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[麦粒灸通过调节凋亡因子半胱氨酸家族增强环磷酰胺对Hepa1-6荷瘤小鼠的抗肿瘤作用]

[Moxibustion with seed-size moxa cone enhances anti-tumor effect of cyclophosphamide on Hepa1-6 hepatoma-bearing mice by regulating apoptosis factor cysteine family].

作者信息

Cheng Yan-Ting, Zhu Tao, Ma Yan-Zhu, Ding Yan-Ze, Meng Li-Qiang, Xue Ling, Ji Lai-Xi

机构信息

The Second Clinical Medicine College, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China.

出版信息

Zhen Ci Yan Jiu. 2023 Sep 25;48(9):914-22. doi: 10.13702/j.1000-0607.20220820.

Abstract

OBJECTIVE

To observe the anti-tumor effect of moxibustion with seed-sized moxa cones on Hepa1-6 liver cancer bearing (HLCB) mice and its regulatory mechanism on cell apoptosis.

METHODS

A total of 40 male C57BL/6 mice were randomly divided into control, moxibustion, cyclophosphamide (CTX) and moxibustion+CTX groups, with 10 mice in each group. The HLCB model was established by subcutaneous inoculation of Hepa1-6 cancer cells into the right armpit. Mice of the CTX and moxibustion+CTX groups were given intraperitoneal injection of CTX (30 mg/kg), once daily for 3 days. Moxibustion with seed-sized moxa cones were applied to "Dazhui" (GV14), bilateral"Zusanli" (ST36) and "Sanyinjiao" (SP6), with 5 moxa cones for each acupoint, once daily for 10 consecutive days. The survival status scores and body weight of HLCB mice were observed, and the tumor weight and tumor inhibition rate were detected. HE staining was used to observe the morphological changes of tumor tissue. ELISA was used to detect the levels of serum interleukin (IL)-2, IL-4 and tumor necrosis factor-α (TNF-α). Western blot and fluorescent quantitative real-time PCR were used to detect the protein and mRNA expressions of cysteine aspartate protease (Caspase) -3 and Caspase-9 in tumor tissues, separately.

RESULTS

Compared with the control group, the survival status scores, body weight, serum IL-2 and TNF-α levels were significantly increased (0.05, 0.01), the tumor weight and serum IL-4 levels were significantly decreased (0.05) in the moxibustion group;while the survival status, body weight, tumor weight, serum IL-2 and IL-4 levels were significantly decreased (0.01, 0.05), the content of TNF-α was significantly increased (0.01) in the CTX group. The protein and mRNA expressions of Caspase-3 and Caspase-9 in the 3 trentment groups were significantly increased (0.05, 0.01). In comparison with the moxibustion group, the survival status scores, body weight and tumor weight, serum content of IL-2 were significantly decreased (0.01, 0.05). In contrast to the CTX group, the survival status scores, body weight, serum IL-2 and TNF-α content, and the expressions of Caspase-3 and Caspase-9 in tumor tissue were significantly increased (0.01, 0.05), and the tumor weight and serum IL-4 content were significantly decreased (0.05) in the moxibustion+CTX group. Results of HE staining showed that the tumor cells in the control group had clear nuclear membranes and nucleoli, with more dividing cells; while less nuclear division and an increase in tumor necrosis areas were found in the 3 treatment groups.

CONCLUSION

Moxibustion with seed-size moxa cone can enhance the anti-tumor effect of CTX and improve the quality of life of HLCB mice, which may be related with its effect in activating the expressions of Caspase-3 and Caspase-9 in tumor tissue.

摘要

目的

观察麦粒灸对荷Hepa1-6肝癌小鼠的抑瘤作用及其对细胞凋亡的调控机制。

方法

将40只雄性C57BL/6小鼠随机分为对照组、艾灸组、环磷酰胺(CTX)组和艾灸+CTX组,每组10只。通过将Hepa1-6癌细胞接种于右侧腋窝皮下建立荷Hepa1-6肝癌模型。CTX组和艾灸+CTX组小鼠腹腔注射CTX(30 mg/kg),每天1次,连续3天。采用麦粒灸对“大椎”(GV14)、双侧“足三里”(ST36)和“三阴交”(SP6)进行艾灸,每个穴位灸5壮,每天1次,连续10天。观察荷Hepa1-6肝癌小鼠的生存状态评分和体重,检测肿瘤重量和抑瘤率。采用HE染色观察肿瘤组织的形态学变化。采用ELISA法检测血清白细胞介素(IL)-2、IL-4和肿瘤坏死因子-α(TNF-α)水平。分别采用Western blot和荧光定量实时PCR法检测肿瘤组织中半胱氨酸天冬氨酸蛋白酶(Caspase)-3和Caspase-9的蛋白和mRNA表达。

结果

与对照组比较,艾灸组小鼠生存状态评分、体重、血清IL-2和TNF-α水平显著升高(P<0.05,P<0.01),肿瘤重量和血清IL-4水平显著降低(P<0.05);CTX组小鼠生存状态、体重、肿瘤重量、血清IL-2和IL-4水平显著降低(P<0.01,P<0.05),TNF-α含量显著升高(P<0.01)。3个治疗组Caspase-3和Caspase-9的蛋白和mRNA表达均显著升高(P<0.05,P<0.01)。与艾灸组比较,艾灸+CTX组小鼠生存状态评分、体重和肿瘤重量、血清IL-2含量显著降低(P<0.01,P<0.05)。与CTX组比较,艾灸+CTX组小鼠生存状态评分、体重、血清IL-2和TNF-α含量以及肿瘤组织中Caspase-3和Caspase-9的表达显著升高(P<0.01,P<0.05),肿瘤重量和血清IL-4含量显著降低(P<0.05)。HE染色结果显示,对照组肿瘤细胞核膜和核仁清晰,分裂细胞较多;3个治疗组核分裂减少,肿瘤坏死面积增加。

结论

麦粒灸可增强CTX的抑瘤作用,提高荷Hepa1-6肝癌小鼠的生活质量,其机制可能与激活肿瘤组织中Caspase-3和Caspase-9的表达有关。

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