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MS14 down-regulates lipocalin2 expression in spinal cord tissue in an animal model of multiple sclerosis in female C57BL/6.

Ebrahimi-Kalan A, Soleimani Rad J, Kafami L, Mohammadnejad D, Habibi Roudkenar M, Khaki AA, Aliyari Serej Z, Mohammadi Roushandeh A - Iran. Biomed. J. (2014)

Bottom Line: MS14 significantly ameliorated EAE symptoms and decreased lymphocyte infiltration into the spinal cord (P<0.05).Our data also revealed that LCN2 expression was significantly down-regulated in acute and chronic phases of EAE both at protein and gene levels after MS14 treatment (P<0.05).The results demonstrated that MS14 regulatory effect on EAE is accompanied by LCN2 down-regulation after treatment with the herb; however, more studies are required for clarifying the other involved mechanisms.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Anatomical Sciences, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran. a.mohammadiroshandeh@umsha.ac.ir.

ABSTRACT

Background: Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis, which is a demyelinating and an inflammatory disease of central nervous system. Recent studies have established that some molecules such as Lipocaline2 (LCN2), which expresses during inflammatory conditions, play an important role in EAE pathogenesis and might involve in its treatment process. Recently, it has been proved that MS14, an herbal-marine drug, has anti-inflammatory properties through reduction of TNF-α and IL-1β. Thus, the present study investigated the effects of MS14 on the course of EAE and its relation to LCN2 expression in both protein and gene levels.

Methods: EAE was induced in female C57BL/6 mice using Hooke kits. Animals were scored for clinical signs of the disease according to a 10-point EAE scoring system. On 21st and 35th days after immunization, mice (n = 4/group) were deeply anesthetized, and the spinal cords were removed. Inflammatory cell infiltration and LCN2 expression in spinal cord were assessed by hematoxylin and eosin staining, immuno-histochemistry, and real-time PCR methods.

Results: MS14 significantly ameliorated EAE symptoms and decreased lymphocyte infiltration into the spinal cord (P<0.05). Our data also revealed that LCN2 expression was significantly down-regulated in acute and chronic phases of EAE both at protein and gene levels after MS14 treatment (P<0.05).

Conclusion: The results demonstrated that MS14 regulatory effect on EAE is accompanied by LCN2 down-regulation after treatment with the herb; however, more studies are required for clarifying the other involved mechanisms.

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Effects of MS14 on LCN2 gene expression. (A) LCN2 gene expression determined by RT-PCR in control and MS14 groups at acute and chronic phases. (A) Lane 1 (normal), lane 2 (control at acute phase), lane 3 (MS14 at acute phase), lane 4 (control at chronic phase), and lane 5 (MS14 at chronic phase). Up-regulation of LCN2 is shown in acute phase of EAE. (B) β-actin expression is shown in all groups. (C) Fold changes of LCN2 expression in both control and MS14 groups at acute and chronic phases. The level of LCN2 expression is higher at acute phase and strongly is affected by MS14 at chronic phase (*P<0.05). Asterisks show significant differences. Our results showed a significant decrease in LCN2 expression in MS14-treated group during acute phase (2.65 ± 0.34) compared to control group (5.32 ± 0.37, P<0.05). Similar results were shown in control and MS14 groups (4.34 ± 0.29 vs. 1.123 ± 0.17) during chronic phase. M, marker
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Figure 4: Effects of MS14 on LCN2 gene expression. (A) LCN2 gene expression determined by RT-PCR in control and MS14 groups at acute and chronic phases. (A) Lane 1 (normal), lane 2 (control at acute phase), lane 3 (MS14 at acute phase), lane 4 (control at chronic phase), and lane 5 (MS14 at chronic phase). Up-regulation of LCN2 is shown in acute phase of EAE. (B) β-actin expression is shown in all groups. (C) Fold changes of LCN2 expression in both control and MS14 groups at acute and chronic phases. The level of LCN2 expression is higher at acute phase and strongly is affected by MS14 at chronic phase (*P<0.05). Asterisks show significant differences. Our results showed a significant decrease in LCN2 expression in MS14-treated group during acute phase (2.65 ± 0.34) compared to control group (5.32 ± 0.37, P<0.05). Similar results were shown in control and MS14 groups (4.34 ± 0.29 vs. 1.123 ± 0.17) during chronic phase. M, marker

Mentions: MS14 down-regulated LCN2 expression. The expression of LCN2 at protein level was determined by immune-histochemical staining. Our findings showed that LCN2 was expressed in both control and MS14-treated groups. Also, LCN2 expression was dramatic-ally decreased in MS14-treated group compared to the control (Fig. 3). To evaluate LCN2 gene expression in spinal cord tissue, RT-PCR and real-time PCR were performed. Semi-quantitative expression of LCN2 was shown in all groups even in a normal mouse without any interference (lane 1, Fig. 4). RT-PCR indicated the alteration of LCN2 gene expression during acute and chronic phases of the disease in control mice. LCN2 was down-regulated in MS14-treated group compared to control in acute phase (lanes 2 and 3, Fig. 4). Similar result was found in chronic phase as well (lanes 4 and 5, Fig. 4), while LCN2 up-regulated at acute phase in both groups.


MS14 down-regulates lipocalin2 expression in spinal cord tissue in an animal model of multiple sclerosis in female C57BL/6.

Ebrahimi-Kalan A, Soleimani Rad J, Kafami L, Mohammadnejad D, Habibi Roudkenar M, Khaki AA, Aliyari Serej Z, Mohammadi Roushandeh A - Iran. Biomed. J. (2014)

Effects of MS14 on LCN2 gene expression. (A) LCN2 gene expression determined by RT-PCR in control and MS14 groups at acute and chronic phases. (A) Lane 1 (normal), lane 2 (control at acute phase), lane 3 (MS14 at acute phase), lane 4 (control at chronic phase), and lane 5 (MS14 at chronic phase). Up-regulation of LCN2 is shown in acute phase of EAE. (B) β-actin expression is shown in all groups. (C) Fold changes of LCN2 expression in both control and MS14 groups at acute and chronic phases. The level of LCN2 expression is higher at acute phase and strongly is affected by MS14 at chronic phase (*P<0.05). Asterisks show significant differences. Our results showed a significant decrease in LCN2 expression in MS14-treated group during acute phase (2.65 ± 0.34) compared to control group (5.32 ± 0.37, P<0.05). Similar results were shown in control and MS14 groups (4.34 ± 0.29 vs. 1.123 ± 0.17) during chronic phase. M, marker
© Copyright Policy
Related In: Results  -  Collection

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Figure 4: Effects of MS14 on LCN2 gene expression. (A) LCN2 gene expression determined by RT-PCR in control and MS14 groups at acute and chronic phases. (A) Lane 1 (normal), lane 2 (control at acute phase), lane 3 (MS14 at acute phase), lane 4 (control at chronic phase), and lane 5 (MS14 at chronic phase). Up-regulation of LCN2 is shown in acute phase of EAE. (B) β-actin expression is shown in all groups. (C) Fold changes of LCN2 expression in both control and MS14 groups at acute and chronic phases. The level of LCN2 expression is higher at acute phase and strongly is affected by MS14 at chronic phase (*P<0.05). Asterisks show significant differences. Our results showed a significant decrease in LCN2 expression in MS14-treated group during acute phase (2.65 ± 0.34) compared to control group (5.32 ± 0.37, P<0.05). Similar results were shown in control and MS14 groups (4.34 ± 0.29 vs. 1.123 ± 0.17) during chronic phase. M, marker
Mentions: MS14 down-regulated LCN2 expression. The expression of LCN2 at protein level was determined by immune-histochemical staining. Our findings showed that LCN2 was expressed in both control and MS14-treated groups. Also, LCN2 expression was dramatic-ally decreased in MS14-treated group compared to the control (Fig. 3). To evaluate LCN2 gene expression in spinal cord tissue, RT-PCR and real-time PCR were performed. Semi-quantitative expression of LCN2 was shown in all groups even in a normal mouse without any interference (lane 1, Fig. 4). RT-PCR indicated the alteration of LCN2 gene expression during acute and chronic phases of the disease in control mice. LCN2 was down-regulated in MS14-treated group compared to control in acute phase (lanes 2 and 3, Fig. 4). Similar result was found in chronic phase as well (lanes 4 and 5, Fig. 4), while LCN2 up-regulated at acute phase in both groups.

Bottom Line: MS14 significantly ameliorated EAE symptoms and decreased lymphocyte infiltration into the spinal cord (P<0.05).Our data also revealed that LCN2 expression was significantly down-regulated in acute and chronic phases of EAE both at protein and gene levels after MS14 treatment (P<0.05).The results demonstrated that MS14 regulatory effect on EAE is accompanied by LCN2 down-regulation after treatment with the herb; however, more studies are required for clarifying the other involved mechanisms.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Anatomical Sciences, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran. a.mohammadiroshandeh@umsha.ac.ir.

ABSTRACT

Background: Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis, which is a demyelinating and an inflammatory disease of central nervous system. Recent studies have established that some molecules such as Lipocaline2 (LCN2), which expresses during inflammatory conditions, play an important role in EAE pathogenesis and might involve in its treatment process. Recently, it has been proved that MS14, an herbal-marine drug, has anti-inflammatory properties through reduction of TNF-α and IL-1β. Thus, the present study investigated the effects of MS14 on the course of EAE and its relation to LCN2 expression in both protein and gene levels.

Methods: EAE was induced in female C57BL/6 mice using Hooke kits. Animals were scored for clinical signs of the disease according to a 10-point EAE scoring system. On 21st and 35th days after immunization, mice (n = 4/group) were deeply anesthetized, and the spinal cords were removed. Inflammatory cell infiltration and LCN2 expression in spinal cord were assessed by hematoxylin and eosin staining, immuno-histochemistry, and real-time PCR methods.

Results: MS14 significantly ameliorated EAE symptoms and decreased lymphocyte infiltration into the spinal cord (P<0.05). Our data also revealed that LCN2 expression was significantly down-regulated in acute and chronic phases of EAE both at protein and gene levels after MS14 treatment (P<0.05).

Conclusion: The results demonstrated that MS14 regulatory effect on EAE is accompanied by LCN2 down-regulation after treatment with the herb; however, more studies are required for clarifying the other involved mechanisms.

Show MeSH
Related in: MedlinePlus