[1]赵东方,尉志强,邢姝琴,等.巴戟天对骨关节炎小鼠软骨细胞自噬、凋亡的影响及机制[J].新乡医学院学报,2023,40(11):1001-1007.[doi:10.7683/xxyxyxb.2023.11.001]
　ZHAO Dongfang,WEI Zhiqiang,XING Shuqin,et al.Effect and mechanism of morinda root on autophagy and apoptosis of chondrocytes in osteoarthritis mice[J].Journal of Xinxiang Medical University,2023,40(11):1001-1007.[doi:10.7683/xxyxyxb.2023.11.001]
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巴戟天对骨关节炎小鼠软骨细胞自噬、凋亡的影响及机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
40卷
期数:
2023年11
页码:
1001-1007
栏目:
基础研究
出版日期:
2023-11-05

文章信息/Info

Title:
Effect and mechanism of morinda root on autophagy and apoptosis of chondrocytes in osteoarthritis mice
作者:
赵东方1; 尉志强1; 邢姝琴2; 祁姣3; 黄江海1; 温鑫柱1; 王亚非1
（1.北京中医药大学东方医院骨科，北京　100078；2.北京中医药大学东方医院肿瘤科，北京　100078；3.北京中医药大学东方医院经开院区门诊护理部，北京　100176）
Author(s):
ZHAO Dongfang1; WEI Zhiqiang1; XING Shuqin2; QI Jiao3; HUANG Jianghai1; WEN Xinzhu1; WANG Yafei1
（1.Department of Orthopedics,Oriental Hospital of Beijing University of Chinese Medicine,Beijing 100078,China;2.Department of Oncology,Oriental Hospital of Beijing University of Chinese Medicine,Beijing 100078,China;3.Department of Outpatient Nursing,Oriental Hospital of Beijing University of Chinese Medicine,Beijing 100176,China）
关键词:
骨关节炎; 巴戟天; 自噬; 凋亡
Keywords:
osteoarthritis; morinda root; autophagy; apoptosis
分类号:
R684.3
DOI:
10.7683/xxyxyxb.2023.11.001
文献标志码:
A
摘要:
目的　探讨巴戟天对骨关节炎小鼠软骨细胞自噬、凋亡的影响及机制。
方法　将40只健康C57BL/6雄性小鼠随机分为对照组、模型组、低剂量巴戟天组、中剂量巴戟天组及高剂量巴戟天组，每组8只。模型组和低、中、高剂量巴戟天组小鼠建立骨关节炎模型，随后模型组小鼠每日给予1 mL生理盐水灌胃，低、中、高剂量巴戟天组小鼠每日分别将100、200、400 mg·kg-1巴戟天溶于1 mL生理盐水灌胃，均持续4周；对照组小鼠制备骨关节炎假模型，随后每日给予1 mL生理盐水灌胃，持续4周。处死各组小鼠，无菌条件下提取右侧膝关节软骨组织，并制备软骨细胞悬液。原位末端转移酶标记染色检测各组小鼠软骨组织中软骨细胞凋亡率，流式细胞术检测各组小鼠软骨细胞悬液中软骨细胞的凋亡情况， Western blot法检测各组小鼠软骨细胞悬液中磷酸化蛋白激酶B（p-Akt）、磷酸化磷脂酰肌醇-3-激酶 （p-PI3K）、自噬效应蛋白1（Beclin1）、人微管相关蛋白轻链3B（LC3B）蛋白表达，实时荧光定量聚合酶链反应法检测各组小鼠软骨细胞悬液中p-PI3K、p-Akt mRNA的表达。
结果　模型组和低、中、高剂量巴戟天组小鼠软骨组织中软骨细胞凋亡率显著高于对照组（P<0.05）；低、中、高剂量巴戟天组小鼠软骨组织中软骨细胞凋亡率显著低于模型组（P<0.05）；中、高剂量巴戟天组小鼠软骨组织中软骨细胞凋亡率显著低于低剂量巴戟天组（P<0.05）；高剂量巴戟天组小鼠软骨组织中软骨细胞凋亡率显著低于中剂量巴戟天组（P<0.05）。模型组和低、中、高剂量巴戟天组小鼠软骨细胞悬液中软骨细胞凋亡率显著高于对照组（P<0.05）；低、中、高剂量巴戟天组小鼠软骨细胞悬液中软骨细胞凋亡率显著低于模型组（P<0.05）；中、高剂量巴戟天组小鼠软骨细胞悬液中软骨细胞凋亡率显著低于低剂量巴戟天组（P<0.05）；高剂量巴戟天组小鼠软骨细胞悬液中软骨细胞凋亡率显著低于中剂量巴戟天组（P<0.05）。模型组和低、中剂量巴戟天组小鼠软骨细胞悬液中Beclin1、LC3B、p-PI3K、p-Akt蛋白相对表达量及p-PI3K mRNA、p-Akt mRNA相对表达量显著低于对照组 (P<0.05)；高剂量巴戟天组与对照组小鼠软骨细胞悬液中Beclin1、LC3B、p-PI3K、p-Akt蛋白相对表达量及p-PI3K mRNA、p-Akt mRNA相对表达量比较差异无统计学意义(P>0.05)；低、中、高剂量巴戟天组小鼠软骨细胞悬液中Beclin1、LC3B、p-PI3K、p-Akt蛋白相对表达量及p-PI3K mRNA、p-Akt mRNA相对表达量显著高于模型组 (P<0.05)；中、高剂量巴戟天组小鼠软骨细胞悬液中Beclin1、LC3B、p-PI3K、p-Akt蛋白相对表达量及p-PI3K mRNA、p-Akt mRNA相对表达量显著高于低剂量巴戟天组（P<0.05）；高剂量巴戟天组小鼠软骨细胞悬液中Beclin1、LC3B、p-PI3K、p-Akt蛋白相对表达量及p-PI3K mRNA、p-Akt mRNA相对表达量显著高于中剂量巴戟天组 (P<0.05)。
结论　巴戟天能通过调控PI3K/Akt信号通路促进骨关节炎小鼠软骨细胞的自噬，抑制软骨细胞凋亡。
Abstract:
Objective　To investigate the effect and mechanism of morinda root on autophagy and apoptosis of chondrocytes in osteoarthritis mice.
Methods　Forty healthy C57BL/6 male mice were randomly divided into the control group,model group,low-dose morinda root group,medium-dose morinda root group and high-dose morinda root group,with 8 mice in each group.The mice in the model group and low-,medium- and high-dose morinda root group were established osteoarthritis models,and then the mice in the model group were given 1 mL of normal saline by gavage daily,while the mice in the low-,medium- and high-dose of morinda root group were given 100,200 and 400 mg·kg-1 morinda root dissolved in 1 mL of normal saline by gavage daily for 4 weeks.The mice in the control group were prepared a fake model of osteoarthritis,and then 1 mL of normal saline was administered daily by gavage for 4 weeks.The mice in each group were killed,and the cartilage tissues of the right knee joint were extracted under aseptic condition,and the chondrocyte suspension was prepared.The apoptosis rate of chondrocytes in the cartilage tissue of mice in each group was detected by TdT-mediated dUTP nick end labeling staining.The apoptosis of chondrocytes in the chondrocytes suspension of mice in each group was detected by flow cytometry.The expressions of phosphorylated protein kinase B (p-Akt),phosphorylated phosphatidylinositol-3-kinase (p-PI3K),autophagy effector protein 1 (Beclin1) and human microtubule associated protein light chain 3B (LC3B) in the chondrocytes suspension of mice in each group were detected by Western blot.The expressions of p-PI3K and p-Akt mRNA in chondrocytes suspension of mice in each group were detected by real time fluorescence quantitative polymerase chain reaction.
Results　The apoptosis rate of chondrocytes in the cartilage tissues of mice in model group and low-,medium- and high-dose morinda root groups was significantly higher than that in the control group (P<0.05).The apoptosis rate of chondrocytes in the cartilage tissues of mice in low-,medium- and high-dose morinda root groups was significantly lower than that in the model group (P<0.05).The apoptosis rate of chondrocytes in the cartilage tissues of mice in medium- and high-dose morinda root groups was significantly lower than that in the low-dose morinda root group (P<0.05).The apoptosis rate of chondrocytes in the cartilage tissues of mice in high-dose morinda root groups was significantly lower than that in the medium-dose morinda root group (P<0.05).The apoptosis rate of chondrocytes in the chondrocytes suspension of mice in the model group and low-,medium- and high-dose morinda root groups was significantly higher than that in the control group (P<0.05).The apoptosis rate of chondrocytes in the chondrocytes suspension in the low-,medium- and high-dose morinda root groups was significantly lower than that in the model group (P<0.05).The apoptosis rate of chondrocytes in the chondrocytes suspension of mice in the medium and high-dose morinda root groups was significantly lower than that in the low-dose morinda root group (P<0.05).The apoptosis rate of chondrocytes in the chondrocytes suspension of mice in the high-dose morinda root group was significantly lower than that in the medium-dose morinda root group (P<0.05).The relative expression levels of Beclin1,LC3B,p-PI3K,p-Akt protein,as well as p-PI3K,p-Akt mRNA in the chondrocytes suspension of mice in the model group and low-,medium-dose morinda root group were significantly lower than those in the control group (P<0.05).There was no statistically significant difference in the relative expression levels of Beclin1,LC3B,p-PI3K,p-Akt protein and p-PI3K,p-Akt mRNA in the chondrocytes suspension of mice between the high-dose morinda root group and the control group (P>0.05).The relative expression levels of Beclin1,LC3B,p-PI3K,p-Akt proteins and p-PI3K,p-Akt mRNA in the chondrocytes suspension of mice in the low-,medium- and high-dose morinda root groups were significantly higher than those in the model group (P<0.05).The relative expression levels of Beclin1,LC3B,p-PI3K,p-Akt proteins and p-PI3K,p-Akt mRNA of chondrocytic in the chondrocyte suspension of mice in the medium- and high-dose morinda root groups were significantly higher than those in the low-dose morinda root group (P<0.05).The relative expression levels of Beclin1,LC3B,p-PI3K,p-Akt proteins,as well as p-PI3K,p-Akt mRNA in the chondrocytes suspension of mice in the high-dose morinda root group were significantly higher than those in the medium-dose morinda root group (P<0.05).
Conclusion　Morinda root can promote autophagy of chondrocytes in osteoarthritis mice and inhibit chondrocyte apoptosis by regulating the PI3K/Akt signaling pathway.

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更新日期/Last Update: 2023-11-05