摘要
Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70(HSP70)in gastric cancer-bearing rats.Methods:A total of 40 healthy Sprague-Dawley(SD)rats were adaptively fed for one week.The gastric cancer model was prepared by Walker-256 cancer tissue transplantation.After 7 d,10 rats were randomly selected to verify the successful modeling,and the remaining 30 rats were divided into a model group,a moxibustion group and an infrared group by the random number table method,with 10 rats in each group.After enrollment,the moxibustion group received suspended moxibustion at Zhongwan(CV 12),Guan yuan(CV 4)and bilateral Zusanli(ST 36),(the first group of acupoints)on the 1st day,and suspended moxibustion at bilateral Pishu(BL 20)and Weishu(BL 21),(the second group of acupoints)on the 2nd day,20 min each time,once a day.Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d.From the day of en roll me nt,rats in the infrared group were irradiated with the infrared radiatio n at the stomach area on the 1st day,and at the T12-Ti3 interspinous region on the 2nd day,20 min each time,once a day,and the two locations were alternately irradiated every other day for 21 d.During the treatment,rats in the model group were intervened by grasping and fixation without treatment.At the end of the treatment,blood was collected from the inner eye orbit,and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay(ELISA).Rats were sacrificed,the tumor volume and growth inhibition rate were measured.The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase(SP);HSP70 protein expression was determined by ELISA;HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction(R「PCR)assay.Results:In comparison of the model group,the volume growth of the gastric cancer in the moxibustion group was significantly restricted(P<0.01);the volume growth inhibition rate in the moxibustion group was 37.93%;the HSP70 expressi on in peripheral blood and the cancer tissues was sign ifica ntly in creased(both P<0.01);the expressio n of HSP70 mRNA and HSP70 content in gastric tumor were both obviously in creased in the moxibustion group(P<0.01);and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group.In comparison of the model group,the volume growth of the gastric cancer in the infrared group was slightly restricted(P<0.05)with a volume growth inhibition rate of 15.89%;the HSP70 expression in the infrared group was increased significantly in peripheral blood(P<0.01)and in the gastric cancer tissues(P<0.05);more HSP70 was released outside of the cancer cells in the infrared group.In comparison of the infrared group,the volume growth of gastric cancer was more restricted in the moxibustion group(P<0.05)z and the HSP70 expression in the gastric cancer tissues was also higher(P<0.05);more HSP70 was released outside of the cancer cells in the moxibustion group.Conclusion:Moxibusti on and in fra red treatme nt inhibit the gastric can cer growth in the gastric cancer-bearing rats,up-regulate the HSP70 expression in gastric cancer tissues,and promote the production and extracellular release of HSP70,and the effect of moxibustion is more obvious.
目的:观察艾灸对胃荷瘤大鼠瘤体内热休克蛋白70(HSP70)mRNA及其蛋白表达的影响。方法:四十只健康Sprague-Dawley(SD)大鼠适应性喂养一周后,采用Walker-256瘤组织胃部移植制备胃荷瘤模型,7 d后随机选取10只验证造模成功,剩余30只采用随机数字表法分入模型组、艾灸组和红外组,每组10只。自入组当日起,艾灸组第1日悬灸中腕、关元、双侧足三里(第一组穴位),第2日悬灸双侧脾俞和胃俞(第二组穴位),每次施灸20 min,每日1次,两组穴位隔日交替施灸,连续21 do自入组当日起,红外组第1日用红外线照射胃腕部,第2日照射背部T12-T13棘突间区域,每次照射20 min,每日1次,两个部位隔日交替照射,连续21 d。治疗期间,模型组动物陪同抓取、固定,不予其他处理。治疗结束后,眶内采血,采用酶联免疫吸附试验(ELISA)测定外周血中HSP70含量。处死动物,测算肿瘤体积及生长抑制率。采用链霉菌抗生物素蛋白-过氧化物酶连结实验(S・P)观测胃瘤体中HSP70的表达位置及变化,ELISA测定HSP70含量,逆转录-聚合酶链反应(RT-PCR)测定瘤体内HSP70 mRNA表达水平。结果:与模型组比较,艾灸组胃部瘤体体积增长明显受限(PV0.01),体积增长抑制率为37.93%,外周血中HSP70含量明显增加(P<0.01)/胃部瘤体内HSP70 mRNA表达及HSP70含量均明显增加(均PV0.01),大量HSP70释放到癌细胞外;与模型组比较,红外组胃部瘤体体积增长稍受限(PV0.05),增长抑制率为15.89%,外周血中HSP70含量明显增加(PV0.01),胃部瘤体内HSP70 mRNA表达及HSP70含量稍增加(均PV0.05),较多HSP70释放到癌细胞外;与红外组比较,艾灸组胃部瘤体体积增长受限更明显(PV0.05),胃部瘤体内HSP70 mRNA的表达量及HSP70含量更高(均PV0.05),释放到癌细胞外HSP70也更多。结论:艾灸及红外线治疗均能一定程度抑制胃荷瘤大鼠的瘤体增长,上调胃部瘤体内HSP70 mRNA的表达,促进HSP70的生成及细胞外释放,且艾灸的作用更明显。
基金
国家自然科学基金项目,No.81574077
湖南省教育厅优秀青年项目,NO.18B257。
