly increased after surgery,the first day to reach the peak,then gradually decreased.The operation destroyed the balance of Th1/Th2 cytokines,resulting in Th polarization drift,resulting in immune disorder.4,After the first day and the third day,lymphocyte proliferation in mice respectively(85.27±0.04)% and(97.54±0.16%)were less than normal mice(1.7238±0.21%),and the difference was statistically significant(P<0.05),but the immune function of mice gradually restored,especially after operation on the seventh day the proliferation ability of lymphocytes(128.51±0.27%),compared to the first day of mice immune function improved.5,In this experiment,the CD4+/CD8+ lymphocytes ratio of the modle mice of the first day group is(1.61±0.33).After three days,the CD4+/CD8+ lymphocytes ratio of modle mouse is(3.21±0.45),after five days the CD4+/CD8+ lymphocytes ratio of modlemouse is(3.40±0.34),compared with normal mice,the differences were statistically significant(P<0.05),and the CD4+/CD8+ lymphocytes ratio of normal group mice is(5.46±1.07).And 7 days after operation,the mice were also higher than the five days after the operation,the difference was statistically significant(P<0.05).Conclusion: compared with normal mice surgery on mice spontaneous activity levels and liver function of greater injury,inhibit the proliferation of lymphocytes,so that the ratio of CD4 + / CD8 + disorders,that orthotopic liver tumor resection operation in mice caused great stress damage and immune suppression.Conclusion:Postoperative proinflammatory cytokine secretion in mice of each level is far higher than that in normal mice,lymphocyte proliferation decreased,CD4+ /CD8+ ratio imbalance,that in situ liver tumor resection caused great stress injury to mice,and the immune suppression in mice is similar to human in the perioperative period after operation,also need some time to gradually return to normal state.Part Two: Effects of exercise on postoperative orthotopic liver tumor mouse lymphocyte proliferation capacity and its mechanismTarget:To observe the effects of different exercise time and exercise intensity effects on orthotopic tumor postoperative lymphocyte proliferation ability and its mechanismMethods:1,Preparing the mouse model after liver tumor in situ,interved the mice with exercise in different time and intensity.2,Preparing of spleen lymphocyte suspension,MTT assay was used to test the movement of model mouse lymphocyte proliferation.3,Using PathScan Intracellular Signaling Array Kit to screen,Western blotting Western blotting(Western Blot WB)to verify,in order to explore the molecular mechanism of effect of exercise on immune function of mice liver after resection of the tumor in situ.Results:1,Early postoperative exercise did not affect the ability of moderate intensity exercise on mouse lymphocyte proliferation,for(258.24±0.64)%,and the normal micelymphocyte proliferation,for(297.91±0.28).There was no significant difference between the two groups.2,High intensity exercise inhibited mice lymphocyte proliferation in to the extent,for(142.03±0.24)%,lower than that of normal mice,and the difference was statistically significant(p<0.01).The lymphocyte proliferation of Moderate intensity excercise with late start time after operation,was(221.76±0.24)%,and the normal mice lymphocyte proliferation was(194.16±0.71)%,there was no significant difference between the two groups,but high intensity exercise enhanced mice lymphocyte proliferation ability to a certain extent.That is(359.87±0.78)%,higher than that in normal mice,and the difference was statistically significant(P<0.01).3,After the PathScan Intracellular Signaling Array Kit were screened and found that the effect of exercise on lymphocyte of mice after resection of liver tumor in situ is mainly through IL6/JAK2/STAT3 and PI3k/AKT/mTOR/STAT3 these two pathways.After verification with WB found late postoperative exercise can improve the expression of phosphorylated STAT3 and AKT key protein,has a positive effect on lymphocyte proliferation.Conclusion:High intensity has negative moderating effect on lymphocyte proliferation in mice early after surgery,postoperative late in intensity and high intensity exercise on lymphocyte proliferation promoted.The movement may have effect on the proliferation of lymphocytes in mice by IL6/JAK2/STAT3 and PI3k/AKT/mTOR/STAT3 these two pathways. |
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