Effect of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury

doi:10.11958/20230374

Abstract

Abstract:

Objective To explore the impact of Jiawei Taoren Chengqi Decoction on autophagy in rats with mechanical ventilation-induced lung injury (VILI) through adenylate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Methods Sixty SPF male rats were randomly grouped into the sham operation group, the model group, the low-dose Jiawei Taoren Chengqi Decoction group (TCM-L group, 2.85 g/kg), the high-dose Jiawei Taoren Chengqi Decoction group (TCM-H group, 8.55 g/kg) and the high-dose Jiawei Taoren Chengqi Decoction + AMPK inhibitor Compound C group (TCM-H+CC group, Jiawei Taoren Chengqi Decoction 8.55 g/kg+ Compound C 250 μg/kg), 12 in each group. Oxygenation index (OI) was measured immediately after intubation and at 1 h, 2 h and 4 h after mechanical ventilation. Bronchoalveolar lavage fluid (BALF) was collected after mechanical ventilation, levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-18 in BALF were detected by enzyme-linked immunosorbent assay (ELISA). Rats were sacrificed, and lung tissue was taken to measure the wet/dry weight (W/D) ratio. HE staining was used to observe pathological changes of lung tissue in each group of rats. Lung injury scores were carried out. Morphology of alveolar epithelial cells was observed by transmission electron microscopy. RT-qPCR was applied to detect mRNA expression levels of AMPK and mTORC1 in rat lung tissue. Western blot assay was applied to detect expression levels of AMPK, p-AMPK, mTORC1, p-mTORC1 and autophagy-related proteins in rat lung tissue. Results Compared with the sham group, the pathological damage of lung tissue was serious, lung W/D, levels of TNF-α, IL-1β and IL-18 in BALF, lung injury score, mTORC1 mRNA expression level, and p-mTORC1 protein expression were increased in the model group (P<0.05). OI values at 2 h and 4 h of mechanical ventilation, AMPK mRNA expression level, p-AMPK, LC3-II/LC3-I and Beclin-1 protein expression in lung tissue were decreased (P<0.05). Compared with the model group, the pathological damage of lung tissue was alleviated in the Chinese medicine-H group, and the trend of changes in related indexes was opposite to the above (P<0.05). Autophagosomes in alveolar epithelial cells were increased. Compound C attenuated the protective effect of Jiawei Taoren Chengqi Decoction on VILI rats (P<0.05). Conclusion Jiawei Taoren Chengqi Decoction may promote autophagy and reduce VILI in rats by activating AMPK/mTOR signaling pathway.

Key words: ventilator-induced lung injury, Tao He Cheng Qi Tang, autophagy, AMP-activated protein kinases, TOR serine-threonine kinases

CLC Number:

R563

Figures/Tables 8

References 18

[1]	BAI Y X, ZHANG J H, ZHAO B C, et al. Dexmedetomidine attenuates one-lung ventilation associated lung injury by suppressing inflammatory responses:A systematic review and meta-analysis[J]. Clin Exp Pharmacol Physiol, 2021, 48(9):1203-1214. doi:10.1111/1440-1681.13525.
[2]	张本旺, 蒋娟, 马福国, 等. 亚甲蓝对大鼠机械通气相关肺损伤中核苷酸结合寡聚化结构域样受体蛋白3炎性小体的影响[J]. 临床麻醉学杂志, 2020, 36(8):793-798.
	ZHANG B W, JIANG J, MA F G, et al. Effect of methylene blue on nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome in rats with ventilator-induced lung injury[J]. Journal of Clinical Anesthesiology, 2020, 36(8):793-798. doi:10.12089/jca.2020.08.015.
[3]	高天巍, 朱克毅, 汪建英. 加味桃仁承气汤辅助机械通气对急性肺损伤/急性呼吸窘迫综合征患儿的疗效观察[J]. 中医儿科杂志, 2015, 11(6):48-52.
	GAO T W, ZHU K Y, WANG J Y. Observation on the therapeutic effect of modified Taoren Chengqi decoction assisted mechanical ventilation on children with acute lung injury/acute respiratory distress syndrome[J]. Journal of Pediatrics of Traditional Chinese Medicine, 2015, 11(6):48-52.
[4]	LIN M, HUA R, MA J, et al. Bisphenol A promotes autophagy in ovarian granulosa cells by inducing AMPK/mTOR/ULK1 signalling pathway[J]. Environ Int, 2021, 147:106298. doi:10.1016/j.envint.2020.106298.
[5]	NGUYEN T N, DUBREUCQ E, PERRIER V, et al. Interactions between trans-resveratrol and CpLIP2 lipase/acyltransferase:Evidenced by fluorescence and in silico[J]. Food Chem, 2020, 318:126482. doi:10.1016/j.foodchem.2020.126482.
[6]	瞿敏, 姚忠岩, 刘天琳, 等. 甲泼尼龙对机械通气相关性肺损伤大鼠肺泡内液体清除的影响[J]. 中国急救医学, 2020, 40(4):343-347.
	QU M, YAO Z Y, LIU T L, et al. Effect of methylprednisolone on alveolar fluid clearance in the rats with mechanical ventilation-related lung injury[J]. Chinese Journal of Critical Care Medicine, 2020, 40(4):343-347. doi:10.3969/j.issn.1002-1949.2020.04.016.
[7]	赵云丽, 袁勇, 马晓莉, 等. 基于AMPK/SIRT1/PGC-1α信号通路研究香青兰总黄酮对大鼠心肌缺血再灌注损伤的保护机制[J]. 中国药房, 2021, 32(3):278-283.
	ZHAO Y L, YUAN Y, MA X L, et al. Study on protective mechanism of dracocephalum moldavica total flavonoids against myocardial ischemia-reperfusion injury in rats based on AMPK/SIRT1/PGC-1α signaling pathway[J]. China Pharmacy, 2021, 32(3):278-283. doi:10.6039/j.issn.1001-0408.2021.03.05.
[8]	孙治霞, 索红亮, 王丽辉. 桃仁承气汤对脓毒症大鼠肠黏膜屏障及TLR9信号通路的影响[J]. 中国实验方剂学杂志, 2019, 25(7):29-35.
	SUN Z X, SUO H L, WANG L H. The effect of Taoren Chengqi decoction on intestinal mucosal barrier and TLR9 signaling pathway in septic rats[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2019, 25(7):29-35. doi:10.13422/j.cnki.syfjx.20190738.
[9]	林新, 徐礼裕, 李小华, 等. 微RNA-146a对机械通气大鼠肺损伤TLR4炎症信号通路的影响及机制[J]. 中华医学杂志, 2018, 98(34):2749-2753.
	LIN X, XU L Y, LI X H, et al. The effect and mechanism of microRNA-146a on TLR4 inflammatory signaling pathway in mechanically ventilated rats with lung injury[J]. Natl Med J China, 2018, 98(34):2749-2753. doi:10.3760/cma.j.issn.0376-2491.2018.34.014.
[10]	王萍, 唐宋琪, 农开旭, 等. 柚皮素预处理通过抑制NLRP3炎症小体通路减轻小鼠机械通气相关性肺损伤[J]. 安徽医科大学学报, 2021, 56(6):871-875.
	WANG P, TANG S Q, NONG K X, et al. Naringenin preconditioning alleviates ventilator-associated lung injury in mice by inhibiting NLRP3 inflammatory body pathway[J]. Acta Universitatis Medicinalis Anhui, 2021, 56(6):871-875. doi:10.19405/j.cnki.issn1000-1492.2021.06.007.
[11]	CHEN H, SUN X, YANG X, et al. Dexmedetomidine reduces ventilator-induced lung injury (VILI) by inhibiting Toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB signaling pathway[J]. Bosn J Basic Med Sci, 2018, 18(2):162-169. doi:10.17305/bjbms.2018.2400.
[12]	HUANG T H, FANG P H, LI J M, et al. Cyclooxygenase-2 activity regulates recruitment of VEGF-secreting Ly6C^high monocytes in ventilator-induced lung injury[J]. Int J Mol Sci, 2019, 20(7):1771. doi:10.3390/ijms20071771.
[13]	刘天华. 中西医结合治疗肝硬化轻微肝性脑病临床疗效观察[J]. 辽宁中医杂志, 2022, 49(1):109-112.
	LIU T H. Clinical observation on the treatment of mild Hepatic encephalopathy due to cirrhosis with integrated traditional Chinese and western medicine[J]. Liaoning Journal of Traditional Chinese Medicine, 2022, 49(1):109-112. doi:10.13192/j.issn.1000-1719.2022.01.031.
[14]	黄耀明, 赵小忠, 章燕清. 桃仁承气汤内服合四黄膏加味外敷治疗脊柱压缩性骨折的临床研究[J]. 现代诊断与治疗, 2020, 31(23):3715-3716.
	HUANG Y M, ZHAO X Z, ZHANG Y Q. Clinical study on treatment of compressibility fracture of spine with taoren chengqi decoction and sihuang ointment[J]. Modern Diagnosis and Treatment, 2020, 31(23):3715-3716.
[15]	马翠芹, 任燕, 马勇. 加味桃仁承气汤联合护理干预对胸腰椎压缩性骨折后腹胀便秘的影响[J]. 中西医结合护理(中英文), 2017, 3(5):66-68.
	MA C Q, REN Y, MA Y. Supplemented Taoren Chengqi decoction and related nursing intervention on abdominal distension and constipation after thoracolumbar compression fracture[J]. Nursing of Integrated Traditional Chinese and Western Medicine, 2017, 3(5):66-68. doi:10.11997/nitcwm.201705020.
[16]	BENEDETTO A, GEMS D. Autophagy promotes visceral aging in wild-type C. elegans[J]. Autophagy, 2019, 15(4):731-732. doi:10.1080/15548627.2019.1569919.
[17]	GUI X, YANG H, LI T, et al. Autophagy induction via STING trafficking is a primordial function of the cGAS pathway[J]. Nature, 2019, 567(7747):262-266. doi:10.1038/s41586-019-1006-9.
[18]	HAN D, JIANG L, GU X, et al. SIRT3 deficiency is resistant to autophagy-dependent ferroptosis by inhibiting the AMPK/mTOR pathway and promoting GPX4 levels[J]. J Cell Physiol, 2020, 235(11):8839-8851. doi: 10.1002/jcp.29727.

基因名称	引物序列（5′→3′）	产物大小/bp
AMPK	上游：GCTGTGGATCGCCAAATTAT	228
AMPK	下游：CACGTGCTCATCATCGAAAG	228
mTORC1	上游：AATACGCCATGAAACACTTCG	214
mTORC1	下游：GGTCTTCCTTGTTTGTGTCCA	214
GAPDH	上游：TGTGTCCGTCGTGGATCTGA	195
GAPDH	下游：TTGCTGTTGAAGTCGCAGGAG	195

基因名称	引物序列（5′→3′）	产物大小/bp
AMPK	上游：GCTGTGGATCGCCAAATTAT	228
AMPK	下游：CACGTGCTCATCATCGAAAG	228
mTORC1	上游：AATACGCCATGAAACACTTCG	214
mTORC1	下游：GGTCTTCCTTGTTTGTGTCCA	214
GAPDH	上游：TGTGTCCGTCGTGGATCTGA	195
GAPDH	下游：TTGCTGTTGAAGTCGCAGGAG	195

组别	插管即刻	通气1 h	通气2 h	通气4 h
Sham组	566.23±34.35	557.68±26.92	548.44±28.48	546.35±25.87
Model组	572.36±32.62	549.73±29.66	265.52±20.31^a	208.94±15.47^a
中药-L组	568.40±33.14	553.17±30.83	286.50±21.22	223.64±16.42
中药-H组	563.83±30.15	555.24±28.35	521.86±27.96^bc	515.30±23.65^bc
中药-H+CC组	568.36±31.07	551.29±29.64	432.64±20.33^d	410.68±23.52^d
F	0.114	0.141	357.164^＊＊	658.911^＊＊

组别	插管即刻	通气1 h	通气2 h	通气4 h
Sham组	566.23±34.35	557.68±26.92	548.44±28.48	546.35±25.87
Model组	572.36±32.62	549.73±29.66	265.52±20.31^a	208.94±15.47^a
中药-L组	568.40±33.14	553.17±30.83	286.50±21.22	223.64±16.42
中药-H组	563.83±30.15	555.24±28.35	521.86±27.96^bc	515.30±23.65^bc
中药-H+CC组	568.36±31.07	551.29±29.64	432.64±20.33^d	410.68±23.52^d
F	0.114	0.141	357.164^＊＊	658.911^＊＊

组别	TNF-α	IL-1β	IL-18
Sham组	16.59±2.48	19.25±2.03	22.34±2.78
Model组	128.45±12.06^a	102.34±11.65^a	139.05±12.59^a
中药-L组	117.63±13.55	96.41±9.50	128.55±11.31
中药-H组	80.27±8.22^bc	68.42±6.22^bc	85.65±8.24^bc
中药-H+CC组	104.20±11.39^d	83.69±8.24^d	116.52±12.35^d
F	223.256^＊＊	197.278^＊＊	257.616^＊＊