Regulatory effect of a Chinese herbal medicine formula on non-alcoholic fatty liver disease

doi:10.3748/wjg.v25.i34.5105

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Basic Study

World J Gastroenterol. Sep 14, 2019; 25(34): 5105-5119
Published online Sep 14, 2019. doi: 10.3748/wjg.v25.i34.5105

Table 1 Primers used for RT-qPCR

Gene	Forward	Reverse
ZO-1	5’- CTGATGGTGTTCTGCCAAATTC – 3’	5' - GTCGCAAACCCACACTATCT - 3'
Occludin	5’- CCATCTGACTATGCGGAAAGAG – 3’	5' - TACCAGAGGCGGTGACTTAT - 3'
β-actin	5’ - ACCGTGAAAAGATGACCCAGAT – 3’	5' - CCAGAGGCATACAGGGACAA - 3'

ZO-1: Zonula occludens-1.

Table 2 Liver coefficient (mean ± SD)

Group	n	Liver weight (g)	Body weight (g)	Liver coefficient (%)
Control	8	11.56 ± 2.42	540.43 ± 90.11	2.13 ± 0.11
Model	8	15.35 ± 2.08	451.09 ± 43.89	3.41 ± 0.43^b
PH	8	10.81 ± 1.00	456.48 ± 42.41	2.37 ± 0.08^d
CHM	8	13.21 ± 2.18	437.81 ± 47.93	3.01 ± 0.21^b^d^f

^aP < 0.05,

^bP < 0.01 vs control group; ^cP < 0.05,

^dP < 0.01 vs model group; ^eP < 0.05,

^fP < 0.01 vs PH group. PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 3 Hepatocyte lipid droplet cavity area ratio (mean ± SD)

Group	n	Area ratio (%)
Control	5	0.00 ± 0.00
Model	5	72.15 ± 1.72
PH	5	31.01 ± 7.53^a
CHM	5	48.69 ± 5.85^a

^aP < 0.05 vs model group. PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 4 Effect of Chinese herbal medicine on blood lipid levels (mean ± SD, n = 8)

Group	TG (mmol/L)	TC (mmol/L)	LDL (mmol/L)	HDL (mmol/L)
Control	0.48 ± 0.20	1.67 ± 0.26	0.24 ± 0.08	0.95 ± 0.10
Model	0.83 ± 0.12^b	3.05 ± 0.67^b	1.38 ± 0.33^b	0.54 ± 0.05^b
PH	0.50 ± 0.08^d	1.69 ± 0.29^d	0.76 ± 0.28^b^d	0.61 ± 0.05^b^c
CHM	0.53 ± 0.12^d	2.04 ± 0.37^d	0.84 ± 0.16^b^d	0.72 ± 0.05^b^d^f

^bP < 0.01 vs control group;

^cP < 0.05,

^dP < 0.01 vs model group;

^fP < 0.01 vs PH group. PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine; TC: Total cholesterol; TG: Triglyceride; HDL: High density lipoprotein; LDL: Low density lipoprotein.

Table 5 Effect of Chinese herbal medicine on triglyceride levels in liver tissue (mean ± SD)

Group	n	TG (mmol/L)
Control	8	1.37 ± 0.67
Model	8	4.81 ± 0.72^b
PH	8	3.41 ± 0.43^b^d
CHM	8	3.41 ± 0.66^b^d

^bP < 0.01 v control group;

^dP < 0.01 vs model group. TG: Triglyceride; PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 6 Effect of Chinese herbal medicine on blood glucose levels (mean ± SD)

Group	n	Blood sugar (mmol/L)
Control	8	4.09 ± 0.26
Model	8	4.90 ± 0.33^b
PH	8	4.11 ± 0.20^d
CHM	8	4.28 ± 0.21^d

^bP < 0.01 vs control group;

^dP < 0.01 vs model group. PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 7 Effect of Chinese herbal medicine on serum liver function markers (mean ± SD)

Group	n	AST (U/L)	ALT (U/L)
Control	8	118.73 ± 13.71	39.91 ± 6.50
Model	8	238.13 ± 39.38^b	59.28 ± 9.69^b
PH	8	137.94 ± 5.83^d	48.84 ± 8.28^a^c
CHM	8	148.19 ± 20.59^a^d	49.36 ± 5.13^a^c

^aP < 0.05,

^bP < 0.01 vs control group;

^cP < 0.05,

^dP < 0.01 vs model group. PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase.

Table 8 Effect of Chinese herbal medicine on liver tissue TNF-α, TGF-β1, NF-kB, and TLR4 levels (mean ± SD)

Group	n	TNF-α (ng/mg)	TGF-β1 (ng/mg)	NF-kB (ng/mg)	TLR4 (ng/mg)
Control	8	5.35 ± 0.80	2.21 ± 0.43	10.87 ± 2.45	21.32 ± 1.87
Model	8	7.22 ± 0.81^b	3.01 ± 0.22^b	15.85 ± 2.50^b	30.28 ± 4.38^b
PH	8	5.48 ± 0.38^d	2.24 ± 0.26^d	12.15 ± 3.13^c	22.56 ± 3.02^d
CHM	8	6.01 ± 0.97^d	2.29 ± 0.40^d	12.57 ± 3.30	23.88 ± 3.21^d

^bP < 0.01 vs control group;

^cP < 0.05,

^dP < 0.01 vs model group. TGF-β1: Transforming growth factor-β1; TNF-α: Tumor necrosis factor-α; TLR4: Toll-like receptor-4; NF-kB: Nuclear factor-kappa B; PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 9 Intestinal sIgA levels (mean ± SD)

Group	n	sIgA (μg/mg)
Control	8	19.80 ± 2.25
Model	8	11.39 ± 1.81^b
PH	8	17.06 ± 2.92^a^d
CHM	8	12.78 ± 2.36^bf

^aP < 0.05,

^bP < 0.01 vs control group;

^dP < 0.01 vs model group; ^efP < 0.01 vs PH group. sIgA: Secreted immunoglobulin A; PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 10 Occludin and zonula occludens-1 protein levels (mean ± SD)

Group	n	Occludin	ZO-1
Control	7	1.12 ± 0.06	1.41 ± 0.03
Model	7	0.66 ± 0.07^b	0.82 ± 0.09^b
PH	7	1.04 ± 0.09^d	1.32 ± 0.04^d
CHM	7	0.68 ± 0.05^b^f	0.84 ± 0.12^b^f

^bP < 0.01 vs control group;

^dP < 0.01 vs model group;

^fP < 0.01 vs PH group. ZO-1: Zonula occludens-1; PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 11 Occludin and zonula occludens-1 mRNA expression levels (mean ± SD)

Group	n	Occludin mRNA	ZO-1 mRNA
Control	6	1.40 ± 0.19	1.31 ± 0.29
Model	6	0.60 ± 0.16^b	0.60 ± 0.11^b
PH	6	1.15 ± 0.24^a^d	1.15 ± 0.23^d
CHM	6	0.70 ± 0.13^b^f	0.79 ± 0.31^b^e

^aP < 0.05,

^bP < 0.01 vs control group;

^dP < 0.01 vs model group;

^eP < 0.05,

^fP < 0.01 vs PH group. ZO-1: Zonula occludens-1; PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Table 12 Levels of plasma endotoxin (mean ± SD)

Group	n	Endotoxin (%)
Control	8	0.028 ± 0.013
Model	8	0.160 ± 0.057^b
PH	8	0.045 ± 0.012^d
CHM	8	0.056 ± 0.018^d

^bP < 0.01 vs control group;

^dP < 0.01 vs model group. PH: Pioglitazone hydrochloride; CHM: Chinese herbal medicine.

Citation: Yang JM, Sun Y, Wang M, Zhang XL, Zhang SJ, Gao YS, Chen L, Wu MY, Zhou L, Zhou YM, Wang Y, Zheng FJ, Li YH. Regulatory effect of a Chinese herbal medicine formula on non-alcoholic fatty liver disease. World J Gastroenterol 2019; 25(34): 5105-5119
URL: https://www.wjgnet.com/1007-9327/full/v25/i34/5105.htm
DOI: https://dx.doi.org/10.3748/wjg.v25.i34.5105