| Objective: To explore, in different drug concentrations, Fungal extracts’s effectson killing mice’s trichinosis, the procedure of killing the parasites and how it will affectthe parasites’ bioactivity. At the same time, to observe the effect on the change of theintestinal mucosa and the host’s body’s immune activity before and after using Fungalextracts to treat the mice’s trichinosis.Methods: Counting method was applied first, to count Trichinellaspiralis’ cysts.Use feeding method to infect mice with Trichinellaspiralis to establish trichinosisanimal models. Infected mice were randomly divided into four groups:3experimentaltreatment groups of different concentrations (A1:50mg/kg; A2:100mg/kg; A3:200mg/kg), a positive model control group (Y group). At the same time, establish anegative control group (Z group). Each group had6mice. After being infected for5days,mice of experimental treatment groups were treated with drugs of differentconcentrations for3days. And2days after treatment, they were all killed. At the sametime, kill the mice of the positive model control group (Y group) and negative normalcontrol group (Z group). Collect all the mature parasites in the mice’s intestinal caecumand count, calculate parasites’ load and reduction rate. Randomly take some of themature Trichinellaspiralis from each experiment group, and use coccinellin to dye theparasites and use transmission electron microscope to inspect, as to observe the externalform, internal structure and the effect of development condition of mice that were fromdifferent groups and treated by different concentrations. And analyze the drug’smechanism on killing the parasites and the place where it affects. In addition, take theileal intestinal tissue, ileum and ileocecal mucosa secretion before and after thetreatment of the mice of each experiment group, and do immunohistochemical stainingobservation respectively, and do evaluation of SIgA test.All the statistics got from each group will be calculated using the method of statistical analysis.Result:1. Observation of drug’s repellent effectThe mature parasites load and reduction rate of the3groups: A1, A2and A3, was57.1%,78.1%and96.2%, respectively. Compare the parasites load and reduction rate ofthese3treatment groups with the positive model control group (Y group), after usingstatistical analysis, it is found out that there were significant differences (P <0.05).There were also significant differences among A1, A2, A3, the3treatment groups onparasites load and reduction rate (P <0.05). And group A3’s killing effect was the best.2. Observation of antiparasitic drugs on parasites’ effectCollect mature parasites from the mice’s intestinal tract in group of A1, A2and A3and the positive model control group (Y group), use morphology measurement softwareto measure the average length of the parasites: group A1’s length and width is1278.95±68.89×49.97±8.83; group A2’s length and width is1213.45±88.04×38.91±5.76; group A3’s length and width is1122.34±42.88×33.80±6.22; group Y’slength and width is1305.55±53.90×57.47±5.76. There were not significant differenceswhich were analyzed by statistical analysis among these4groups or in each group (P>0.05). But under light microscope’ observation, because of the antiparasitic drugs’effect, parasites among these4groups and in each group were not as big as they shouldbe or were not grown balanced.3. Observation of drugs’ effect under light microscopeEpidermises of Trichinellaspiralis in the positive model control group are complete,smooth, and intact. Intestines’ epithelial cell’s top’s microvilli of mature parasites arecomplete, tidily arranged and forming brush border. Because of Fungal extracts’ effect,epidermises of mature parasites in group A1, A2, A3became rough and uneven.Intestines’ epithelial cell’s top’s microvillus of mature parasites in group A1’s integrityare damaged, untidily arranged and of different lengths, some of the microvillus werelodging. Microvilli of group A2not only became lodging but also became short andsmall, their growth was limited. Microvilli of group A3were badly damaged and therewere serious pathological changes like defection and merge.4. Mice’s intestinal mucosa’s laminae propria’s FAS protein’s expressionsAll the experimental mice’s intestinal mucosa’s laminae propria’s FAS protein’sexpressions are dyed by immunohistochemical staining, and analyzed by Gray analysissoftware, the results are as follows: group Y’s mice’s intestinal mucosa’s laminae propria’s FAS protein’s expressions’ average optical density value is0.2195±0.0160,compared with group Z’ s0.1099±0.0042, group Y had raised obviously.After the experimental mice were treated with drugs, and along with the density’s riseof the drugs, FAS protein’s expressions descended gradually. Group A1is0.1541±0.0055; group A2is0.1268±0.0074; group A3is0.0944±0.0046. Afterstatistical analysis, the FAS protein’s expressions among these3groups had obviousdifferences (P<0.05). After being treated with mass quantity of pesticide, groupA3’mice’s FAS protein’s expressions level recovered, and were near to the level ofgroup Z, but there were still obvious differences on statistical treatment (P<0.05).5. Mice’s level of SIgA secretionAfter being tested, all the experimental mice’s intestinal mucosa’s level of SIgAsecretions were as follows: group Y’s host intestinal fluid’s level of SIgA secretion was897.2473±20.3669, compared with group Z’s79.0611±4.9235, it obviously raised(P<0.05). After the3experimental treatment groups were been given drugs and as theconcentration of the drug raised, intestinal mucosa’s level of SIgA secretions descendedgradually. A1was140.0946±14.5452; A2was100.1102±7.5841and A3was63.8602±8.7385. There were also obvious differences on statistical analysis of these3groups (P<0.05). After group A3’s mice were being killed by large quantity of pesticide,Small intestinal mucosa’s level of SIgA secretion recovered and was near the level ofthe negative normal control group (Z group). There were also obvious differences onstatistical analysis (P<0.05).Conclusion:(A) Fungal extracts has fairly good effect on killing mature Trichinellaspiralis andhas obvious damaging effects on mature Trichinellaspiralis’ surface and their digestivesystems. The reason why the killing mechanism works maybe that the drug canrestrain the Trichinellaspiralis’ NFRD’s biological activity in, and lead to Eliminatingthe disturbance of energy metabolism, eventually lead to the gradual death becauseexhausting of energy depletion, and excrete out of the body. Fungal extracts’s bestconcentration for killing parasites is200mg/kg.(B) Through direct or indirect ways, Fungal extracts can reduce Mice’s intestinalmucosa’s laminae propria’s FAS protein’s expressions; restore the pathophysiologychange of lagre quantity of programmed cell death caused by Trichinellaspiralis’infection on Intestinal epithelial cells.(C) Through direct or indirect ways, Fungal extracts can reduce intestinal mucosa’s level of SIgA secretions, stimulate hosts to generate adaptive immunity, and assist hoststo generate immune response, so as to remove the Trichinellaspiralis’ living andplanting in the intestinal and defense the parasites’ infection.Fungal extracts has been proved as a new type of antiparasitic, it has good effect onkilling Trichinellaspiralis and the treatment of trichinosis. Further development andresearch of this drug will help our country’s medication industry’s independentdevelopment and it has good market development prospects. |
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