DNA methylation of skeletal muscle function-related secretary factors identifies FGF2 as a potential biomarker for sarcopenia.

BACKGROUND: Sarcopenia is characterized by progressive loss of muscle mass and function due to aging. DNA methylation has been identified to play important roles in the dysfunction of skeletal muscle. The aim of our present study was to explore the whole blood sample-based methylation changes of skeletal muscle function-related factors in patients with sarcopenia. METHODS: The overall DNA methylation levels were analysed by using MethlTarget™ DNA Methylation Analysis platform in a discovery set consistent of 50 sarcopenic older adults (aged ≥65 years) and 50 age- and sex-matched non-sarcopenic individuals. The candidate differentially methylated regions (DMRs) were further validated by Methylation-specific PCR (MSP) in another two independent larger sets and confirmed by pyrosequencing. Receiver operating characteristic (ROC) curve analysis was used to determine the optimum cut-off levels of fibroblast growth factor 2 (FGF2)_30 methylation best predicting sarcopenia and area under the ROC curve (AUC) was measured. The correlation between candidate DMRs and the risk of sarcopenia was investigated by univariate analysis and multivariate logistic regression analysis. RESULTS: Among 1149 cytosine-phosphate-guanine (CpG) sites of 27 skeletal muscle function-related secretary factors, 17 differentially methylated CpG sites and 7 differentially methylated regions (DMRs) were detected between patients with sarcopenia and control subjects in the discovery set. Further methylation-specific PCR identified that methylation of fibroblast growth factor 2 (FGF2)_30 was lower in patients with sarcopenia and the level was decreased as the severity of sarcopenia increased, which was confirmed by pyrosequencing. Correlation analysis demonstrated that the methylation level of FGF2_30 was positively correlated to ASMI (r = 0.372, P < 0.001), grip strength (r = 0.334, P < 0.001), and gait speed (r = 0.411, P < 0.001). ROC curve analysis indicated that the optimal cut-off value of FGF2_30 methylation level that predicted sarcopenia was 0.15 with a sensitivity of 84.6% and a specificity of 70.1% (AUC = 0.807, 95% CI = 0.756-0.858, P < 0.001). Multivariate logistic regression analyses showed that lower FGF2_30 methylation level (<0.15) was significantly associated with increased risk of sarcopenia even after adjustment for potential confounders including age, sex, and BMI (adjusted OR = 9.223, 95% CI: 6.614-12.861, P < 0.001). CONCLUSIONS: Our results suggest that lower FGF2_30 methylation is correlated with the risk and severity of sarcopenia in the older adults, indicating that FGF2 methylation serve as a surrogate biomarker for the screening and evaluation of sarcopenia.

[Mechanism of Fuke Qianjin Capsules in treating intrauterine adhesion in rats through TGF-ß1-PI3K/Akt signaling pathway].

To explore the effect of Fuke Qianjin Capsules on anti-endometrial fibrosis in intrauterine adhesion(IUA) rats through TGF-ß1-PI3 K/Akt signaling pathway. With female SD rats as the object, IUA rat models were established through mechanical injury and infection, and they were randomly divided into normal group, sham operation group, Bujiale group(0.63 mg·kg~(-1)·d~(-1)), and high-dose Fuke Qianjin Capsules group(1.008 g·kg~(-1)·d~(-1)), medium-dose Fuke Qianjin Capsules group(0.504 g·kg~(-1)·d~(-1)), low-dose Fuke Qianjin Capsules group(0.252 g·kg~(-1)·d~(-1)). The rats were sacrificed 21 days after drug administration, and the uterus and liver were removed after blood collection from the abdominal aorta. The morphology of the uterus was observed with the naked eyes; the pathological and morphological changes of the uterine tissue and liver were observed by HE staining; the degree of fibrosis of the uterine tissue was observed by Masson staining; the expressions of TGF-ß1, TNF-α and IL-6 in serum were detected; the expressions of TGF-ß1, PI3 K, Akt, p-Akt protein in uterine tissue were detected by Western blot. The results showed that Fuke Qianjin Capsules could improve the pathological changes of uterine tissues in IUA rats, without damage to liver tissues, and reduce the expressions of TGF-ß1, TNF-α and IL-6 in serum(P<0.01); significantly reduce TGF-ß1, PI3 K, p-Akt protein expression in uterine tissues(P<0.05, P<0.01). It is indicated that Fuke Qianjin Capsules could exert the anti-endometrial fibrosis effect by regulating the TGF-ß1-PI3 K/Akt signal pathway, so as to achieve the effect in treating IUA rats, especially with the best effect in medium-dose Fuke Qianjin Capsules group.

Reduction of fibrosis in dibutyltin dichloride-induced chronic pancreatitis using rat umbilical mesenchymal stem cells from Wharton's jelly.

OBJECTIVES: The objective of this study was to investigate the effects of rat umbilical cord mesenchymal stem cells (UCMSCs) from Wharton's jelly on dibutyltin dichloride (DBTC)-induced chronic pancreatitis (CP) and subsequent pancreatic fibrosis in rats. METHODS: A rat model of CP induced by DBTC was used. Male Sprague-Dawley rats were randomly divided into 4 groups: the control, DBTC, DBTC + UCMSCs, and control + UCMSC groups. Umbilical cord mesenchymal stem cells were administered intravenously on day 5 after the administration of DBTC. On days 14 and 28, the rats were evaluated morphologically and biochemically. The expression levels of inflammatory cytokines and chemokines in the pancreatic tissues of different groups were evaluated using quantitative real-time polymerase chain reaction. The activation of pancreatic stellate cells was estimated by immunochemistry and Western blot analysis of α-smooth muscle actin. RESULTS: Umbilical cord mesenchymal stem cells were detected in inflamed pancreatic tissues. Umbilical cord mesenchymal stem cell treatment improved the histological scores and alleviated the fibrosis of pancreas samples, The expression of cytokines in the DBTC + UCMSC group was significantly lower than that in the DBTC group. Also, pancreatic stellate cell activation was inhibited by UCMSC treatment. CONCLUSIONS: Xenogeneic transplantation of UCMSCs is a novel approach for the treatment of CP and subsequent fibrosis. Umbilical cord mesenchymal stem cells may be a promising therapeutic intervention for human CP in the future.