Experimental Study on the Protective Effects of Ziyin Mingmu Decoction on Retinal Injury in Dry AMD Rats via Regulation of Inflammation and Autophagy
OBJECTIVE To investigate the protective effects of Ziyin Mingmu Decoction(ZYMM)on retinal injury in dry age-related macular degeneration(dAMD)rats through the regulation of inflammation and autophagy.METHODS A dAMD rat model was established using a light damage device.Thirty-six successfully modeled rats were randomly divided into the model group(MG),vitamin E group(VE),and ZYMM group,with a control group(CG)set up separately,each consisting of 12 rats.Rats in the VE group were administered vitamin E solution intragastrically,while the ZYMM group received ZYMM Decoction intragastrically.The CG and MG groups were administered an equivalent volume of 0.9% sodium chloride injection intragastrically,continuously for two weeks.After the last administration,the rats'serum and retina were collected.Hematoxylin-eosin(HE)staining was used to observe retinal morphology;Enzyme-linked immunosorbent assay(ELISA)was used to detect serum levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α);Real-time fluorescent quantitative PCR(qRT-PCR)was used to detect mRNA expressions of nuclear factor-kappa B(NF-κB),microtubule-associated protein light chain 3B(LC3B),and Beclin-1 in the retina.Immunohistochemistry and Western Blot were used to detect retinal protein expressions of NF-κ B,LC3B,and Beclin-1.RESULTS(1)Retinal morphology:MG rats exhibited blurred boundaries between retinal layers,reduced thickness,unclear outer nuclear layer structure,and disordered cell arrangement.The ZYMM and VE groups showed reduced disorder in retinal structure and less edema compared to the MG group.(2)Inflammatory factors:Serum IL-1β and TNF-α levels in the MG group were higher than those in the CG group(tIL-1β=6.277,tTNF-α=9.085,both P=0.000).The VE and ZYMM groups showed lower serum IL-1β and TNF-α levels compared to the MG group(VE:tIL-1β=5.821,tTNF-α=7.574,both P=0.000;ZYMM:tIL-1β=4.528,tTNF-α=5.996,both P=0.000).(3)mRNA:Retinal mRNA expressions of NF-κB,LC3B,and Beclin-1 in the MG group were higher than those in the CG group(tNF-κB=4.776,tLC3B=5.423,tBeclin-1=4.854,all P=0.000).The VE and ZYMM groups had lower retinal mRNA expressions of NF-κB,LC3B,and Beclin-1 compared to the MG group(VE:tNF-κB=3.444,P=0.002;tLC3B=4.092,P=0.000;tBeclin-1=2.841,P=0.008;ZYMM:tNF-κB=2.655,P=0.013;tLC3B=3.539,P=0.001;tBeclin-1=2.481,P=0.009).(4)Protein:Retinal protein expressions of p-NF-κB,LC3B,and Beclin-1 in the MG group were higher than those in the CG group(tp-NF-κB=6.446,P=0.000;tLC3B=5.543,P=0.001;tBeclin-1=9.075,P=0.000).VE group retinal LC3B and Beclin-1 expressions were lower than those in the MG group(tLC3B=2.717,P=0.026;tBeclin-1=5.936,P=0.000).The ZYMM group retinal p-NF-κB and Beclin-1 protein expressions were higher than those in the MG group(tp-NF-κB=4.754,P=0.001;tBeclin-1=7.394,P=0.000).CONCLUSIONS Ziyin Mingmu Decoction can protect the retina in dry AMD rats by reducing inflammation and autophagy levels.
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