Chinese Medicine Scutellaria Barbata 5 - Fluorouracil Against Hepatocellular Carcinoma Enhanced The Role Of Experimental Research

Objective:This experiment used the 3～4 week male BALB/C nude mice as research sample.Establishing the model of hepatocarcinoma HepG2 cell xenografts in nude mice. To investigate the anti-toumor activity of Barbated Skullcup Herb in vivo.To observe the enhancement of BarbatedSkullcup Herb to the antitumor effect of 5-FU in nude mice inoculated hepatocarcinoma HepG2 cell xenografts.And discussing the antitumor mechanism caused by Barbated Skullcup Herb and 5-FU.So as to offer new thoughts for hepatocarcinoma therapy.Methods:HepG2 cell was dissociated by 0.125%trypsin which contains 0.02%EDTA and washed twice by PBS.Transfer the cell to a tube the centrifuge at 1000rpm for 3min,adjusting cell suspension to 1×10~8/ml with Sodium Chloride.The 60 nude mices to which injected the suspension were divided into six groups at radom in next day,namely the positive control group(5-fluorouracil25mg/kg),A group(Barbated Skullcup Herb 50mg/kg+5-fluorouracil 25mg/kg),B group(Barbated Skullcup Herb 30mg/kg+5-fluorouracil 25mg/kg )and C group(Barbated Skullcup Herb10mg/kg+5-fluorouracil 25mg/kg),Barbated Skullcup Herb control group(100mg/kg).5-fluorouracil(25mg/kg.d) inject to intraperitoneal twice a week in positive control group.The same volume Sodium Chloride administer in negative control group.Respectively Barbated Skullcup Herb(50mg/kg,30mg/kg,10mg/kg) in A group,B group and C group with intragastric administration.And 5-fluorouracil(25mg/kg.d) inject to intraperitoneal twice a week in the three groups.Barbated Skullcup Herb(100mg/kg) in Barbated Skullcup Herb control group with intragastric administration negative control group(normal sodium group),General state of health and tumor growth were observed every day.Draw the tumor volume time-growth curve according to measured the length(a),width(b)and highly(c) of tumor wit vernier caliperbare which on the basisi of formula(V=ab~2/2)every week.All the nude mices were killed after 5 weeks,separation blood serum -20℃to save.Strip and weigh the lump tissue,liver,and spleen.To calculate average weight of the lump tissue and the inhibition rate of tumor growth..AFP in blood serum was tested with radio-immunity.Beta-Endorphin in blood serum and PPARγin lump tissue were tested with enzyme linked immunosorbent assay.Testing the content of Cytochrome C and mitochondrial calcium in lump tissue.Results:The model of hepatocarcinoma HepG2 cell xenografts in nude mice was preparated succedly.The tumor-growth curve showed that negative control group was increased obviously,meanwhile,the tumor growth of the Barbated Skullcup Herb control group and positive control group were slowly.The tumor inhibition rate of Skullcup Herb control group was 26.4%and 5-Fu control group was 24.4%which contrast to the negative group(p＜0.05).The tumor inhibition rate of Barbated Skullcup Herb large dosage group, Barbated Skullcup Herb middle dosage group and Barbated Skullcup Herb small dosage group respectively was 30%,39.4%,33.1%which contrast to the positive control group(p＜0.05).B group contrasts obviously to A group and C group(p＜0.01).The weight of spleen and liver was increased in A group,B group,C group and Skullcup Herb control group,which contrast to positive control group(p＜0.05 or p＜0.01).It showed that Barbated Skullcup Herb can enhance the immunity of nude mice and reduce the liver injury induced by 5-FU.In positive control group or Barbated Skullcup Herb control group,the level of Cyt C was increased and mitochondrial calcium of lump tissue was lowered.And this change especially showed in B group.Meanwhile,PPARγin lump tissue and Beta-Endorphin in blood serum were increased obviously in Barbated Skullcup Herb middle dosage group.Conclusion:The Barbated Skullcup Herb has marked effect of anti-tumor in vivo and inhibit the tumor of nude mice.Especially,the B group(30 mg/kg) and 5-FU(25mg/kg) can markedly inhibit the tumor growth of hepatocarcinoma HepG2 cell xenografts in nude mice but does not decrease the weight of spleen and liver,suggesting that the combination may decrease the side effect produced by 5-FU.Cyt-C and calcium ion participated lump tissue apoptosis.The combination of Barbated Skullcup Herb(30 mg/kg) and 5-FU (25mg/kg) can increase the expression of PPARγin lump tissue and Beta-Endorphin in blood serum to reduce inflammation injury and cancerous pain.Therefore,Barbated Skullcup Herb can enhance the antitumor effect of 5-FU.