Effects Of Direct Moxibustion "Yuyu" On T Lymphocyte Subsets, NK Cells And IL - 2 In HCC Precancerous Lesion Rats

Objective:To observe the effects of direct moxibustion and ginger moxibustion at "Ganshu"(BL18) on the content of T cells、NK cells and the expression of IL-2 in rat with precancerous lesion of liver cancer(HCC), induced by intraperitoneal injection of DEN, and then to explore the mechanism of enhance immunity function by moxibustion.Methods:1. Grouping and modeling methods:A totle of 70 male Wistar rats were randomly divided into control group(n=10),model group(n:=15),direct moxibustion-single group(n=15), direct moxibustion-double group(n=15)and ginger moxibustion-single group(n=15). HCC model group was established by intraperitoneal injection of diethylnitrosamine(DEN,50mg/kg), once every 3 days for 10 weeks. Direct moxibustion-single group and direct moxibustion-double group were respectively applied to bilateral "Ganshu"(BL 18) with 3 seed-sized-moxa cones or 6 seed-sized-moxa cones, ginger moxibustion-single group was treated with 3 seed-sized moxa cones,once every other day, the same as the model group’s treatment. Rats in the normal group received no treatment.2. Index detection:The contents of CD3+、CD4+、CD8+T cells and NK cells were examined by flow cytometry(FCM); using enzyme-linked immunosorbent assay to detect the expression of IL-2 in serum of each groups; using HE staining to observe rat liver tissue morphology.Results:1. The liver pathological changes:normal group:normal structure of hepatic lobules, hepatic cell degeneration and e necrosis, the nucleus clear and hepatic sinusoid without coronary stenosis; model group: Central vein distracted or absent, hepatic cords derangement or disappears, Hepatic blood sinus stenosis, obvious increase of collagenous fiber, with a sign of pseudolobuli formation, cell size inequality with hyperchromatic nuclei, higher cell atypia, Megakaryocytes tumor cells; moxibustion on each intervention group:high degree of differentiation of cancer cells under microscope, low cell atypia with some visible steatosis and cellular inflammation, it suggested that direct moxibustion and indirect moxibustion can inhibit the formation and development of hepatocellular carcinoma. In the pathological changes, each moxibustion intervention group showed high degree of differentiation of cancer cells under microscope and low cell atypia.2. Changes of T lymphocyte subsets and NK cells in peripheral blood of rats in each group:(1) Compared with the control group, the content of CD3+T cell and CD4+T cell in model groups were decreased (P<0.05,P<0.01), there was no significant difference in the other three groups; compared with model group, the content of CD3+T cell and CD4+T cell in direct moxibustion 1 groups were increased significantly (P<0.05, P<0.01), the content of CD3+T and CD4+T cell in direct moxibustion 2 groups were increased significantly (P<0.01, P<0.01), ginger moxibustion group compared with the control group, the content of CD4+T cell was increased significantly (p<0.01), while there was no significant difference in CD3+T cell content (P>0.05)(2) Compared with the control group, the content of CD8+T cell was increased (P<0.01), there was no difference between three moxibustion intervention groups and control group; compared with model group, the content in direct moxibustion 1 group, direct moxibustion 2 group and ginger moxibustion group were significantly reduced (P<0.01,P<0.01,P<0.01), there was no difference among the three groups.(3) Compared with the control group, the ratio of CD4+/CD8+ became significantly decreased (P<0.01), the ratio of three moxibustion intervention groups were signigicantly increased (P<0.01,P<0.01,P<0.01); compared with model group, after the moxibustion intervention in three groups, the ratio of ratio of CD4+/CD8+ became significantly increased (P<0.01,P<0.01,P<0.01), there was no difference among the three groups.(4) Compared with the control group, the content of NK cell was signigicantly increased in model group、direct moxibustion-single group and ginger moxibustion-single group (P<0.01,P<0.01,P<0.05,); compared with model group, the content in direct moxibustion 1 group, direct moxibustion 2 group and ginger moxibustion group were significantly decreased (P<0.05,P<0.01,P<0.01); compared with direct moxibustion-single group, the content in direct moxibustion-double group were decreased (P<0.05).3. Expression changes of IL-2 in serum of rats in each group:Compared with the control group, the expression of IL-2 in model group was significantly decreased (P<0.01), it also decreased in direct moxibustion 1 group and ginger moxibustion group (P<0.05, P<0.05), there was no difference between the direct moxibustion two groups and the control group; compared with model group, the expression of IL-2 in direct moxibustion 1 group, direct moxibustion 2 group and ginger moxibustion group were was significantly (P<0.05, P<0.01, P<0.05); there was no difference between the groups.Conclusion:1. The establishment of HCC precancerous lesion model was successful induced by intraperitoneal injection of DEN.2. Although there were natural anti-tumor immune responses in HCC precancerous lesion model of rats, while anti-tumor immune function decreased and disorder.3. Direct moxibustion and ginger moxibustion on Ganshu point can impove the content of CD3+T cell and CD4+T cell,decrease regulated cytotoxic CD4+T cell content, make the ratio of CD4+/CD8+tend to normal range, regulate the immune reaction and improve the anti-tumor immune function.4. The expression of IL-2 in rat serum was decreased in HCC precancerous lesion model, direct moxibustion and ginger moxibustion on Ganshu point can enhance the expression of IL-2, it is suggested that impoving the expression of IL-2 may one of the mechanisms of anti tumor by direct moxibustion and ginger moxibustion.In conclusion, direct moxibustion and ginger moxibustion can improve the pathological morphological changes of precancerous lesion of hepatocellular carcinoma, reduce the content of CD8+T, NK cell in the peripheral blood, improve the content of CD3+, CD4+T cell, increase the ratio of CD4+/CD8+ and enhance the expression of IL-2, so as to enhance the effect sufficiently immune response, and thus advance the anti-tumor immunity and immune surveillance, avoiding the escape the tumor cell.