Heat-Killed Lacticaseibacillus paracasei GMNL-653 Exerts Antiosteoporotic Effects by Restoring the Gut Microbiota Dysbiosis in Ovariectomized Mice

Abstract

Osteoporosis is a metabolic inflammatory disease, an imbalance occurs between bone resorption and formation, leading to bone loss. Anti-inflammatory diet is considered having the potential to ameliorate osteoporosis. Heat-killed probiotics exhibit health benefits in relation to their immunomodulatory effects, but the detail mechanism involved in gut microbiota balance, host metabolism, immunity, and bone homeostasis remains unclear. In this study, we evaluated the antiosteoporotic effects of heat-killed Lacticaseibacillus paracasei GMNL-653 in vitro and in ovariectomized (OVX) mice. Furthermore, whole-genome sequencing and comparative genomics analysis demonstrated potentially genes involved in antiosteoporotic activity. The GMNL-653 exerts anti-inflammatory activity which restored gut microbiota dysbiosis and maintained intestinal barrier integrity in the OVX mice. The levels of IL-17 and LPS in the sera decreased following GMNL-653 treatment compared with those of the vehicle control; mRNA levels of RANKL were reduced and TGF-β and IL-10 enhanced in OVX-tibia tissue after treatment. The levels of IL-17 were significantly associated with gut microbiota dysbiosis. Gut microbial metagenomes were further analyzed by PICRUSt functional prediction, which reveal that GMNL-653 intervention influence in several host metabolic pathways. The analysis of whole-genome sequencing accompanied by comparative genomics on three L. paracasei strains revealed a set of GMNL-653 genes that are potentially involved in antiosteoporotic activity. Our findings validated antiosteoporotic activity of heat-killed GMNL-653 using in vitro and in vivo models, to whole-genome sequencing and identifying genes potentially involved in this gut microbiota–bone axis.

Suppressive Effects of Lactobacillus on Depression through Regulating the Gut Microbiota and Metabolites in C57BL/6J Mice Induced by Ampicillin

“…In this study, 16S ribosomal RNA gene amplicon sequencing was performed as previously described [ 32 ]. Ileum samples were collected from each group after 12 days, and all specimens were extracted using a Qiagen DNA kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.…”

“…The processing and statistical analysis of meta-taxonomic data were performed as previously described [ 32 ]. To determine the difference in microbial composition among groups, the Shannon diversity index was used to analyze the alpha diversity of taxonomic composition.…”

“…To specifically measure the expression of intestinal tight junction protein-related genes, inflammatory cytokine genes, and gut-derived 5-HT biosynthesis genes, we collected data from 4–5 mice in each group. The front section of the ileum samples obtained from the experimental mice was used for RNA extraction [ 32 ]. After washing with sterilized cold PBS to remove stool particles, the mucosa and intestinal wall of the ileum were collected by a scraper and stored at −80 °C and used as samples for RNA isolation.…”

New Advances in Improving Bone Health Based on Specific Gut Microbiota

“…After excluding differences related to BMI and medications, the study suggested an adverse effect of Lactobacillus on osteoporosis (Das et al, 2019). In contrast, part of Lactobacillus belonging to this genera, such as Lactobacillus acidophilus, Lacticaseibacillus paracasei GMNL-653, and Lactobacillus reuteri et al, were thought to positively regulate bone health (Dar et al, 2018;Lee et al, 2021;Jhong et al, 2022). Britton et al showed that Lactobacillus reuteri suppressed ovariectomyinduced increases in bone marrow CD4+ T lymphocytes and inhibited estrogen deficiency-induced bone resorption and bone loss (Britton et al, 2014).A study using a mouse model with gut microbial dysbiosis after antibiotic treatment found that Lactobacillus reuteri prevented osteoblastopenia and increased osteoclast activity, thereby preventing trabecular bone loss in the femur and spine and exerting a positive skeletal effect.…”

“…After excluding differences related to BMI and medications, the study suggested an adverse effect of Lactobacillus on osteoporosis ( Das et al., 2019 ). In contrast, part of Lactobacillus belonging to this genera, such as Lactobacillus acidophilus, Lacticaseibacillus paracasei GMNL-653, and Lactobacillus reuteri et al, were thought to positively regulate bone health ( Dar et al., 2018 ; Lee et al., 2021 ; Jhong et al., 2022 ). Britton et al.…”