Zuogui Pill Promotes Neurite Outgrowth by Regulating OPN/ IGF-1R/PTEN and Downstream mTOR Signaling Pathway

Title:Zuogui Pill Promotes Neurite Outgrowth by Regulating OPN/ IGF-1R/PTEN and Downstream mTOR Signaling Pathway

Volume: 27

Author(s): Yan Liu, Dan Wu, Xiaohui Yan, Xinyu Xu, Jian Zhu, Changyin Li, Qinghua Feng, Li Li, Minghua Wu*Wenlei Li*

Affiliation:

• Department of Neurology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, China
• The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, 210029, China

• Department of Neurology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, China
• The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, 210029, China

Abstract: Aim and Objective: Zuogui pill (ZGP) is the traditional Chinese medicine for tonifying kidney yin. Clinical and animal studies have shown that ZGP effectively enhances neurologic impairment after ischemic stroke, which may be related to promoting neurite outgrowth. This investigation aimed to prove the pro-neurite outgrowth impact of ZGP and define the underlying molecular pathway in vitro.

Materials and Methods: The major biochemical components in the ZGP were investigated using UPLC-QTOF-MS. All-trans retinoic acid (ATRA) was employed to stimulate SH-SY5Y cells to develop into mature neurons, followed by oxygen-glucose deprivation and reoxygenation damage (OGD/R). Then the cells were supplemented with different concentrations of ZGP, and cell viability was identified by CCK-8. The neurites' outgrowth abilities were detected by wound healing test, while immunofluorescence staining of β-III-tubulin was used to label neurites and measure their length. Western blot was employed to discover the changes in protein levels.

Results: ZGP improved the cell viability of differentiated SH-SY5Y cells following OGD/R damage, according to the CCK-8 assay. Concurrently, ZGP promoted neurite outgrowth and improved neurite crossing and migration ability. Protein expression analysis showed that ZGP upregulated the expression of GAP43, OPN, p-IGF-1R, mTOR, and p-S6 proteins but downregulated the expression of PTEN protein. Blocking assay with IGF-1R specific inhibitor Linstinib suggested IGF-1R mediated mTOR signaling pathway was involved in the pro-neurite outgrowth effect of ZGP.

Conclusion: This work illustrated the molecular mechanism underpinning ZGP's action and offered more proof of its ability to promote neurite outgrowth and regeneration following ischemic stroke.

Cite this article as:

Liu Yan, Wu Dan, Yan Xiaohui, Xu Xinyu, Zhu Jian, Li Changyin, Feng Qinghua, Li Li, Wu Minghua*, Li Wenlei*, Zuogui Pill Promotes Neurite Outgrowth by Regulating OPN/ IGF-1R/PTEN and Downstream mTOR Signaling Pathway, Combinatorial Chemistry & High Throughput Screening 2024; 27 () . https://dx.doi.org/10.2174/0113862073295309240214060857