Experimental Study On The Effect Of Invigorating Qi And Spleen And Anti - Cancer On Spleen Essence Of Colorectal Cancer And Anti - Tumor Effect

Purpose：One common malignant tumors of colorectal cancer is still a serioushazard to human health, the most common malignancy in the world is located inthe first three colorectal cancer, colorectal cancer resulted in deathsaccounted for10%of all cancer deaths. Cancer patients performed significantlyspleen deficiency, on the one hand to reduce colorectal cancer itself spleenand up-cleaning function, on the other hand after surgery and chemotherapy,the spleen and clean up a further decline in function, so throughout the colonSpleen the occurrence and development of the whole process. Control the progressof colon cancer, improve the quality of life of cancer patients, improveimmunity should Spleen. Preliminary clinical studies have demonstrated thatprescription in the " Qi and spleen cancer," the principle of the formationof the treatment of digestive cancer, anti-cancer, improve symptoms, reducethe role of radiotherapy and chemotherapy toxicity. Vitro studies havedemonstrated that anti-cancer medicine Spleen Qi has the role ofapoptosis-related gene expression to promote apoptosis and inhibition ofcolorectal cancer, but the anti-tumor mechanisms at the molecular level researchis not deep enough. This study aims to influence the mechanism of Spleen Qispleen colorectal cancer on the nature of law and anti-tumor research, analysisof colorectal cancer diagnosis and treatment law; further evidence of thetherapeutic effect of anti-cancer medicine Spleen Qi method, in-depth studyof Qi intrinsic relationship between the spleen and cancer, at the cellularlevel, the molecular level, the level of depth to reveal the protein levelspleen Qi anticancer mechanism of law, this experiment has a more importantclinical significance.Material and method：1. Colon cancer cell line： Human colon carcinoma cellline HT-29， provided by bacterial libraries in cell biology laboratory of Chinese Medical Sciences University.2. Experimental animal：40nude mice.3.Drugs： Medicine of replenishing Qi and tonifying Spleen， referred to as“Xiangsha Liujunzi” soup， includes padix pseudostellariae， poria cocos，ptractylodes，liquorice，radices saussureae，fructus amomi，pinellia ternate，and dried orange peel， each50gram. Anticancer medicine， includes Fritillariathunbergii Miq.， rhizoma smilacis glabrae， pseudobulbus cremastrae seupleiones and Oldenlandia diffusa， each40gram. All drugs are provided byofficinal of Liaoning College of traditional Chinese Medicine. The medicine isboiled in common method into200ml liquid， and is concentrated into85ml liquid.The liquid is poured into small bottles，10ml each bottle. After autoclaving，the medicine is reserved at a temperature of4℃.To establish the model of humancolon cancer cell HT29subcutaneous xenotransplanted tumor in nude mice， amodeling method is taken from the literature. The logarithmic growth phase tumorcells are chosen and the consistence is107/ml. The mice are vaccinated eachwith0.1ml under arms. The whole process is required to be under axenic condition.The growth conditions of the tumor are observed every day. After15days， ifthere is tumor whose diameter is no less than0.8cm where the tumor cells arevaccinated， and the texture is hard， the tumor is formation is affirmed.The40mice are divide into groups randomly， named blank group， model group，replenishing Qi and tonifying Spleen group and anticancer group，10mice ineach group.All mice are intragastric administrated for14days， blank groupand model with normal saline， replenishing Qi and tonifying Spleen group with“Xiangsha Liujunzi” soup， and anticancer group with both “XiangshaLiujunzi” soup and anticancer medicine. Related physiological index of thespleen should be observed in detail.The first part of the experiment is toresearch the effects of the replenishing Qi and tonifying Spleen anticancermethod on digestive and absorptive functions. The measured data includes thecommon condition such as the weight， eating capacity and water intake， thegastric emptying rate and the intestinal propulsive rate.The second part of the experiment is to research the effects of the replenishing Qi and tonifying Spleen anticancer method on spleen PKC. The measured dataincludes the spleen coefficient and the spleen PKC protein expression fromWestern blot method.The third part of the experiment is to research the effectof the replenishing Qi and tonifying Spleen anticancer method on tumor EGF，VEGF expression. The measured data includes the tumor weight， the tumorinhibiting rates， apoptosis of tumor cells from TUNEL method， EGF and VEGFexpression from Western-blot method.Results：1. The increase of body weight：The maximum increase appears in the blank group，which is6.33±2.4g， higher than the other three groups. The minimum increaseappears in the model group，which is2.48±2.1g. The increase of the replenishingQi and tonifying Spleen group is4.78±2.9g. The increase of the anticancer groupis5.03±3.2g. The latter two groups are both better than the model group. Forthe replenishing Qi and tonifying Spleen group and the anticancer group， thecomparison has no statistical significance， but the latter is better on thetrend. The eating capacity and water intake show the same results with theincrease of the body weight.2. The gastric residual rate and the intestinal propulsive rate：For the blankgroup， the gastric residual rate is29.4±13.2%， and the intestinal propulsiverate is68.4±9.4%. It shows that the gastric residual rate is the lowest， andthe intestinal propulsive rate is the highest， which is the best among the fourgroups. For the model group， the gastric residual rate is47.5±12.3%， andthe intestinal propulsive rate is49.3±10.2%， which is the worst among thefour groups. For the replenishing Qi and tonifying Spleen group， the gastricresidual rate is35.7±14.1%，and the intestinal propulsive rate is62.5±11.3%.For the anticancer group， the gastric residual rate is40.2±12.5%， and theintestinal propulsive rate is58.7±10.6%. The two groups have almost nostatistical difference， but the latter is better than on the trend.3. The spleen coefficient：The maximum value appears in the blank group， whichis1.19±0.28%. The minimum value appears in the model group， which is 0.76±0.19%. The value of the replenishing Qi and tonifying Spleen group is1.07±0.22%. The value of the anticancer group is1.07±0.35%， a little largerthan the replenishing Qi and tonifying Spleen group， without statisticalsignificance. The coefficients of both the model group and the replenishing Qiand tonifying Spleen group are smaller than the blank group， with statisticalsignificance (p<0.05). Compared with the model group， the other three groupshave larger values， with statistical significance (p<0.05).4. Spleen PKC expression：The maximum value appears in the blank group， whichis0.567±0.011. The anticancer group has a value of0.480±0.014， which isa little lower than the blank group (p<0.05). The replenishing Qi and tonifyingSpleen group has a value of0.442±0.019， and the model group has a value of0.422±0.008， ever lower than the anticancer group (p<0.01). Compared with themodel group，the other three groups have higher values，with obvious statisticalsignificance (p<0.01).5. The weight of the tumor and the tumor inhibiting rates：For the weight ofthe tumor， the model group， the replenishing Qi and tonifying Spleen groupand the anticancer group have values of3.94±0.81g，3.01±0.45g， and2.43±0.36g respectively. The latter two groups are obviously smaller than themodel group， with statistical significance (p<0.05). And the anticancer groupis smaller than the replenishing Qi and tonifying Spleen group， with statisticalsignificance (p<0.05).For the tumor inhibiting rates， the replenishing Qi andtonifying Spleen group and the anticancer group have values of24%and38%respectively， which are obviously different， with statistical significance(p<0.05).6. The number of cancer cell apoptosis：The replenishing Qi and tonifying Spleengroup， the anticancer group and the model group have the values of3.5±1.08，8.70±1.34and0.60±0.70respectively， which show obviously difference， withstatistical significance (p<0.01). The replenishing Qi and tonifying Spleengroup has higher value than that of the anticancer group， with statisticalsignificance (p<0.01). 7. Tumor EGF， VEGF testing result：The model group， the replenishing Qi andtonifying spleen group and the anticancer group and have the values of0.574±0.020，0.596±0.014，0.506±0.023，0.533±0.016，0.368±0.021，0.419±0.012respectively， which show obviously difference， with statisticalsignificance (p<0.01)， which show that the model group has the maximum values，and the replenishing Qi and tonifying spleen group has lower values， but withoutstatistical significance. The anticancer group has lower values than the formertwo groups，with statistical significance (p<0.05). The model group has highervalues than the other two groups (p<0.01) and the anticancer is higher than thereplenishing Qi and tonifying spleen group (p<0.01).Conclusion：1.Intestinal cancer cause spleen deficiency, affecting digestion and absorptionfunctions of the spleen is significantly; Spleen Qi herbs can improve the generalstate of tumor-bearing mice, weight loss caused by decreased colorectal spleen;Spleen Qi herbs can enhance gastrointestinal motility, improve the function ofthe spleen and stomach.2. Analysis of colorectal tumors in rats cellular level, leading to objectivereasons Analysis spleen, colon cancer can inhibit the liver, spleen, kidneygrowth itself; Spleen Qi method can improve the experimental anti-cancer coloncancer mouse spleen function, and its mechanism is by increasing spleen weightand spleen index, spleen enhanced expression of PKC activity achieved; PKCactivity of each organization spleen Qi Recipe for colorectal cancer spleendeficiency mice expressing a significant influence. This may be one of itsmechanism of action.3．Spleen Qi and Spleen Qi group for cancer HT-29human colon cancer xenograftsin mice significantly inhibited, but the latter than the former anti-cancereffect; its mechanism of inhibition of tumor growth by inducing cell may beapoptosis and reduced tumor epidermal growth factor EGF, the expression ofvascular endothelial growth factor (VEGF) implementations. This method isapplied for Spleen Qi colorectal cancer provides a comprehensive treatment of laboratory evidence.