扁蒴藤素调节ROS/ASK1/JNK信号通路对二乙基亚硝胺诱导大鼠肝癌的抑制作用

doi:10.3969/j.issn.1006-5725.2023.20.007

Abstract

Abstract:

Objective To investigate the influence of pristimerin （Pris） on rat hepatocellular carcinoma （HCC） induced by diethylnitrosamine （DEN） by regulating ROS/ASK1/JNK signal pathway. Methods Six SD rats were randomly selected as the control group， and the remaining rats were injected with DEN to construct the rat model of HCC. The rats successfully modeled were randomly grouped into HCC group， Pris group （0.8 mg/kg Pris）， Vaccarin group （100 mg/kg ROS/ASK1/JNK signal pathway inhibitor Vaccarin）， and Pris+Vaccarin group （0.8 mg/kg Pris+100 mg/kg Vaccarin）， with 6 rats in each group， and were injected continuously for one week. HCC group and control group were injected with the same amount of normal saline. Body weight， liver weight and liver weight ratio were measured； liver function， inflammatory factors， antioxidant index and ROS level were detected by ELISA； HE staining was applied to detect the pathological changes of liver； western blot was applied to detect the expression of apoptotic markers （Bax， Bcl-2 and cleaved-Caspase-3） and ROS/ASK1/JNK signal pathway protein. Results The rats in the control group showed normal liver tissue structure. In HCC group， necrosis of some hepatocytes and focal nodular hyperplasia were observed， body weight， Bax， cleaved-Caspase-3， ROS， p-ASK1/ASK1， and p-JNK/JNK level in HCC group were obviously lower than those in the control group （P < 0.05）， but liver weight， liver weight ratio， ALT， AST， ALP， LDH contents， IL-6， TNF-α， CCL-2 contents， SOD， GR， GPx， CAT contents， and Bcl-2 protein level were obviously higher （P < 0.05）. Hepatocyte necrosis and focal nodular hyperplasia in Pris group were improved， and body weight， Bax， cleaved-Caspase-3， ROS， p-ASK1/ASK1， and p-JNK/JNK level in Pris group were obviously higher than those in HCC group （P<0.05）； liver weight， liver weight ratio， ALT， AST， ALP， LDH contents， IL-6， TNF-α， CCL-2 contents， SOD， GR， GPx， CAT contents， and Bcl-2 protein level were obviously lower （P<0.05）. The opposite trend was observed in Vaccarin group； Vaccarin reversed the anticancer effect of Pris on HCC rats. Conclusion Pris may play a certain role in inhibiting HCC rats by activating ROS/ASK1/JNK signal pathway.

Key words: pristimerin, ROS/ASK1/JNK signal pathway, diethylnitrosamine, hepatocellular carcinoma

CLC Number:

S852

Yan LIU,Donghua ZHANG,Yuqian HUANG,Youshun LIU,Ji. HUANG. Inhibitory effect of pristimerin on diethylnitrosamine⁃induced hepatocellular carcinoma in rats by regulating ROS/ASK1/JNK signal pathway[J]. The Journal of Practical Medicine, 2023, 39(20): 2597-2602.

Figures/Tables 9

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References 23

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组别	体质量（g）	肝脏质量（g）	肝脏体质量比（%）
对照组	410.28 ± 55.34	12.49 ± 1.28	3.04 ± 0.24
HCC组	281.14 ± 33.54^a	23.35 ± 2.14^a	8.31 ± 0.63^a
Pris组	389.21 ± 46.83^b	13.34 ± 1.26^b	3.42 ± 0.27^b
Vaccarin组	200.45 ± 24.37^b	34.73 ± 3.05^b	17.32 ± 1.25^b
Pris+Vaccarin组	298.16 ± 35.36^c	21.46 ± 2.14^c	7.20 ± 0.61^c
F值	26.566	112.499	296.136
P值	< 0.001	< 0.001	< 0.001

组别	ALT（U/mL）	AST（U/mL）	ALP（U/mL）	LDH（U/mL）
对照组	105.35 ± 11.82	136.23 ± 18.64	140.89 ± 17.09	311.57 ± 36.18
HCC组	279.58 ± 30.97^a	354.14 ± 40.42^a	313.43 ± 38.46^a	591.08 ± 63.12^a
Pris组	134.57 ± 21.57^b	186.07 ± 20.63^b	191.34 ± 20.11^b	421.44 ± 45.16^b
Vaccarin组	416.46 ± 54.64^b	532.48 ± 57.26^b	436.56 ± 49.69^b	930.61 ± 95.07^b
Pris+Vaccarin组	210.21 ± 21.13^c	304.57 ± 38.43^c	283.01 ± 30.32^c	501.17 ± 53.12^c
F值	93.460	101.324	70.912	86.871
P值	< 0.001	< 0.001	< 0.001	< 0.001

组别	CCL-2（pg/mL）	IL-6（pg/mL）	TNF-α（pg/mL）
对照组	103.64 ± 14.15	22.32 ± 2.73	0.76 ± 0.10
HCC组	251.35 ± 25.37^a	54.34 ± 5.54^a	3.47 ± 0.44^a
Pris组	123.98 ± 13.26^b	33.59 ± 3.37^b	1.27 ± 0.16^b
Vaccarin组	368.48 ± 47.58^b	69.57 ± 6.37^b	6.57 ± 0.68^b
Pris+Vaccarin组	205.24 ± 24.24^c	49.34 ± 4.29^c	3.00 ± 0.35^c
F值	88.266	93.392	193.237
P值	< 0.001	< 0.001	< 0.001

组别	SOD（U/mL）	GR（U/mL）	GPx（U/mL）	CAT（U/mL）
对照组	14.35 ± 2.37	16.35 ± 1.63	9.58 ± 0.97	31.14 ± 2.42
HCC组	23.64 ± 3.15^a	28.78 ± 2.91^a	15.35 ± 1.82^a	46.23 ± 6.64^a
Pris组	16.48 ± 1.58^b	18.21 ± 1.78^b	10.46 ± 1.24^b	33.07 ± 4.63^b
Vaccarin组	33.98 ± 5.26^b	37.45 ± 4.75^b	34.57 ± 4.57^b	61.48 ± 7.26^b
Pris+Vaccarin组	19.24 ± 2.04^c	24.67 ± 2.56^c	13.21 ± 1.56^c	43.57 ± 4.43^c
F值	15.618	72.724	84.278	84.278
P值	< 0.001	< 0.001	< 0.001	< 0.001

组别	Bax/GAPDH	Bcl-2/GAPDH	cleaved-Caspase-3/GAPDH
对照组	1.26 ± 0.13	0.74 ± 0.08	1.37 ± 0.16
HCC组	0.68 ± 0.05^a	1.33 ± 0.15^a	0.96 ± 0.10^a
Pris组	1.13 ± 0.09^b	0.89 ± 0.17^b	1.31 ± 0.14^b
Vaccarin组	0.39 ± 0.02^b	1.82 ± 0.09^b	0.48 ± 0.05^b
Pris+Vaccarin组	0.71 ± 0.06^c	1.30 ± 0.15^c	1.00 ± 0.12^c
F值	120.314	55.055	80.433
P值	< 0.001	< 0.001	< 0.001

Inhibitory effect of pristimerin on diethylnitrosamine⁃induced hepatocellular carcinoma in rats by regulating ROS/ASK1/JNK signal pathway

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组别	ROS（mg/mL）	p-ASK1/ASK1	p-JNK/JNK
对照组	25.00 ± 2.89	0.95 ± 0.05	0.78 ± 0.09
HCC组	10.66 ± 1.05^a	0.52 ± 0.04^a	0.44 ± 0.05^a
Pris组	21.91 ± 2.07^b	0.83 ± 0.07^b	0.72 ± 0.09^b
Vaccarin组	5.34 ± 0.52^b	0.38 ± 0.03^b	0.27 ± 0.04^b
Pris+Vaccarin组	13.70 ± 1.46^c	0.66 ± 0.05^c	0.50 ± 0.06^c
F值	116.200	158.283	54.753
P值	< 0.001	< 0.001	< 0.001

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