雷公藤内酯醇对类风湿性关节炎模型大鼠血管新生和PTEN/PI3K/AKT通路的影响

doi:10.13481/j.1671-587x.20200619

Abstract

Abstract:

Objective To investigate the effect of triptolide on the angiogenesis and phosphatase and tensin homolog delected on chromosome/ phosphoinositide-3-kinase / protein kinase B （PTEN / PI3K / AKT） pathway in the rats with rheumatoid arthritis，and to elucidate the possible mechanism of triptolide in the treatment of rheumatoid arthritis.

Methods

A total of 80 rats were randomly divided into control group， model group， positive drug group and treatment group；there were 20 rats in each group.The rheumatoid arthritis model was established with type Ⅱ collagen（Col Ⅱ） induction.The rats in positive drug group were treated with diclofenac sodium sustained-release tablets，and the rats in treatment group were treated with triptolide.After treatment，the body weights， paw thickness and arthritis index scores of the rats in various groups were measured；HE staining was used to determine the pathomorphology of joint synovial tissue of the rats in various groups；immunohistochemical staining was used to determine the microvessel density（MVD） of vascular endothelial growth factor （VEGF） expression level in synovial tissue of the rats in various groups； ELISA method was used to determine the serum levels of VEGF of the rats in various groups；Western blotting method was used to determine the expression levels of PTEN， PI3K， AKT and phosphorylated-AKT （p-AKT） proteins in synovial tissue of the rats in various groups.

Results

Compared with control group， the body weight of rats in model group was decreased （P<0.05）， the paw thickness was increased （P<0.05）， the MVD and expression level of VEGF in synovial tissue were increased （P<0.05）， the expression level of PTEN protein in synovial tissue was decreased （P<0.05）， and the expression levels of PI3K， AKT and p-AKT proteins were increased （P<0.05）.Compared with model group， the body weights of the rats in positive drug group and treatment group were increased （P<0.05），the paw thickness was decreased （P<0.05），the MVD and the expression levels of VEGF in synovial tissue were decreased （P<0.05）， the expression levels of PTEN protein were decreased （P<0.05）， and the expression levels of PI3K， AKT and p-AKT proteins were decreased （P<0.05）.Compared with positive drug group， the MVD and expression levels of VEGF in synovial tissue of the rats in treatment group were decreased （P<0.05）， the expression level of PTEN protein in synovial tissue was increased （P<0.05）， and the expression levels of PI3K， AKT and p-AKT proteins were decreased （P<0.05）.The paw thickness and arthritis index of rheumatoid arthritis rats were positively correlated with VEGF（r=0.564， r =0.492） and p-AKT（r=0.561， r=0.468）（P<0.05），and negatively correlated with PTEN （r=－0.437， r=－0.521）（P<0.05）；MVD was positively correlated with VEGF（r =0.587）， PI3K（r =0.567）， and p-AKT（r=0.601）（P<0.05）， and negatively correlated with PTEN （r=－0.502）（P<0.05）.

Conclusion

Triptolide can inhibit the angiogenesis and PTEN / PI3K / AKT pathway activation in the rheumatoid arthritis rats， thereby exerting its therapeutic effect on rheumatoid arthritis.

Key words: triptolide, rheumatoid arthritis, blood vessels, phosphatase and tensin homolog deleted on chromosome ten/phosphoinositide-3-kinase/protein kinase B

CLC Number:

R684.3

Jing LIU,Lijun YAN. Effects of triptolide on angiogenesis and PTEN / PI3K / AKT pathway in model rats with rheumatoid arthritis[J].Journal of Jilin University(Medicine Edition), 2020, 46(6): 1227-1233.

Figures/Tables 6

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References 24

1	武占成，赵晶，朱秀惠，等. 类风湿关节炎的骨病变从络病论治［J］. 中国组织工程研究， 2020， 24（5）： 760-765.
2	BANERJEE S， SANGWAN V， MCGINN O， et al. Triptolide-induced cell death in pancreatic cancer is mediated by O-GlcNAc modification of transcription factor Sp1［J］. J Biol Chem， 2013， 288（47）： 33927-33938.
3	LI X， LU Q， XIE W， et al. Anti-tumor effects of triptolide on angiogenesis and cell apoptosis in osteosarcoma cells by inducing autophagy via repressing Wnt/β-Catenin signaling［J］. Biochem Biophys Res Commun， 2018， 496（2）： 443-449.
4	陈颖婷，何柯新，王云秀，等. 雷公藤内酯醇对类风湿关节炎大鼠TLR4/NF-κB信号通路的调控作用研究［J］. 国际检验医学杂志， 2019， 40（17）： 2053-2057.
5	陈柏松，徐玉东，钟淑琦，等. 大鼠佐剂性关节炎模型的建立与评价［J］. 哈尔滨医科大学学报， 2005， 39（6）： 489-491.
6	陈颖婷，何柯新，王云秀，等. 雷公藤内酯醇对类风湿关节炎大鼠TLR4/NF-κB信号通路的调控作用研究［J］. 国际检验医学杂志， 2019， 40（17）： 2053-2057.
7	陈璐璐，钟晓岚，朱颖敏，等. 雌性Lewis大鼠佐剂诱导关节炎模型的建立与评价［J］. 山西医科大学学报， 2019， 50（10）： 1453-1460.
8	CHEN Z， WANG H， XIA Y， et al. Therapeutic potential of mesenchymal cell-derived miRNA-150-5p-expressing exosomes in rheumatoid arthritis mediated by the modulation of MMP14 and VEGF［J］. J Immunol， 2018， 201（8）： 2472-2482.
9	LEE Y H， BAE S C. Correlation between circulating VEGF levels and disease activity in rheumatoid arthritis： A meta-analysis［J］. Z Rheumatol， 2018， 77（3）： 240-248.
10	SHEVCHENKO AV， PROKOFYEV VF， KOROLEV MA，et al.Polymorphism in the regulatory regions -С2578A and +C936T of the vascular endothelial growth factor （VEGF-A） gene in Russian women with rheumatoid arthritis［J］.Ter Arkh，2017，89（5）：60-64.
11	CHEN C Y， SU C M， HSU C J， et al. CCN₁ promotes VEGF production in osteoblasts and induces endothelial progenitor cell angiogenesis by inhibiting miR-126 expression in rheumatoid arthritis［J］. J Bone Miner Res， 2017， 32（1）： 34-45.
12	郑栩良，张郜晨茜，张烁. 鲁贤昌教授治疗类风湿性关节炎经验［J］. 浙江中医药大学学报， 2018， 42（5）： 370-373.
13	曾克勤，武剑，郭雨凡，等. 雷公藤治疗类风湿关节炎的临床疗效与安全性的Meta分析［J］. 中国血液流变学杂志， 2017， 27（3）： 284-287.
14	ZHANG F Z， HO D H， WONG R H. Triptolide， a HSP90 middle domain inhibitor， induces apoptosis in triple manner［J］. Oncotarget， 2018， 9（32）： 22301-22315.
15	胡伟锋，王昌兴. 雷公藤内酯醇对类风湿关节炎大鼠的治疗作用及其网络药理学研究［J］. 中国全科医学， 2016， 19（12）： 1408-1413.
16	王进，李森，张友朋，等. 雷公藤内酯醇联合顺铂对荷膀胱癌T24细胞裸鼠免疫功能及血管生长作用的影响［J］. 中华实验外科杂志， 2016， 33（6）： 1448-1450.
17	LIU Y， PAN Y F， XUE Y Q， et al. uPAR promotes tumor-like biologic behaviors of fibroblast-like synoviocytes through PI₃K/Akt signaling pathway in patients with rheumatoid arthritis［J］. Cell Mol Immunol， 2018， 15（2）： 171-181.
18	JIAO Y， DING H， HUANG S， et al. Bcl-XL and Mcl-1 upregulation by calreticulin promotes apoptosis resistance of fibroblast-like synoviocytes via activation of PI₃K/Akt and STAT3 pathways in rheumatoid arthritis［J］. Clin Exp Rheumatol， 2018， 36（5）： 841-849.
19	LIN J， HE Y， WANG B， et al. Blocking of YY₁ reduce neutrophil infiltration by inhibiting IL-8 production via the PI₃K-Akt-mTOR signaling pathway in rheumatoid arthritis［J］. Clin Exp Immunol， 2019， 195（2）： 226-236.
20	ZHANG L， LIU X， LIU J， et al. miR-26a promoted endometrial epithelium cells （EECs） proliferation and induced stromal cells （ESCs） apoptosis via the PTEN-PI₃K/AKT pathway in dairy goats［J］. J Cell Physiol， 2018， 233（6）： 4688-4706.
21	DAI Y X， WAN Y K， QIU M K， et al. lncRNA MEG3 suppresses the tumorigenesis of hemangioma by sponging miR-494 and regulating PTEN/PI₃K/AKT pathway［J］. Cell Physiol Biochem， 2018， 51（6）： 2872-2886.
22	LI N N， MENG X S， BAO Y R， et al. Evidence for the involvement of COX-2/VEGF and PTEN/Pl3K/AKT pathway the mechanism of oroxin B treated liver cancer［J］. Pharmacogn Mag， 2018， 14（54）： 207-213.
23	刘斐，谭布珍. 雷公藤内酯醇联合紫杉醇对COC1/DDP细胞凋亡的影响及其对PI₃K/AKT/GSK3β信号通路的调控［J］. 现代妇产科进展， 2013， 22（12）： 945-950.
24	戴小娜，林进.老年起病类风湿关节炎研究进展[J].中国实用内科杂志,2019,39(6):562-567.

Group	n	Body weight（m/g）	Paw thickness（l/mm）	Arthritis index score
Control	20	10.84±0.92	5.78±0.24	－
Model	17	8.11±0.87^*	8.65±0.33^*	7.32±0.94
Positive drug	18	9.10±0.86^Δ	6.93±0.31^Δ	6.38±0.92^Δ
Treatment	19	9.02±0.91^Δ	6.91±0.28^Δ	6.47±0.89^Δ
F		30.881	302.636	5.591
P		0.000	0.000	0.006
“－”:No data. *P<0.05 compared with control group；^ΔP<0.05 compared with model group.

Group	n	MVD（S/mm^－2）	VEGF
Control	20	9.35±1.02	18.21±1.97
Model	17	104.36±3.57^*	143.24±2.76^*
Positive drug	18	66.29±3.18^Δ	96.35±2.64^Δ
Treatment	19	48.77±3.25^Δ#	81.02±2.51^Δ#
F		3431.055	8125.035
P		0.000	0.000

Group	n	VEGF
Control	20	38.67±4.21
Model	17	73.26±4.85^*
Positive drug	18	53.24±4.98^Δ
Treatment	19	46.23±5.02^Δ＃
F		174.773
P		0.000

Group	n	PTEN	PI3K	AKT	p-AKT
Control	20	0.96±0.15	0.09±0.03	0.08±0.02	0.08±0.03
Model	17	0.37±0.11^*	0.73±0.12^*	0.43±0.05^*	0.47±0.09^*
Positive drug	18	0.52±0.14^Δ	0.69±0.11^Δ	0.39±0.03^Δ	0.36±0.04^Δ
Treatment	19	0.83±0.15^Δ＃	0.35±0.08^Δ＃	0.17±0.06^Δ＃	0.22±0.05^Δ＃
F		70.187	211.478	291.861	169.944
P		0.000	0.000	0.000	0.000

Index	Foot claw thickness		Arthritis index		MVD
Index	r	P	r	P	r	P
VEGF	0.564	0.000	0.492	0.000	0.587	0.000
PTEN	－0.437	0.000	－0.521	0.000	－0.502	0.000
PI3K	0.234	0.097	0.246	0.089	0.567	0.000
AKT	0.182	0.113	0.195	0.102	0.196	0.113
p-AKT	0.561	0.000	0.468	0.000	0.601	0.000

Effects of triptolide on angiogenesis and PTEN / PI3K / AKT pathway in model rats with rheumatoid arthritis

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