Normal Diet Group Research Articles

Time-restricted feeding improves metabolic syndrome by activating thermogenesis in brown adipose tissue and reducing inflammatory markers

BackgroundObesity and metabolic syndrome (MetS) have become increasingly significant global health issues. Time-restricted feeding (TRF), as a novel dietary intervention, has garnered attention in recent years. However, there is limited research focusing on the effects of TRF on energy expenditure and systemic low-grade inflammation. This study aims to investigate the impact of TRF on weight management, glucose metabolism, insulin resistance, and lipid metabolism in male C57BL/6J mice, particularly in the context of metabolic disorders induced by a high-fat diet (HFD).MethodsC57BL/6J mice were divided into two groups: a normal diet (ND) group and a high-fat diet (HFD) group. The study duration was 12 weeks. Key parameters observed included body weight, glucose tolerance (via glucose tolerance tests), insulin resistance (HOMA-IR), and insulin secretion under glucose stimulation. Additionally, liver tissue was subjected to Oil Red O staining to assess lipid accumulation, and white and brown adipose tissues were stained with hematoxylin and eosin (HE) to evaluate adipocyte size. The expression of hepatic lipogenesis-related genes (Srebp-c, Chrebp, Fasn, and Acc1) and thermogenic genes in brown adipose tissue (UCP1 and PGC-1α) were also measured. Furthermore, temperature changes in the interscapular brown adipose tissue (BAT) were monitored.ResultsIn the ND group: TRF improved insulin resistance and reduced circulating levels of the pro-inflammatory cytokine IL-6, with a slight reduction in body weight.In the HFD group: TRF significantly mitigated weight gain, improved glucose tolerance and insulin resistance, and enhanced insulin secretion under glucose stimulation. Additionally, TRF reduced hepatic steatosis by downregulating the expression of lipogenesis-related genes in the liver. TRF also increased thermogenesis by upregulating the expression of thermogenic genes (UCP1 and PGC-1α) in BAT, while lowering serum levels of pro-inflammatory cytokines IL-6 and TNF-α, though IL-1β levels remained unchanged.ConclusionThis study demonstrates that TRF can activate thermogenesis in brown adipose tissue and reduce inflammation maker, leading to an improvement in hepatic steatosis and a reduction in white adipose tissue accumulation. These findings suggest that TRF may be a promising intervention for mitigating metabolic disturbances associated with obesity and metabolic syndrome. The study provides mechanistic insights into the beneficial effects of TRF, highlighting its potential in modulating lipid metabolism and exerting anti-inflammatory effects.

Saffron's protective role against atherosclerosis-induced cataract progression in New Zealand white rabbits with phytochemical analysis of saffron's extract.

Cataracts are significant causes of blindness, closely linked to prolonged hypercholesterolemia. While saffron has the potential for eye health, its effects on lens lesions remain understudied. This study aimed to investigate the effect of saffron on the lens changes in atherosclerotic-induced New Zealand white rabbits (NZWR). Thirty-five NZWRs were subjected to four to eight weeks of high-cholesterol diet to induce atherosclerosis, resulting in cataractous lens changes. The rabbits were categorised randomly into three groups: normal diet group, pre-treated group and treated group. The pre-treated group was divided into early atherosclerosis(HC4) and established atherosclerosis (HC8). The saffron-treated group was fed with the HCD diet followed by saffron treatment of 50mg/kg/day (TG450, TG840) and 100mg/kg/day (TG4100, TG8100) of saffron ethanolic extract (SEE) respectively. The normal diet group was given a normal diet over the 8 weeks. After completing the 16-week experimental protocol, the NZWR were euthanized, and their lenses were extracted for histopathological evaluation. The pre-treated group exhibited cataractous lens changes of grade 2, characterized by increased homogenisation, swollen lens fibers, and intracellular vacuolisation. Interestingly, these cataract changes showed a positive trend from grade 2 to grade 1 post-treatment with SEE. In the saffron-treated group, vacuoles and pinkish homogenised areas were reduced. Additionally, a uniform layer of anterior epithelium and decreased non-swollen lens fibers indicated significant cataract lesion improvement. The normal diet group displayed minimal to zero cataractous changes (Grade 0). HPLC analysis demonstrated the presence of crocin, crocetin, and picocrocin in the saffron ethanolic extract, with peak absorptions at 440nm (12.817min), 440nm (1.620min), and 254nm (6.553min) respectively. The phytochemical screening of saffron ethanolic extract was conducted and showed the presence of phytochemical compounds including saponins, flavonoids, tannins, and steroids. The positive effects on lenses in the TG groups could be due to crocin and crocetin, bioactive components of saffron, and its phytochemical compounds. This study highlights saffron's potential in managing cataract-induced conditions, emphasizing the importance of further research for its full therapeutic potential in cataract management.

Royal jelly reduced non-rapid eye movement sleep fragmentation and restored sleep stability in diet-induced obese mice.

Royal jelly (RJ) is recognized due to its high nutritional value and potential health benefits. Previous research showed that RJ supplementation decreased fat accumulation, resulting in weight loss and improvements in hyperglycemia and insulin resistance in high-fat diet (HFD)-induced obese mice. To expand the weight-reducing properties of RJ, this study aimed to investigate the effects of RJ supplementation on HFD-induced obese mice with impaired sleep stabilization. Over a 20-week period, the C57BL/6J mice were divided into the following dietary groups: normal diet (ND), ND supplemented with 5% lyophilized RJ powder (ND + RJ), HFD, and HFD supplemented with 5% lyophilized RJ powder (HFD + RJ) groups. Compared with the HFD group, the HFD + RJ group exhibited a significant reduction in body weight via a decrease in fat mass. Moreover, much like the ND group, the HFD + RJ group demonstrated improvements in the fragmentation of non-rapid eye movement (NREM) sleep and wakefulness. These processes contributed to the reestablishment of sleep/wake continuity and restored the overall stability of sleep. In contrast, the ND + RJ and ND groups exhibited a similar sleep/wake architecture. Thus, RJ supplementation in the ND demonstrated no substantial effect on sleep/wake. According to these findings, dietary RJ improves the sleep/wake architecture and restores sleep stability. Hence, RJ is a promising dietary component for addressing obesity and restoring sleep stability.

Effect of ketosis induced by on delayed-onset muscle soreness, inflammation and redox status: a randomized, open-label, crossover pilot study.

Previous studies show that ketosis caused by the consumption of low-carbohydrate diets improves cognitive functions and that ketogenic diets can be used to treat epilepsy. In vivo and in vitro experiments have shown that ketosis regulates pain, inflammation, and oxidative stress. Thus, we investigated the effects of ketosis induced by a low-carbohydrate diet on muscle soreness, inflammation, and redox status in human subjects. The research method was an open-label, crossover, pilot study. The study included eight men with no exercise habits associated with muscle soreness and consumed a low-carbohydrate and a normal diet for 6 days. Each dietary intake was for 3 days, and the participants performed the isotonic exercise on the fourth day. Before and after the exercise (immediately after, 24 h later, and 48 h later), the subjective value of muscle soreness, interleukin-6 level, tumor necrosis factor-alpha level, total ketone bodies, and redox status biomarkers were measured. The results revealed that the low-carbohydrate-diet group showed no significant difference in the subjective value of muscle soreness, whereas the normal diet group showed a significant increase in the subjective muscle soreness scale after 24 h. There were no significant changes in biomarkers of inflammation and redox status in either group. This result suggests that ketosis caused by consuming a low-carbohydrate diet suppresses delayed-onset muscle soreness. However, the ketosis state did not suppress inflammation or oxidative stress markers.

Effects of Bifidobacterium and rosuvastatin on metabolic-associated fatty liver disease via the gut–liver axis

Background/aimsResearch has indicated that treatment with rosuvastatin can improve liver pathology in metabolic-associated fatty liver disease (MAFLD) patients and that treatment with Bifidobacterium can improve MAFLD. Therefore, the effects of Bifidobacterium, rosuvastatin, and their combination on related indices in a rat model of diet-induced MAFLD need to be investigated.MethodsForty rats were divided into five groups: the normal diet group (N), high-fat diet (HFD) model group (M), HFD + probiotic group (P), HFD + statin group (S), and HFD + probiotic + statin group (P-S). To establish the MAFLD model, the rats in Groups M, P, S, and P-S were fed a HFD for 8 weeks. The treatments included saline in Group N and either Bifidobacterium, rosuvastatin, or their combination in Groups P, S, and P-S by intragastrical gavage. After 4 weeks of intervention, the rats were euthanized, and samples were harvested to analyze gastrointestinal motility and liver function, pathological changes, inflammatory cytokine production, and the expression of proteins in key signaling pathways.ResultsHFD feeding significantly increased the body weight, liver index, and insulin resistance (IR) index of the rats, indicating that the MAFLD model was successfully induced. Bifidobacterium reduced the liver of MAFLD rats, while Bifidobacterium with Rosuvastatin decreased the liver index, IR index, and levels of aspartate aminotransferase and alanine aminotransferase in MAFLD rats. The MAFLD model showed altered expression of proteins in signaling pathways that regulate inflammation, increased production of inflammatory cytokines, an elevated MAFLD activity score (MAS), and pathological changes in the liver. The MAFLD model also showed reduced relative counts of intestinal neurons and enteric glial cells (EGCs), altered secretion of gastrointestinal hormones, and slowed gastrointestinal emptying. Bifidobacterium, rosuvastatin, or their combination inhibited these various changes. HFD feeding changed the rats’ gut microbiota, and the tested treatments inhibited these changes. These results suggest that the gastrointestinal motility disorder and abnormal liver function in MAFLD rats may be related to a reduction in Escherichia-Shigella bacteria and an increase in Asticcacaulis bacteria in the gut microbiota and that the improvement in liver function induced by Bifidobacterium plus rosuvastatin may be related to increases in Sphingomonas and Odoribacter bacteria and a decrease in Turicibacter bacteria in the gut microbiota.ConclusionsThe combined use of Bifidobacterium and rosuvastatin could better regulate the gut microbiota of MAFLD model rats, promote gastrointestinal emptying, and improve liver pathology and function than single treatment with Bifidobacterium or rosuvastatin. This provides a better strategy for the treatment of MAFLD.

The possible antioxidative effects of ketogenic diet by modifying brain klotho expression: a rat model study

ABSTRACT Objectives: The ketogenic diet (KD) has long been used as an alternative nonpharmacological therapy to manage pharmacoresistant epilepsy. The anticonvulsant mechanisms of KD have yet to be fully elucidated. The present study explored whether a KD could exert antioxidative effects by altering brain Klotho (Kl) gene expression. Methods: Thirty male rats were divided into three groups: the normal diet (ND) group received standard rat chow; the calorie-restricted diet (CRD) group was maintained at 90% of the calculated energy need; and the KD group received a diet composed of 8% protein, 2% carbohydrates, and 90% fat (per calorie macronutrient). The levels of β-hydroxybutyrate (BHB) in the serum, Kl gene expression in the brain, and Kl protein, malondialdehyde (MDA), and protein carbonyl (PC) levels in the serum and brain were evaluated by standard methods. Results: The serum BHB levels in the KD group were significantly greater than those in the ND and CRD groups (p < 0.001). The Kl expression in the brain was significantly greater in the KD group than in the ND group (p = 0.028). The brain MDA levels in the KD group were significantly lower than those in the ND group (p = 0.006). Elevated BHB was positively correlated with brain Kl expression (r = 0.668, p < 0.001). The brain MDA levels were negatively correlated with brain Kl expression (r = −0.531, p = 0.003) and serum BHB levels (r = 0.472, p = 0.020). Discussion: KD might exert antioxidative effects by increasing BHB and upregulating Kl in the brain. This could be considered a possible anticonvulsant mechanism of KD.

BPA Exacerbates Zinc Deficiency-Induced Testicular Tissue Inflammation in Male Mice Through the TNF-α/NF-κB/Caspase8 Signaling Pathway.

Bisphenol A (BPA) is an endocrine-disrupting chemical that is toxic to reproduction. Zinc (Zn) plays an important role in male reproductive health. Zn deficiency (ZD) can co-exist with BPA. In order to investigate the specific mechanism of reproductive damage caused by BPA exposure in ZD male mice, a mouse model of ZD, BPA exposure, and their combined exposure was established in this study. Forty 4-week-old SPF male ICR mice with an average body weight of 31.7 ± 4.2g were divided into four groups including normal Zn diet group 30mg/(kg•d), BPA exposure group 150mg/(kg•d), zinc deficiency diet group 7.5mg/(kg•d), and BPA + ZD combined exposure group (BPA 150mg/(kg•d) + ZD 7.5mg/(kg•d)). The mice were kept for 8weeks. The results showed that the testicular tissue structure was disturbed, and semen quality, serum Zn, testicular tissue Zn, and testicular tissue free Zn ions were decreased in the BPA-exposed and ZD groups. The expression of zinc transporters (ZIP7, ZIP8, ZIP13, and ZIP14) in testicular tissue was changed. The expressions of pro-inflammatory cytokines including TNF-α and IL-1β as well as inflammatory pathway-related proteins (IKB-α, p-IKB-α, NF-κB, p-NF-κB, Caspase8, and Caspase3) were increased, while the expressions of anti-inflammatory cytokines (TGF-β and IL-10) were decreased. The changes in the above indexes in the BPA + ZD group were more obvious. Both BPA exposure and ZD can induce testicular tissue inflammation through the TNF-α/NF-κB/Caspase8 signaling pathway, and BPA further aggravates zinc deficiency-induced testicular tissue inflammation and apoptosis damage.

The effect of oyster mushroom (Pleurotus ostreatus) beta-glucan extract on brain -derived neurotrophic factor and neuron in high-fat-high-fructose diet-induced male Sprague Dawley rats

Obesity is a condition where there is an accumulation of excess body fat, which can increase oxidative stress either in the blood or in tissues, such as brain tissue. This situation can trigger atrophy in neurons and reduce brain-derived neurotrophic factor (BDNF) levels, which can increase the risk of impaired cognitive function. This study aimed to ascertain the effect of oyster mushroom (Pleurotus ostreatus) β-glucan extract on the number of neurons and levels of BDNF in rats. The study design was a true experimental with a randomized control group post-test only with white male Sprague Dawley rats as the subjects. A total of twenty-four rats were divided into four groups, namely the normal diet group fed AIN-93M standard diet (KN), the high-fat-high-fructose (HFHF) diet group (KP), and the HFHF diet group with the addition of β-glucan extract dose of 125 mg/kg body weight (P1), and dose of 375 mg/kg body weight (P2). The BDNF was measured by ELISA, and the number of neurons was by histological staining approach. Although there was no significant difference in the number of neurons (p = 0.692), the data revealed a significant difference in BDNF levels (p = 0.005). There was no relationship between the number of neurons and BDNF levels (p = 0.874) and between the β-glucan dose and the number of neurons (p = 0.796). However, there was a relationship between β-glucan dose and BDNF levels (p = 0.001; r = 0.695). This study concludes that the dose of β-glucan from oyster mushroom extract positively affects BDNF levels but not the number of neurons. There was no relationship between the number of neurons with BDNF levels and β-glucan dose. It was presumable because the staining technique was less specific, so the possibility of other cells counted as neurons may occur

A094: The Impact of Aerobic Exercise Combined with Intermittent Fasting on Alleviating Chronic Inflammation in Obesity

Research Aim: This study utilized aerobic exercise and intermittent fasting as intervention strategies to investigate their effects on the polarization of visceral fat macrophages and the amelioration of chronic fat inflammation in rats with obesity. Research Methods: Sixty male Sprague Dawley (SD) rats, aged 3 weeks, were randomly assigned to either a normal diet group (ND) or a high-fat diet group (HFD). Following a 12-week intervention period, the groups were further subdivided into the normal diet control group (ND), high-fat diet sedentary group (HC), high-fat diet intermittent fasting group (IF), high-fat diet aerobic exercise group (HE), and aerobic exercise combined intermittent fasting group (HIE). The aerobic exercise group underwent 10 weeks of medium-intensity training at 65% of their VO2max. The fasting group adhered to a 5:2 pattern, involving a 24-hour fasting period every Wednesday and Saturday commencing at 6 pm. During post-intervention, epididymal fat samples were collected, and wet weights were recorded. HE staining facilitated the observation of fat histopathology, while immunofluorescence staining was employed to identify the expression and localization of macrophage polarization-related proteins CD86 and CD206. Protein expression levels of IL-6, IL-10, TNF-Α, iNOS, and Arg-1 in adipose tissue were determined through Western blot analysis. Research Results: 1) In comparison to the NC group, the HC group exhibited a significant increase in epididymal adipose tissue weight and adipocyte surface area. 2) Following a 10-week intervention, the epididymal adipose tissue weight and adipocyte surface area in the HE, IF, and HIF groups showed a significant decrease when compared to the HC group. Notably, the HIF group demonstrated a more pronounced reduction. 3) Furthermore, in comparison to the HC group, the HIF group displayed a notable decrease in inflammation cells with a corona-like structure infiltrating the stroma of adipocytes. The protein levels of IL-6 and TNF-Α significantly decreased, while the IL-10 protein level exhibited a significant increase. Moreover, the expression of CD86 and iNOS showed a significant decrease, whereas CD206 and Arg-1 expression exhibited a significant increase. Conclusion: High-fat diet induces an increase in M1 polarization and a decrease in M2 polarization in rat adipose tissue, triggering the onset of chronic inflammation. Following a ten-week intervention, moderate-intensity aerobic exercise combined with intermittent fasting proves was more effective in ameliorating chronic inflammation in the adipose tissue of rats with obesity.

Chemopreventive Effects of Piper betle (Sirih) on High-Fat Diet-Induced and Azoxymethane-Induced Colon Cancer in Male Sprague-Dawley Rats.

A high-fat diet could lead to obesity, increasing colorectal cancer risk due to dyslipidemia and chronic inflammation, while Piper betle (PB) exhibits anti-tumor, anti-inflammation, and anti-oxidant benefits. This study aimed to determine whether PBpossesses chemopreventive effects on high-fat diet (HFD)-induced and azoxymethane (AOM)-induced colon cancer. Male Sprague-Dawley rats receiving either a normal diet orHFD were divided into control, PB, AOM, and AOM+PB subgroups which were then sacrificed after 24 weeks. The lipid profile, leptin, and inflammatory markers were measured from serum, and aberrant crypt foci (ACF) in the colon were detected by methylene blue staining. Cellular proliferation was identified through immunohistochemical staining of antigen Kiel 67 (Ki67) and beta-catenin. There were significant differences in serum total cholesterol, low-density lipoprotein, triglycerides, and high-density lipoproteinin the HFD compared to the normal diet group. The AOM group for normal diet and HFD exhibited significantly increased serum leptin, interleukin-6, IL-12p70, tumor necrosis factor-α, and nuclear factor-κB, with overexpression of Ki67 and beta-catenin. These changes were reversed by PB supplementation. In conclusion, PB demonstrated lipid-modifying and chemopreventive effects against HFD and AOM-induced colon cancer in rats.

The Effect of Standardized Mangosteen Skin Extraction, Nano-emulsion, Nano-chitosan and Treadmill Exercise on Atherogenic Rat Model

Background: Atherosclerosis, the most common cause of CVD, is associated with oxidative stress and cholesterol. Antioxidant and regular physical exercise have been considered as the probable interventions to dampen the process. The pericarp of mangosteen (Garcinia mangostana Linn) is known for containing a high amount of xanthones including α-mangostin with antioxidant effects. The current study investigates the anti-atherogenic potential of mangosteen skin extract in nano-emulsion and nano-chitosan formulations, singly and in combination with treadmill exercise, versus the statin, Atorvastatin. Methods: A randomized controlled trial was conducted on 30 male Wistar rats, divided into six groups: normal diet control (C1), atherogenic diet control (C2), and four treatment groups receiving an atherogenic diet plus treadmill exercise combined with Atorvastatin (T1), standardized mangosteen extract (T2), mangosteen nano-emulsion (T3), or mangosteen nano-chitosan (T4). The aortic tunica intima and tunica intima-media thickness were measured histologically after 56 days. Results: The aortic intimal thickness was noticeably higher in the atherogenic diet group (C2: 12.13±1.87 mm) compared to the normal diet group (C1: 4.27±0.75 mm). In the treatment groups, the intimal thickness ranged from 2.97±0.45 mm to 4.17±1.70 mm and showed no significant differences from the normal diet group. A similar pattern was seen in the intima-media thickness, with 145.63±17.12 mm recorded for the normal diet group and values ranging from 106.90±10.41 mm to 135.90±12.63 mm in the treatment groups, again without significant differences. Survival analysis using Kaplan-Meier curves showed clear differences between the groups (p &lt; 0.001). The untreated atherogenic diet group (C2) had the poorest survival, with no rats surviving until the study's conclusion. In contrast, survival improved in all treatment groups, with the Mangosteen Nano-chitosan (T4) group and the normal diet group (C1) achieving the best outcomes, as all rats in these groups survived. Conclusion: Mangosteen skin extracts, whether in nano-emulsion or nano-chitosan forms, combined with treadmill exercise, showed significant differences in maintaining the survivability of rat with atherogenic-induced diet despite no significant differences in preventing atherogenesis compared to Atorvastatin or a normal diet. Further research is needed to confirm these potential therapeutic effects.

Impact of daily fasting duration on body composition and cardiometabolic risk factors during a time-restricted eating protocol: a randomized controlled trial

BackgroundTime-restricted eating (TRE) is a dietary regimen that limits food intake for at least 12 h daily. Unlike other fasting protocols, TRE does not dictate what or how much to eat but rather focuses on the timing of meals. This approach has been previously demonstrated to improve body composition in individuals with obesity or metabolic impairments. However, its impact on body composition and cardiometabolic factors in healthy individuals remains unclear. Furthermore, the optimal fasting duration is still debated. Thus, we aimed to compare the effects of 8 weeks of different fasting durations on body composition and biochemical parameters in metabolically healthy, non-trained individuals using a parallel randomized controlled trial.MethodsForty-one volunteers were randomly assigned to one of the four experimental groups: TRE 16:8 (16 h of fasting,8 h of eating), TRE 14:10 (14 h of fasting,10 h of eating), TRE 12:12 (12 h of fasting,12 h of eating) or a normal diet group (ND; no dietary restriction). Participants underwent body composition measurements and blood tests for lipid profiles (i.e., total cholesterol, LDL, HDL, and triglycerides), fasting glucose, leptin, and anabolic hormones (i.e., insulin and testosterone) levels. Data were analyzed using both intention-to-treat (ITT) and per-protocol (PP) analysis to account for compliance. A two-way ANOVA for repeated measures was employed to assess interactions between time and group.ResultsIn the ITT analysis, TRE 16:8 reduced body mass (-2.46%, p = 0.003) and absolute fat mass (-8.65%, p = 0.001) with no changes in lean soft tissue and in calorie intake. These results were consistent with the PP analysis which included 8 participants in TRE 16:8, 5 in TRE 14:10, 9 in TRE 12:12, and the entire ND group. Participants in the TRE 16:8 group spontaneously reduced their total caloric intake, although this reduction was not statistically significant. None of the other measurements significantly changed after 8 weeks.ConclusionsOur results suggest that a 16-hour fasting window, even without caloric restriction, may be a viable strategy for improving body composition in healthy and non-trained individuals, whereas a shorter fasting period may be insufficient to produce significant changes in a healthy population.Trial registrationNCT, NCT04503005. Registered 4 August 2020, https://clinicaltrials.gov/study/NCT04503005.

Gut microbiota and mycobiota change with feeding duration in mice on a high-fat and high-fructose diet

BackgroundMetabolic dysfunction-associated fatty liver disease (MAFLD), formerly known as nonalcoholic fatty liver disease (NAFLD), is becoming the most common chronic liver disease. The gut microbiome is regarded to play a crucial role in MAFLD, but the specific changes of gut microbiome, especially fungi, in different stages of MAFLD are not well understood. This study aimed to observe the longitudinal changes of colon bacteria and fungi of mice at different feeding duration of a high-fat and high-fructose diet (HFHFD), and explore the association between the changes and the progression of MAFLD.MethodsTwenty-eight male C57BL6J mice were randomly assigned to the normal diet (ND) group and HFHFD group. At the 8th and 16th weeks, mice were sacrificed to compare the diversity, composition, and co-abundance network of bacteria and fungi in colon contents among groups.ResultsHFHFD-8W mice exhibited increases in Candida and Dorea, and decreases in Oscillospira and Prevotella in comparison to ND-8W mice, HFHFD-16W mice had increases in Bacteroides, Candida, Desulfovibrio, Dorea, Lactobacillus, and Rhodotorula, and decreases in Akkermansia, Aspergillus, Sterigmatomyces, and Vishniacozyma in comparison to ND-16W mice. And compared to HFHFD-8W mice, HFHFD-16W mice had increases in Desulfovibrio, Lactobacillus, Penicillium, and Rhodotorula, and decreases in Talaromyces and Wallemia. Spearman and GEE correlation analysis revealed that Bacteroides, Candida, Desulfovibrio, and Lactobacillus positively correlated with NAFLD activity score (NAS).ConclusionGut microbiota and mycobiota undergo diverse changes at different stages of MAFLD.Clinical trial numberNot applicable.

HM-chromanone attenuates obesity and adipose tissue inflammation by downregulating SREBP-1c and NF-κb pathway in high-fat diet-fed mice

Background: Obese adipose tissue produces various pro-inflammatory cytokines that are major contributors to adipose tissue inflammation. Objective: The present study aimed to determine the effects of HM-chromanone (HMC) against obesity and adipose tissue inflammation in high-fat diet-fed mice. Materials and methods: Twenty-four C57BL/6J male mice were divided into three groups: ND (normal diet), HFD (high-fat diet), and HFD + HMC. The ND group was fed a normal diet, whereas the HFD and HFD + HMC groups were fed a high-fat diet. After 10 weeks of feeding, the animals were orally administered the treatments daily for 9 weeks. The ND and HFD group received distilled water as treatment. The HFD+HMC group was treated with HM-chromaone (50 mg/kg). Results: HM-chromanone administration decreased body weight, fat mass, and adipocyte diameter. HM-chromanone also improved plasma lipid profiles, decreased leptin levels, and increased adiponectin levels. The inhibiting effect of HM-chromanone on SREBP-1c, PPARγ, C/EBPα, and FAS decreased adipogenesis, thereby alleviating lipid accumulation. Furthermore, HM-chromanone administration exhibited a reduction in macrophage infiltration and the expression of pro-inflammatory cytokines. HM-chromanone suppressed the phosphorylation of IκBα and NF-κB, leading to the inhibition of iNOS and COX2 expressions, resulting in decreased inflammation in adipose tissue. Discussion and conclusion: These results highlight the anti-obesity and anti-inflammatory properties of HM-chromanone, achieved through the downregulation of the SREBP-1c and NF-κB pathway in high-fat diet-fed mice.

Cholesterol-Lowering Activity of Lactiplantibacillus pentosus KS6I1 in High-Cholesterol Diet-Induced Hypercholesterolemic Mice.

Hypercholesterolemia is a risk factor of coronary heart disease and cholesterol-lowering probiotics are seen as alternative to drugs for the management of this condition. In the present study, we evaluated the cholesterol-lowering activity of Lactiplantibacillus pentosus KS6I1 in high-cholesterol diet-induced hypercholesterolemic mice. The mice were fed with high-cholesterol diet (HCD) and were divided into three groups: HCD group, KS6I1 group (fed with HCD + 200 μl of 1010 CFU/ml L. pentosus KS6I1), and L.ac group (fed with HCD + 200 μl of 1010 CFU/ml L. acidophilus ATCC 43121 as the positive control). Simultaneously, a normal control diet (NCD) group was maintained. After 6 weeks, the low-density lipoprotein (LDL)-cholesterol and total cholesterol levels were significantly reduced in the blood plasma of KS6I1 group mice. Analysis of liver tissue showed a decrease in total cholesterol and LDL-cholesterol and increase in triglyceride levels in KS6I1 compared to those in HCD group. Fecal total cholesterol and total bile acid content was significantly increased in the KS6I1 group than in other groups. Additionally, gene expression analysis showed that LDLR, SREBF2, CYP7A1 genes were significantly upregulated in KS6I1 group compared to the HCD group, while the expression of NPC1L1 gene was significantly reduced in KS6I1 group compared to HCD group. These observations show that the cholesterol-lowering effect of L. pentosus KS6I1 could be attributed to increased excretion of bile acids and cholesterol in the feces of mice. These results indicate that L. pentosus KS6I1 could be developed into a potential probiotic for hypercholesterolemia.

Mechanism of vitamin D deficiency involvement in the development of pulmonary arterial hypertension related to monocrotaline-induced connective tissue disease in rats

Objective: To investigating the impact of vitamin D (VitD) deficiency on the jagged 1 protein (Jagged1)/Notch3 signaling pathway in the pulmonary arteries of rats with monocrotaline (MCT)-induced connective tissue disease (CTD)-related pulmonary arterial hypertension (PAH) and to explore the pathological and molecular mechanisms of VitD involvement in the development of CTD-PAH. Methods: Twenty-four 7-week-old male Wistar rats were divided into a normal diet group and a VitD-free diet group using random number table, with 12 rats in each group. After 5 weeks of feeding, the rats were further randomly divided into saline and MCT groups, forming group A (normal diet+saline), group B (normal diet+MCT), group C (VitD-free+saline), and group D (VitD-free+MCT), with 6 rats in each group, and the rats were continued to be fed for another 4 weeks. The MCT group was injected with MCT solution subcutaneously on the back of the neck to construct the CTD-PAH model, and the saline group was injected with an equal amount of saline as a control. At the end of the experiment, blood samples were collected from all rats, and the serum 1, 25-dihydroxy vitamin D levels were determined by enzyme-linked immunosorbent assay. Hemodynamic measurements were performed on all rats, recording and calculating the mean values of right ventricular pressure and pulmonary artery pressure. Lung tissues and pulmonary arteries of all rats were stained with hematoxylin-eosin staining, the inner and outer diameters of pulmonary vessels were measured, and the percentage of pulmonary artery medial layer thickness was calculated. The right ventricular hypertrophy index was determined by weighing the parts of the heart and calculating the ratio. The apoptosis of pulmonary artery smooth muscle cells was assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. The expression levels of Jagged1, Notch3, and hairy and enhancer of split-1 (Hes1) mRNA and protein in the pulmonary arteries were analyzed by reverse transcription quantitative polymerase chain reaction and Western blotting. Results: The serum 1, 25-dihydroxy vitamin D levels in group D were significantly lower than those in groups A, B, and C [(51.01±0.96) μg/L, (65.15±1.83), (57.49±0.28), (54.52±2.87) μg/L, respectively, all P<0.05]. The right ventricular pressure, pulmonary artery systolic pressure, pulmonary artery diastolic pressure, and mean pulmonary artery pressure in Group D rats were significantly higher than those in groups A, B and C (all P<0.05). The percentage of pulmonary artery medial layer thickness in group D (56.28%±9.51%) was significantly higher than that in Groups A (21.28%±1.89%), B [22.72% (22.16%, 30.10%)], and C (38.73%±7.34%) (all P<0.05); the right ventricular hypertrophy index in group D (45.74%±12.68%) was significantly higher than that in groups A (21.78%±2.80%), B (40.93%±9.10%), and C (23.71%±1.22%) (all P<0.05). The apoptosis of pulmonary artery smooth muscle cells in group D (0.07±0.01) was significantly lower than that in groups A (1.00±0.08), B (0.17±0.02), and C (0.49±0.07) (all P<0.05). The levels of Jagged1, Notch3, and Hes1 mRNA and protein in Group D were all significantly higher than those in Groups A, B, and C (all P<0.05). Conclusions: VitD deficiency is involved in the pathophysiological mechanisms of CTD-PAH development. This occurs by upregulating the expression of the Jagged1/Notch3/Hes1 pathway in the pulmonary arteries of rats with MCT-induced PAH. The consequences include pulmonary artery thickening, increased pulmonary artery pressure, increased right ventricular pressure, right ventricular hypertrophy, and decreased apoptosis of pulmonary artery cells.

FSCN1 is a Potential Therapeutic Target for Atherosclerosis Revealed by Single-Cell and Bulk RNA Sequencing.

Atherosclerosis (AS) is the major cause of cardiovascular disease. Using integrated single-cell and bulk RNA sequencing data of atherosclerosis, we aimed to investigate the cell phenotype, intercellular communication, and potential therapeutic target in AS. Single-cell sequencing data from aortic arch of Apoeko mice in normal diet (ND) and high fat diet (HFD) groups (obtained from GSE206239) were analyzed by Seurat, singleR, ReactomeGSA, and cellchat package. scRNA-seq dataset GSE159677 from the carotid artery of the patients with carotid endarterectomy were used to validate the distribution of fascin actin-bundling protein 1 (FSCN1) in cell populations. Bulk RNA sequencing data (GSE43292 and GSE28829) were used to analyzed the expression of FSCN1in AS. A cross-sectional clinical study was utilized to examine the association between FSCN1 and AS. Circulating concentrations of FSCN1 were measured using ELISA kits and assessed using logistic regression analysis and receiver operating characteristic (ROC) curves. Apoeko mice fed with HFD and MAECs treated with oxidized low-density lipoprotein (ox-LDL) were established to detect the expression of FSCN1. Furthermore, we knocked down FSCN1 in MAECs to observe its influence on pyroptosis and migration. The HFD group had a significantly lower percentage of T cells, fibroblasts, and B cells and a significantly higher percentage of monocytes/macrophages cells. Strong interactions between endothelial cell (EC) and fibroblast in ND groups, while EC interactions with smooth muscle cells (SMC) and T cells were stronger in HFD groups. Semaphorin 7 (SEMA7) mediated signaling pathways were enriched in HFD groups and targeted EC driving by SMC. FSCN1was mainly expressed in EC and had a high expression in human AS samples. The cross-sectional study identified that high level of FSCN1 was associated with increased risk of AS. We also observed that high expression of FSCN1 in ox-LDL-induced MAECs and Apoeko mice fed with HFD. Knockdown of FSCN1 reduced pyroptosis and increased the migration in MAECs. Knockdown of FSCN1 in EC could alleviate the development and progression of AS. FSCN1 may be a potential prognostic biomarker and therapeutic target in AS.

Shen-Bai-Jie-Du decoction suppresses the progression of colorectal adenoma to carcinoma through regulating gut microbiota and short-chain fatty acids

BackgroundShen-Bai-Jie-Du decoction (SBJDD), a traditional Chinese herb formula developed based on evidence-based medicine, is efficacy to reduce the recurrence and carcinogenesis of colorectal adenoma. However, the mechanism of SBJDD to treat colorectal adenoma remains unclear. The present study aims to investigate the efficacy and mechanism of SBJDD on colorectal adenoma carcinogenesis from the aspects of regulating gut microbiota and short-chain fatty acids (SCFAs).MethodsTwenty-one patients diagnosed with colorectal adenoma were recruited in the study and required to take SBJDD for four consecutive weeks. Analysis of gut microbiota was conducted using 16S rRNA gene amplicon sequencing, while levels of SCFAs in fecal and serum samples were determined through HPLC–MS/MS. Additionally, twenty-four Apcmin/+ mice were randomly assigned to normal diet (ND), high-fat diet (HFD), and SBJDD groups. The pharmacological effects and mechanism of SBJDD on colorectal adenoma carcinogenesis were assessed using RT-qPCR, HE staining, IHC staining, Western blot, IF staining, and Flow cytometry assays.ResultsOur clinical study has shown that SBJDD can regulate the gut microbiota composition and enhance SCFAs production in patients with colorectal adenoma. SBJDD alleviated colorectal adenoma formation and carcinogenesis, as well as protected the integrity of the intestinal barrier in the Apcmin/+ mice model compared to the HFD group. Additionally, SBJDD was found to regulate gut microbiota capable of producing SCFAs. G protein-coupled receptors GPR43, GPR41, and GPR109a were effectively activated in the SBJDD group, while HDAC1 and HDAC3 were inhibited. Furthermore, decreased expression levels of interleukin 1 beta (IL-1β) and interleukin 6 (IL-6), along with elevated expression level of interleukin 10 (IL-10), were observed in the colorectal tissue of the SBJDD group. Finally, SBJDD exhibited the ability to reduce the proportion of M1-type macrophages while increasing the proportion of M2-type macrophages.ConclusionsOur study objectively demonstrated the pharmacological effects of SBJDD in inhibiting the progression of colorectal adenoma and investigated its mechanisms in terms of regulating gut microbiota, increasing SCFAs, and reducing colorectal inflammation.

Acupoint catgut embedding: a potential intervention strategy for obesity-related precocious puberty.

Obesity-related precocious puberty is induced by obesity, and acupoint catgut embedding (ACE) therapy is known to treat obesity. This study aims to validate the hypothesis that ACE can delay the onset of obesity-related precocious puberty. Female Sprague-Dawley rats, 21 days old, were randomly divided into three groups: the high-fat diet combined with ACE treatment group (ACE), the high-fat diet group (HFD), and the normal control diet group (NCD), with 8 rats in each group. The vaginal opening (VO) time was monitored, and serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and total estradiol (E2) were measured, followed by statistical analysis. Kaplan-Meier survival curves, with VO as the endpoint, showed that vaginal opening was delayed in the ACE group compared to the HFD group, with a statistically significant difference (p < 0.05). The changes in levels of FSH, LH, and E2 indicated that sexual development was delayed in the ACE group compared to the HFD group and was more similar to the NCD group. Combining the vaginal opening time and changes in hormone levels, this study confirms the potential role of ACE in delaying the onset of obesity-related precocious puberty.

Aerobic Exercise Prevents High-Fat-Diet-Induced Adipose Tissue Dysfunction in Male Mice

Background/Objectives: This study aimed to assess the effect of aerobic exercise on capillary density and vascular smooth muscle cell (VSMC) phenotype in the visceral and subcutaneous adipose tissue of high-fat-diet (HFD) mice in order to understand the mechanisms underlying improvements in insulin resistance (IR) and chronic inflammation in adipose tissue (AT). Methods: Male C57BL/6J mice were divided into HFD and normal diet groups for 12 weeks and then further split into sedentary and aerobic exercise subgroups for an additional 8 weeks. Various parameters including body weight, fat weight, blood glucose, lipid profile, insulin levels, glucose tolerance, and inflammatory cytokines were evaluated. Results: Aerobic exercise reduced HFD-induced weight gain, IR, and improved lipid profiles. HFD had a minimal effect on inflammatory cytokines except in visceral adipose tissue (VAT). IR was associated with capillary density in subcutaneous adipose tissue (SAT) and VSMC phenotype in VAT. Aerobic exercise promoted anti-inflammatory responses in VAT, correlating with VSMC phenotype in this tissue. Conclusions: Aerobic exercise can alleviate HFD-induced IR and inflammation through the modulation of VSMC phenotype in AT.