猪囊尾蚴的能量代谢变化规律和药物作用机理研究
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摘要
本研究首次系统地测定了体内发育过程中、体外培养条件下及抗囊药物作用下猪囊尾蚴能量代谢的有关物质含量和酶活性的变化，阐明了猪囊尾蚴的能量代谢变化规律和苯并咪唑氨基甲酸酯类药物作用机理，为筛选有效的抗囊药物以及为药物治疗提供了重要的实验和理论依据。
通过观察试验猪以六钩蚴人工感染后，体内囊尾蚴发育过程中的形态学变化和测定以糖代谢为主的能量代谢的有关物质含量(Glc、LAC)和酶活性(PEPCK、PK、LDH、SDH、ICD、XOD、ME、GDH、FR、ATPase、ACP、AKP、G6Pase、FE)的变化及进行LDH、SDH、ICD同工酶电泳观察，阐明了猪囊尾蚴未成熟期和成熟期的能量代谢变化规律。结果表明猪囊尾蚴具有与其它寄生蠕虫类似的物质转运、糖有氧分解、糖无氧分解、局部的逆向三羧酸循环、三羧酸循环、脂类分解、氨基酸分解、嘌呤分解等能量代谢途径，由未成熟期到成熟期，能量代谢逐渐活跃，但糖无氧酵解生成乳酸的途径减弱。
观察了抗囊药物NX0、NX1、NX2对感染猪体内未成熟期和成熟期猪囊尾蚴治疗效果和对体外培养条件下未成熟期和成熟期猪囊尾蚴的杀伤作用，结果表明NX0和NX1均具有显著的疗效，NX1的药效优于NX0，NX1对未成熟期猪囊尾蚴的杀灭作用优于成熟期。本试验成功地筛选了对猪囊尾蚴未成熟期和成熟期均有效的药物NX1。研究了抗囊药物作用下猪囊尾蚴能量代谢的有关物质含量和酶活性的变化，结果表明NX0和NX1通过引起猪囊尾蚴能量耗竭而杀死虫体。
测试了NX0和NX1对猪囊尾蚴组织匀浆延胡索酸还原酶活性的抑制作用，结果表明两种苯并咪唑氨基甲酸酯类药物非竞争性抑制延胡索酸还原酶复合体活性。
药物对猪囊尾蚴能量代谢的影响、药物对延胡索酸还原酶活性的抑制作用和药物构效关系分析结果表明，苯并咪唑氨基甲酸酯类药物通过抑制虫体葡萄糖摄取及抑制延胡索酸还原酶复合体活性，致使虫体能量耗竭而死亡。
依据药物作用机理，进行了苯并咪唑氨基甲酸酯类药物的类型衍化和新药设计。进一步的研究可为猪囊尾蚴病等寄生蠕虫疾病的治疗提供更好的药物。猪囊尾蚴可成为研究寄生蠕虫重要的实验动物模型，而有关药物则可成为研究寄生蠕虫生物化学与分子生物学的重要研究工具。
In this test, metabolites and enzyme activities involved in energy metabolism in Cysticercus cellulosae were determined systematicly in vivo and in vitro and under action of drugs.The principles of variation of energy metabolism in Cysticercus cellulosae and mechanism of action of the benzimidazol carbamate drugs were illustrated so as to select an effective drug against immature and mature cysticerci and to provide a theoretical and experimental chemotherapy.
Changes of morphology of cysticerci were observed in swines infected with oncospheres and metabolites and enzyme activities involved in energy metabolism in Cysticercus cellulosae were determined systematicly including the content of Glc and LAC and the activities of PEPCK. PK LDH SDH ICD, XOD ME, GDH, PR, ATPase, ACP,AKP,G6Pase,FE,MDH,HK and the isoenzymes of LDH,SDH,ICD were observed. Priciples of variation of energy metabolism in immature and mature cysticerci were illustrated.The results showed that transport of substances anaerobic glycolysis aerobic glycolysis, partial inversed tricarboxylic acid cycle. tricarboxylic acid cycle, fat decomposing, amino acid decomposing, xanthine decomposing metabolism were almost the same as those in other helminthes, and the pathways of energy metabolism enhanced from immature state to mature state while the PK activities declined.
Effect of drugs against cysticercosis including NX0 NX1 and NX2 on immature and mature cysticerci in swines were observed. The results showed that NX0 NX1 both had obvious effect, NX1 was better than NXO and had better effect on immature cysticerci than mature ones. Thus a kind of drugs agaist immature and mature cysticerci was selected successfully. The testing results of indexes involved in energy metabolism in Cysticercus cellulosae after action of drugs showed that the two kinds of drugs against cysticercosis resulted in exhaustion of energy and death of cysticerci.
Inhabition of NXO and NX1 on the activities of fumaric reductase in tissue homogenate of cysticerci was tested.The results showed that the two kinds of benzimidazol carbamate drugs could inhabite the activity of fumaric reductase complex noncompetently.
The results of influences of drugs on energy metabolism in Cysticercus cellulosae direct inhabition of drugs on the activity of fumaric reductase and structure - activity relation analysis of drugs showed that benzimidazol carbamate drugs could inhabite the
absorbtion of glucose and inhabite the activity of fumaric reductase complex.
Depending on the mechanism of action of drugs, type derivation of benzimidazol carbamate drugs and new drug designing were discussed.Next step of reseach will provide better drugs against cysticercosis and other helminth diseases. Cysticercus cellulosae may become an important kind of experimental model for helminth study, while drugs involved may become an important tool in biochemistry and molecular biology study of helminth.
通过观察试验猪以六钩蚴人工感染后，体内囊尾蚴发育过程中的形态学变化和测定以糖代谢为主的能量代谢的有关物质含量(Glc、LAC)和酶活性(PEPCK、PK、LDH、SDH、ICD、XOD、ME、GDH、FR、ATPase、ACP、AKP、G6Pase、FE)的变化及进行LDH、SDH、ICD同工酶电泳观察，阐明了猪囊尾蚴未成熟期和成熟期的能量代谢变化规律。结果表明猪囊尾蚴具有与其它寄生蠕虫类似的物质转运、糖有氧分解、糖无氧分解、局部的逆向三羧酸循环、三羧酸循环、脂类分解、氨基酸分解、嘌呤分解等能量代谢途径，由未成熟期到成熟期，能量代谢逐渐活跃，但糖无氧酵解生成乳酸的途径减弱。
观察了抗囊药物NX0、NX1、NX2对感染猪体内未成熟期和成熟期猪囊尾蚴治疗效果和对体外培养条件下未成熟期和成熟期猪囊尾蚴的杀伤作用，结果表明NX0和NX1均具有显著的疗效，NX1的药效优于NX0，NX1对未成熟期猪囊尾蚴的杀灭作用优于成熟期。本试验成功地筛选了对猪囊尾蚴未成熟期和成熟期均有效的药物NX1。研究了抗囊药物作用下猪囊尾蚴能量代谢的有关物质含量和酶活性的变化，结果表明NX0和NX1通过引起猪囊尾蚴能量耗竭而杀死虫体。
测试了NX0和NX1对猪囊尾蚴组织匀浆延胡索酸还原酶活性的抑制作用，结果表明两种苯并咪唑氨基甲酸酯类药物非竞争性抑制延胡索酸还原酶复合体活性。
药物对猪囊尾蚴能量代谢的影响、药物对延胡索酸还原酶活性的抑制作用和药物构效关系分析结果表明，苯并咪唑氨基甲酸酯类药物通过抑制虫体葡萄糖摄取及抑制延胡索酸还原酶复合体活性，致使虫体能量耗竭而死亡。
依据药物作用机理，进行了苯并咪唑氨基甲酸酯类药物的类型衍化和新药设计。进一步的研究可为猪囊尾蚴病等寄生蠕虫疾病的治疗提供更好的药物。猪囊尾蚴可成为研究寄生蠕虫重要的实验动物模型，而有关药物则可成为研究寄生蠕虫生物化学与分子生物学的重要研究工具。
In this test, metabolites and enzyme activities involved in energy metabolism in Cysticercus cellulosae were determined systematicly in vivo and in vitro and under action of drugs.The principles of variation of energy metabolism in Cysticercus cellulosae and mechanism of action of the benzimidazol carbamate drugs were illustrated so as to select an effective drug against immature and mature cysticerci and to provide a theoretical and experimental chemotherapy.
Changes of morphology of cysticerci were observed in swines infected with oncospheres and metabolites and enzyme activities involved in energy metabolism in Cysticercus cellulosae were determined systematicly including the content of Glc and LAC and the activities of PEPCK. PK LDH SDH ICD, XOD ME, GDH, PR, ATPase, ACP,AKP,G6Pase,FE,MDH,HK and the isoenzymes of LDH,SDH,ICD were observed. Priciples of variation of energy metabolism in immature and mature cysticerci were illustrated.The results showed that transport of substances anaerobic glycolysis aerobic glycolysis, partial inversed tricarboxylic acid cycle. tricarboxylic acid cycle, fat decomposing, amino acid decomposing, xanthine decomposing metabolism were almost the same as those in other helminthes, and the pathways of energy metabolism enhanced from immature state to mature state while the PK activities declined.
Effect of drugs against cysticercosis including NX0 NX1 and NX2 on immature and mature cysticerci in swines were observed. The results showed that NX0 NX1 both had obvious effect, NX1 was better than NXO and had better effect on immature cysticerci than mature ones. Thus a kind of drugs agaist immature and mature cysticerci was selected successfully. The testing results of indexes involved in energy metabolism in Cysticercus cellulosae after action of drugs showed that the two kinds of drugs against cysticercosis resulted in exhaustion of energy and death of cysticerci.
Inhabition of NXO and NX1 on the activities of fumaric reductase in tissue homogenate of cysticerci was tested.The results showed that the two kinds of benzimidazol carbamate drugs could inhabite the activity of fumaric reductase complex noncompetently.
The results of influences of drugs on energy metabolism in Cysticercus cellulosae direct inhabition of drugs on the activity of fumaric reductase and structure - activity relation analysis of drugs showed that benzimidazol carbamate drugs could inhabite the
absorbtion of glucose and inhabite the activity of fumaric reductase complex.
Depending on the mechanism of action of drugs, type derivation of benzimidazol carbamate drugs and new drug designing were discussed.Next step of reseach will provide better drugs against cysticercosis and other helminth diseases. Cysticercus cellulosae may become an important kind of experimental model for helminth study, while drugs involved may become an important tool in biochemistry and molecular biology study of helminth.
引文
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