Notoginsenoside R1 protects human keratinocytes HaCaT from LPS-induced inflammatory injury by downregulation of Myd88
Authors: Qibing Zheng
Jingqun Zhang
Ying Li
Fang Bi
Fangfang Jin
Jiahong Xu
Haiqiang Lu
Journal: International Journal of Immunopathology and Pharmacology (2019)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: MyD88; NF-κB; burn injury; inflammation; notoginsenoside R1
Affiliations: Department of Burns and Plastic Surgery, Affiliated Hospital of Jining Medical University, Jining, China

Abstract:  Burn injury is a gigantic challenge in public health which brings multiple negative effects to patients both in physical and spiritual aspects. Inflammation plays vital roles in the progression of burn injury, and our study investigated whether notoginsenoside R1 (NGR1) alleviated lipopolysaccharide (LPS)-induced human keratinocyte HaCaT cell inflammatory injury. Inflammatory injury was induced by LPS in HaCaT cells. Stimulated cells were then treated by NGR1 in different concentrations. Cell viability and cell apoptosis were detected by Cell Counting Kit-8 and flow cytometry, respectively. The concentration of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) was measured by enzyme-linked immunosorbent assay (ELISA). The accumulated levels of apoptosis-related proteins (caspase-3 and caspase-9), nuclear factor κB (NF-κB), p38 mitogen-activated protein kinase (p38MAPK) signal pathways-related proteins (p65, IκBα, and p38MAPK), and myeloid differentiation primary response 88 (MyD88) were examined by western blot. Transfection was used to alter the expression of MyD88. We found that LPS stimulated HaCaT cells and induced cell inflammation, evidenced by decreasing cell viability, increasing cell apoptosis, and elevating TNF-α and IL-6 expressions. Then, we found that NGR1 reversed the results by enhancing cell viability, inhibiting cell apoptosis, and reducing TNF-α and IL-6 expressions. In addition, NGR1 decreased the phosphorylation of p65, IκBα, and p38MAPK, which increased by LPS. Moreover, NGR1 negatively regulated the expression of MyD88, and transfection with pMyD88 led to the opposite results with what showed by NGR1 in LPS-stimulated HaCaT cells. To sum up, NGR1 alleviates LPS-induced HaCaT cell inflammatory injury by downregulation of MyD88, as well as inactivation of NF-κB and p38MAPK signal pathways. 

Retracted: No

PubPeer comments:
 The Western blots and other figures in this paper have an interesting layout that appears to be shared with many other papers, as discussed in a January 2020 post on Leonid Schneider s blog: https: forbetterscience.com 2020 01 24 the-full-service-paper-mill-and-its-chinese-customers The list of these papers is available here: https: docs.google.com spreadsheets d 1KXqTAyl4j-jVorFPMD2XRpr76LcIKJ0CVyIvRj0exYQ edit?usp=sharing In almost all papers, Western blot panels within the same figure, and across figures and papers, appear to share the same background, while the bands are regularly spaced, all have similar rounded edges without the usual smudges and specks, and with some bands showing a recognizable  jumping sardine shape. Also in almost all papers, bar graphs show a characteristic layout with bars in solid shades of grey, and the left-most bar in black, as well as double-edged error bars. Many papers have  cell viability and  apoptotic cells graphs. In some of these papers, including this one, flow cytometry plot show a unnaturally dense and well defined  hairball of events in Q3 with a few regularly spaced events in Q1 and Q2, and a rectangular or crab-shaped block of events in Q4. To illustrate the recognizable layout of the graphs in this paper, here is Figure 1. ![file] , 
 February 2021 Expression of Concern found here: https: journals.sagepub.com doi full 10.1177 20587384211000847 From the Notice:  *It has been brought to SAGEu2019s attention that this article may have been written (and possibly submitted) by a third-party agency, and that the contents of the article may be partially or wholly fabricated. SAGE is currently conducting a thorough investigation into this to establish the most appropriate way forward.* , 
 Retracted [2 September 2021] . H t [RetractionWatch] . ------------------------- At the request of the Journal Editor(s) the following article has been retracted. Zhang J, Zheng Q, Lu H, Jin F, Li Y, Bi F, Xu J (2019) Notoginsenoside R1 protects human keratinocytes HaCaT from LPS-induced inflammatory injury by downregulation of Myd88. International Journal of Immunopathology and Pharmacology 33: 1-10. DOI: 10.1177 2058738419857550. Significant similarities between this paper and a number of other papers have been identified, as well as indication of image and data manipulation suggestive of the use of a u201cpaper millu201d. These papers share common structures and wording yet do not share any common authors. The authors did not provide a sufficient response or evidence to refute these claims. As a result, the scientific accuracy of this paper is not reliable. Adhering to the international guidelines established by the Committee on Publication Ethics, the Journal has determined these are grounds for retraction. , 
 A [report from the Affiliated Hospital of Jining Medical College] reads as follows (thanks Google Translate): ------------------------ 9. Xu Jiahong from the Affiliated Hospital of Jining Medical College is the corresponding author of the paper  Nodoginsenoside R1 protects human keratinocytes HaCaT from LPS-induced inflammatory injury by downregulation of Myd88 . After investigation, Xu Jiahong of the Affiliated Hospital of Jining Medical College is the corresponding author, Zhang Jingqun is the first author, Zheng Qibing is the second author, Lu Haiqiang is the third author, Jin Fangfang is the fourth author, Li Ying is the fifth author, and Bi Fang is the fifth author. For the sixth author s paper  Nodoginsenoside R1 protects human keratinocytes HaCaT from LPS-induced inflammatory injury by downregulation of Myd88 , there are problems of ghostwriting and investment and paper fraud. The Affiliated Hospital of Jining Medical College will deal with the relevant responsible persons as follows: 1. Give the corresponding author Xu Jiahong (1) order him to make a deep inspection; (2) order him to withdraw the manuscript of the paper involved; (3) cancel the five-year application for scientific and technological plan projects (special projects, funds, etc.), scientific and technological awards, and titles of scientific and technological talents, etc. Eligibility, from March 2, 2020 to March 1, 2025; (4) Cancellation of the five-year postgraduate tutor application and appointment qualifications from March 2, 2020 to March 1, 2025 ; (5) Cancellation of three-year professional title promotion from March 2, 2020 to March 1, 2023; (6) Criticism is reported in the hospital. 2. Zhang Jingqun, the first author, will be (1) ordered to make an in-depth inspection; (2) the five-year application for scientific and technological plan projects (special projects, funds, etc.), scientific and technological awards, and titles of scientific and technological talents will be cancelled, starting from March 2, 2020 Until March 1, 2025; (3) Cancellation of five-year postgraduate tutor application and appointment qualifications from March 2, 2020 to March 1, 2025; (4) Cancellation of three-year professional title promotion qualifications, The time will be from March 2, 2020 to March 1, 2023; (5) Criticism is notified in the hospital. 3. The second author, Zheng Qibing, the third author, Lu Haiqiang, the fourth author, Jin Fangfang, the fifth author, Li Ying, and the sixth author, Bi Fang, will be criticized in the hospital. ,