麦粒灸对荷瘤小鼠肿瘤组织中Bax和Bcl-2表达的影响

Effect of Moxibustion with Seed-Sized Moxa Cone on Expressions of Bax and Bcl-2 in Tumor Tissues of Hepa1-6 Tumor-Bearing Mice

目的:观察麦粒灸对荷瘤小鼠肿瘤组织中B细胞淋巴相关X蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)表达水平的影响,从肿瘤细胞凋亡角度探讨麦粒灸治疗肿瘤的作用机制。方法:将40只C57BL/6小鼠皮下接种Hepa1-6肿瘤细胞,建立荷瘤小鼠模型,随机分为对照组、麦粒灸组、环磷酰胺(CTX)组和麦粒灸+CTX组,每组10只。对照组给予腹腔注射生理盐水0.2 mL/d,1次/d,连续3 d;CTX组、麦粒灸+CTX组按荷瘤小鼠体质量给予腹腔注射CTX 30 mg/kg,1次/d,连续3 d;麦粒灸组、麦粒灸+CTX组给予麦粒灸“足三里”(双侧)、“三阴交”(双侧)和“大椎”,每穴5壮,1次/d,连续10 d。每日观察并记录小鼠干预前后的一般情况;检测瘤体质量及抑瘤率;HE染色法观察肿瘤形态变化;实时荧光定量PCR法和Western blot法检测肿瘤组织中Bax、Bcl-2 mRNA及蛋白表达水平。结果:经治疗后麦粒灸组、CTX组和麦粒灸+CTX组抑瘤率分别为23.9%、45.9%和67.1%。与对照组比较,麦粒灸组、CTX组与麦粒灸+CTX组肿瘤组织Bax mRNA及蛋白表达水平均升高,差异具有统计学意义(P<0.01或P<0.05),Bcl-2 mRNA及蛋白表达水平均降低,差异具有统计学意义(P<0.01或P<0.05),麦粒灸组肿瘤坏死区明显增多;与CTX组比较,麦粒灸+CTX组肿瘤组织Bax mRNA及蛋白表达水平显著升高,差异具有统计学意义(均P<0.05),Bcl-2 mRNA及蛋白表达水平显著降低,差异具有统计学意义(均P<0.05),肿瘤细胞核分裂明显减少。结论:麦粒灸能够明显减轻Heap1-6荷瘤小鼠的瘤体质量,对肿瘤的生长有抑制作用;与CTX合用,抑瘤作用更有效,其机制可能与调节Bax、Bcl-2的表达水平有关。

Objective:To observe the effects of moxibustion with seed-sized moxa cone on the expressions of Bax and Bcl-2 in tumor tissues of Hepa1-6 tumor-bearing mice,and to explore its action mechanism in treating tumors from the perspective of tumor cell apoptosis.Methods:A total of 40 C57BL/6 mice were subcutaneously inoculated with Hepa1-6 tumor cells to establish tumor-bearing model,and were randomly divided into the control group,the moxibustion group,the cyclophosphamide(CTX)group,and the moxibustion plus CTX group,with 10 mice in each group.The control group was given intraperitoneal injection of normal saline(0.2 mL/d,once a day for 3 days).The CTX group and the moxibustion plus CTX group were intraperitoneally injected CTX(30 mg/kg,once a day for 3 days).The moxibustion group and the moxibustion plus CTX group were given moxibustion with seed-sized moxa cone at bilateral Zusanli(ST36),bilateral Sanyinjiao(SP6)and Dazhui(DU14),5 cones each acupoint,once a day for 10 days.The changes of mice before and after intervention were observed and recorded daily,the tumor mass and the tumor inhibition rate were detected,the morphological changes of tumor were observed by HE staining,and the mRNA and protein expressions of Bax and Bcl-2 in tumor tissues were detected by real-time fluorescence quantitative PCR and Western blot.Results:After the treatment,the tumor inhibition rate was 23.9%in the moxibustion group,45.9%in the CTX group and 67.1%in the moxibustion plus CTX group,respectively.Compared with those in the control group,the mRNA and protein expressions of Bax were significantly increased(P<0.01,P<0.05),and the mRNA and protein expressions of Bcl-2 were significantly decreased in the moxibustion group,the CTX group and the moxibustion plus CTX group(P<0.01,P<0.05);the area of tumor necrosis increased significantly in the moxibustion group.Compared with those in the CTX group,the mRNA and protein expressions of BAX were significantly increased(P<0.05),and the mRNA and protein expressions of Bcl-2 were significantly decreased in the moxibustion plus CTX group(P<0.05);tumor nuclear division was decreased obviously.Conclusion:Moxibustion with seed-sized moxa cone can significantly reduce the tumor mass of Hepa1-6 tumor-bearing mice and inhibit tumor growth.Combined with CTX,the tumor inhibition effect was more significant,and the mechanism may be related to regulating the expressions of Bax and Bcl-2.