健脾消癌方对结直肠癌移植模型裸鼠NLRP3炎性小体表达及免疫功能的影响

Effects of Jianpi Xiaoai Formula on expression of NLRP3 inflammasomes and immunity in nude mice transplanted with colorectal cancer

目的:探讨健脾消癌方防治结直肠癌转移可能的作用机制。方法:随机抽取10只BALB/C-nu裸鼠作为空白组,其余裸鼠采用盲肠黏膜内注射HCT116细胞方法建立人结直肠癌裸鼠移植模型,建模3d后取存活裸鼠30只,随机分为模型组、健脾消癌方高、低剂量组。健脾消癌方高、低剂量组分别予健脾消癌方41.6、10.4g·kg-1·d-1灌胃;空白组与模型组以0.9%氯化钠溶液2mL/100g灌胃,连续给药4周,每日给药1次。Western Blot检测肝转移瘤组织NLRP3、Caspase-1蛋白表达量;流式细胞术检测血液NK细胞数量,CSFE/7-AAD法检测NK细胞的杀伤活性;比较各组裸鼠肝、肺、淋巴转移情况。结果:中药高、低剂量组肝、肺、淋巴结转移率均低于模型组(P<0.05),健脾消癌方高剂量组肝、肺转移率低于低剂量组(P<0.05)。与空白组比较,各组裸鼠肝转移瘤组织NLRP3、Caspase-1蛋白表达均下降(P<0.05),NK细胞数量及活性均下降;与模型组比较,健脾消癌方高、低剂量组肝转移瘤组织NLRP3、Caspase-1蛋白表达均有升高(P<0.05,P<0.01),NK细胞数量及活性均有升高(P<0.05,P<0.01)。结论:NLRP3炎性小体表达下调可能参与结直肠癌病情进展,健脾消癌方可拮抗结直肠癌转移,提高结直肠癌移植模型裸鼠血液NK细胞数量及活性,其作用机制可能与升高NLRP3、Caspase-1蛋白表达有关。

Objective:To explore the possible mechanism of Jianpi Xiaoai Formula in preventing and treating metastasis of colorectal cancer.Methods:Ten BALB/C-nu nude mice were randomly selected as the control group.The remaining mice were used to establish the model of uude mice transplanted with colorectal cancer.Three days after modeling,30 survived nude mice were randomly collected and divided into model group,high and low dose group of Jianpi Xiaoai Formula.The high and low dose groups of Jianpi Xiao Formula were given Jianpi Xiao Formula 41.6,10.4 g·kg-1·d-1 respectively,while the blank group and the model group were given 2 mL/100 g saline orally for 4 weeks,once a day.The protein expressions of NLRP3 and Caspase-1 in liver metastasis tumor tissues were evaluated by Western Blot,flow cytometry was used to detect the number of NK cells in blood,CSFE/7-AAD was used to detect the killing activity of NK cells,liver,lung and lymphatic metastasis of nude mice in each group were compared.Results:The metastasis rate of liver,lung and lymph node in high and low dose group of Jianpi Xiao Formula were lower than that in model group(P<0.05),while that in high dose group was lower than that in low dose group(P<0.05).Compared with the blank group,the expression of NLRP3 and Caspase-1 protein in liver metastasis tumor tissues in each group significantly decreased(P<0.05),and the number and activity of NK cells decreased.Compared with the model group,the expression of NLRP3 and Caspase-1 protein in liver metastasis tumor tissues in the high and low dose group of Jianpi Xiao Formula significantly increased(P<0.05,P<0.01),and the number and activity of NK cells in each group significantly increased(P<0.05,P<0.01).Conclusion:The down-regulation of NLRP3 expression may be involved in the progress of colorectal cancer.Jianpi Xiaoai Formula can antagonize the metastasis of colorectal cancer and increase the number and activity of NK cells in nude mice transplanted with colorectal cancer.The mechanism may be related to the increase of NLRP3 and Caspase-1 protein expression.