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The research method of sirtuin 1 regulating the mechanism of senile alveolar bone loss mediated by Bmi1

A mechanism of action, technology of alveolar bone, applied in the field of Sirtuin1-mediated regulation of senile alveolar bone loss through Bmi1, can solve the problems of lack of in vivo research, unexplained mechanism, lack of alveolar bone research, etc.

Active Publication Date: 2020-12-29
AFFILIATED STOMATOLOGICAL HOSPITAL OF NANJING MEDICAL UNIV
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Problems solved by technology

[0005] The current reports on Sirt1 in the skeletal system mostly focus on long bones and vertebrae, so what is the role of Sirt1 in alveolar bone? Few literatures mentioned: Sirt1- / - mice have abnormal craniofacial bone structure and delayed mineralization, but did not clarify the specific abnormalities and mechanisms; human dental pulp cells express Sirt1, and overexpression of Sirt1 can promote mineralization. However, there is a lack of in vivo studies, and the mechanism has not been elucidated; Sirt1 may mediate the inflammatory pathway and oxidative stress pathway in the development of periodontal soft tissue inflammation, but there is a lack of research on alveolar bone

Method used

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Embodiment 1

[0054] The research method for Sirtuin 1 regulating the mechanism of senile alveolar bone loss mediated by Bmi1 includes the following steps:

[0055] a. Experimental animal breeding and genotype identification, select 4-week-old mice with WT, Sirt1TG, Bmi1KO, Sirt1TG / Bmi1KO genes respectively;

[0056] b. Alveolar bone phenotype analysis related index detection;

[0057] c. Mouse jawbone M-MSCs culture and in vitro experiments;

[0058] d. Detection of indicators related to oxidative stress;

[0059] e. Sirt1 activator treatment of natural aging mice;

[0060] f. Clinical alveolar bone sample collection, jaw bone M-MSCs culture and treatment with Sirt1 activator.

[0061] Said step b comprises the following sub-steps:

[0062] 1) Raw material: Mice were sacrificed by neck dislocation after anesthesia. The mandible was removed and fixed with PLP (2% paraaldehyde, 75mM lysine, 10mM sodium periodate) fixative that can better preserve enzyme activity and tissue resistance, f...

Embodiment 2

[0091] First, the role of Sirt1 overexpression in mesenchymal cells in preventing senile alveolar bone loss was investigated:

[0092] 1. In order to clarify whether the overexpression of Sirt1 in mesenchymal cells (MSCs) has the effect of increasing alveolar bone mass and preventing alveolar bone loss, the wild-type (WT) littermates aged 3, 9, and 18 months were observed by imaging methods and MSC overexpression of Sirt1 transgenic (Sirt1TG) mice mandibular morphology and bone density changes; using histological and histochemical staining methods to observe the changes in WT and Sirt1TG mice mandibular alveolar bone mass.

[0093] 2. In order to clarify whether the effect of MSC overexpression of Sirt1 on mouse mandibular alveolar bone mass is related to the increase of osteoblast bone formation and the decrease of osteoclast bone resorption, through histology, histochemical staining, immunohistochemical staining and Real -time RT-PCR and Western blot methods were used to obs...

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Abstract

The invention discloses a research method for the mechanism of Sirtuin 1 regulating senile alveolar bone loss through Bmi1, which includes the following steps: a. Experimental animal cultivation and genotype identification, selecting 4-week-old animals with WT, Sirt1TG, and Mice with Bmi1KO and Sirt1TG / Bmi1KO genes; b. Detection of indicators related to alveolar bone phenotyping analysis; c. Culture and in vitro experiments of mouse jaw M-MSCs; d. Detection of indicators related to oxidative stress; e. Sirt1 activator Processing of naturally aging mice; f. Clinical alveolar bone sample collection, jaw M‑MSCs culture and treatment with Sirt1 activator. This research method is expected to elucidate the role and mechanism of Sirt1 in preventing alveolar bone loss, and provide theoretical and experimental basis for the clinical application of Sirt1 activator resveratrol in preventing and treating senile alveolar bone loss.

Description

technical field [0001] The invention relates to the technical field of biological genes, in particular to a research method for Sirtuin 1 regulating the action mechanism of senile alveolar bone loss mediated by Bmi1. Background technique [0002] Oral diseases have been listed by the World Health Organization and my country's Ministry of Health as one of the three major non-communicable diseases that seriously threaten human health. Among oral diseases, the incidence rate of tooth and jaw-related diseases is as high as more than 80%. According to NIH statistics in the United States: 94% of women over the age of 65 suffer from loose teeth and loss; while those with bone loss, the incidence of tooth loss is three times higher than those without bone loss. According to the third national epidemiological survey of oral diseases in 2005, people over the age of 50 in my country lost an average of 12 teeth, and the number of patients over 65 years of age was as high as 30%. Tooth ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/50G01N33/569G01N33/94G01N15/00G01N15/10G01N15/14C12Q1/6883C12Q1/02
CPCG01N15/00G01N15/10G01N15/1434G01N33/5044G01N33/56966G01N33/94C12Q1/6883G01N2015/1006G01N2800/18C12Q2600/158G01N15/01
Inventor 孙雯苗登顺王华
Owner AFFILIATED STOMATOLOGICAL HOSPITAL OF NANJING MEDICAL UNIV