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Òâ¼ûÈçÏ£º Reviewer: 1 Comments to the Author In this paper, Yu et al. demonstrated that leonurine could protect PC12 cells from Na2S2O4- and glucose deprivation-induced apoptosis. The authors found that leonurine reduced Na2S2O4- and glucose deprivation-induced ROS production and restored the loss of mitochondria membrane potential. They also found that leonurine increased VEGF, Ras and Raf expression. The findings are of potential significance. Nevertheless, several issues preclude the acceptance of authors' work for publication in JPP at this stage. Major comments: 1. The manuscript is full of typo-grammatical mistakes. 2. The author did not explain why they chose PC12 cells as the cell model and Na2S2O4 as hypoxia-inducing agent. 3. The author did not explain why the protein levels of Ras and Raf were increased after leonurine treatment. VEGF is supposed to increase the activity but not the protein levels of Ras and Raf. 4. The author did not demonstrate that leonurine actually protected PC12 cells from apoptosis "through" VEGF/Ras/Raf/Erk pathway as described in the title. The authors should neutralize VEGF with antibody and determine if such manipulation could affect the protective effect of leonurine. Minor comments: 1. ¡°OGD¡± is not a standard abbreviation and shall be fully spelled out in the title Reviewer: 2 Comments to the Author Manuscript Id: JPP-13-0647 Manuscript Title: Leonurine protects OGD-induced PC12 cells apoptosis through activation of VEGF/Ras/Raf/Erk signaling pathway. The current manuscript by Yu et al the showed the protective role of Leonurine against OGD induced PC12 cells apoptosis through activation of signaling pathway. However there are major concerns:- Authors mainly focussed to antiapoptotic activity of Leonurine. On the hand authors themselves mentioned in introduction section that leonurine, was reported to have cardioprotective and neuroprotective effect via increasing the antioxidantive and antiapoptotic effects. What difference this manuscript offer for readers? Authors used in vitro setup to show Leonurine effects and concluded that it might be applied as a potential therapeutic drug for ischemic injury treatment in abstract section. Is it justified to extrapolate in vitro preliminary results? Can leonurine cross BBB? Authors used vitamin E as positive control in some experiments but did not used for mechanistic studies though Vitamin E has reported to upregulate VEGF and other HIF-1 dependent genes in previous studies. Why authors used variable dose(s) of LN in different experiments. In Annexin why authors choosed only 25 uM and then again for caspase-3 author authors used 12.5 and 25 uM and for western blot 25 uM? Signalling pathway data has been not explained properly in result as well as discussion section which is supposed to be strength of manuscript. Minor comments:- References are not uniform, please check for format. Check for speling mistakes, typographical mistakes and please check for overall English. (Page 5:- 4th paragraph photoes spelling;Page 6 2nd paragraph Annexin ...kit was using to assess...Page 7 under result section 7th line was used is coming twice, 13th line showed no significant between..... Page 8 line 16; authors used only one concentration 25 uM but they mention concentration dependent manner? Date Sent: 28-Oct-2013 |
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