| Objective 1)To observe the inhibitory effect of moxibustion on the growth of gastric cancer in rats;2)To explore the effect of moxibustion on the synthesis and release of HSP70 in gastric cancer tissue;3)To clarify the promoting effect of moxibusiton on immune function of gastric cancer rats based on the effect of HSP70on immune function.Methods Sixty healthy Sprague-Dawley(SD)rats were adaptively fed for one week.By random number table method,10 rats were randomly selected as a normal group(group A),and 12 rats were randomly selected to simulate the tumor transplantation process;after transplantation,10 rats were randomly selected as a sham operation group(group B).The remaining 38 rats were used to prepare a gastric cancer model by gastric transplantation of the Walker-256 tumor tissue;8 rats were randomly selected to verify the successful modeling after 7 d;the remaining 30 rats were randomly divided into a model group(group C),a moxibustion group(group D)and an infrared group(group E)by random number table method,with 10 rats in each group.From the first day of enrollment,the rats in the moxibustion group received mild moxibustion at Zhongwan(CV 12),Guanyuan(CV 4)and bilateral Zusanli(ST 36)(the first group)and bilateral Pishu(BL 20)and Weishu(BL 21)(the second group),the two groups of acupoints were alternated every other day.Rats in the infrared group received infrared radiation on the stomach and the area on the back between the T12-T13 spinous process,the two areas were alternated every other day.Rats in the moxibustion group and the infrared group were treated for 20 min each time,once a day for 21 d.Rats in the blank group,the sham operation group,and the model group were simultaneously grasped and fixed,and no other treatment was performed.After 21 d of intervention,the rats in each group were fasted for 12 h,and blood was collected from the orbits.The T cell apoptosis,T cell activation,T cell subsets CD3~+CD4~+,CD~+3CD8~+,CD4~+CD25~+,CD8~+CD28~+,CD8~+CD28~-ratio in peripheral blood were determined by flow cytometry.After sacrificing,thoracic cavity and abdominal cavity were dissected under aseptic condition.Thymus and spleen index were weighed and calculated.Tumor growth and metastasis were observed.Proliferation and apoptosis of tumor cells were measured by PCNA and TUNEL.HSP70 and HSP70 expression in gastric cancer tissues and serum were determined by immunohistochemistry,ELISA and RT-PCR.Cytokines expressed in gastric cancer tissues were screened by cytokine protein chip and verified by ELISA.Results 1)Compared with the model group,the survival status of rats in the moxibustion group was improved,and the survival status score increased significantly after 15 days of treatment(P<0.01);the growth of gastric tumors was inhibited(the volume growth inhibition rate was 41.89%),the invasion of cancer cells to gastric mucosa and abdominal organs was less;the proliferation of blood vessels in gastric cancer tissue was less,the proliferation of cancer cells was less and apoptosis was more(all P<0.01).Compared with the model group,the survival status of rats in the infrared group was slightly improved,the survival status score was slightly higher after 15 days of treatment(P<0.05),the growth of gastric tumors was slightly inhibited(volume growth inhibition rate was 28.09%),the cancer cells moved to various abdominal organs,a large number of blood vessels grew in gastric cancer tissue,less proliferation of cancer cells,and more apoptosis(all P<0.01).Compared with the infrared group,after 15 days of moxibustion treatment,the survival state score of rats in the moxibustion group increased more significantly(P<0.05),the proliferation of cancer cells in gastric tumors was less and the apoptosis was more(all P<0.05).2)Compared with the model group,the content of HSP70 in serum increased in the moxibustion group(P<0.01),the expression of HSP70 and HSP70 mRNA in gastric cancer tissues increased(all P<0.01),HSP70 was mostly expressed in extracellular interstitial space.Compared with model group,the content of HSP70 in serum of rats in infrared group increased significantly(P<0.01),the expression of HSP70 mRNA and the content of HSP70 in gastric cancer tissue increased(all P<0.05).Compared with the infrared group,the content of HSP70 and the expression of HSP70 mRNA in gastric cancer tissues of the moxibustion group were higher(all P<0.05),and most of HSP70 was in extracellular(P<0.05).3)Compared with the blank group,the index of thymus and spleen in the model group increased significantly(all P<0.01),the proportion of activated lymphocyte in peripheral blood increased,the proportion of apoptotic lymphocyte decreased(all P<0.05),the proportion of CD3~+,CD3~+CD8~+,CD8~+CD28~-T cells in peripheral blood increased(all P<0.01),the ratio of CD4~+CD25~+T cells increased slightly(all P<0.05),and the ratio of CD3~+CD4~+/CD3~+CD8~+T cells decreased significantly(P<0.01).Compared with the model group,the index of thymus and spleen in moxibustion group increased significantly(all P<0.01),the proportion of activated lymphocyte in peripheral blood increased,the proportion of apoptotic lymphocyte decreased(all P<0.05),the proportion of CD3~+,CD3~+CD4~+,CD8~+CD28~+T cells in peripheral blood increased significantly(all P<0.01),the ratio of CD3~+CD8~+T cells,CD3~+CD4~+/CD3~+CD8~+T cells increased(all P<0.05);the percentage of CD4~+CD25~+,CD8~+CD28~-T lymphocyte decreased(all P<0.05);the content of TNF-?and IFN-?increased(P<0.01,P<0.05),and the content of IL-6 and IL-22decreased(all P<0.05).Compared with the model group,the spleen index increased(P<0.05),the ratio of CD8~+CD28~+T cells in peripheral blood increased(P<0.05),the ratio of CD4~+CD25~+T cells decreased(P<0.01),the content of Th1 cytokine TNF-?increased(P<0.05),and the content of Th2 cytokine IL-6 decreased(P<0.05).Compared with the infrared group,the thymus index of the moxibustion group increased(P<0.05),the proportion of activated lymphocyte in peripheral blood increased,the proportion of apoptotic lymphocyte decreased(P<0.05),the proportion of CD3~+,CD3~+CD4~+T cells increased significantly(all P<0.01),the proportion of CD3~+CD8~+,CD8~+CD28~+T cells increased slightly(all P<0.05),and the content of Th1 cytokines TNF-?and IFN-?in gastric cancer tissue increased slightly(all P<0.05).Conclusion 1)Moxibustion can inhibit the growth and metastasis of gastric cancer in rats,reduce the proliferation of cancer cells and promote their apoptosis.2)Moxibustion can induce the synthesis and extracellular release of HSP70 in gastric cancer cells of rats.3)Moxibustion can enhance immune fuction by increasing the index of immune organs,activatting T cell and adjusting the distribution of it,adjusting the imbalance of Th1/Th2 cytokines and activate the immune function of gastric cancer rats. |
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