LncRNA Meg3 expression level is negatively correlated with liver fibrosis severity in patients with Wilson disease

doi:10.12122/j.issn.1673-4254.2025.11.09

Abstract

Abstract:

Objective To investigate the expression of the long non-coding RNA maternally expressed gene 3 (LncRNA Meg3) in patients with the Wilson disease (WD) and its correlation with the severity of liver fibrosis and autophagy-related markers. Methods A total of 100 WD patients and 50 healthy individuals were enrolled from the First Affiliated Hospital of Anhui University of Chinese Medicine. Serum biomarkers, including platelet count, hyaluronic acid (HA), laminin (LN), type III procollagen N-terminal peptide (PIIINP), type IV collagen (C‑IV), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured, and the non-invasive indices APRI and FIB-4 were calculated. Peripheral blood levels of LncRNA Meg3, Beclin-1 and LC3B were detected using RT-qPCR, and liver stiffness (LSM) and shear wave velocity (SWV) were evaluated using two-dimensional shear wave elastography (2D-SWE). The liver tissues from 10 WD patients and 10 patients with hepatic hemangioma were examined using histochemical staining, transmission electron microscopy, and RT-qPCR. Results The expression level of LncRNA Meg3 was significantly lower, while the levels of AST, ALT, HA, LN, PIIINP, C‑IV, APRI, FIB-4, LSM and SWV were significantly higher in WD patients than in the healthy individuals (all P<0.01). LncRNA Meg3 was negatively correlated with LSM, SWV, APRI, FIB-4, Beclin-1 and LC3B (P<0.05). ROC analysis demonstrated that LncRNA Meg3 effectively discriminated >F4 stage fibrosis (AUC=0.902) with a sensitivity of 92.9% and a specificity of 83.7% at the optimal cut-off value, outperforming APRI (AUC=0.746) and FIB-4 (AUC=0.661). The liver tissues from WD patients exhibited characteristic histopathological changes and lowered expression of LncRNA Meg3, which was negatively correlated with Beclin-1 and LC3B expressions (P<0.05). Liver fibrosis staging (7 S4 cases and 3 S3 cases) was significantly associated with LSM and SWV levels (P<0.05). Conclusion The expression level of LncRNA Meg3 is significantly decreased in WD patients, which is negatively correlated with the severity of liver fibrosis and closely related to the level of autophagy.

Key words: Wilson disease, LncRNA Meg3, liver fibrosis, autophagy

Daiping HUA, Qiaoyu XUAN, Lanting SUN, Qingsheng YU, Qin WANG, Tao WANG, Qiyan MA, Wenming YANG, Han WANG. LncRNA Meg3 expression level is negatively correlated with liver fibrosis severity in patients with Wilson disease[J]. Journal of Southern Medical University, 2025, 45(11): 2365-2374.

Figures/Tables 11

Tab.1 Baseline data and observation indexes in WD and control groups

Variable	WD group (n=100)	Control group (n=50)	χ² /z/t	P
Male [n (%)]	61 (61.00)	29 (58.00)	0.125	0.724
Age (year)	26.00 (22.00, 35.00)	25.00 (23.00, 31.00)	-0.647	0.518
BMI (kg/m²)	21.62 (20.21, 22.69)	21.05 (20.28, 22.64)	-0.451	0.652
ALT (U/L)	30.00 (20.45, 54.20)	16.70 (14.40, 22.40)	-5.132	<0.001
AST (U/L)	30.40 (22.15, 42.15)	19.00 (16.40, 22.60)	-6.127	<0.001
APRI	0.53 (0.33, 0.78)	0.32 (0.28, 0.39)	-5.482	<0.001
FIB-4	0.98 (0.56, 1.44)	0.52 (0.39, 0.58)	-5.753	<0.001
HA (ng/mL)	108.07 (66.65, 175.39)	67.47 (54.74, 83.28)	-5.116	<0.001
LN (ng/mL)	112.12 (84.56, 160.35)	80.77 (76.59, 90.46)	-4.956	<0.001
PⅢNP (ng/mL)	16.20 (10.01, 26.36)	9.62 (7.90, 10.33)	-5.759	<0.001
C-Ⅳ (ng/mL)	72.31 (53.44, 100.65)	48.70 (38.52, 59.83)	-5.512	<0.001
LSM (kPa)	10.17 (7.93, 13.82)	5.20 (3.81, 5.77)	-9.217	<0.001
SWV (m/s)	1.85 (1.62, 2.11)	1.26 (1.09, 1.32)	-9.603	<0.001
LncRNA Meg3	0.59 (0.11, 1.42)	1.20 (0.56, 1.84)	3.700	<0.001
Beclin-1	1.18 (0.72, 1.61)	1.11 (0.95, 1.30)	-0.339	0.735
LC3B	1.35 (0.54, 2.02)	1.17 (0.85, 1.45)	-0.797	0.425

Fig.1 Flow chart of the study.

Fig.2 Scatter plot diagram showing the correlation of LncRNA Meg3 in peripheral blood of WD patients with liver fibrosis indexes and Beclin-1 and LC3B. LncRNA Meg3 in peripheral blood was negatively correlated with HA (A), LN (B), PⅢNP (C), C-Ⅳ (D), APRI (E), FIB-4 (F), LSM (G), SWV (H), Beclin-1 (I) and LC3B (J).

Tab.2 Multiple linear regression analysis of peripheral blood LncRNA Meg3 and the continuous variables in WD patients

Variable	β	t	P
HA	-0.140	-1.376	0.172
LN	-0.072	-0.710	0.480
PⅢNP	-0.032	-0.308	0.758
C-Ⅳ	-0.125	-1.240	0.218
APRI	-0.231	-2.340	0.021
FIB-4	-0.276	-2.566	0.012
LSM	-0.569	-6.441	<0.001
SWV	0.572	-6.500	<0.001
Beclin-1	-0.402	-4.321	<0.001
LC3B	-0.278	-2.821	0.006

Fig.3 ROC curves for peripheral blood LncRNA Meg3, FIB-4, and APRI in WD patients. The AUC for LncRNA Meg3 was 0.902 (95% CI: 0.835-0.969), significantly higher than that of FIB-4 (AUC=0.661) and APRI (AUC=0.746).

Fig.4 2D-SWE liver fibrosis stage and observation index analysis box plot of WD patients.Histograms are shown for HA (A), LN (B), PⅢNP (C), C-Ⅳ (D), APRI (E), FIB-4 (F), LncRNA Meg3 (G), Beclin-1 (H) and LC3B (I) for different 2D-SWE liver fibrosis stages. *P<0.05, **P<0.01.

Fig.5 HE (A), Masson (B) and copper staining (C) of the liver tissues from WD and healthy control (HC) groups (scale bar=50 μm).

Fig.6 Transmission electron microscopy of the liver tissues in WD and HC groups (A: original magnification: ×10 000; B: ×25 000). Red arrows indicate the autophagosomes.

Fig.7 Box plots of the expression levels of LncRNA Meg3 (A), Beclin-1 (B) and LC3B (C) in the liver tissues in WD and HC groups. *P<0.05.

Fig.8 Scatter plots for correlation analysis of LncRNA Meg3 with Beclin-1 (A) and LC3B (B) in the liver tissue of WD group.

Fig.9 Box plot of LSM (A) and SWV (B) levels in WD patients with different liver fibrosis stages. *P<0.05

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