The Preliminary Research Of Anti-tumor Activity Extracts And Their Metal Complexes Of Zanthoxylum Nitidum And Plumbago Zeylanica

Objective:to detect the anti-tumor activity of forty compounds in vitro and screen out the compounds with cytoctony against tumor cell,observe the effects of the compounds with cytoctony against tumor cell on the primary mice hepatocyte,establish the method for evaluating the drug-induced hepatotoxicity in vitro in our labs,study the effects of the compounds with cytoctony against tumor cell and low hepatotoxicity in vivo preliminarilyMethod:(1)The cytotoxicity of forty compounds against seven tumor cell lines was measured by MTT assay,the compounds with cytoctony anainst tumor cell in vitro were screened out and the forty compounds' IC₅₀values against the seven tumor cell lines were calculated respectively.(2)The primary mice hepatocyte was isolated by two-step collagenase perfusion method.The total growth process of the hepatacyte was observed and the content of ALT,ALB, LDH,BUN,in the supernatants of culture conditional media were detected all through the process to determine the optimal experimental session and the experimental project.The cytotoxic active compounds' IC₅₀values against the primary mice hepatocyte were measured by MTT assay respectively.The hepatocellular viability was detected through the indexes above to learn the effect of the cytotoxic active compounds on the primary mice hepatocyte.The method for evaluating the drug-induced hepatotoxicity in vitro in our labs was established through the above means.(3)The compounds with cytoctony against tumor cell and low hepatotoxicity were screened out and the compounds' LD₅₀ were obtained by acute toxicology experiment in vivo.Mice with S180 were established to observe the antitumor activity and the effect on the weight of main organs after ip N-17.The content of SOD,MDA,NO,GSH-PX in mice live homogenates were measured,the tumor issue were analyzed by histopathological observation.Results:(1)Among forty compounds,ten compounds were screened out for their good cytotoxicity on the seven cell lines,and got the ten compounds' IC50 values against the seven cell lines respectively.(2)The primary mice hepatocyte isolated by two-step collagenase perfusion method was attached to the wall 24 hours after isolation and developed successfully in 2 to 5 days,the activities of ALT,LDH were at a stable level during the period and the levels of ALB,BUN were at their activity peaks.The primary mice hepatocyte were died quickly at the fifth day after isolation,meanwhile the indexes of hepatocyte viability were also changed.So the optimal experimental session to investigate the drug-induced hepatotoxicity by the primary mice hepatocyte was determined at the second to the fifth days after isolation.The ten compounds' IC₅₀values against the primary mice hepatocyte and their effects to hepatocyte viability were taken into account,the N-1,N-3 and the N-17 compounds of the ten were screened out for their high IC₅₀values and little effects to hepatocyte viability. Further investigation of the three compounds was deserved.(3)The N-17 compound showed a dose-dependent inhibitory activity against mice bearing S180.The inhibition rate of N-17 against the growth of S180 was 42.9%at the dose of 10mg/kg,within the dose of 2.5-10mg/kg;the inhibition rate of N-17 against the growth of S180 was 1.95-42.9%.N-17 at different dosages had no marked effect on liver and spleen indexes of mice bearing S180.N-17 showed a dose-dependent inhibitory activity on thymus in mice bearing S180,within the dosage of 2.5-10mg/kg,the inhibition rate of N-17 against the thymus was 5.69-18.51%.N-17 could significantly increase the activities of SOD and GSH-PX;decrease the contents of MDA and NO.Different extents of tumor cytonecrosis at different dosage groups were observed by using pathological section of the tumor tissue,and obvious tumor cytonecrosis could be found at the high dosage group.Conclusion:Ten compounds were screened out from the forty compounds for their cytotoxic activities.By the establishment of the screen model of hepatotoxicity,the N-1,N-3,N-17 were screened out from the ten compounds for their cytotoxic activity and low hepatotoxicity.In the pharmacology experiment in vivo,N-17 shows a marked dose-dependent inhibitory activity against mice bearing S180 and immune toxicity.The anti-tumor activity of N-17 is related to its cytotoxicity on the tumor cell,meanwhile,N-17 can improve the activities of antioxidases,and it revealed that its anti-tumor activity may also relate to its antioxidation.