| Objective:Establish Henan strains of Trichinella spiralis infection kunming mice experimental animal model, By pathology and molecular biology means of experimental to detect the cell apoptosis in mouse model of intestinal and muscle tissue of Trichinella spiralis infection at different time. Discussion on Trichinella spiralis infection status of host cell apoptosis by Habitat under the mechanism, for prevention, treatment of trichinellosis, kill the Trichinella spiralis provide experimental basisMethods:80 mice were randomly divided into Group A(Adult stage) and Group B(Larval stage).4O rats in each group. A group randomly divided into infected with Trichinella 3、5、7 d group and normal control group,10 rats in each group. B group randomly divided into infected with Trichinella 3、4、5 w group and normal control group,10 rats in each group. Each group of mice infected with Trichinella larvae each 200, The control group received normal feeding, take small intestinal tissue of adult period Trichinella spiralis mice in the time of 3、5、7days.Take the muscle tissue of larval stage Trichinella spiralis mice in the time of 3、4、5weeks.Routine pathological section, Hematoxylin- eosin staining (HE) observe the histopathological changes of the small intestinal and muscle tissue. Western-Blot method to detect Caspase-3 expression level changes in the intestinal tissue of mice of different infection periods. Situ end labeling (TUNEL) method to detect the small intestine apoptotic index. To make a comprehensive evaluation of adults cause the intestinal tissue and larvae cause muscle tissue apoptosis by Pathology and molecular biology methods, Local mucosal epithelial cell is edema.Results:1 Adult:1.1 HE staining results:Intestinal tissue of normal control mice did not show any pathological changes, Intestinal villi is normal, Mucosal layer without change, no change in mucosal epithelium also, Intrinsic membrane is normal. Infection of 3d group, Intestinal mucosa epithelial cells edema was not obvious, The mucosal layer was slightly bleeding. Intrinsic membrane vacuole change is not obvious, Infection of 5d group, Intestinal mucosal epithelial cells showed slight edema, The mucosa of intestinal gland showed obvious hemorrhage, Fewer inflammatory cells, the lamina propria has obvious vacuolization change. Infection of 7d group, Small intestinal mucosa has inflammatory cells infiltration and with massive hemorrhage,Intrinsic membrane vacuole changed more significantly, intestinal villi impairment.1.2 The results of Western-blot:Caspase-3 protein average gray value of normal control group (0.79±0.03).During the development of adult Trichinella spiralis, Caspase-3 protein mean gray ratio significantly increased with the prolongation of Trichinella infection(P<0.05). The gray value of 3,5,7d group were (0.86±0.03) (1.04±0.02) and (1.34±0.02). Compared with the normal control group, Infected with Trichinella spiralis 3,5,7 d group Caspase-3 protein the average gray were significantly increase(P<0.05), differences between any two groups also(P<0.05).1.3 The results of TUNEL method:Normal control group, the small intestinal mucosa layer can be seen scattered in the nucleus was dark brown or brown apoptotic cells, the apoptotic index (0.08 ± 0.02)%. With the growth in time of infection, the apoptotic index was significantly increased. The Trichinella infection apoptotic index of 3,5,7d group were (0.26 ± 0.03)% (0.38 ± 0.03)%and (0.73 ± 0.09)%. Compared with the normal control group(P<0.05), differences between any two groups also(P<0.05).2 larval stage:2.1 HE staining results:Muscle tissue of normal control mice did not show any pathological changes, Muscle fiber thickness and uniform law neatly arranged. As the infection time to 3w,4w, HE staining results manifested as muscle fibrosis swelling, Patchy, disorganized, and stripes disappear, reticular structure. The parasite resides Muscular pulp dissolved, Worms near the muscle cell necrosis, disintegration, Muscle interstitial edema and inflammatory cell infiltration. Inflammatory cells mainly lymphocytes, large mononuclear cells and eosinophils. Infection of 5w, The larvae formed around encapsulated, as the larvae grow up, curl, the muscle tissue of the parasitic parts gradually increasing spindle-shaped, the formation of fusiform muscle cavity wrapped around the body, acute inflammation subsides.2.2 The results of Western-blot:Caspase-3 protein average gray value of normal control group (0.23±0.02)., During the development of larval Trichinella spiralis, Infection 3,4 w of Caspase-3 protein mean gray ratio significantly increased with the prolongation of Trichinella infection(P<0.05). Various gray values were (0.31 ± 0.02), (0.54 ± 0.02). Infection of 5w, the average gray value of Caspase-3 protein was (0.36 ±0.03). Compared with the normal control group, infected with Trichinella spiralis 3,4,5 w group Caspase-3 protein the average gray were significantly increase (P<0.05). 3,4,5w group group between any two differences (P<0.05).2.3 The results of TUNEL method:Dark brown or brown apoptotic cells and nuclei were visible in normal control mice muscle tissue patchy. The apoptosis index was (0.12 ± 0.03)%. With the growth in time of infection, the apoptotic index was significantly increased. Trichinella infection 3,4,5w group apoptotic index were (0.32±0.05)%, (0.68 ± 0.03)%and (0.47 ±0.05)%. Compared with the control group (P<0.05).3,4,5w between groups, pairwise comparisons were significant differences (P<0.05).Conclusion:1 Apoptosis phenomenon is exist during the interaction with the host of Trichinella spiralis, Trichinella spiralis infection caused by parasite tissue cell apoptosis and pathological injury has certain relation.2 Adult of Trichinella spiralis parasites in the intestine of mice can induce apoptosis in the intestinal tissue. In a certain period of time, with the prolongation of infection time, also increased the tissue levels of apoptosis, the 7d infection reach apoptosis peak.3 Larvae of Trichinella spiralis parasites in the masule of mice can induce apoptosis in host tissue, and the level of apoptosis and infection time has close correlation.3,4 w Infection, the apoptosis significantly increased, at the 5w, Capsule formed, the level of apoptosis decreased. |
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