E a week. At the end of the 10-week treatment period, mice were killed by cervical dislocation after blood gathering. Liver, spleen, kidney, and adipose tissues (mesenteric, subcutaneous, epididymal, perirenal) were dissected precisely and weighed. An in vivo CT analysis (InveonTM, Siemens Medical Solutions USA Inc.) was carried out prior the killing of animals under 1.5? isoflurane in O2 anesthesia.Statistical AnalysisThe data were expressed as mean values with their standard errors. Analyses were performed using pairwise t-tests and Wilcoxon rank sum tests. Differences were considered to be statistically significant at values of P,0.05.Results L. gasseri BNR17 get Anlotinib Inhibits High-sucrose Diet-induced Body Weight Gain and Fat Weight AccumulationHigh-sucrose diet feeding induced significant body weight gain throughout the study period compared to the ND group (Figure 1A and Table 2). The administration of BNR17 induced less body weight gain than the HSD group, although not in a dosedependent manner. Food intake differed significantly between the ND and HSD groups (Figure 1B and Table 2); whereas no significant differences were observed in daily food intake between the HSD and HSD+BNR17 groups. Moreover, energy intake were similar for all groups. This suggests that BNR17 contributed to the reduced body weight gain. Total cholesterol and LDLcholesterol in the HSD group and HSD+BNR17 groups increased compared to the ND group; however, no significant reduction was caused by BNR17 administration (Table 2). In addition, glucose levels did not change with BNR17 administration. High-sucrose diet also induced increased adipose tissue weight as compared with normal diet feeding (Table 2). BNR17 administration significantly suppressed the increase of fat mass in all white adipose tissues, including mesenteric, subcutaneous, epididymal and A 196 perirenal adipose tissue (Table 2). Further, CT imaging showed a significant reduction in body fat profile with BNR17 treatment (Figures 1C and D). Moreover, HE staining of white adipose tissues revealed that supplementation with BNR17 was associated with a significant reduction in average adipocyte size in mesenteric, subcutaneous, epididymal and perirenal adipose tissues, as compared with the HSD group (Figures 1E and F).Real-time PCR AnalysisRNA was extracted from ,0.1 g of tissues using the RNeasy Mini kit (Qiagen) for liver and RNeasy Lipid Tissue Mini kit (Qiagen) for white adipose tissue, according to the manufacturer’s protocols. cDNA was synthesized using the AccupowerH RocketscriptTM Cycle RT Premix kit from Bioneer (Daejeon, South Korea). qPCR was performed using an Exicycler (Bioneer) with AccupowerH 26 Greenstar qPCR Master Mix (Bioneer). Primer sequences for the targeted mouse genes are listed in Table 1.Biochemical AnalysesEndocrine peptides (ghrelin, GIP, GLP-1, glucagon, insulin, leptin) were determined using a Bio-Plex suspension array system (Luminex, Austin, USA). Metabolic parameters including glucose, total cholesterol, HDL-cholesterol, and LDL-cholesterol levels in serum were analyzed using a Clinical Analyzer 7020 (HITACHI, Tokyo, Japan).Measurement of Adipocyte SizeAdipocyte sizes in the mesenteric, subcutaneous, epididymal and perirenal adipose tissue were measured in paraffin-embedded tissue. Briefly, adipocyte tissues were fixed in 10 neutral formalin solution, embedded in paraffin, cut into 4-mm sections, and stained with hematoxylin and eosin. Cell sizes were measured using a DIXI3000 (L.E a week. At the end of the 10-week treatment period, mice were killed by cervical dislocation after blood gathering. Liver, spleen, kidney, and adipose tissues (mesenteric, subcutaneous, epididymal, perirenal) were dissected precisely and weighed. An in vivo CT analysis (InveonTM, Siemens Medical Solutions USA Inc.) was carried out prior the killing of animals under 1.5? isoflurane in O2 anesthesia.Statistical AnalysisThe data were expressed as mean values with their standard errors. Analyses were performed using pairwise t-tests and Wilcoxon rank sum tests. Differences were considered to be statistically significant at values of P,0.05.Results L. gasseri BNR17 Inhibits High-sucrose Diet-induced Body Weight Gain and Fat Weight AccumulationHigh-sucrose diet feeding induced significant body weight gain throughout the study period compared to the ND group (Figure 1A and Table 2). The administration of BNR17 induced less body weight gain than the HSD group, although not in a dosedependent manner. Food intake differed significantly between the ND and HSD groups (Figure 1B and Table 2); whereas no significant differences were observed in daily food intake between the HSD and HSD+BNR17 groups. Moreover, energy intake were similar for all groups. This suggests that BNR17 contributed to the reduced body weight gain. Total cholesterol and LDLcholesterol in the HSD group and HSD+BNR17 groups increased compared to the ND group; however, no significant reduction was caused by BNR17 administration (Table 2). In addition, glucose levels did not change with BNR17 administration. High-sucrose diet also induced increased adipose tissue weight as compared with normal diet feeding (Table 2). BNR17 administration significantly suppressed the increase of fat mass in all white adipose tissues, including mesenteric, subcutaneous, epididymal and perirenal adipose tissue (Table 2). Further, CT imaging showed a significant reduction in body fat profile with BNR17 treatment (Figures 1C and D). Moreover, HE staining of white adipose tissues revealed that supplementation with BNR17 was associated with a significant reduction in average adipocyte size in mesenteric, subcutaneous, epididymal and perirenal adipose tissues, as compared with the HSD group (Figures 1E and F).Real-time PCR AnalysisRNA was extracted from ,0.1 g of tissues using the RNeasy Mini kit (Qiagen) for liver and RNeasy Lipid Tissue Mini kit (Qiagen) for white adipose tissue, according to the manufacturer’s protocols. cDNA was synthesized using the AccupowerH RocketscriptTM Cycle RT Premix kit from Bioneer (Daejeon, South Korea). qPCR was performed using an Exicycler (Bioneer) with AccupowerH 26 Greenstar qPCR Master Mix (Bioneer). Primer sequences for the targeted mouse genes are listed in Table 1.Biochemical AnalysesEndocrine peptides (ghrelin, GIP, GLP-1, glucagon, insulin, leptin) were determined using a Bio-Plex suspension array system (Luminex, Austin, USA). Metabolic parameters including glucose, total cholesterol, HDL-cholesterol, and LDL-cholesterol levels in serum were analyzed using a Clinical Analyzer 7020 (HITACHI, Tokyo, Japan).Measurement of Adipocyte SizeAdipocyte sizes in the mesenteric, subcutaneous, epididymal and perirenal adipose tissue were measured in paraffin-embedded tissue. Briefly, adipocyte tissues were fixed in 10 neutral formalin solution, embedded in paraffin, cut into 4-mm sections, and stained with hematoxylin and eosin. Cell sizes were measured using a DIXI3000 (L.
