Experimental Study On Effect Of Yiqi Jianpi Anti - Cancer Traditional Chinese Medicine On Protein Kinase C In Liver And Kidney Of Gastric Cancer

Purpose：To study the effects of replenishing spleen Qi anticancer traditional Chinesemedicine on protein kinase C activity in the liver and kidney of nude mice bearinggastric cancer by observing the change of weight of mice, liver weight, kidneyweight, liver coefficient, kidney coefficient and PKC activities in liver andkidney.Material and method：1. Laboratory animal and cell line40nude mice, clean grade, each weights20±2g, bought from laboratory animalcenter of China medical university, animal certification number: SCXK (辽)2008-0005. Fodder is SPF grade, bought from laboratory animal center of Chinamedical university, along with Aseptic packing. Human gastric cancer SGC-7901cell line, bought from cell biological laboratory library of China medicaluniversity.2. Experimental method(1).Laboratory animal grouping and treatmentRandomly pick30out of40nude mice and armpit inoculated with human gastriccancer cell SGC-7901. Then nude mice bearing gastric cancer are randomly assignedto model group, tonifying spleen control group and tonifying spleen anticancergroup, with10mice in each group. The rest10non-inoculated nude mice areassigned to a blank control group.The blank control group is dosed with0.9%NaCl; the model group is dosed with0.9%NaCl; the tonifying spleen control group is dosed with replenishing spleenQi traditional Chinese medicine; the tonifying spleen anticancer group was dosedwith replenishing spleen Qi and anticancer traditional Chinese medicinecombination. The blank control group and model group are dosed with0.4ml0.9% NaCl per day, and for14days. Each drug group is treated with0.4ml correspondingdrug per day, and for14days by gavage.(2).Gastric cancer nude mice model preparationPick logarithmic phase tumor cells, adjust the cell concentration to107/ml,inoculate the nude mice with0.1ml tumor cells, equivalent to106cells/mouse.The whole operation requires aseptic technique. Observe the tumor growth on dailybasis.15days after inoculation, the tumor nodules with a diameter of0.8cm,hard texture and other indicators which appear in the inoculate position areidentified as tumors.(3).Sample collection and index detection(a).By the end of the experiment, all animals required to be executed, open theabdominal cavity, strip the peritoneum, take fresh tissue, wash away the bloodwith a0.9%NaCl solution, observe animal liver, kidney tissue general conditionand weight it, calculate the organ coefficient. Organ coefficient=organ weight(g)/body weight (g)×100%.(b).Liver and kidney PKC test: using Western blot method to exam the proteinexpression.Results：1. Nude mice weight, liver weight and kidney weight resultComparing with the black control group, the model group’s weight, liver weightand kidney weight all clearly dropped. Comparing with the model group, the bodyweight, liver weight and kidney weight of both tonifying spleen control groupand tonifying spleen anticancer group increased significantly. However, thereis no clear difference between tonifying spleen control group and tonifyingspleen anticancer group on body weight, liver weight and kidney weight2. Nude mice liver coefficient and kidney coefficient calculation resultLiver coefficient and kidney coefficient of the model group are lower than theblank control group. The difference is statistically significant (P <0.05).Liver coefficient and kidney coefficient of both tonifying spleen control groupand tonifying spleen anticancer group increased，comparing to the model group. The difference is statistically significant (P <0.05). However, there is nostatistically significant difference between tonifying spleen control group andtonifying spleen anticancer group (P>0.05). The PKC activities in liver andkidney of the model group was significantly higher than the blank control group.After treatment， the PKC activities in the nude mice’s liver and kidney intonifying spleen group and tonifying spleen anticancer group were bothdecreased， especially significant in the tonifying spleen anticancer group，compared with the model group (P <0.01).3. Test result of PKC expression in nude mice liver and kidneyComparing to the blank control group, the PKC activities in liver and kidneyof the model group has increased. The difference is statistically significant(P <0.01). Comparing to the model group, the PKC activities in the nude miceliver and kidney in tonifying spleen group and tonifying spleen anticancer groupwere both significantly decreased (P <0.01). The liver PKC activity drop moresignificantly in the tonifying spleen anticancer group than the tonifying spleengroup，and has almost recovered to the level of the blank control group. Thekidney PKC activity drop more significantly in the tonifying spleen anticancergroup than the tonifying spleen group，but there are still differences comparingto the blank control group (P <0.05).Conclusion：1. The nude mice bearing gastric cancer have symptoms of spleen deficiency.2. Both replenishing spleen Qi traditional Chinese medicine and replenishingspleen Qi anticancer traditional Chinese medicine have regulating effect on theweight， liver weight， kidney weight， liver coefficient， kidney coefficientto the gastric cancer bearing nude mice.3. Both replenishing spleen Qi traditional Chinese medicine and replenishingspleen Qi anticancer traditional Chinese medicine have regulating effect on PKCactivities in liver and kidney to the gastric cancer bearing nude mice.4. The replenishing spleen Qi anticancer traditional Chinese medicine has a moresignificant regulating effect on PKC activities over replenishing spleen Qi traditional Chinese medicine.