Effect of Xiao Chaihutang and Its Cold and Hot Herb-subtracted Formulae on Human Colon Carcinoma Cell Proliferation, Lipid Accumulation and Alkaline Phosphatase

XU Bin; LIANG Xiao-feng; LU Jia; LIU Xiao-qiu

doi:10.13422/j.cnki.syfjx.20181209

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Effect of Xiao Chaihutang and Its Cold and Hot Herb-subtracted Formulae on Human Colon Carcinoma Cell Proliferation, Lipid Accumulation and Alkaline Phosphatase

更新时间：2024-01-04

- Effect of Xiao Chaihutang and Its Cold and Hot Herb-subtracted Formulae on Human Colon Carcinoma Cell Proliferation, Lipid Accumulation and Alkaline Phosphatase
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 14, Pages: 134-141(2018)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20181209
  CLC： R285.5
- Published：2018
- 稿件说明：
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XU Bin, LIANG Xiao-feng, LU Jia, et al. Effect of Xiao Chaihutang and Its Cold and Hot Herb-subtracted Formulae on Human Colon Carcinoma Cell Proliferation, Lipid Accumulation and Alkaline Phosphatase[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(14): 134-141.
DOI：

XU Bin, LIANG Xiao-feng, LU Jia, et al. Effect of Xiao Chaihutang and Its Cold and Hot Herb-subtracted Formulae on Human Colon Carcinoma Cell Proliferation, Lipid Accumulation and Alkaline Phosphatase[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(14): 134-141. DOI： 10.13422/j.cnki.syfjx.20181209.

摘要

目的：观察小柴胡汤及其寒热减方对HCT116结肠癌细胞增殖、脂肪及碱性磷酸酶的影响，探讨其作用机制。方法：分别取小柴胡汤、小柴胡汤去柴胡、小柴胡汤去黄芩、小柴胡汤去半夏、小柴胡汤去生姜煎液不同质量浓度(0.5，1.0，1.5 g·L-1)培养HCT116细胞，另设不加药空白组和秋水仙碱(1.0，2.5，10.0 μmol·L-1)组。采用噻唑蓝(MTT)比色法检测HCT116细胞的增殖活力，苏丹红Ⅳ染色法检测脂肪，碱性磷酸酶试剂盒检测酶活性，热稳定实验区分组织特异性碱性磷酸酶和组织非特异性碱性磷酸酶(TNAP)，肠型碱性磷酸酶特异抑制剂(L-苯丙氨酸)抑制实验，区分组织特异性碱性磷酸酶中的肠碱性磷酸酶(IAP)。结果：与空白组比较，小柴胡汤及其寒热减方对HCT116结肠癌细胞增殖均有不同程度的抑制作用，小柴胡汤的抑制作用更明显，秋水仙碱组细胞抑制作用最为明显。空白组脂肪累积细胞率显著高于小柴胡汤及其寒热减方5组(P<0.01)，小柴胡汤组脂肪累积细胞率最低(P<0.05)，小柴胡汤去柴胡组脂肪累积细胞率显著低于小柴胡汤去半夏组和小柴胡汤去生姜组(P<0.01)，小柴胡汤去黄芩组脂肪累积细胞率显著低于小柴胡汤去半夏组和小柴胡汤去生姜组(P<0.01)，秋水仙碱组无脂肪累积。小柴胡汤及其寒热减方5组细胞内IAP酶活性高于TNAP活性(P<0.01)；培养液中TNAP酶活性高于IAP酶活性(P<0.01)。小柴胡汤去柴胡组细胞内IAP酶活性高于小柴胡汤去半夏组和小柴胡汤去生姜组，小柴胡汤去黄芩组细胞内IAP酶活性高于小柴胡汤去半夏组和小柴胡汤去生姜组(P<0.01)；在培养液中，小柴胡汤去半夏组和小柴胡汤去生姜组TNAP酶活性均高于小柴胡汤去柴胡组(P<0.05)和小柴胡汤去黄芩组。结论：小柴胡汤及其寒热减方均抑制HCT116癌细胞增殖，小柴胡汤寒热并用全方抑制作用最强。寒药减方与热药减方对脂肪累积和碱性磷酸酶的影响不同。小柴胡汤中寒药和热药并用，分别作用于TNAP和IAP，调节脂肪转运代谢，从而抑制癌细胞生长。

Abstract

Objective: To explore the effect of Xiao Chaihutang (XCHT) and its cold and hot herb-subtracted formulae on HCT116 human colon carcinoma cell proliferation

lipid accumulation and alkaline phosphatase

and investigate its mechanism. Method: HCT116 cells were cultured with XCHT

XCHT-Bupleuri Radix

XCHT-Scutellariae Radix

XCHT-Pinelliae Rhizoma

and XCHT-Zingiberis Rhizoma Recens at final drug concentrations of 0.5

1.0

1.5 g·L-1

and in addition

another blank control group and colchicine positive control groups (1.0

2.5

10.0 μmol·L-1) were set in this study. The proliferation viability of HCT116 cells was detected by 3-(4

5-dimethylthiazol-2-yl) -2

5-diphenyltetrazolium bromide (MTT) assay; the lipid accumulation was detected by Sudan red Ⅳ staining;alkaline phosphatase kit was used to detect the enzyme activity; heat stable test was used to distinguish tissue specific alkaline phosphatase and tissue non-specific alkaline phosphatase (TNAP)

and intestinal alkaline phosphatase specific inhibitor (L-phenylalanine) inhibition test was used to distinguish intestinal alkaline phosphatase (IAP) in tissue specific alkaline phosphatase. Result: As compared with blank control group

XCHT and its cold and hot herb-subtracted formulae all had obvious inhibitory effect on the cell proliferation activity of HCT116 human colon carcinoma cells and the inhibitory effect of XCHT was more obvious. The inhibitory effect was most obvious in colchicine positive control groups. The cell percentage of lipid accumulation in blank control group was significantly higher than that in XCHT and its cold and hot herb-subtracted formulae groups (P<0.01)

and the cell percentage of lipid accumulation was lowest in XCHT group (P<0.05). The cell percentage of lipid accumulation in XCHT-Bupleuri Radix group was significantly lower than that in XCHT-Pinelliae Rhizoma group and XCHT-Zingiberis Rhizoma Recens group (P<0.01). The cell percentage of lipid accumulation in XCHT-Scutellariae Radix was significantly lower than that in XCHT-Pinelliae Rhizoma group and XCHT-Zingiberis Rhizoma Recens group (P<0.01)

and there was no lipid accumulation in the colchicine positive control groups.The IAP activity was higher than TNAP activity in all the five XCHT and its cold and hot herb-subtracted formulae groups (P<0.01)

and the TNAP activity was higher than IAP activity in culture medium (P<0.01). The IAP activity in cells of XCHT-Bupleuri Radix group was higher than that in XCHT-Pinelliae Rhizoma group and XCHT-Zingiberis Rhizoma Recens group

and the IAP activity in XCHT-Scutellariae Radix group was higher than that in XCHT-Pinelliae Rhizoma group and XCHT-Zingiberis Rhizoma Recens group (P<0.01). In the culture medium

the TNAP activity in XCHT-Pinelliae Rhizoma group and XCHT-Zingiberis Rhizoma Recens group was higher than that in XCHT-Bupleuri Radix group (P<0.05) and XCHT-Scutellariae Radix. Conclusion: XCHT and its cold and hot herb-subtracted formulae can effectively inhibit HCT116 human colon carcinoma cell proliferation and XCHT (a combination use of cold and hot herbs) exhibited the strongest inhibitory effect. There was a significant difference between the effect of cold herb-subtracted formulae and hot herb-subtracted formulae on lipid accumulation and alkaline phosphatase. A combination use of cold and hot herbs in XCHT

could act on TNAP and IAP respectively

and regulate fat transport and metabolism

thus inhibiting the growth of cancer cells.

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