ABSTRACT
Aim To explore the effect of berberine (B E) on RSV infected HEp-2 cells and the related mechanism. Methods HEp-2 cells were infected with RSV and treated with BE. Cell viability was assessed using the CCK-8 assay. Protein expression levels of NLRP3, ASC, caspase-1, PINK1, Parkin, Beclinl, p62, LC3 I,LC3 II,and BNIP3 in HEp-2 cells were detected by Western blot. The secretion level of IL-1 p in HEp-2 cells was measured using ELISA. Apoptosis rate and mitochondrial membrane potential of HEp-2 cells were examined by flow cytometry. Mitochondrial ROS (mtROS) in HEp-2 cells was detected through MitoSOX staining. Colocalization of mitochondria and autophagosomes in HEp-2 cells was investigated using immunofluorescence staining. Cyclosporin A was used for validation experiments. Results BE could significantly improve the activity of RSV-infected HEp-2 cells,reduce the apoptosis rate (P < 0. 05), and decrease the activation level of NLRP3 inflammasomes and IL-lp level (P <0. 05); BE improved mitochondrial function by increasing mitochondrial membrane potential and ATP levels,and reduced mtROS. BE significantly promoted the colocalization of mitochondria-autophagosome in RSV infected cells, inducing PINK1/ Parkin and BNIP3 to mediate mitochondrial autophagy; cyclosporine A aggravated RSV infection. Conclusions BE has protective effects on HEp-2 cells infected by RSV. The mechanism may be related to the inhibitory effect of BE on the production of mtROS and the activation of NLRP3 inflammasomes by inducing PINK1/ Parkin and BNIP3-mediated mitochondrial autophagy.
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Aim To discuss the effect of berberine ( BE) on the activity of HSV-1 virus infected HEp-2 cells and its related molecular mechanisms.Methods Hie infected cell model was constructed and divided into control group, infection group, low concentration group ( 5 (xmol • L 1 -BE) , medium concentration group ( 10 (xmol • L '-BE) and high concentration group ( 15 (xmol • L '-BE) ) , and then incubated for 24 hours.qRT-PCR was used to determine HSV-1 infection-related genes ( gD, ICP-4, ICP-8, ICP-27 ) and mRNA expression levels of LncBNA NRAV, miR- 299-3p, RAB5C.CCK-8 method and flow cytometry were applied to detect cell viability and apoptotic rate.The expression levels of PI3K/AKT signaling pathway and JNK/p38 MAPK signaling pathway related protein were analysed by WB.Results It was found that BE j j reduced the mRNA expression of gD, ICP-4, ICP-8, anrl ICP-27, improved cell viability, and inhibited eell apoptosis.BE promoted the expression of miR-299-3p by inhibiting LncRNA NRAV and RAB5C.BE inhibited the protein expression levels of PBK/AKT signaling pathway and JNK/p38 MAPK signaling pathway proteins PI3K, AKT, JNK, and P38.Conclusions The mechanism that BE enhances the activity of HEp-2 cells after HSV-1 infection and suppresses its apoptosis may be related to LncRNA NRAV and RAB5C targeting competitive binding to miH-299-3p, inhibiting the activation of PBK/AKT signaling pathway and JNK/ p38 MAPK signaling pathway.
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Aim To investigate the potential protective function of nervonic acid (NA) on the motor disorder in mice subjected to MPTP and the underlying mechanism. Methods The PD mice model was constructed and divided into control group, model group, nervonic acid (20 mg • k g
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Aim To explore the effects of linoleic acid on the joint swelling in rats caused by rheumatoid arthritis (RA), and to reveal possible mechanism underlying the inhibitory effect of linoleic acid on TLR4/NF-κB signaling pathway in RA. Methods The RA rat model was constructed and divided into control group, model group, linoleic acid (0. 1 mL) group, linoleic acid (0.2 mL) group, linoleic acid (0.4 mL) group and methotrexate (MTX) group. After the model was successfully established, the corresponding drugs were given by gavage for seven days. The control group and the model group were given normal saline. The changes of rat joint swelling were measured; joint pathological damage was assessed by HE staining; the protein expression levels and the mRNA expression levels of TNF-α, IL-1β, IL-6 and IL-10 were determined by ELISA and qRT -PCR; the protein expression levels of TLR4 and p-p65 were determined by immunohisto-chemistry and Western blot. Results Compared with model group, linoleic acid significantly alleviated the joint swelling of RA rats; linoleic acid significantly inhibited the joint pathological damage, and markedly reduced the protein expression levels and mRNA expression levels of TNF-α, IL-1β, IL-6 and IL-10; linoleic acid inhibited the protein expression levels of TLR4 and p-p65 in the TLR4/NF-κB signaling pathway. Conclusions Linoleic acid inhibits the protein expression levels of TLR4 and p-p65 in the TLR4/NF-κB signaling pathway, thereby inhibiting the expression of inflammatory factors TNF-α, IL-1β, IL-6 and IL-10.