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Paeoniflorin Suppressed High Glucose-Induced Retinal Microglia MMP-9 Expression and Inflammatory Response via Inhibition of TLR4/NF-κB Pathway Through Upregulation of SOCS3 in Diabetic Retinopathy

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Abstract

Diabetic retinopathy (DR) is a serious-threatening complication of diabetes and urgently needed to be treated. Evidence has accumulated indicating that microglia inflammation within the retina plays a critical role in DR. Microglial matrix metalloproteinase 9 (MMP-9) has an important role in the destruction of the integrity of the blood-retinal barrier (BRB) associated with the development of DR. MMP-9 was also considered important for regulating inflammatory responses. Paeoniflorin, a monoterpene glucoside, has a potent immunomodulatory effect on microglia. We hypothesized that paeoniflorin could significantly suppress microglial MMP-9 activation induced by high glucose and further relieve DR. BV2 cells were used to investigate the effects and mechanism of paeoniflorin. The activation of MMP-9 was measured by gelatin zymography. Cell signaling was measured by western blot assay and immunofluorescence assay. High glucose increased the activation of MMP-9 in BV2 cells, which was abolished by HMGB1, TLR4, p38 MAPK, and NF-κB inhibition. Phosphorylation of p38 MAPK induced by high glucose was decreased by TLR4 inhibition in BV2 cells. Paeoniflorin induced suppressor of cytokine signaling 3 (SOCS3) expression and reduced MMP-9 activation in BV2 cells. The effect of paeoniflorin on SOCS3 was abolished by the TLR4 inhibitor. In streptozotocin (STZ)-induced diabetes mice, paeoniflorin induced SOCS3 expression and reduced MMP-9 activation. Paeoniflorin suppressed STZ-induced IBA-1 and IL-1β expression and decreased STZ-induced high blood glucose level. In conclusion, paeoniflorin suppressed high glucose-induced retinal microglia MMP-9 expression and inflammatory response via inhibition of the TLR4/NF-κB pathway through upregulation of SOCS3 in diabetic retinopathy.

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Acknowledgments

The authors acknowledge grants from the National Natural Science Foundation of China (Nos. 81173502, 30973534, and 81571069) and the Foundation of Nanjing Medical University (No. 2014NJMUZD015). Clearly state if you received funds for covering the costs to publish in open access.

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  1. Su-Hua Zhu, Bing-Qian Liu, and Mao-Juan Hao contributed equally to this work.

Authors and Affiliations

  1. Jiangsu Provincial Key Lab of Cardiovascular Diseases and Molecular Intervention, Department of Pharmacology, Nanjing Medical University, Nanjing, 210029, People’s Republic of China

    Su-Hua Zhu, Mao-Juan Hao, Cheng Qian, Peng Teng, Xiao-Wei Zhou, Liang Hu, Wen-Tao Liu & Qing-Ping Li

  2. Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu, 210029, People’s Republic of China

    Bing-Qian Liu & Zhi-Lan Yuan

  3. Department of Pharmacology, China Pharmaceutical University, Nanjing, Jiangsu, 211198, People’s Republic of China

    Yi-Xin Fan

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  1. Su-Hua Zhu
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Contributions

Qing-ping Li, Zhi-lan Yuan, Su-hua Zhu and Bing-Qian Liu designed and performed the experiments. Maojuan Hao, Yi-Xin Fan performed the immunoassays. Bingqian Liu, Cheng Qian and Wen-Tao Liu performed the western blotting analysis. Peng Teng, Xiao Wei Zhou and Liang Hu carried out the cell cultures. Zhi-lan Yuan analyzed the results. Qing-ping Li, Su-hua Zhu and Bing-Qian Liu drafted the manuscript. Qing-ping Li secured funding for the project. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Zhi-Lan Yuan or Qing-Ping Li.

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The authors declare that they have no conflicts of interest.

Funding

This work was supported by the National Natural Science Foundation of China (Nos. 81173502, 30973534, and 81571069) and the Foundation of Nanjing Medical University (No. 2014NJMUZD015).

Ethics Approval and Consent to Participate

All procedures were strictly performed in accordance with the regulations of the ethics committee of the International Association for the Study of Pain and the Guide for the Care and Use of Laboratory Animals (The Ministry of Science and Technology of China, 2006). All animal experiments were approved by the Nanjing Medical University Animal Care and Use Committee and were designed to minimize suffering and the number of animals used.

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Zhu, SH., Liu, BQ., Hao, MJ. et al. Paeoniflorin Suppressed High Glucose-Induced Retinal Microglia MMP-9 Expression and Inflammatory Response via Inhibition of TLR4/NF-κB Pathway Through Upregulation of SOCS3 in Diabetic Retinopathy. Inflammation 40, 1475–1486 (2017). https://doi.org/10.1007/s10753-017-0571-z

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