龟甲水提液调控NF-<italic style="font-style: italic">κ</italic>B炎症微环境对MC3T3-E1成骨分化的影响

张玉卓; 尚奇; 沈耿杨; 任辉; 敖海清; 江晓兵; 周本根

doi:10.13422/j.cnki.syfjx.20200838

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龟甲水提液调控NF-κB炎症微环境对MC3T3-E1成骨分化的影响

药理 | 更新时间：2021-02-09

- 龟甲水提液调控NF-κB炎症微环境对MC3T3-E1成骨分化的影响
- Effect of Testudinis Carapax et Plastrum Aqueous Extract Regulating NF-κB Inflammation Microenvironment on Osteogenic Differentiation of MC3T3-E1
- 中国实验方剂学杂志 2020年26卷第8期页码：89-94
- 作者机构：
  
  1.广州中医药大学，广州　 510006
  2.广州中医药大学　第一附属医院，广州　 510405
- 作者简介：
  
  [第一作者]　张玉卓，在读博士，从事藏象理论与中医药防治疾病研究，E-mail：852649371@qq.com
  *江晓兵，博士，副教授，从事中医药防治骨质疏松脊柱伤病研究，E-mail：spinedrjxb@163.com
  *周本根，博士，副教授，从事中医药防治骨质疏松脊柱伤病研究，E-mail：zbg810@sina.com；
- 基金信息：
  
  国家自然科学基金项目(81904225);广东省普通高校特色创新项目(2018KTSCX041);广东省自然科学基金博士科研启动项目(2018A030310615);广东省医学科研基金项目(A2019167)
- DOI：10.13422/j.cnki.syfjx.20200838
  中图分类号： R2-0; R289; R285.5
- 收稿日期：2019-09-14，
  
  网络出版日期：2020-01-07，
  
  纸质出版日期：2020-04-20
- 稿件说明：
移动端阅览
张玉卓, 尚奇, 沈耿杨, 等. 龟甲水提液调控NF-κB炎症微环境对MC3T3-E1成骨分化的影响[J]. 中国实验方剂学杂志, 2020,26(8):89-94.

Yu-zhuo ZHANG, Qi SHANG, Geng-yang SHEN, et al. Effect of Testudinis Carapax et Plastrum Aqueous Extract Regulating NF-κB Inflammation Microenvironment on Osteogenic Differentiation of MC3T3-E1[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(8): 89-94.
张玉卓, 尚奇, 沈耿杨, 等. 龟甲水提液调控NF-κB炎症微环境对MC3T3-E1成骨分化的影响[J]. 中国实验方剂学杂志, 2020,26(8):89-94. DOI： 10.13422/j.cnki.syfjx.20200838.

Yu-zhuo ZHANG, Qi SHANG, Geng-yang SHEN, et al. Effect of Testudinis Carapax et Plastrum Aqueous Extract Regulating NF-κB Inflammation Microenvironment on Osteogenic Differentiation of MC3T3-E1[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(8): 89-94. DOI： 10.13422/j.cnki.syfjx.20200838.

摘要

目的：

探讨龟甲水提液调控核转录因子-

B(NF-

B)炎症微环境促小鼠前成骨细胞系(MC3T3-E1)成骨分化的作用及机制。

方法：

体外培养MC3T3-E1细胞，并进行成骨诱导，龟甲水提液干预。实验分为空白组、成骨诱导组、龟甲水提液(20 mg·L

-1

)联合成骨诱导组。细胞增殖毒性检测试剂盒(CCK-8)法检测MC3T3-E1增殖能力，并确定最佳干预浓度；碱性磷酸酶(ALP)染色和茜素红(ARS)染色检测MC3T3-E1成骨分化能力；实时荧光定量PCR(Real-time PCR)检测NF-

B p65，NF-

B p105，白细胞介素-6(IL-6)，ALP和Ⅰ型胶原(COL-Ⅰ) mRNA的表达。

结果：

CCK-8结果显示，随着时间的增长，MC3T3-E1增殖虽无统计学差异，但呈上升趋势，而在20 mg·L

-1

质量浓度时增殖较其他组明显；ALP和ARS染色结果显示，成骨诱导组和龟甲水提液联合成骨诱导组染色阳性率较空白组明显升高；Real-time PCR结果显示，龟甲干预第7天，与空白组比较，龟甲水提液联合成骨诱导组NF-

B p105和IL-6 mRNA的表达明显降低(

0.01)，ALP和COL-Ⅰ mRNA的表达明显升高(

0.05)；龟甲干预第14天，与空白组比较，龟甲水提液联合成骨诱导组NF-

B p65，NF-

B p105和IL-6 mRNA的表达明显降低(

0.01)，ALP和COL-Ⅰ mRNA的表达明显升高(

0.05，

0.01)。

结论：

龟甲水提液能够促进MC3T3-E1成骨分化，其机制可能与抑制NF-

B炎性微环境有关。

Abstract

Objective:

To investigate the role and mechanism of Testudinis Carapax et Plastrum aqueous extract in promoting osteogenic differentiation of mouse preosteoblast cell line(MC3T3-E1) by regulating nuclear transcription factor-

B(NF-

B) inflammation microenvironment.

Method:

MC3T3-E1 cells were cultured

in vitro

and osteogenic induction (OI) was performed. Testudinis Carapax et Plastrum was prepared and treated the cells. Cells were devided into control group

osteogenic induction group and Testudinis Carapax et Plastrum (20 mg·L

-1

)with osteogenic induction group. The proliferation of MC3T3-E1 was detected by cell counting kit-8 (CCK-8)

and the optimum concentration of intervention was determined. MC3T3-E1 differentiation and osteogenic mineralization were assayed using alkaline phosphatase (ALP) and Alizarin red staining (ARS)

respectively. The expressions of NF-

B p65

NF-

B p105

interleukin-6(IL-6)

ALP and Collagen-Ⅰ(COL-Ⅰ) mRNA were detected by Real-time PCR.

Result:

The results of CCK-8 showed that the proliferation of MC3T3-E1 did not change statistically with time

but it showed an upward trend

while the proliferation at 20 mg·L

-1

was more obvious than other groups. The ALP and ARS showed that the positive staining rate of osteogenic induction group and Testudinis Carapax et Plastrum with osteogenic induction group were higher than control group.Real-time PCR results showed that on the 7

day in culture

the expression of NF-

B p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (

0.01)

and the expression of ALP and COL-Ⅰ mRNA was significantly upregulated(

0.05)

on the 14

day

the expression of NF-

B p65

NF-

B p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (

0.01). The expression of ALP and COL-Ⅰ mRNA was significantly increased (

0.05

0.01).

Conclusion:

Testudinis Carapax et Plastrum aqueous extract can promote osteogenic differentiation of MC3T3-E1 via a mechanism associated with the regulation of inhibition of NF-

B inflammatory microenvironment.

关键词

Keywords

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龟甲水提液调控NF-κB炎症微环境对MC3T3-E1成骨分化的影响

Effect of Testudinis Carapax et Plastrum Aqueous Extract Regulating NF-κB Inflammation Microenvironment on Osteogenic Differentiation of MC3T3-E1

DOI：10.13422/j.cnki.syfjx.20200838

摘要

Abstract

关键词

Keywords

references