健脾化瘀方促进小鼠肝癌休眠的作用及其免疫机制研究

JIANPI HUAYU FORMULA ON THE PROMOTION OF TUMOR DORMANCY AND THE IMMUNE MECHANISM IN MICE WITH HEPATIC CARCINOMA

目的通过建立小鼠肝癌休眠模型,并予健脾化瘀法干预,研究其促进肿瘤休眠的作用及其免疫机制。方法建立小鼠肝癌休眠模型。并随机分为4组:空白对照组、化疗对照组、中药低剂量组、中药高剂量组,分别给予生理盐水、卡莫氟、中剂量健脾化瘀方及高剂量健脾化瘀方进行干预。处死小鼠后,接种部位取材,免疫组化染色检测Ki-67、TGF-β、IL-10等相关指标的表达。结果 1肿瘤组织休眠情况:在反复外伤刺激下,空白对照组休眠率为0。化疗对照组休眠率为80.0%。中药低剂量组和高剂量组休眠率均为86.7%;2对Ki-67增殖指数的影响:化疗对照组、中药低剂量组、中药高剂量组与空白对照组两两比较,其Ki-67增殖指数均低于空白对照组(p<0.05);中药低剂量组、中药高剂量组与化疗对照组两两比较,均低于化疗对照组(p<0.05);而中药低剂量组及高剂量组间两两比较无明显差异(p>0.05);3对TGF-β表达水平的影响:化疗对照组、中药低剂量组、中药高剂量组与空白对照组两两比较,空白对照组TGF-β表达水平明显高于前三组(p<0.05);但化疗对照组、中药低剂量组与中药高剂量治疗组间两两比较,TGF-β表达水平无明显差异(p>0.05);4对IL-10表达水平的影响:化疗对照组、中药低剂量组、中药高剂量组与空白对照组两两比较,空白对照组IL-10表达水平明显高于前三组(p<0.05),但化疗对照组、中药低剂量治疗组及高剂量治疗组间两两比较,IL-10表达水平无明显差异(p>0.05)。结论健脾化瘀方有促进小鼠肝癌休眠的作用,其作用机制可能与其通过调节免疫抑制肿瘤细胞的TGF-β、IL-10表达有关。

Objective To study the effects of Jianpi Huayu formula on the promotion of tumor dormancy and the immune mechanism in mice with hepatic carcinoma by building tumor dormancy model of hepatoma in mouse in- tervened with Jianpi Huayu formula. Methods A tumor dormancy model of hepatoma in mouse was established. Mice were randomly divided into four groups： blank control group, chemotherapy control group, low -dosage Tradi- tional Chinese medicine （TCM） group, high- dosage TCM group, intervened with normal saline, carmofur, middle- dosage Jianpi Huayu formula and high - dosage Jianpi Huayu formula respectively. After the mice were sacrificed, immunohistocbemical staining was performed in the vaccinated parts extracted from mice to detect the expression of relevant indicators including ki - 67, TGF - β and IL - 10. Results ①Tumor dormancy situations ： After repeated traumatic stimulation, tumor dormancy rates of blank control group and chemotherapy control group were 0 and 80.0 % respectively. The tumor dormancy rates of low - dosage and high - dosage TCM groups were both 86.7%. ②Effects on Ki - 67 proliferation index： Ki - 67 proliferation indexes of chemotherapy control group, low - dosage and high - dos- age TCM groups were lower than that of blank control group （p 〈 0. 05 ）. Ki -67 proliferation indexes of low -dosage and high -dosage TCM groups were lower than that of chemotherapy control group （p 〈 0.05 ）. There was no signifi- cant difference between Ki - 67 proliferation indexes of low - dosage and high - dosage TCM groups （p 〉 0.05 ）. ③ Effects on expressions of TGF - β ： The expressions of TGF - β in chemotherapy control group, low - dosage and high -dosage TCM groups were significantly lower than that in blank control group （p 〈 O. 05 ）. There was no signifi-cant difference in expressions of TGF - β between chemotherapy control group, low - dosage and high - dosage TCM groups （p 〉 0.05 ）. ④ Effects on the expressions of IL - 10 ： The expressions of IL - 10 in chemotherapy control group, low - dosage and high - dosage TCM groups were significantly lower than that in blank control group （p 〈 0. 05）. There was no significant difference in expressions of IL- 10 between chemotherapy control group, low- dos- age and high - dosage TCM groups （p 〉 0.05）. Conclusion Jianpi Huayu formula plays a role in promoting tumor dormancy in mice with hepatic carcinoma by adjusting the immunity and suppressing the expressions of TGF -β, IL - 10 in tumor cells.