Ethyl acetate fraction of Huogu formula inhibits adipogenic differentiation of bone marrow stromal cells via the BMP and Wnt signaling pathways
Authors: Xiangying Kong
Weiheng Chen
Qianqian Wang
Chunfang Liu
Liuluan Zhu
Hongye Wan
Cuiling Liu
Xiaomin Li
Qingxia Qin
Jia Zhu
Na Lin
Cun Zhang
Xuan Wu
Journal: International journal of biological sciences (2017)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Adipogenic differentiation; Bone marrow stromal cells; Chinese medicine; Huogu Formula; Osteonecrosis.
Affiliations: Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, 100700 Beijing, China., Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, China., Wangjing Hospital, China Academy of Chinese Medical Sciences, 100102 Beijing, China.

Abstract:  Elevated adipogenesis of bone marrow stromal cells (BMSCs) is closely associated with non-traumatic osteonecrosis of femoral head (ONFH). Our previous studies have shown that Huogu (HG) formula was effective both in clinic experience and experimental ONFH. How HG impacts the differentiation of BMSCs and what is the underlying molecular mechanism remain largely unknown. Our results showed that ethyl acetate extract of HG (HGE) significantly decreased the adipocyte differentiation as determined by oil red staining, while slightly increased the ALP activity. Investigation of the molecular mechanism revealed that HGE could inhibit the mRNA and protein expression of peroxisome proliferators-activated receptor (PPAR)γ, lipoprotein lipase (LPL) and adipocyteprotein2 (AP2). Interestingly, the inhibition of adipogenic differentiation in BMSCs by HGE could be restored by DKK-1, an inhibitor of Wnts. However, Noggin (an inhibitor of BMPs) displayed an additive role with HGE in suppressing the expression of PPARγ, LPL, and AP2. Furthermore, the bone marrow fat formation, as well as the expression of Wnt3a and PPARγ, was effectively regulated by HGE in the steroid-induced ONFH rats. Our results demonstrated that HGE treatment significantly inhibited adipogenesis and slightly promoted osteogenesis of BMSCs through regulating the BMP and Wnt pathways. The findings shed lights on the molecular mechanism of HGE in the inhibition of adipogenesis and provide scientific rationale for its clinical application of HGE in the treatment of ONFH. 

Retracted: No

PubPeer comments:
 Like the other [three RT-PCR tests] , this will also give +ve results for SARS . See this [White House briefing] (min 24) Here is a much [better biomarker] - 500bp found only in Wuhan nCoV See the similarity of the PCR s designed here to SARS... ![file] , 
 #1 From a public health perspective, if that patient has either SARS-1 or SARS-2, that requires hospitalization and quarantine. We should be focusing on high sensitivity without as much concern for specificity. , 
 Dear Dr. Dorsten, I would like to ask how your laboratory tested the Clinical samples used to extimate the specificity of your assay (section u201cExclusivity of 2019 novel coronavirus based on clinical samples pre-tested positive for other respiratory virusesu201d). Did you checked whether the clinical samples sent by the biobanks were still containing integer viral genetic material which could be amplified? This control could be done by qPCR with specific primers for that viruses, ideally in parallel with the RT-PCR done with the primers used for SARS-COV2. Did you make this control or you relied on the previous tests done in the past before storing samples in the biobanks? Iu2019m asking this because it does not seem it was done, according to the protocol. I think this kind of control could be important in the optics of extimating the assay specificity, because the virus samples could be degraded due to the storage in the biobanks, the shipping from the biobanks to your laboratory and the thawing of the samples. In this case there is the risk that the control viruses were not able to be amplified anymore in the clinical samples, making not feasible checking the specificity of your diagnostic test for the viruses indicated in table 2 of your protocol (the number of test you did in the part u201cCross-reactivity with other coronavirusesu201d could be not enough to validate the assay, considering also that you checked the presence of coronaviruses by RT-PCR in 5 7 coronavirus-related viral isolates). This could potentially lead to a not representative estimation of assay specificity (now is 100%) This issue could be relevant when you are testing people in areas with low percentage of infections of SARS-COV2. Kind regards, , 
 See http: doi.org 10.5281 zenodo.4298004 for comments and see https: www.pepijnvanerp.nl 2020 12 some-considerations-on-the-retraction-paper-for-the-corman-drosten-pcr-test for backgrounds about this comment. ,