| Part 1 Observation on the killing effect of sodium abietate upon adult worms of Schistosoma japonicum in vitroObjective: To observe the killing effect of sodium abietate upon adult male and female worms of Schistosoma japonicum in vitro.Method: Mice infected with cercariae of Schistosoma japonicum were sacrificed and perfused five weeks later, obtained adult worms by the portal perfusion method.Adult worms of Schistosoma japonicum were cultivated in DMEM medium containing different concentrations of sodium abietate and artesunate,added control group,then observed the death and motility reduce rate. The adult worms were collected to detected the protein contents using Bradford method. Observed after they were stained by hydrochloric acid carmine and were investigated by routine pathological techniques preparation and HE staning.Result: These results showed that the death and motility reduce rate of adult worms were raised significantly after drugs'treatment;the activity of sodium abietate on adult male worms was more obvious than on adult female worms. At the same time the high and middle concentrations of sodium abietate's killing effect on adult male worms were significantly better than artesunate's;But artesunate's killing effect on adult female worms were significantly better than sodium abietate's.Compared with control group,the protein contents of adult male and female worms were reduced(P<0.05).The adult male worms'intestinal canal enlarged and appeared black or brown bands or spots after treatment.The content of adult female worms was distributed asymmetrically.The shape of some worms'ovaries were anomalism and coloring was asymmetrical.Conclusion: To Speculate from the result, sodium abietate could kill adult worms of Schistosoma japonicum in vitro and affected worms'protein metabolism. Part 2 Study on acute toxicity of sodium abietate to miceObjective: The acute toxicity of sodium abietate to mice was studied and observed its security.Method: The mice were randomly divided into groups: A1 group(200mg/kg), A2 group (400mg/kg),A3 group(800mg/kg)and A4 group(Normal sodium).Every group were given sodium abietate of the corresponding dose by intragastric administration one time.To observe changes of the ingestation, drinking and body weight.Then the mice were sacrificed,perfused,weighed these import organs and calculated the organ/body ratio.These organs were investigated by routine pathological techniques preparation and HE staning.The mice in addition were randomly divided into groups: B1 group(200mg/kg),B2 group (400mg/kg),B3 group(800mg/kg)and B4 group (Normal sodium). Every group were given sodium abietate of the corresponding dose by intragastric administration three times.The method of study was same with A group.Result: After single administration and repeated administration,the ingestation, drinking and body weight of mice were changed in different degrees.The ingestation and drinking were restored to or higer than the level of before administration at the third day after administration.The body weight of mice had increased in different degrees.The organ/body ratio of every experimental groups haven't significant difference with control group(P>0.05). No markedly abnormal change were seen in the organizational structure of every organ.Conclusion: In addition to influence the appetite and body weight temporarily after administration, no significant toxicity was observed.Part 3 Influence of immunosupression on the effect of artesunate against Schistosoma japonicumObjective : To observe the influence of immunosupression on artesunate killing larvae and adult worms of Schistosoma japonicum.Method: The mice infected with Schistosoma japonicum were randomly divided into groups:A1 control group,A2 artesunate group(Art group),A3 hydrocortisone group(HC group) and A4 hydrocortisone plus artesunate group(HC+Art group).HC was given to the mice by the way of intraperitoneal injection from 1st day after infection,continued 7 days.Then Art was given by intragastric administration at 8th day after infection. The mice were sacrificed and perfused seven weeks after infection with Schistosoma japonicum; the worm reduction rates were calculated; right lobe of liver were fixed and the liver paraffin section were prepared to measure of the diameter of the egg granuloma in liver;the remain liver were dissolved to count eggs.The mice in addition were deal with the same method, intraperitoneal injection from 28th day after infection, intragastric administration at 35th day after infection. Obstaining blood from some mice of every group 24h later and detected the level of specific anti-SWAP antibody. Obstaining worms and observed by SEM. The method of deal with the other mice was same with A group.Result: Compared with Art group, the worm reduction rate of HC+Art group were decreased significantly and the egg reduction rate and mean granuloma size also existed some difference;compared with control group, the worm reduction rate, egg reduction rate and mean granuloma size of HC group all reduced more or less. The level of specific anti-SWAP antibody of HC group(B3) and HC+Art group(B4) were reduced after hydrocortisone treatmentl(P<0.05). The result of SEM implied that body wall damage of Art group(B2) were more serious than HC+Art group(B4).Conclusion : To Speculate from the result,immunosupression can lessen the effect of artesunate killing larvae and adult worms of Schistosoma japonicum. |
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