LncRNA TUG1 Knockdown Reduces Cardiomyocyte Damage in Viral Myocarditis by Targeting the miR-140-3p/CXCL8 Axis

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  Figure 1.. High expression of LncRNA TUG1 in VMC. A: HE staining of heart tissue (×200). B: TUNEL staining of heart tissue (×200). C: CCK-8 measured cell proliferation. D: Flow cytometry measured apoptosis rate. E, F: MDA, SOD, and ROS in heart tissue of mice and HL-1 cells. G, H: ELISA checked IFN-γ, IL-6, and IL-1β in mouse heart tissue and HL-1 cells. I, J: Western blot evaluated p65 and p-p65 in mouse heart tissue and HL-1 cells. K, L: Western blot evaluated Bcl-2, Bax, and Cleaved Caspase-3 in mouse heart tissue and HL-1 cells. M: RT-qPCR measured LncRNA TUG1 in heart tissue. N: RT-qPCR measured LncRNA TUG1 expression in HL-1. Data were expressed as mean ± SD (n = 3). *P < .05. *P < .05 vs. control mice.

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  Figure 2.. Silencing LncRNA TUG1 alleviates VMC. A, B: RT-qPCR measured LncRNA TUG1 expression in heart tissue and HL-1 cells after LncRNA TUG1 knockdown. C: HE staining of heart tissue (×200). D: TUNEL staining of heart tissue (×200). E: ELISA checked IFN-γ, IL-6, and IL-1β in mouse heart tissue. F: Western blot evaluated p65 and p-p65 in mouse heart tissue. G: Western blot evaluated Bcl-2, Bax, cleaved caspase-3 in mouse heart tissue. H: MDA, SOD, and ROS in heart tissue of mice. I: CCK-8 measured cell proliferation. J: Flow cytometry measured apoptosis rate. E, F: MDA, SOD, and ROS in heart tissue of mice and HL-1 cells. K: ELISA checked IFN-γ, IL-6, and IL-1β in HL-1 cells. L: Western blot evaluated p65 and p-p65 in HL-1 cells. M: Western blot evaluated Bcl-2, Bax, and Cleaved Caspase-3 in HL-1 cells. N: MDA, SOD, and ROS in HL-1 cells. Data were expressed as mean ± SD (n = 3). *P < .05. *P < .05 vs. control mice.

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  Figure 3.. LncRNA TUG1 negatively regulates miR-140-3p expression. A: Binding sites between LncRNA TUG1 and miR-140-3p predicted by StarBase. B, C: RT-qPCR measured miR-140-3p in the myocardium of VMC mice. D, E: RNA pull-down and dual luciferase reporter assay measured the targeting relationship between LncRNA TUG1 and miR-140-3p. *P < .05 vs. NC mimic. F: RT-qPCR measured miR-140-3p in cells. Data were expressed as mean ± SD (n = 3). *P < .05.

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  Figure 4.. LncRNA TUG1 is involved in VMC by regulating miR-140-3p. A: RT-qPCR measured LncRNA TUG1 and miR-140-3p in VMC cells. B: CCK-8 measured cell proliferation. C: Flow cytometry measured apoptosis rate. D: ELISA checked IFN-γ, IL-6, and IL-1β in HL-1 cells. E: Western blot evaluated p65 and p-p65 in HL-1 cells. F: Western blot evaluated Bcl-2, Bax, and Cleaved Caspase-3 in HL-1 cells. G: MDA, SOD, and ROS in HL-1 cells. Data were expressed as mean ± SD (n = 3). *P < .05.

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  Figure 5.. Knocking down LncRNA TUG1 can up-regulate the inhibition of CXCL8 expression by miR-140-3p. A: Binding sites between CXCL8 and miR-140-3p predicted by StarBase. B: RT-qPCR measured CXCL8 in mouse heart tissue and HL-1 cells. C: Western blot evaluated CXCL8 in mouse heart tissue and HL-1 cells. D: Targeting relationship between CXCL8 and miR-140-3p in the dual luciferase reporter gene assay. E, F: RT-qPCR detection and Western blot evaluated CXCL8 after altering miR-140-3p. G: RT-qPCR measured miR-140-3p and CXCL8 in cells after si-TUG1 treatment. H: Western blot evaluated CXCL8 after si-TUG1 treatment. Data were expressed as mean ± SD (n = 3). *P < .05.

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  Figure 6.. LncRNA TUG1 influences VMC by regulating the miR-140-3p/CXCL8 axis. A: RT-qPCR measured LncRNA TUG1 and CXCL8 in VMC cells. B: CCK-8 measured cell proliferation. C: Flow cytometry measured apoptosis rate. D: ELISA checked IFN-γ, IL-6, and IL-1β in HL-1 cells. E: Western blot evaluated p65 and p-p65 in HL-1 cells. F: Western blot evaluated Bcl-2, Bax, and Cleaved Caspase-3 in HL-1 cells. G: MDA, SOD, and ROS in HL-1 cells. Data were expressed as mean ± SD (n = 3). *P < .05.