Effects Of Mulberry Leaf Extract Research Articles

Fortification of yogurt with mulberry leaf extract: Effects on physicochemical, antioxidant, microbiological and sensory properties during 21-days of storage

Mulberry leaves are medicinal and edible, with many physiological functions. To improve the potential function of yogurt, the effects of mulberry leaf extract (MLE) incorporation on the fermentation kinetics, physicochemical, antioxidant properties, and sensory parameters of yogurt were evaluated. The results showed that 0.1–0.3 % MLE improved the acidification rate and shortened the fermentation process. The addition of MLE significantly increased the values of total titratable acids, water holding capacity (WHC), total phenolic content and antioxidant capacities of the yogurt (p＜0.05). Specifically, the WHC values of 0.1 % MLE added yogurt were 1.33–1.41 times that of the control over 21 days of storage. In addition, MLE changed the texture and sensory quality of yogurt, resulting in light green, more stable products. Compared to the control, the yogurt with an appropriate concentration of MLE (0.1 % and 0.2 %) showed stable microbiological properties, and the survival of lactic acid bacteria in the yogurt was able to maintain a stable probiotic count of 108 CFU/g over 21 days of shelf life. The yogurt containing 0.1 % MLE achieved a good balance between the physicochemical and sensory qualities of the yogurt, and the use of MLE as an ingredient in yogurt production was a step towards the development of healthier dairy products.

Positive effects and mechanism of mulberry leaf extract on alleviating fatty liver hemorrhagic syndrome in laying hens

This experiment was conducted to investigate the effects of mulberry leaf extract (MLE) on alleviating fatty liver hemorrhagic syndrome (FLHS) in laying hens. The 576 Jing Fen laying hens of 56 weeks of age with good health and similar weights (1.76 ± 0.17 kg) were randomly divided into 6 groups, with 8 replicates in each group and 12 chickens in each replicate. The experiment lasted 56 d. The control group was fed a corn-soybean meal diet. The FLHS group was fed a high energy-low protein (HELP) diet, and the other four experimental groups were fed HELP diets supplemented with 0.04, 0.40, 0.80, and 1.20% MLE, respectively. The results showed that HELP treatment significantly induced liver injury, which indicated that the FLHS model was successfully established. MLE supplementation could alleviate the FLHS by reducing the liver index, abdominal fat percentage, total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL) in the serum (P < 0.05), and subsequently increase the egg production rate (P < 0.05). The laying hens fed 0.8% MLE exhibited the greatest production performance (P < 0.05) and could improve serum lipid levels. In addition, the genes associated with fatty acid synthesis (ACC, HMGR and SREBP-1C) were downregulated (P < 0.05), and genes related to fatty acid oxidation (CPT1A, AMPK, and ATGL) were found to be upregulated (P < 0.05). Supplementation with 1.2% MLE significantly reduced the relative abundance of Firmicutes and Desulfurized Bacillus (P < 0.05) and significantly increased the relative abundance of Fecal Bacillus (P < 0.05). In conclusion, MLE may regulate the mRNA expression of lipid metabolism-related genes through the AMPK signaling pathway and improve cecal microbiota balance and serum lipid levels to alleviate FLHS in laying hens and subsequently improve egg production performance.

Effects of ultrasound waves on rat liver mitochondria

Succinate dehydrogenase has a wide range of properties in plant and animal cells and is distributed throughout the inner membrane of mitochondria, therefore it is the enzyme without which energy metabolism cannot occur. The goal of our research was to investigate the effect of ultrasonic waves on the intensity of lipid peroxidation, activity of succinate dehydrogenase and cytochrome c oxidase in rat liver mitochondria, as well as antioxidant effect of mulberry leaf extract and biosep oil extract. Study involved white lab rats weighing 180-200 grams. The ultrasound device used was Mindrey DP-50 Vet. The work was based on the following methods: Schneider differential centrifugation, spectrophotometry, chromatography, pH-metry, photometry. The results of the study explain the elucidation of the mechanisms of damage to liver tissue by ultrasonic waves and the correction of the mitochondrial dysfunction caused by it with the extract of mulberry leaves and the oil extract of biosep. It was revealed that changes in lipid peroxidation led to disruption of the functional activity of membrane-bound cytochrome c oxidase and succinate dehydrogenase.

Effects of Mulberry Leaf Extract on the Liver Function of Juvenile Spotted Sea Bass (Lateolabrax maculatus).

In order to explore the effect of mulberry leaf extract (ELM) on the liver function of spotted sea bass, 360 fish with healthy constitution (average body weight 9.00 ± 0.02 g) were selected and randomly divided into six groups with three repetitions, and six groups of fish were randomly placed into 18 test tanks (200 L) with 20 fish per tank for the 52-day feeding test. Every day, the fish were fed the experimental feed with different concentrations (0, 3, 6, 9, 12, 15 g/kg) to the level of apparent satiation, with a crude protein content of 48.0% and a crude fat content of 8.6%. And the water temperature was maintained at 25-28°C with a salinity of 0.5%-1‰. After feeding, five fish were randomly selected to collect their livers and serum for detection of indicators. The results showed that, compared with the control group, ELM significantly increased the activities of lipase (LPS) and trypsin (TRS) in the liver, and reached the highest level when the amount of ELM added was 6 g/kg (P < 0.05). ELM significantly increased the activities of lactate dehydrogenase (LDH) and glutamic-oxaloacetic transaminase (GOT) involved in the metabolic process in liver tissue, and GOT activity reached the highest when ELM was added at 9 g/kg, and LDH activity reached the highest when ELM was added at 15 g/kg (P < 0.05). ELM had no significant effect on liver antioxidant enzymes (P > 0.05), but the content of malondialdehyde was significantly reduced (P < 0.05). Compared with the control group, ELM significantly increased the activities of AKP and ACP in the liver, and the AKP activity reached the highest when the ELM addition amount was 3 g/kg, and the ACP activity reached the highest when the ELM addition amount was 9 g/kg (P < 0.05). Through comparative transcriptomic analysis, it was indicated that ELM enhanced the hepatic lipids and carbohydrates metabolism ability, as manifested in the upregulation of expression of phosphatidate phosphatase, glucuronosyltransferase, inositol oxygenase, carbonic anhydrase, and cytochrome c oxidase subunit 2. ELM can also increase the expression of signal transducer and activator of transcription 1, ATP-dependent RNA helicase and C-X-C motif chemokine 9 involved in the immune process. The above results show that the ELM can enhance the digestion, metabolism, and immunity of the liver by increasing the activity of digestive enzymes, metabolic enzymes, and the expression of metabolism and immune regulation genes. This study provides a theoretical basis for the application of ELM in the cultivation of spotted sea bass by exploring the effect of ELM on the liver function of spotted sea bass.

Dietary Morus alba L. leaf supplementation improves hepatic lipid accumulation of laying hens via downregulating CircACACA

Fatty liver hemorrhagic syndrome (FLHS) is the most common metabolic disease in laying hens. Morus alba L. (mulberry) leaf has the effect of regulating lipid metabolism. We evaluated the effects of dietary 3% mulberry leaf (MUL) supplementation in production performance, egg quality, and liver lipid deposition in laying hens. Differentially expressed genes and circRNAs in the liver were identified using whole-transcriptome sequencing. We also evaluated the effects of the MUL extract using an in vitro model of primary hepatocytes induced by free fatty acids and explored the role of key circRNAs in this process. Dietary supplementation with 3% MUL alleviated liver steatosis in laying hens, as shown by decreased fatty liver color score, relative liver weight (P < 0.01), and triglyceride levels (P < 0.05), and showed a tendency to reduce the mortality rate of laying hens (P=0.09). In addition, mulberry leaf supplementation significantly reduced cholesterol content in egg yolk (P < 0.01). Dietary mulberry leaf supplementation downregulated the expression of genes involved in fatty acid and cholesterol biosynthesis, and upregulated the expression of fatty acid oxidation-related genes in the liver. CircACACA, which is derived from exons 2 and 3 of the acetyl-CoA carboxylase alpha (ACACA) pre-mRNA, was significantly reduced in the MUL group (P < 0.01). Upregulation of circACACA expression reversed the lipid-lowering effect of mulberry leaf extract by upregulating sterol regulatory element-binding proteins 1 c (SREBP-1c) and fatty acid synthase (FASN) (P < 0.05). Overall, mulberry leaf is an effective therapeutic strategy for FLHS in hens and can improve liver lipid metabolism by downregulating circACACA.

Larvacidal Activity of the Mulberry (Morus alba L.) Leaf Extract Against Larvae of Aedes aegypti

Dengue Haemorrhagic Fever (DHF) is one of the major public health problems in Indonesia. As the population density increases, the number of sufferers increases. Aedes aegypti mosquitoes are vectors for the disease. The absence of drugs make the best prevention effort by eradicating mosquito nests, killing larvae and adult mosquitoes. Mulberry leaves (Morus alba L.) may be used as larvacides in the presence of chemical compounds of flavonoids and saponins that inhibit feeding and disrupt the process of insect metabolism. The purpose of this research has to determine the effect of mulberry leaf extract (Morus alba L.), to determine the larvicide effect of mulberry leaf extract (Morus alba L.) and to determine the concentration of mulberry leaf extract (Morus alba L.) which is optimal in killing third instar Aedes aegypti larvae. This research used Randomized Design Group (RDG) method with treatment consisted 4 concentrations (0.25%, 0.5%, 0.75%, and 1%), negative control and positive control (ABATE) with 6 repetitions. The results of probit analysis showed that LC50 values were 1.124% and LC90 was 4.413%. From the one way ANOVA test at each concentration of 0.25%, 0.5%, 0.75%, and 1%, the F count result is 208.331, the value was greater than F table which is 2.53 and the significant value is 0.000 (sig &lt;0.05) then mulberry leaf extract (Morus alba L.) has a affected to eliminated of Aedes aegypti larvae. Conclusion from the results of the one way ANOVA test of mulberry leaf extract (Morus alba L.) was affected to eliminated third instar Aedes aegypti larvae

Mulberry leaves attenuate D-galactose-induced aging in vivo and in vitro.

Mulberry leaves contain many bioactive compounds and have been widely used in traditional medicines and functional foods for prevention and treatment of age-related diseases, such as diabetes, cognitive impairment and obesity-mediated liver cancer. Aging has an irreversible negative impact on human health for many years, even decades, before death, which is a social and economic burden on society. The objective of this study was to investigate the antioxidant and anti-aging effects of mulberry leaf extract (MLE) in vivo and in vitro. The Caenorhabditis elegans (C. elegans) was used as a model organism to observe the effects of different concentrations of MLE (1, 2, 4, 8 mg/mL) on nematodes' healthy lifespan, reproductive capacity, locomotion, stress resistance, and antioxidation. In addition, D-galactose (D-gal) induced liver aging in mice and L-02 cells were established. The antioxidant and anti-aging effects of MLE were evaluated by body weight, organ indexes, malondialdehyde (MDA), total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC), aspartate and alanine aminotransferases (AST and ALT), reactive oxygen species (ROS), mitochondrial membrane potential (MMP), hematoxylin and eosin (H&E), senescence-associated β-galactosidase (SA-β-Gal). Besides, the expressions of AMPK/SIRT1/PGC-1α and Nrf2-Keap1 were detected by Western blotting. MLE could significantly prolonged nematodes' average life span and improved most physiological indicators related to aging of C. elegans. Moreover, Treatment with MLE ameliorated the decreased body weight and organ index (weight of organ/body weight) in model mice, and protected against oxidative stress in mice and liver cells, in a dose-dependent manner, up-regulating T-SOD and T-AOC, while reducing ROS and MDA levels. MLE decreased both liver and cell levels of AST and ALT, and enhanced the mitochondrial membrane potential. MLE activated the AMPK/SIRT1/PGC-1α pathways, participated in mitochondrial biosynthesis and oxidative metabolism and delayed D-gal-induced aging. MLE promoted the accumulation of Nrf2 in the nucleus, indicating that the improved oxidative stress response was mediated by the Nrf2-Keap1 pathway in vivo and in vitro. MLE appeared to have great potential for stimulating the oxidative stress response and attenuating the aging process of in vivo and in vitro, and provide a novel health-promoting resource against aging and aging-related diseases.

Pharmacological and Pathological Effects of Mulberry Leaf Extract on the Treatment of Type 1 Diabetes Mellitus Mice.

This study investigated the pharmacological and pathological effects of aqueous mulberry leaf extract on type 1 diabetes mellitus mice induced with an intraperitoneal injection of streptozotocin (STZ). Diabetic mice were randomized into six groups: control (normal group), model, metformin-treated mice, and high-dose, medium-dose, and low-dose mulberry. The mulberry-treated mice were divided into high-, medium-, and low-dose groups based on the various doses of aqueous mulberry leaf extract during gavage. The efficacy of the six-week intervention was evaluated by measuring levels of fasting plasma glucose, alkaline phosphatase, alanine aminotransferase, aspartate transaminase, blood urea nitrogen, gamma-glutamyl transferase, glucose, high-density lipoprotein cholesterol, lactate dehydrogenase, and low-density lipoprotein cholesterol and recording body weight. Results revealed that mulberry leaf extract exhibited an ideal hypoglycemic effect, and the high-dose group was the most affected. Histology analysis, glycogen staining and apoptosis detection were used to study the extract's effects on the liver, kidney, and pancreatic cells of diabetic mice, enabling the assessment of its effectiveness and complications on a clinical and theoretical basis. It was shown that a certain concentration of aqueous mulberry leaf extract repaired the islet cells of type 1 diabetes mellitus mice, promoting normal insulin secretion. Herein, it was confirmed that mulberry leaf could be used to develop new hypoglycemic drugs or functional health food with broad applicability.

Effects of mulberry leaf extract on the intestinal health of spotted sea bass (Lateolabrax maculatus)

Mulberry leaf extract (ELM) has the functions of promoting growth, antioxidant, improving intestinal microbial composition, thus providing a potential solution the occurrence of fish intestinal diseases. Therefore, this experiment was conducted to explore the effects of ELM on intestinal health of spotted sea bassLateolabrax maculatus. A total of 360 spotted sea bass (9.00 ± 0.02 g) were selected and randomly divided into 6 groups. Fish in each group were given feed with varying ELM concentration (0, 3, 6, 9, 12, 15 g/kg) for 52 days, respectively. Results show, dietary intake of 9 g/kg ELM increased the weight gain, specific growth ratio and feed intake of the spotted sea bass (P&amp;lt;0.05). Meanwhile, dietary intake of 9 g/kg ELM increased the activity of enteric trypsin, amylase and lipase (P&amp;lt;0.05). The enteric catalase activity was improved in fish fed with 3 g/kg ELM (P&amp;lt;0.05), while a limited effect of ELM on the activity of enteric superoxide dismutase, glutathione, and content of malonaldehyde was observed (P&amp;gt;0.05). ELM improved the morphology of fish intestine, as manifested in significant improvement in the length of intestinal villi, thereby increasing the surface area of the intestinal tract (P&amp;lt;0.05). Compared with the control group, dietary intake of ELM significantly increased the intestinal microbial ACE, Chao1, and Shannon indexes of fish (P&amp;lt;0.05), indicated that the intestinal microbial composition and the abundance of the dominant flora of fish were improved. The above results suggested that the dietary supplementation of about 9 g/kg ELM can improve the growth performance, enteric antioxidant capacity, and intestinal morphology and microbial composition, therefore improving the intestinal health of spotted sea bass. The research results provide a theoretical basis for the application of ELM in improving the enteric health of spotted sea bass, and providing a potential solution the occurrence of fish intestinal diseases.

A Randomized, Placebo-Controlled Crossover Study to Evaluate Postprandial Glucometabolic Effects of Mulberry Leaf Extract, VitaminD, Chromium, and Fiber in People with Type2 Diabetes.

Reducing postprandial (PP) hyperglycemia and PP glucose excursions is important for overall glycemic management. Although most therapeutic lifestyle interventions that reduce caloric intake would affect this, there is no particular nutritional intervention favored. We evaluated the effects of a novel natural food adjuvant combining mulberry leaf extract (MLE) with other bioactive ingredients, in people with type2 diabetes (T2D) originating from Asia, on improving PP glucometabolic response in a randomized controlled exploratory crossover, two-center study (USA, Singapore). A 2-g blend of 250mg MLE [containing 12.5mg of 1-deoxynojirimycin (DNJ)], fiber (1.75g), vitamin D3 (0.75μg), and chromium (75μg), compared with a similar blend without the MLE, was sprinkled over a 350-kcal breakfast meal (55.4g carbs) and PP blood glucose (primary exploratory endpoint), insulin, and incretin hormones (GLP-1, GIP) were evaluated in blood samples over 3h. Changes in incremental areas under the concentration curve (iAUC) and maximum concentrations (Cmax) were compared. Thirty individuals (12 women, mean age 59years, HbA1c 7.1%, BMI 26.5kg/m2) were enrolled and the MLE-based blend relative to the blend without MLE significantly reduced glucose iAUC at 1h (- 20%, p < 0.0001), 2h (- 17%, p = 0.0001), and 3h (- 15%, p = 0.0032) and Cmax [mean (95% CI) difference - 0.8 (- 1.2, - 0.3) mmol/L, p = 0.0006]. A statistically significant reduction in 1h insulin iAUC (- 24%, p = 0.0236) was observed, but this reduction was no longer present at either 2h or 3h. No difference in GLP-1 was seen, but GIP response (iAUC and Cmax) was less with the MLE-based blend. The observation of a significant glucose reduction paralleled with a significant lower insulin response supports a reduced gastrointestinal glucose absorption. These results support the use of a 2-g natural blend of MLE, fiber, vitaminD, and chromium in T2D as a convenient dietary adjuvant to improve PP glucometabolic response. gov identifier NCT04877366.

Mulberry leaf extract improves non-specific immunity and antioxidant capacity of largemouth bass (Micropterus salmoides) fed a high-starch diet

A 70-day experiment was conducted to investigate the effects of mulberry leaf extract (MLE) on growth, proximate compositions, antioxidant and plasma biochemical parameters, and indices of non-specific immunity of largemouth bass (Micropterus salmoides) fed a high-starch diet. Two hundred eighty largemouth bass (initial body weight, 68.00 ± 0.19 g) were randomly fed seven diets: basal-starch diet (BSD; 8.88% starch), high-starch diet (HSD; 11.42% starch), and HSD diets supplemented with 0.05%, 0.10%, 0.20%, 0.50%, and 1.00% MLE (MLE1, MLE2, MLE3, MLE4, and MLE5, respectively). The results indicated that HSD and MLE did not significantly affect the growth performance of largemouth bass during the experimental period compared with that of the BSD, but the supplementation with more than 0.20% (MLE3, MLE4, and MLE5) MLE significantly decreased the hepatosomatic index (HSI) values, and 1.00% (MLE5) MLE significantly decreased the viscerosomatic index (VSI) values. The whole-body moisture of fish fed an HSD decreased significantly, while the whole-body lipid content increased significantly in the HSD group compared with the BSD group. Compared with HSD, MLE supplementation significantly decreased the moisture and lipid contents in the whole body. Supplementation with more than 0.20% MLE (MLE3, MLE4, and MLE5) significantly decreased the moisture content in the muscle. Supplementation with 1.00% MLE significantly decreased the content of hepatic and muscle glycogen. The malondialdehyde (MDA) content of the HSD group was significantly increased compared to that of the BSD group, whereas more than 0.10% (MLE2, MLE3, MLE4, and MLE5) MLE significantly decreased the MDA content. Additionally, the total antioxidant capacity (T-AOC), catalase (CAT), and glutathione peroxidase (GSH-Px) activities of MLE5 were significantly higher than those of the HSD group. The complement-3 (C3) content and globulin (GLB) in the plasma of the HSD group were significantly lower than those of the BSD group. Plasma C3 levels in the MLE3, MLE4, and MLE5 groups were significantly higher than those in the HSD group. In addition, glucose (GLU) levels in the MLE3, MLE4, and MLE5 groups were significantly lower than those in the HSD group. Supplementation with 0.50% (MLE4) MLE significantly increased the lysozyme (LYZ) content and decreased the activities of alanine transaminase (ALT) and aspartate transaminase (AST). Supplementation with 1.00% MLE significantly increased complement-4 (C4) and GLB contents and alkaline phosphatase (ALP) activity. Overall, these findings suggest that MLE could improve antioxidant capacity, immune function, and glycolipid metabolism, thereby alleviating the negative effects of a high-starch diet in M. salmoides.

Positive effects of Mulberry leaf extract on egg quality, lipid metabolism, serum biochemistry, and antioxidant indices of laying hens.

Plant extracts are becoming a hot topic of research by animal husbandry practitioners following the implementation of a global policy to restrict antibiotic use in animal production. Mulberry leaf extract has received considerable attention as a new plant extract. Mulberry leaf polysaccharides and flavonoids are its main constituents, and these substances possess immunoregulatory, hypoglycemic, antioxidant, and anticoagulant properties. It is however less common to use them in poultry production. Therefore, we investigated the effects of adding MLE to the diet of laying hens on egg quality, lipid metabolism, serum biochemistry, and antioxidant indices in this study. A total of 288 Lohmann Silber layers, aged 38 weeks, were randomly assigned to four groups (six replicates of 12 hens each). Hens were fed a basal diet supplemented with 0 (control diet), 0.4, 0.8, or 1.2% MLE for 56 d. Results showed that the addition of 0.4–1.2% MLE to the diet improved aspartate transaminase (AST) activity in the serum of laying hens, reduced low-density lipoprotein (LDL-C) content in the serum, and significantly decreased yolk triglyceride (TG) and total cholesterol (TC) contents (P < 0.05). No adverse effects were observed on production performance (P > 0.10). MLE (0.4 and 1.2%) significantly reduced the TG and TC levels in the liver (P < 0.05). MLE (0.8 and 1.2%) significantly increased glutathione peroxidase (GSH-Px) activity in the serum, decreased alanine transaminase (ALT) activity, TG and TC content in the serum, and improved egg yolk color (P < 0.05). MLE (1.2%) significantly increased high-density lipoprotein (HDL-C) content and superoxide dismutase (SOD) activity in the serum and enhanced eggshell strength (P < 0.05). The liver-related lipid metabolism gene assay revealed that the relative mRNA expression of PPARα and SIRT1 in the liver was significantly upregulated and that of FASN and PPARγ was significantly decreased after the addition of MLE. In contrast, the relative mRNA expression of SREBP-1c in the liver dramatically decreased after the addition of 0.8 and 1.2% MLE (P < 0.05). The addition of MLE to the diet improved egg quality and the economic value of hens by increasing antioxidant capacity and lipid metabolism. The most appropriate amount of MLE to be added to the diet of laying hens was 0.8%. Our study provides a theoretical reference for the application of MLE in egg production and to promote the healthy and sustainable development of the livestock and poultry industry under the background of antibiotic prohibition.

Mulberry Leaf and Neochlorogenic Acid Alleviates Glucolipotoxicity-Induced Oxidative Stress and Inhibits Proliferation/Migration via Downregulating Ras and FAK Signaling Pathway in Vascular Smooth Muscle Cell.

Mulberry leaf (Morus alba L.) has been used as a health food and in traditional medicine to treat several metabolic diseases, including diabetes, hypertension, and hyperlipidemia. However, the mechanism by which mulberry leaf and its functional components mediate atherosclerosis remains unclear. This study aimed to determine the effect of mulberry leaf extract (MLE) and its major component, neochlorogenic acid (nCGA), on the proliferation and migration of rat aortic vascular smooth muscle cells (VSMCs, A7r5 cell line) under diabetic cultured conditions (oleic acid and high glucose, OH). Our findings showed that MLE and nCGA significantly inhibited cell proliferation and migration in A7r5 cells as determined by a scratch wound assay and a Transwell assay. Furthermore, we observed MLE and nCGA inhibited cell proliferation and migration, such as reducing the phosphoinositide 3-kinases (PI3K)/protein kinase B (Akt), focal adhesion kinase (FAK), and small GTPase proteins using Western blot analysis. In conclusion, we confirmed the anti-atherosclerotic effects of MLE and nCGA in reducing vascular smooth muscle cell (VSMC) migration and proliferation under diabetic cultured conditions via inhibition of FAK/small GTPase proteins, PI3K/Akt, and Ras-related signaling.

Mulberry Leaf Extract Alleviates Staphylococcus aureus-Induced Conjunctivitis in Rabbits via Downregulation of NLRP3 Inflammasome and Upregulation of the Nrf2 System and Suppression of Pro-Inflammatory Cytokines

Introduction: Mulberry (Morus alba L.) leaves are widely used in traditional Chinese medicine for their antioxidant, anti-inflammatory, antibacterial, anti-obesity, antidiabetic, antiatherosclerotic, and anticancer properties. The current study aimed to investigate the effect of mulberry leaf extract (MLE) on Staphylococcus aureus (S. aureus)-induced conjunctivitis (5 × 10⁹ colony-forming units, 0.5 mL/eye) in a rabbit model. Methods: Rabbits were treated with MLE (5 mL/kg·d⁻¹ and 10 mL/kg·d⁻¹), 0.9% saline, pearl bright eye (PBE) drops, or erythromycin eye ointment (EEO) group for 5 days. The ocular infection symptoms, bacterial negative conversion rate, and conjunctival histopathological changes of rabbits in each group were observed. The expression of caspase-1, apoptosis-associated speck-like protein containing a caspase recruitment domain, NOD-like receptor leucine-rich pyrin domain-containing protein 3 (NLRP3), interleukin (IL)-18, IL-6, IL-1β, TNFα, Keap1, and nuclear factor erythroid 2-related factor 2 (Nrf2) in conjunctival tissue of rabbits were detected by quantitative real-time reverse transcription PCR and/or Western blot analysis. Results: The results showed that MLE treatment significantly reduced the clinical sign scores of conjunctivitis, alleviated clinical signs, and decreased bacterial load, and histological damage in a time- and dose-dependent manner was compared to that in the control group. The antibacterial and anti-inflammatory activities of MLE (10 mL/kg·d⁻¹) were similar to those of the positive control drug PBE and EEO. In addition, MLE significantly decreased the levels of pro-inflammatory cytokines, downregulated the NLRP3 inflammasome, and upregulated the Nrf2 system. Conclusions: MLE is effective in alleviating S. aureus-induced conjunctivitis in rabbits, and this mechanism is associated with the inhibition of the NLRP3 inflammasome and activation of the Nrf2 system to regulate pro-inflammatory signaling.

Uji Sitotoksik Ekstrak Daun Murbei (Morul Alba L) Terhadap Sel Kanker Mcf-7 Dan Selektifitasnya

Mulberry, Morus alba L., is traditionally used to treat various diseases such as bacterial, fungal, and protozoal. In addition, the society in South Sulawesi uses mulberry leaves as an anti-cancer. The present study aimed to investigate the cytotoxic effect of mulberry leaf extract on breast cancer cells MCF-7 and their selectivity against Vero normal cells. The dried mulberry leaves were extracted by maceration using 96% ethanol. The cytotoxic activity was determined using MTT method, while the selectivity index (SI) was calculated by comparing IC50 of MCF-7 cells against Vero cells. The results showed that the ethanol extract of mulberry leaves was toxic to MCF-7 cells with an IC50 of 75.32±2.78 µg/mL and also toxic to Vero cells (71.73±4.39 µg/mL). However, ethanol extract of mulberry leaves was not selective on MCF-7 cells with an SI value of 0.95. Our study indicates that mulberry could be considered an effective agent in the prevention of cancer, but further studies are necessary needed for safety assessment.

Network pharmacology approach to evaluate the therapeutic effects of mulberry leaf components for obesity.

Obesity is a chronic condition that has become a serious public health challenge globally due to the association with a high incidence of complications. Mulberry leaf is one of the most commonly used medicinal and herbal medicines that has been reported to ameliorate obesity and hyperlipidemia. However, the mechanism remains unclear. In the present study, a network pharmacology approach was used to explore the potential mechanism underlying the effects of mulberry leaf extract on obesity. First, the potential targets of mulberry leaf and obesity were predicted using SwissTargetPrediction, Online Mendelian Inheritance in Man, GeneCards and Comparative Toxicogenomics Database databases, which were then used to construct the protein-protein interaction networks. Gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyzes were performed using R version 3.6.3. Finally, results of this network analysis were verified by using the mulberry leaf extract to treat high-fat diet-induced obese mice. In total, 24 target genes associated with obesity that could potentially be affected by mulberry leaf treatment were predicted by network pharmacology, using which top seven related pathways were determined by KEGG enrichment analysis. Triglyceride (TG) and total cholesterol (TC) levels in mice serum were detected using TG and TC assay kits. Hepatic fat accumulation was detected by H&E staining whereas liver lipid droplets were detected by Oil red O staining in mice tissues. The expression of IL-1β, NF-κB inhibitor α, inducible nitric oxide synthase, AMP-activated protein kinase (AMPK), sterol regulatory element-binding proteins and fatty acid synthase in the visceral white adipose tissues of mice was analyzed by western blotting. The expression of TNF-α, peroxisome proliferator activated receptor (PPAR)D, PPARG, fatty acid amide hydrolase (FAAH) and hydroxysteroid 11-β dehydrogenase 1 (HSD11B1) in the visceral white adipose tissues of mice was detected by reverse transcription-quantitative PCR. Mulberry leaf extract was found to reduce fat accumulation and hepatic lipid droplet formation. Mulberry leaf also alleviated inflammation and lipogenesis whilst promoting lipid catabolism and fatty acid oxidation by promoting the AMPK signaling pathway. The possible anti-obesity effects of mulberry leaf on the mice may be due to the downregulation of TNF-α, PPARD and PPARG and the upregulation of FAAH and HSD11B1. These results were consistent with the GO enrichment analysis and suggested that mulberry leaf may regulate lipid metabolism and catabolism, fatty acid metabolism and biosynthesis and the inflammatory response to reduce obesity.

Study on Hypoglycemic Effect of Mulberry Leaf Extract Based on Big Data analysis

With the improvement of people’s living standards and the emergence of aging society, the incidence of diabetes is increasing year by year, and diabetes and its complications have become the third killer threatening human health. There are many drugs for treating diabetes, but their toxic and side effects are all great. Therefore, using big data analysis to find and develop natural products with hypoglycemic effect has high practical value. Therefore, the purpose of this paper is to study the hypoglycemic effect of mulberry leaf extract based on big data analysis. Firstly, this paper studies the medicinal value of mulberry leaves and related application methods by consulting relevant data. Through big data analysis, it is found that mulberry leaves are rich in alkaloid component 1-deoxynojirimycin (DNJ), accounting for more than 50% of alkaloid content. Therefore, this paper takes mulberry leaf extract DNJ as the research object, and firstly prepares mulberry leaf extract. Mulberry leaf water extract was perfused into stomach for 15 consecutive days, and fasting blood glucose of diabetic mice was measured 2 hours after eating by glucose oxidase method at specified time. The hypoglycemic effect of mulberry leaf water extract was analyzed by big data. The results showed that mulberry leaf extract could effectively inhibit the enzyme activity of decomposing sugar and prevent glucose absorption. Therefore, it can significantly reduce the blood sugar level of diabetic mice by about 50%. And the higher the dose of mulberry leaf extract, the better the hypoglycemic effect, showing a dose effect.

Hot Water Extract of Mulberry Leaf Ameliorates Hepatic Fat Accumulation and Inflammation in Rats Fed a High-Fat Diet

Abstract Objectives This study was conducted to investigate the effect of mulberry leaf extract on hepatic fat accumulation and inflammation in rats fed a high-fat diet. Methods Male Sprague–Dawley rats were randomly divided into three groups. Each group fed normal diet (NOR), high-fat diet (HF), or HF supplemented with 0.8% (w/w) hot water extract of mulberry leaf (HF + ME) for 14 weeks. Results The mulberry extract (ME) supplementation reduced body weight and white adipose tissues (epididymal, retroperitoneal, and mesenteric) weights. Serum levels of triglyceride (TG), total cholesterol (TC), free fatty acids (FFAs), and low-density lipoprotein cholesterol (LDL-C) were lower, while high-density lipoprotein cholesterol (HDL-C) level was higher in the HF + ME group compared to the HF group. The ME reduced the hepatic total lipid, TG, and TC levels compared to the HF group. The mRNA levels of genes related to fatty acid synthesis, such as CD36, sterol regulatory element binding protein 1c (SREBP-1c), and fatty acid synthase (FAS) were down-regulated by the ME supplementation. In addition, the ME lowered the mRNA levels of pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6), compared to the HF group. The serum TNF-α level of the HF + ME group was significantly lower than that of the HF group. Conclusions These results suggested that the ME attenuated high-fat diet-induced hepatic fat accumulation and inflammation via regulating gene expression related to hepatic lipid metabolism and pro-inflammatory mediators. Therefore, it is postulated that the ME might be useful as a functional food ingredient to prevent obesity-induced hepatic fat accumulation and inflammation. Funding Sources None.

Mulberry leaf extract displays antidiabetic activity in db/db mice via Akt and AMP-activated protein kinase phosphorylation.

BackgroundAugmenting glucose utilization in skeletal muscle via the phosphatidylinositol-3 kinase (PI3 kinase)/protein kinase B (Akt) pathway or the adenosine monophosphate (AMP)-activated protein kinase (AMPK) pathway is necessary to regulate hyperglycemia in patients with type 2 diabetes mellitus.ObjectiveWe investigated the effect of mulberry leaf extract (MLE) on glucose uptake in skeletal muscle cells and explored its in vivo antidiabetic potential.DesignMale db/db mice were treated with either MLE (50 mg/kg, 100 mg/kg, and 250 mg/kg) or metformin (100 mg/kg) for 8 weeks.ResultsMLE treatment stimulated glucose uptake, driven by enhanced translocation of glucose transporter 4 to cell membranes in L6 myotubes. These effects of MLE were synergistic with those of insulin and were abolished in the presence of PI3K inhibitor or AMPK inhibitor. In db/db mice, supplementation with MLE decreased fasting blood glucose and insulin levels and enhanced insulin sensitivity, with increases of p-Akt and p-AMPK in skeletal muscle. Moreover, MLE improved blood lipid parameters and attenuated hepatic steatosis in diabetic db/db mice.DiscussionThese findings suggest that MLE exerts antidiabetic activity through stimulating glucose disposal in skeletal muscle cells via the PI3K/Akt and AMPK pathways.ConclusionsMLE can potentially improve hyperglycemia and hepatic steatosis in patients with type 2 diabetes.

Vasodilatory effects of mulberry (Morus spp.) leaf extract on porcine cerebral arteries in vitro: Possible underlying mechanisms

Vasodilatory effects of mulberry leaf extract (MLE) and its possible underlying mechanisms on porcine basilar artery (PBA) were investigated by measuring changes in isometric tension in vitro. MLE slightly relaxed 60mMKCl pre-contracted PBA rings (9.4% at 540μg/mL). A treatment with MLE induced extensive relaxation under pre-contraction with 0.1μM U-46619 (EC50=49.4μg/mL). Neither endothelial denudation nor treatment with atenolol (10μM) affected MLE-induced relaxation. Relaxation induced by MLE (540μg/mL) was significantly attenuated by cimetidine (10μM). MLE inhibited angiotensin I-induced contraction in endothelium-intact PBA rings (EC50=25μg/mL), similar to captopril. Therefore, the possible mechanisms underlying MLE-induced relaxation in PBA might be mediated through: (1) functional antagonism of thromboxane A2 receptors; (2) suppression of angiotensin I-induced contraction; (3) Ca2+ influx inhibition; (4) stimulation of H2-receptors. Moreover, MLE contained significant amounts of nitrite, nitrate and several phenolic compounds, possibly associated with MLE-induced relaxation.