Study on the molecular mechanism of dihydromyricetin in alleviating liver cirrhosis based on network pharmacology.

Dihydromyricetin (DHM) is a bioactive flavonoid extracted from Hovenia dulcis, which has various activities. In the present study, the molecular mechanism of dihydromyricetin (DHM) in relieving liver cirrhosis was investigated through network pharmacology and experimental verification. The cell model was induced by TGF-ß1 activating the human hepatic stellate cell line (HSC; LX-2). The protein levels of α-SMA, collagen I, and collagen III and pathway-related proteins within LX-2 cells were detected using Western blot. EdU staining was conducted to detect cell proliferation. Immunofluorescence staining was performed to detect the expression levels of α-SMA and collagen I. Next, the drug targets of DHM were screened from the PubChem database. The differentially expressed genes in the liver cirrhosis dataset GSE14323 were identified. The expression of the identified drug targets in LX-2 cells was verified using qRT-PCR. The results showed that TGF-ß1 treatment notably increased LX-2 cell viability, promoted cell proliferation, and elevated α-SMA, collagen I, and collagen III protein contents. DHM treatment could partially eliminate TGF-ß1 effects, as evidenced by the inhibited cell viability and proliferation and reduced α-SMA, collagen I, and collagen III contents. After network pharmacology analysis, nine differentially expressed target genes (MMP2, PDGFRB, PARP1, BCL2L2, ABCB1, TYR, CYP2E1, SQSTM1, and IL6) in liver cirrhosis were identified. According to qRT-PCR verification, DHM could inhibit the expression of MMP2, PDGFRB, PARP1, CYP2E1, SQSTM1, and IL6, and enhance ABCB1 expression levels within LX-2 cells. Moreover, DHM inhibited mTOR and MAPK signaling pathways in TGF-ß1-induced HSCs. In conclusion, DHM could inhibit HSC activation, which may be achieved via acting on MMP2, PDGFRB, PARP1, CYP2E1, SQSTM1, IL6, and ABCB1 genes and their downstream signaling pathways, including mTOR and MAPK signaling pathway.

A putative pyruvate transporter TaBASS2 positively regulates salinity tolerance in wheat via modulation of ABI4 expression.

BACKGROUND: High salinity adversely affects crop production. Pyruvic acid is the precursor of abscisic acid (ABA) and other chemicals that are synthesized in chloroplast, some of which are involved in the response to salt. The transportation of pyruvic acid into chloroplast is mediated by pyruvate transporters. However, whether pyruvate transporters are involved in salt response has not been studied so far. Here, we answered this issue by assessing the function of a wheat pyruvate transporter in salt response. RESULTS: A pyruvate transporter TaBASS2 was isolated from salt-tolerant wheat cultivar Shanrong 3. The expression of TaBASS2 was induced by NaCl stress as well as H2O2 and ABA treatments. Constitutive expression of TaBASS2 in Arabidopsis bass2-1 mutant complemented the mevastatin-sensitive phenotype that reflects the deficiency of transporting pyruvic acid into chloroplast. Overexpression of TaBASS2 enhanced salinity tolerance and reactive oxygen species scavenging in wheat. Arabidopsis constitutively expressing TaBASS2 also exhibited enhanced tolerance to salinity and oxidative stress. In Arabidopsis, TaBASS2 repressed the expression of ABA INSENSITIVE 4 (ABI4), a node linking ABA signaling and plastid retrograde signaling pathways. However, the enhanced salinity tolerance of TaBASS2 overexpression Arabidopsis was abolished when ABI4 expression was restored to the level of wild-type through overexpressing ABI4. CONCLUSIONS: Our work demonstrates that TaBASS2 enhances salinity tolerance of plants via modulating ABI4 expression. This indicates that pyruvate transporters indeed participate in the interaction of plants with environmental stimuli.

A wheat allene oxide cyclase gene enhances salinity tolerance via jasmonate signaling.

One of the two branches of the α-linolenic acid metabolism pathway is catalyzed by 12-oxo-phytodienoic acid reductase I, and the other is involved in jasmonic acid (JA) synthesis. The former is known to be active in the response to salinity tolerance in wheat (Triticum aestivum), but the participation of the latter in this response has not been established as yet. Here, the salinity-responsive bread wheat gene TaAOC1, which encodes an allene oxide cyclase involved in the α-linolenic acid metabolism pathway, was constitutively expressed in both bread wheat and Arabidopsis (Arabidopsis thaliana). In both species, transgenic lines exhibited an enhanced level of tolerance to salinity. The transgenic plants accumulated a higher content of JA and developed shorter roots. Both the shortened roots and the salinity tolerance were abolished in a background lacking a functional AtMYC2, a key component of the JA and abscisic acid signaling pathway, but were still expressed in a background deficient with respect to abscisic acid synthesis. We provide the first evidence, to our knowledge, suggesting that JA is also involved in the plant salinity response and that the α-linolenic acid metabolism pathway has a regulatory role over this response.

Isolation and characterization of a bread wheat salinity responsive ERF transcription factor.

A screen conducted on both a suppression subtractive hybridization and a full length cDNA library made from a salinity tolerant bread wheat cultivar SR3 (Triticum aestivum cv. SR3) resulted in the recognition of TaERF4, a gene including both an AP2/ERF domain and a nuclear localization signal. The 982 bp TaERF4 cDNA comprised a 582 bp open reading frame, encoding a 193 residue polypeptide of molecular weight 20 kDa and calculated pI 8.48. A TaERF4-GFP fusion protein localized preferentially to the nuclei of Arabidopsis thaliana protoplasts. TaERF4 is a member of the B-1 group within the ERF sub-family and was not transactivatable in yeast. The presence of an ERF-associated amphiphilic repression (EAR) motif at its C-terminus suggests that TaERF4 is probably a transcription repressor. TaERF4 was inducible by exposure to salinity and osmotic stresses, but not to exogenously supplied abscisic acid (ABA). The heterologous constitutive expression of TaERF4 in Arabidopsis enhanced the level of sensitivity to salinity stress, possibly via the repression of tonoplast Na(+)/H(+) antiporter activity. There was no phenotype associated with the transgene's presence when plants were subjected to either osmotic stress or ABA treatment. TaERF4 appears to be a transcription repressor acting within the ABA-independent response to salinity stress.

TaCHP: a wheat zinc finger protein gene down-regulated by abscisic acid and salinity stress plays a positive role in stress tolerance.

The plant response to abiotic stresses involves both abscisic acid (ABA)-dependent and ABA-independent signaling pathways. Here we describe TaCHP, a CHP-rich (for cysteine, histidine, and proline rich) zinc finger protein family gene extracted from bread wheat (Triticum aestivum), is differentially expressed during abiotic stress between the salinity-sensitive cultivar Jinan 177 and its tolerant somatic hybrid introgression cultivar Shanrong No.3. TaCHP expressed in the roots of seedlings at the three-leaf stage, and the transcript localized within the cells of the root tip cortex and meristem. TaCHP transcript abundance was higher in Shanrong No.3 than in Jinan 177, but was reduced by the imposition of salinity or drought stress, as well as by the exogenous supply of ABA. When JN17, a salinity hypersensitive wheat cultivar, was engineered to overexpress TaCHP, its performance in the face of salinity stress was improved, and the ectopic expression of TaCHP in Arabidopsis (Arabidopsis thaliana) also improved the ability of salt tolerance. The expression level of a number of stress reporter genes (AtCBF3, AtDREB2A, AtABI2, and AtABI1) was raised in the transgenic lines in the presence of salinity stress, while that of AtMYB15, AtABA2, and AtAAO3 was reduced in its absence. The presence in the upstream region of the TaCHP open reading frame of the cis-elements ABRE, MYBRS, and MYCRS suggests that it is a component of the ABA-dependent and -independent signaling pathways involved in the plant response to abiotic stress. We suggest that TaCHP enhances stress tolerance via the promotion of CBF3 and DREB2A expression.