Inhibition of miRNA-1-Mediated Inflammation and Autophagy by Astragaloside IV Improves Lipopolysaccharide-Induced Cardiac Dysfunction in Rats

Figures & data

Figure 1 AS-IV chemical structure.

Table 1 List of Primers Used for Real-Time RT-PCR

Bax	Forward: TTTTTGCTACAGGGTTTCATCC
	Reverse: CCAGTTCATCGCCAATTCG
Bcl-2	Forward: GGGATGCCTTTGTGGAACTATA
	Reverse: CTTTTGCATATTTGTTTGGGGC
Beclin 1	Forward: AGGAGTTGCCGTTGTACTGTTCTG
	Reverse: TGCCTCCAGTGTCTTCAATCTTGC
P62	Forward: ATCCGCTACCTCATCTCCTCCAAC
	Reverse: GCAGACCATCCGTCACCAGATTG
LC3-II	Forward: CGTCCTGGACAAGACCAAGTTCC
	Reverse: CAGGAGGAAGAAGGCTTGGTTAGC
LC3-I	Forward: TCGCCGACCGCTGTAAGGAG
	Reverse: CGCCGGATGATCTTGACCAACTC
HSP70	Forward: ACAAGGCGGAGTCTGAGGAGATG
	Reverse: AGGTCCACGGTGCTGGTCTTC
IKBα	Forward: GGTGTTTGAATGTATTGCTGG
	Reverse: AGGCTGTTTGGCTGAGGT
IKKβ	Forward: GACTTCACCATGGAACCCGT
	Reverse: GGAGACTGCCCATTCTCGAC
TRL4	Forward: GCTCTCAACCTTGGTACTGACAGG
	Reverse: GTCTCCACAGCCACCAGATTCTC
P65	Forward: GGTGCCTCCAGTGAGAAGAACAAG
	Reverse: GCCGTCCTCACAGTGCTTGC

Figure 2 AS-IV decreases miR-1 mRNA expression in the heart tissue of LPS-treated rats.

Notes: The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 3 AS-IV improves heart dysfunction and attenuates the pathological changes of heart tissue in LPS-treated rats.

Notes: (A) Representative photograph of echocardiography. (B) EF, FS, LVIDd, and LVIDs. (C) Pathological changes, the arrows indicate inflammatory cell infiltration. The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^#p<0.05, ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 4 AS-IV reduces the levels of inflammatory cytokines in the serum of LPS-treated rats.

Notes: The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 5 AS-IV regulates the protein expression of inflammation signaling molecules in the heart tissue of LPS-treated rats.

Notes: (A and B) Protein expression of HPS70, TLR4, IKBα, IKKβ, and p65 determined by Western blotting. (C) protein expression of HPS70 and NF-κB determined by immunofluorescence (400×). The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 6 AS-IV regulates autophagy in the heart tissue of LPS-treated rats.

Notes: (A and B) protein expression of Bax, Bcl-2, Beclin 1, LC3-I, LC3-II, and P62 determined by Western blotting. (C) protein expression of LC3-I and LC3-II determined by immunofluorescence (400×). The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 7 AS-IV decreases miR-1 mRNA expression in LPS-treated heart cells.

Notes: The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 8 AS-IV regulates the protein expression of inflammation signaling molecules in LPS-treated heart cells.

Notes: (A) mRNA expression of HPS70, TLR4, IKBα, IKKβ, and p65. (B and C) Protein expression of HPS70, TLR4, IKBα, IKKβ, and p65. The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^#p<0.05, ^##p<0.01 vs LPS group; ^▲▲p<0.01 vs LPS + AS-IV group.

Figure 9 AS-IV regulates autophagy in LPS-treated heart cells.

Notes: (A) mRNA expression of Bax, Bcl-2, Beclin 1, LC3-II/LC3-I, and P62. (B and C) Protein expression of Bax, Bcl-2, Beclin 1, LC3-II/LC3-I, and P62. (D) Autophagosome found under the electron microscope as indicated of the red arrowheads. The data are presented as mean±SD. n=6. **p<0.01 vs control group; ^##p<0.01 vs LPS group; ^▲p<0.05, ^▲▲p<0.01 vs LPS + AS-IV group.