TY - JOUR
T1 - Neuroprotective effect of He-Ying-Qing-Re formula on retinal ganglion cell in diabetic retinopathy
AU - Zhang, Cheng
AU - Xu, Yu
AU - Tan, Hor Yue
AU - Li, Sha
AU - Wang, Ning
AU - Zhang, Yinjian
AU - Feng, Yibin
N1 - Funding Information:
The study was partially financial supported by “Fan Ophthalmology of Traditional Chinese Medicine” (Project code: ZY3-CCCX-1-2004), scientific and technological research project of Shanghai Municipal Science and Technology Commission (Project code: 17401970700), grants from the research council of the University of Hong Kong (Project codes: 104003422, 104004092, 104004460), the Research Grants Committee (RGC) of Hong Kong, HKSAR (Project codes: 766211, 17152116), Gala Family Trust (Project code: 200007008), and Government-Matching Grant Scheme (Project code: 207060411).
Publisher Copyright:
© 2017 The Authors.
PY - 2018/3/25
Y1 - 2018/3/25
N2 - Ethnopharmacology relevance: He-Ying-Qing-Re Formula (HF) was empirically modified from Si-Miao-Yong-An Decoction (SD), which was recorded in the literature of Divine Doctor's Secret Transmission, and has been utilized for centuries to treat vasculopathy through clearing heat and accelerating bloodstream. HF has been used as an effective holistic treatment of diabetic retinopathy (DR) for decades and experimentally reported to ameliorate retinal condition in diabetic mice.Aim of the study: Our study aims to investigate the effect of HF in preventing sustained hyperglycemia and hyperlipidemia-associated retinal ganglion cell (RGC) cell death and its possible mechanism.Materials and methods: Chromatographic fingerprint of HF was obtained upon the UPLC-based analytic system; Diabetic retinopathy was established in streptozotocin (STZ) injection-induced hyperglycemic mice; Alterations of retinal structure was assayed by H&E staining. Expression of PSD-95 and CHOP in retinae was assessed by immunofluorescence; RGC cell line (mRGC) was used for in vitro study. Cell death was analyzed by flow cytometry; Intracellular reactive oxygen species (ROS) was measured by 2′,7′-dichlorofluorescin diacetate (DCFDA); Apoptosis-related proteins and signaling were monitored with immunoblotting and colorimetric assay.Results: Chlorogenic acid, ferulic acid, and rutin were identified in HF. HF attenuates the loss of RGCs, thinning of inner retinal layers in diabetic mice. Furthermore, expressions of Brn3a and PSD-95 were restored while CHOP level was downregulated upon HF treatment. In vitro study, HF alleviates H2O2-induced apoptosis of mRGCs and loss of postsynaptic protein via scavenging ROS and suppressing ATF4/CHOP-mediated endoplasmic reticulum stress and mitochondria-related pro-apoptotic factors, probably as cleaved-caspase-3, and phospho-p38 mitogen-activated protein kinase (MARK). Meanwhile, both pro-survival protein levels like Bcl-2/Bcl-xL and postsynaptic protein of PSD-95 were upregulated upon HF treatment.Conclusion: HF administration was a valid therapeutic approach for DR treatment, oriented at the blockade of endoplasmic reticulum- and mitochondria-dependent oxidative stress-induced retinal neurodegeneration including RGC apoptosis.
AB - Ethnopharmacology relevance: He-Ying-Qing-Re Formula (HF) was empirically modified from Si-Miao-Yong-An Decoction (SD), which was recorded in the literature of Divine Doctor's Secret Transmission, and has been utilized for centuries to treat vasculopathy through clearing heat and accelerating bloodstream. HF has been used as an effective holistic treatment of diabetic retinopathy (DR) for decades and experimentally reported to ameliorate retinal condition in diabetic mice.Aim of the study: Our study aims to investigate the effect of HF in preventing sustained hyperglycemia and hyperlipidemia-associated retinal ganglion cell (RGC) cell death and its possible mechanism.Materials and methods: Chromatographic fingerprint of HF was obtained upon the UPLC-based analytic system; Diabetic retinopathy was established in streptozotocin (STZ) injection-induced hyperglycemic mice; Alterations of retinal structure was assayed by H&E staining. Expression of PSD-95 and CHOP in retinae was assessed by immunofluorescence; RGC cell line (mRGC) was used for in vitro study. Cell death was analyzed by flow cytometry; Intracellular reactive oxygen species (ROS) was measured by 2′,7′-dichlorofluorescin diacetate (DCFDA); Apoptosis-related proteins and signaling were monitored with immunoblotting and colorimetric assay.Results: Chlorogenic acid, ferulic acid, and rutin were identified in HF. HF attenuates the loss of RGCs, thinning of inner retinal layers in diabetic mice. Furthermore, expressions of Brn3a and PSD-95 were restored while CHOP level was downregulated upon HF treatment. In vitro study, HF alleviates H2O2-induced apoptosis of mRGCs and loss of postsynaptic protein via scavenging ROS and suppressing ATF4/CHOP-mediated endoplasmic reticulum stress and mitochondria-related pro-apoptotic factors, probably as cleaved-caspase-3, and phospho-p38 mitogen-activated protein kinase (MARK). Meanwhile, both pro-survival protein levels like Bcl-2/Bcl-xL and postsynaptic protein of PSD-95 were upregulated upon HF treatment.Conclusion: HF administration was a valid therapeutic approach for DR treatment, oriented at the blockade of endoplasmic reticulum- and mitochondria-dependent oxidative stress-induced retinal neurodegeneration including RGC apoptosis.
KW - Diabetic retinopathy
KW - Endoplasmic reticulum stress
KW - He-Ying-Qing-Re formula
KW - Retinal degeneration
KW - RGC apoptosis
UR - http://www.scopus.com/inward/record.url?scp=85038384685&partnerID=8YFLogxK
U2 - 10.1016/j.jep.2017.12.018
DO - 10.1016/j.jep.2017.12.018
M3 - Journal article
C2 - 29253613
AN - SCOPUS:85038384685
SN - 0378-8741
VL - 214
SP - 179
EP - 189
JO - Journal of Ethnopharmacology
JF - Journal of Ethnopharmacology
ER -
