中药复方防感汤对SARS-CoV-2刺突蛋白引起的肺和结肠上皮细胞损伤的保护作用与TGF-β/Smad2/3和NF-κB通路的调节有关

doi:10.24920/004198

Abstract

Objective To explore the effects and mechanisms of a traditional Chinese medicine (TCM) prescription, “Fang-gan Decoction” (FGD), in protecting against SARS-CoV-2 spike protein-induced lung and intestinal injuries in vitro and in vivo.
Methods Female BALB/c mice and three cell lines pretreated with FGD were stimulated with recombinant SARS-CoV-2 spike protein (spike protein). Hematoxylin-eosin (HE) staining and pathologic scoring of tissues, cell permeability and viability, and angiotensin-converting enzyme 2 (ACE2) expression in the lung and colon were detected. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of inflammatory factors in serum and cell supernatant. The expression of NF-κB p65, p-NF-κB p65, p-IκBα, p-Smad2/3, TGF-β1, Caspase3, and Bcl-2 was evaluated by Western blotting.
Results FGD protected against the damage to the lung and colon caused by the spike protein in vivo and in vitro according to the pathologic score and cell permeability and viability (P<0.05). FGD up-regulated ACE2 expression, which was reduced by the spike protein in the lung and colon, significantly improved the deregulation of inflammatory markers caused by the spike protein, and regulated the activity of TGF-β/Smads and NF-κB signaling.
Conclusion Traditional Chinese medicine has a protective effect on lung and intestinal tissue injury stimulated by the spike protein through possible regulatory functions of the NF-κB and TGF-β1/Smad pathways with tissue type specificity.

Key words: COVID-19, SARS-CoV-2, traditional chinese medicine, prevention, transforming growth factor-β

Funding:

Chao Huang, Hao-Sheng Liu, Bing-Jun Liang, Sheng-Rong Liao, Wei-Zeng Shen. A Chinese Herb Prescription “Fang-gan Decoction” Protects Against Damage to Lung and Colon Epithelial Cells Caused by the SARS-CoV-2 Spike Protein by Regulating the TGF-β/Smad2/3 and NF-κB Pathways[J].Chinese Medical Sciences Journal, 2023, 38(3): 206-217.

Figures/Tables 7

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Markers	Time	Normal group	Model Gorup	NS group	TCM group	P value
IL-6	3d	644.61	717.36^*	696.89^*	702.85^*	0.009
	5d	268.02	342.09^*	344.88^*	298.89^#?	<0.001
IL-10	3d	25.14	82.12^*	74.86^*	44.7^#	0.009
	5d	87.61	145.79^*	123.84^*	121.52	0.019
IL-13	3d	352.69	344.69	362.46	351.09	0.998
	5d	91.9	102.87	92.42	100.23	0.931
TNF-α	3d	566.19	853.87	840.04	633.38	0.141
	5d	575.77	1042.36^*	1236.73^*#	864.43^*#?	<0.001
CRP	3d	18175.8	21129.27^*	21550.72^*	19786.15	0.042
	5d	5933.09	7413.08^*	6567.13^#	6093.33^#	0.001

Markers	Control group	S model group	S model+10% TCM group	S model+25% TCM group	S model+50% TCM group	P value
IL-6 (ng/L)	20.24	33.01^*	29.81^*#	27.95^*#	20.01^#	<0.001
IL-10 (ng/L)	428.32	435.40	435.54	435.38	418.36	0.762
IL-13 (ng/L)	41.33	27.17^*	30.75^*#	33.67^*#	35.04^*#	<0.001
TNF-α (ng/L)	212.14	309.29^*	277.51^*	263.84	222.83	<0.001
NF-κB (ng/L)	215.69	187.85	183.57	206.98	213.29	0.154
TGF-β1 (ng/L)	285.49	156.97^*	185.03^*#	196.94^*#	272.26^#	<0.001

Cells	Markers	Control group	S model group	S model+10% TCM group	S model+25% TCM group	S model+50% TCM group	P value
HsIMECs	IL-6 (ng/L)	22.40	30.49^*	27.62^*	25.39^#	22.11^#	<0.001
	IL-10 (ng/L)	348.14	307.22^*	318.12^*	327.84^*#	339.72^#	0.001
	IL-13 (ng/L)	30.47	30.59	28.49	31.09	29.16	0.757
	TNF-α (ng/L)	121.90	273.29^*	271.30^*	234.02^*	202.70^*#	<0.001
	NF-κB (ng/L)	178.52	177.08	175.88	178.74	180.86	0.859
	TGF-β1 (ng/L)	149.71	146.78	143.00	141.65	145.37	0.986
HCoEpiCs	IL-6 (ng/L)	24.13	28.28^*	25.26^#	23.78^#	21.95^#	0.001
	IL-10 (ng/L)	372.10	384.02	378.86	388.56	388.92	0.910
	IL-13 (ng/L)	23.69	15.99^*	20.01	22.77^#	21.53^#	0.021
	TNF-α (ng/L)	168.63	286.16^*	281.62^*	230.50^*#	221.83^*#	<0.001
	NF-κB (ng/L)	133.02	131.88	134.09	126.03	134.28	0.201
	TGF-β1 (ng/L)	177.31	173.80	176.35	177.77	171.37	0.452

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A Chinese Herb Prescription “Fang-gan Decoction” Protects Against Damage to Lung and Colon Epithelial Cells Caused by the SARS-CoV-2 Spike Protein by Regulating the TGF-β/Smad2/3 and NF-κB Pathways

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