Abelmoschus Manihot Research Articles

Effects and mechanisms of total flavones of Abelmoschus manihot in inhibiting podocyte necroptosis and renal fibrosis in diabetic kidney disease

Previous studies have shown that high blood glucose-induced chronic microinflammation can cause inflammatory podocyte injury in patients with diabetic kidney disease(DKD). Therein, necroptosis is a new form of podocyte death that is closely associated with renal fibrosis(RF). To explore the effects and mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese herbal medicine Abelmoschus manihot for treating kidney diseases, on podocyte necroptosis and RF in DKD, and to further reveal its scientific connotation with multi-pathway and multi-target, the authors randomly divided all rats into four groups: a namely normal group, a model group, a TFA group and a rapamycin(RAP) group. After the modified DKD rat models were successfully established, four group rats were given double-distilled water, TFA suspension and RAP suspension, respectively by gavage every day. At the end of the 4th week of drug treatment, all rats were sacrificed, and the samples of their urine, blood and kidneys were collected. And then, the various indicators related to podocyte necroptosis and RF in the DKD model rats were observed, detected and analyzed, respectively. The results indicated that, general condition, body weight(BW), serum creatinine(Scr), urinary albumin(UAlb), and kidney hypertrophy index(KHI) in these modified DKD model rats were both improved by TFA and RAP. Indicators of RF, including glomerular histomorphological characteristics, fibronectin(FN) and collagen type Ⅰ(collagen Ⅰ) staining extent in glomeruli, as well as the protein expression levels of FN, collagen Ⅰ, transforming growth factor-β1(TGF-β1) and Smad2/3 in the kidneys were improved respectively by TFA and RAP. Podocyte damage, including foot process form and the protein expression levels of podocin and CD2AP in the kidneys was improved by TFA and RAP. In addition, tumor necrosis factor-α(TNF-α)-mediated podocyte necroptosis in the kidneys, including the morphological characteristics of podocyte necroptosis, the extent and levels of the protein expression of TNF-α and phosphorylated mixed lineage kinase domain like pseudokinase(p-MLKL) was improved respectively by TFA and RAP. Among them, RAP had the better effect on p-MLKL. More importantly, the activation of the receptor interacting serine/threonine protein kinase 1(RIPK1)/RIPK3/MLKL signaling axis in the kidneys, including the expression levels of its key signaling molecules, such as phosphorylated receptor interacting serine/threonine protein kinase 1(p-RIPK1), p-RIPK3, p-MLKL and cysteinyl aspartate specific proteinase-8(caspase-8) was improved respectively by TFA and RAP. Among them, the effect of TFA on p-RIPK1 was superior. On the whole, in this study, the authors demonstrated that TFA alleviates podocyte necroptosis and RF in DKD through inhibiting the activation of the TNF-α-mediated RIPK1/RIPK3/MLKL signaling axis in diabetic kidneys. The authors' findings provide new pharmacological evidence to reveal the scientific connotation of TFA in treating RF in DKD in more depth.

Huangkui capsule attenuates diabetic kidney disease through the induction of mitophagy mediated by STING1/PINK1 signaling in tubular cells

BackgroundMitochondria is critic to tubulopathy, especially in diabetic kidney disease (DKD). Huangkui capsule (HKC; a new ethanol extract from the dried corolla of Abelmoschus manihot) has significant clinical effect on DKD. Previous studies have shown that HKC protects kidney by regulating mitochondrial function, but its mechanism is still unclear. The latest research found that the stimulator of interferon genes (STING1) signal pathway is closely related to mitophagy. However, whether HKC induces mitophagy through targeting STING1/PTEN-Induced putative kinase (PINK1) in renal tubular remains elusive. ObjectiveThis study aims to clarify the therapeutic effect of HKC on renal tubular mitophagy in DKD and its potential mechanism in vivo and in vitro. MethodsForty male C57BL/6 mice were randomly divided into 5 groups: CON group, DKD group, HKC-L (1.0 g/kg/day, by gavage), HKC-H (2.0 g/kg/day), and LST group. Diabetes model was induced by high-fat diet (HFD) combined with intraperitoneal injection of Streptozotocin (STZ). LST (losartan) is used as a positive control drug. Then, the glomeruli, renal tubular lesions, mitochondrial morphology and function of renal tubular cells and mitophagy levels were detected in mice. In addition, a high glucose injury model was established using HK2 human renal tubular cells. Pretreate HK2 cells with HKC or LST and detect mitochondrial function, mitophagy level, and autophagic flux. In addition, small interfering RNAs (siRNAs) of STING1 and PINK1 and overexpressing pcDNA3.1 plasmids were transfected into HK-2 cells to validate the mitophagy mechanism regulated by STING1/PINK1 signaling. ResultsThe ratio of urinary albumin to creatinine (ACR), fasting blood glucose, body weight in the early DKD mice model was increased, with damage to the glomerulus and renal tubules, mitochondrial structure and dysfunction in the renal tubules, and inhibition of STING1/PINK1 mediated mitophagy. Although the fasting blood glucose, body weight and serum creatinine levels were hardly ameliated, high dose HKC (2.0 g/kg/day) treatment significantly reduced ACR in the DKD mice to some extent, improved renal tubular injury, accurately upregulated STING1/PINK1 signaling mediated mitophagy levels, improved autophagic flux, and restored healthy mitochondrial pools. In vitro, an increase in mitochondrial fragments, fusion to fission, ROS and apoptosis, and a decrease in respiratory function, mtDNA, and membrane potential were observed in HK2 cells exposed to high glucose. HKC treatment significantly protected mitochondrial dynamics and function, which is consistent with in vivo results. Further research has shown that HKC can increase the level of mitophagy mediated by STING1/PINK1 in HK2 cells. ConclusionsOur results suggest that HKC ameliorates renal tubulopathy in DKD and induces mitophagy partly through the up-regulation of the STING1/PINK1 pathway. These findings may provide an innovative therapeutic basis for DKD treatment.

11-LB: The Efficacy and Safety of Abelmoschus manihot for Reduction of Microalbuminuria in Patients with Type 2 Diabetes and Diabetic Nephropathy—A Multicenter, Randomized, Open-Label, Parallel-Controlled Clinical Trial

Objective: Huangkui capsule, an extract from the flowers of Abelmoschus Manihot, has been proved clinically effective in improving chronic kidney disease. This study aimed to evaluate the efficacy and safety of Huangkui capsule in patients with stage 3 diabetic nephropathy. Methods: This multicenter, randomized, open-label, parallel-controlled trial was conducted in six hospitals. The inclusion criteria were 30 mg/g ≤ albumin creatinine ratio (ACR) <300 mg/g, eGFR ≥ 60 mL/min/1.73 m2, and type 2 diabetes with glycosylated hemoglobin ≤ 9%. We enrolled 239 patients who were randomly assigned to the two groups in a 1:1 ratio. They were treated with Huangkui capsule 2.15 g three times a day, or losartan potassium tablets 100 mg/day for 24 weeks. The primary outcome was the change in ACR from baseline and the secondary outcome was the change in eGFR. Results: The ACR in the losartan group was 119.47±78.16 mg/g at baseline and 90.51±136.12 mg/g after 24 weeks of treatment, with a decrease of 30.46±134.16 mg/g (P=0.049). In the Huangkui group, the ACR decreased from 121.98±84.31 mg/g to 88.13±76.66 mg/g, with a drop of 31.94±91.66 mg/g (P=0.002). The change in eGFR was -3.58±10.35 mL/min/1.73 m2 and -3.82±13.05 mL/min/1.73 m2 in the losatan and Huangkui groups, respectively. In subgroup analysis of patients with baseline ACR ≥ 100 mg/g, losatan and Huangkui lowered ACR by 56.83 mg/g (P=0.044) and 78.72 mg/g (P<0.001), respectively. There were no significant differences between the two groups (P>0.05). Conclusions: Huangkui capsule significantly reduced ACR of patients with type 2 diabetes and microalbuminuria and is a promising medicine for diabetic nephropathy. Disclosure J. Yin: None. S. Chen: None. L. Wei: None. W. Jia: None. Funding National Natural Science Foundation of China (82070885); Shanghai Municipal Education Commission (20172025); Xuhui Joint Research Projects on Important Diseases (XHLHGG202110)

Study on the lipid and flavor compounds of Hibiscus manihot L. oil induced by different pressing method

Hibiscus manihot L. seed oil is an emerging high-value edible vegetable oil that also has a variety of important health functions, and concerns about its pressing method have great significance for commercial and consumer health. This study compared triacylglycerols (TAGs), diacylglycerol (DAG), free fatty acids (FFAs), and volatile flavor compounds for discriminating cold-pressed oil from hot-pressed oil. The profiles of TAGs, DAGs, FFAs, and flavor compounds were established by a lipidomic approach and electronic-nose (E-nose). Our results demonstrated that a slight decrease in TAG content and a significant increase in FFA and DAG contents were observed after roasting. The E-nose results showed that sulfides, terpenes, methane, compounds containing oxygen and nitrogen, alcohol and aromatic compounds greatly influenced the oil flavor. The PCA result of the E-nose data indicated that flavor compounds showed great promise for distinguishing the two samples. The quality indices and the oxidative stability of oil samples were detected to identify their nutritional value and edible quality. This study revealed that roasting greatly influenced the lipid and flavor substances of the Hibiscus manihot L. seed oil. Our findings could provide useful information on the impact of the pressing method for Hibiscus manihot L. seed oil.

Effects and mechanisms of total flavones of Abelmoschus manihot in attenuating diabetic tubulopathy by targeting endoplasmic reticulum stress-induced cell apoptosis

Renal tubular injury in patients with diabetic kidney disease(DKD) may be accompanied by glomerular and microvascular diseases. It plays a critical role in the progression of renal damage in DKD, and is now known as diabetic tubulopathy(DT). To explore the multi-targeted therapeutic effects and pharmacological mechanisms in vivo of total flavones of Abelmoschus manihot(TFA), an extract from traditional Chinese medicine for treating kidney disease, in attenuating DT, the authors randomly divided all rats into four groups: a normal control group(normal group), a DT model group(model group), a DT model+TFA-treated group(TFA group) and a DT model+rosiglitazone(ROS)-treated group(ROS group). The DT rat model was established based on the DKD rat model by means of integrated measures. After successful modeling, the rats in the four groups were continuously given double-distilled water, TFA suspension, and ROS suspension, respectively by gavage every day. After 6 weeks of treatment, all rats were sacrificed, and the samples of their urine, blood, and kidneys were collected. The effects of TFA and ROS on various indicators related to urine and blood biochemistry, renal tubular injury, renal tubular epithelial cell apoptosis and endoplasmic reticulum stress(ERS), as well as the activation of the protein kinase R-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α)-activating transcription factor 4(ATF4)-C/EBP homologous protein(CHOP) signaling pathway in the kidney of the DT model rats were investigated. The results indicated that hypertrophy of renal tubular epithelial cells, renal tubular hyperplasia and occlusion, as well as interstitial extracellular matrix and collagen deposition occurred in the DT model rats. Moreover, significant changes were found in the expression degree and the protein expression level of renal tubular injury markers. In addition, there was an abnormal increase in tubular urine proteins. After TFA or ROS treatment, urine protein, the characteristics of renal tubular injury, renal tubular epithelial cell apoptosis and ERS, as well as the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney of the DT model rats were improved to varying degrees. Therein, TFA was superior to ROS in affecting the pathological changes in renal tubule/interstitium. In short, with the DT model rats, this study demonstrated that TFA could attenuate DT by multiple targets through inhibiting renal tubular ERS-induced cell apoptosis in vivo, and its effect and mechanism were related to suppressing the activation of the PERK-eIF2α-ATF4-CHOP signaling pathway in the kidney. These findings provided preliminary pharmacological evidence for the application of TFA in the clinical treatment of DT.

Effects and mechanisms of total flavones of Abelmoschus manihot in improving insulin resistance and podocyte epithelial-mesenchymal transition in diabetic kidney disease based on IRS1/PI3K/Akt pathway

This study aimed to explore the effects and mechanisms of total flavones of Abelmoschus manihot(TFA), the extracts from traditional Chinese medicine indicated for kidney diseases, on insulin resistance(IR) and podocyte epithelial-mesenchymal transition(EMT) in diabetic kidney disease(DKD), and further to reveal the scientific connotation. Thirty-two rats were randomly divided into a normal group, a model group, a TFA group, and a rosiglitazone(ROS) group. The modified DKD model was induced in rats by methods including high-fat diet feeding, unilateral nephrectomy, and streptozotocin(STZ) intraperitoneal injection. After modeling, the rats in the four groups were given double-distilled water, TFA suspension, and ROS suspension correspondingly by gavage every day. At the end of the 8th week of drug administration, all rats were sacrificed, and the samples of urine, blood, and kidney tissues were collected. The parameters and indicators related to IR and podocyte EMT in the DKD model rats were examined and observed, including the general condition, body weight(BW) and kidney weight(KW), the biochemical parameters and IR indicators, the protein expression levels of the key signaling molecules and structural molecules of slit diaphragm in the renal insulin receptor substrate(IRS) 1/phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(Akt) pathway, foot process form and glomerular basement membrane(GBM) thickness, the expression of the marked molecules and structural molecules of slit diaphragm in podocyte EMT, and glomerular histomorphological characteristics. The results showed that for the DKD model rats, both TFA and ROS could improve the general condition, some biochemical parameters, renal appearance, and KW. The ameliorative effects of TFA and ROS were equivalent on BW, urinary albumin(UAlb)/urinary creatinine(UCr), serum creatinine(Scr), triglyceride(TG), and KW. Secondly, they could both improve IR indicators, and ROS was superior to TFA in improving fast insulin(FIN) and homeostasis model assessment of insulin resistance(HOMA-IR). Thirdly, they could both improve the protein expression levels of the key signaling molecules in the IRS1/PI3K/Akt pathway and glomerulosclerosis in varying degrees, and their ameliorative effects were similar. Finally, both could improve podocyte injury and EMT, and TFA was superior to ROS. In conclusion, this study suggested that podocyte EMT and glomerulosclerosis could be induced by IR and the decreased activation of the IRS1/PI3K/Akt pathway in the kidney in DKD. Similar to ROS, the effects of TFA in inhibiting podocyte EMT in DKD were related to inducing the activation of the IRS1/PI3K/Akt pathway and improving IR, which could be one of the scientific connotations of TFA against DKD. This study provides preliminary pharmacological evidence for the development and application of TFA in the field of diabetic complications.

In-Depth Insight into the Mechanism of Incorporation of Abelmoschus manihot Gum on the Enhancement of Gel Properties and In Vitro Digestibility of Frankfurters.

This study aimed to investigate the effects of different concentrations (0.1, 0.2, 0.3, 0.4, and 0.5% w/w) of Abelmoschus manihot gum (AMG) on the gel properties and in vitro digestibility of frankfurters. The results indicated that AMG incorporation significantly enhanced the emulsion stability and texture of frankfurters, as well as the dynamic rheological characteristics of raw meat batter, with the optimal concentration being 0.3% (p < 0.05). Furthermore, hydrogen bonds and disulphide bonds were the main molecular forces of the frankfurters in the presence of AMG. Microstructural images showed that more uniform and dense microstructures of frankfurters were formed due to AMG supplementation. In addition, AMG incorporation significantly increased the in vitro protein digestibility of frankfurters as the level of addition increased (p < 0.05). In conclusion, our results provided critical information for the practical application of AMG in the production of emulsified meat products.

Aktivitas Antihiperglikemik Ekstrak Daun Gedi Merah (Abelmoschus manihot L.Medik) pada Tikus Model Diabetes Melitus Tipe II

Type II diabetes mellitus is a metabolic disorder characterized by increased blood glucose levels due to damage to pancreatic β cells which causes reduced insulin production or abnormal insulin action. The leaves of Abelmoschus manihot L.Medik as an antidiabetic have been used empirically by the community. This study aims to determine the activity of the ethanol extract of Abelmoschus manihot L.Medik leaves as an antihyperglycemic agent in male rats fed a high-fat diet and experiencing type 2 DM induced by streptozotocin. The test animals were divided into six groups: the normal control group (KN) without treatment; the negative control group (K-), which was treated with 0.5% NaCMC, the positive control group (K+), which was treated with metformin 9 mg and the test group of extracts. With doses varying from 75 mg/kg, 150 mg/kg, and 300 mg/kg, rats were first given a high-fat diet, then induced by streptozotocin at a 30 mg/kg dose extract was given for seven days. The results of the phytochemical screening showed that the ethanol extract of red gedi leaves contained alkaloids, flavonoids, tannins, saponins, and steroids. The measurement results showed that administering ethanol extract of red gedi leaves (Abelmoschus manihot L.Medik) doses of 75 mg/kg, 150 mg/kg, and 300 mg/kg affected reducing blood glucose levels in male rats of the Wistar strain type II diabetes mellitus model. Significant based on the One-way ANOVA test and the Least Significance Difference (LSD) post hoc test.

Standardized Identification of Compound Structure in Tibetan Medicine Using Ion Trap Mass Spectrometry and Multiple-Stage Fragmentation Analysis.

Tibetan medicines are complex and contain numerous unknown compounds, making in-depth research on their molecular structures crucial. Liquid chromatography-electrospray ionization time-of-flight mass spectrometry (LC-ESI-TOF-MS) is commonly used to extract Tibetan medicine; however, many unpredictable unknown compounds remain after using the spectrum database. The present article developed a universal method for identifying components in Tibetan medicine using ion trap mass spectrometry (IT-MS). The method includes standardized and programmed protocols for sample preparation, MS setting, LC prerun, method establishment, MS acquisition, multiple-stage MS operation, and manual data analysis. Two representative compounds in the Tibetan medicine Abelmoschus manihot seeds were identified using multiple-stage fragmentation, with a detailed analysis of typical compound structures. In addition, the article discusses aspects such as ion mode selection, mobile phase adjustment, scanning range optimization, collision energy control, collision mode switchover, fragmentation factors, and limitations of the method. The developed standardized analysis method is universal and can be applied to unknown compounds in Tibetan medicine.

An efficient two-step approach for the preparative separation and purification of eight polyphenols from Hibiscus manihot L. flower with high-speed countercurrent chromatography

In the present study, a green and efficient ultrasonic-assisted extraction method of polyphenols from Hibiscus manihot L. flowers (HMLF) was performed, and the corresponding experimental parameters were optimized by Taguchi OA Design. Meanwhile, a two-step high-speed counter-current chromatography coupled with macroporous resin was successfully established with the optimal two-phase solvent systems comprising EtOAc-MeOH-H2O (100:3:100, v/v/v) and Hex-EtOAc-MeOH-H2O (1:5:1:5, v/v/v/v) to isolate and prepare eight polyphenols with purities exceeding 95%, including two phenolic acids and six flavonoids, respectively. The yield of the isolated polyphenols was determined to be 50.4 mg from the enriched extract in a single run. Moreover, the interactions of the eight isolated compounds with AChE and BuChE were explicitly evaluated and described by molecular docking. This study confirms that the proposed method is a cost-effective and powerful strategy for the scaled-up extraction and preparative separation of the high-purity polyphenols from HMLF, and provides an opportunity for automation and systematic preparation of natural products used in the food and pharmaceutical industries.

Effects of Abelmoschus manihot gum content, heating temperature and salt ions on the texture and rheology properties of konjac gum/Abelmoschus manihot gum composite gel

To improve the gelling property of konjac gum (KGM) and enhance the application value of Abelmoschus manihot (L.) medic gum (AMG), a novel type of gel was prepared using KGM and AMG in this study. The effects of AMG content, heating temperature and salt ions on the characteristics of KGM/AMG composite gels were studied by Fourier transform infrared spectroscopy (FTIR), zeta potential, texture analysis and dynamic rheological behavior analysis. The results indicated that the AMG content, heating temperature and salt ions could affect the gel strength of KGM/AMG composite gels. Hardness, springiness, resilience, G′, G* and η* of KGM/AMG composite gels increased when AMG content increased from 0 to 2.0 %, but they decreased when AMG increased from 2.0 % to 3.5 %. High-temperature treatment significantly enhanced the texture and rheological properties of KGM/AMG composite gels. The addition of salt ions reduced the zeta potential absolute value and weakened the texture and rheological properties of KGM/AMG composite gels. Furthermore, the KGM/AMG composite gels could be classified as non-covalent gels. The non-covalent linkages included hydrogen bonding and electrostatic interactions. These findings would help understand the properties and formation mechanism of KGM/AMG composite gels and help improve the application value of KGM and AMG.

Total flavone of flowers of Abelmoschus manihot (L.) Medic inhibits the expression of adhesion molecules in primary mesenteric arterial endothelial cells and ameliorates dextran sodium sulphate-induced ulcerative colitis in mice

BackgroundFlowers of Abelmoschus manihot (L.) medic (AM) is a traditional Chinese medicine used to treat chronic nephritis, nephrotic syndrome, diabetic nephropathy, and colonic inflammation. PurposeThis study aimed to explore the influence of the total flavone of AM flowers (TFA) on acute ulcerative colitis (UC) and the potential underlying mechanism. MethodsEfficacy of TFA (30, 60, 120 mg/kg) on UC was evaluated in a dextran sodium sulphate (DSS)-induced colonic inflammatory mouse model by analyzing disease activity index (DAI), histopathological score, colon length, and cytokine expression. Expression levels of critical adhesion molecules and nuclear factor kappa B (NF-κB) were examined by qRT-PCR, Western blotting, or immunofluorescence labeling. Myeloperoxidase activity was examined using ELISA. In vitro THP-1 adhesion assay was used to evaluate monocyte adhesion. ResultsTFA significantly reduced DAI score, prevented colon shortening, and ameliorated histological injuries of colons in DSS-treated mice. TFA inhibited the expression of cytokines (IL-1β and TNF-α) and adhesion molecules (ICAM-1, VCAM-1, and MAdCAM-1) in colon tissues of DSS mice. In vitro studies on mesenteric arterial endothelial cells (MAECs) showed that TFA attenuated TNF-α-induced upregulation of ICAM-1, VCAM-1, and MAdCAM-1, as well as THP-1 cell adhesion to MAECs. TFA also suppressed the phosphorylation and nuclear translocation of NF-κB in MAECs. ConclusionTFA efficaciously ameliorates UC possibly by inhibiting monocyte adhesion through blocking TNF-α-induced NF-κB activation, which in turn suppresses the upregulation of adhesive molecules in colon endothelial cells. Inhibiting the expression of adhesion molecule in MAECs may represent a useful strategy for therapeutic development to treat UC, with TFA being a safe and efficacious therapeutic agent.

Transcriptome and Metabolome Analysis Reveal the Flavonoid Biosynthesis Mechanism of Abelmoschus manihot L. at Different Anthesis Stages.

Abelmoschus manihot L. (HSK) is a rare and endangered species in the wild that grows on the cliffs of deep mountains. As a natural plant, the chemical composition of HSK is relatively complex, which mainly includes flavonoids, organic acids, polysaccharides, and various trace elements with good effects of clearing away heat, anti-inflammatory, analgesic, and calming nerves, and inhibiting tumor cells. In this experiment, different developmental stages of HSK flowers were used for optimization of the flavonoid extraction and determining method. The antioxidant activities, flavonoid accumulation pattern, and synthesis regulatory network were analyzed using biochemistry, RNA-seq, and UPLC-MS/MS. The total content of flavonoids, vitexin rhamnoside, hyperoside, and rutin in HSK flowers at T3 stage (flower wilting) was significantly higher than in T2 (full flowering) and T1 (bud) stages. Compared with T1 and T2, the antioxidant capacity of the T3 flower alcohol extract was also the strongest, including the total reducing ability, DPPH clearance, OH clearance, O2- clearance, and total antioxidant capacity. A total of 156 flavonoids and 47,179 unigenes were detected by UPLC-MS/MS and RNA-Seq, respectively. The candidate genes and key metabolites involved in flavonoid biosynthesis were identified and the regulatory networks were also analyzed in this study. qRT-PCR test further proved that the gene expression level was consistent with the results of RNA sequence data. The relationship between the gene expression and flavonoid accumulation network provides a theoretical basis for the mining and regulation of functional genes related to the flavonoid biosynthesis and metabolism in Abelmoschus manihot L.

Characteristics of Soil Selenium Enrichment And Its Effects on the Aboveground Parts Of Abelmoschus Manihot (L.) Medic

Pot experiments were carried out to study characteristics of soil selenium (Se) enrichment and its influence on Chinese medicinal plant Abelmoschus manihot, and selenium contents in aboveground parts of A. manihot under conditions of natural Se-rich, low soils and exogenous Se addition were analyzed. Results indicated that contents of total Se and various Se fractions of natural Se-rich soil were significantly higher than that of Se-low soil (P &lt; 0.05), and Se content in aboveground parts of A. manihot grown in natural Serich soil was significantly higher than that in natural Se-low soil (P &lt; 0.05). The Se content in aboveground parts of A. manihot increased with the increase of soil total Se content, soil pH and contents of various Se fractions, showing significant tolerance to high Se. Moreover, it was initially found that A. manihot could effectively absorb soil Se even in relatively low Se soil conditions (soil Se content of 0.11~0.12 mg/kg) and thus reaching the standard of Se-rich crops. Bangladesh J. Bot. 51(4): 961-970, 2022 (December) Special

Fermentation of Abelmoschus manihot Extract with Halophilic Bacillus licheniformis CP6 Results in Enhanced Anti-Inflammatory Activities.

Microbial fermentation provides a valorization strategy, through biotransformation, to convert plant-derived raw materials into health-promoting agents. In this study, we have investigated the antioxidative activity of Abelmoschus manihot fermented with various Bacillaceae strains from specific environments and demonstrated the anti-inflammatory effects of Bacillus licheniformis CP6 fermented A. manihot extract (FAME) in lipopolysaccharide (LPS)-stimulated Raw264.7 macrophages. Of 1500 bacteria isolated from various specific environments, 47 extracellular protease- and amylase-producing strains with qualified presumption safety status, belonging to the family Bacillaceae, were selected for A. manihot fermentation. Among them, strain CP6, a halophilic bacterium isolated from Tongyeong seawater in Korea and identified as B. licheniformis, showed the highest antioxidant activity. In particular, FAME exerted anti-inflammatory effects on LPS-stimulated Raw264.7 macrophages. Consequently, FAME had a potent inhibitory effect on nitric oxide (NO) production in LPS-stimulated macrophages, without cytotoxicity. Moreover, FAME downregulated LPS-induced pro-inflammatory mediator and enzyme levels in LPS-induced Raw264.7 cells, including IL-1β, IL-6, TNF-α, iNOS, and COX-2, compared to levels when cells were incubated in A. manihot extract (IAME). Further detailed characterization indicated that FAME suppresses inflammation by blocking NF-κB via IKK phosphorylation inhibition and IκB-α degradation and by downregulating NO production, and inflammatory mediators also decreased NF-κB translocation. Furthermore, FAME inhibited LPS-stimulated activation of MAPKs, including ERK1/2, JNK, and p38, compared to that with either IAME. Therefore, we suggest that FAME could be used for inflammation-related disorders.

IL-10-dependent Effect of Chinese Medicine Abelmoschus manihot on Alleviating Intestinal Inflammation and Modulating Gut Microbiota.

Inflammatory bowel disease (IBD) is a recurrent disease associated with a potential risk of colorectal cancer. Abelmoschus manihot (AM), a Chinese herbal medicine, is known to alleviate IBD. However, its mechanism of action requires further clarification. Here, we focused on the role of IL-10 and the gut microbiota in the mechanism of action of AM. The effects of AM on intestinal inflammation, mucus production, and gut microbes were evaluated in dextran sodium sulfate (DSS)-induced acute and chronic IBD models and in IL-10-deficient mice (IL-10[Formula: see text]). AM exhibited protective effects on acute and chronic models of IBD in wild-type mice by restoring body weight and colon length, promoting IL-10 secretion, and decreasing TNF-[Formula: see text] levels. Moreover, AM alleviated inflammatory infiltration, increased mucin 2 transcription, and increased the number of goblet cells in the colon. On the contrary, these effects were diminished in IL-10[Formula: see text] mice, which implied that the effect of AM on intestinal inflammation is IL-10-dependent. A gut microbial sequencing analysis showed that gut microbial dysbiosis was modulated by AM intervention. The regulatory effects of AM on Eggerthellaceae, Sutterellaceae, Erysipelotrichaceae, Burkholderiaceae, Desulfovibrionaceae, and Enterococcaceae were dependent on IL-10. These results revealed that AM ameliorated IBD and modulated gut microbes by promoting IL-10 secretion, indicating that AM has the potential to improve IBD and that AM is IL-10-dependent.

Efficient, rapid and incremental extraction of bioactive compounds from the flowers of &lt;i&gt;Hibiscus manihot&lt;/i&gt; L.

Flavonoids are the primary functional components in the flower of <italic>Hibiscus manihot</italic> L. (HMLF). In this work, an efficient and green ionic liquid-high-speed homogenization coupled with the microwave-assisted extraction (IL-HSH-MAE) technique was firstly established and implemented to extract seven target flavonoids from HMLF. Single-factor experiments and Box-Behnken design (BBD) were utilized to maximize the extraction conditions of IL-HSH-MAE, which were as follows: 0.1 M of [C₄mim]Br, homogenate speed of 7000 rpm, homogenate time of 120 s, liquid/solid ratio of 24 mL/g, extraction temperature of 62 °C and extraction time of 15 min. The maximal total extraction yield of seven target flavonoids attained 22.04 mg/g, which was considerably greater than the yields obtained by IL-HSH, IL-MAE, 60% ethanol-HSH-MAE and 60% ethanol-MAE. These findings suggested that the IL-HSH-MAE method can be exploited as a rapid and efficient approach for extracting natural products from plants. Meanwhile, it possesses outstanding superiority of environmentally friendly and high extraction efficiency that is expected to be a luciferous prospect extraction technology.

MICROENCAPSULATION OF FLAVONOIDS FROM GEDI LEAVES (Abelmoschus manihot L.) IN β-CYCLODEXTRIN USING COPRECIPITATION AND KNEADING METHODS

The protection of active compounds such as flavonoids as candidate drug compounds has been the focus of several studies in recent years. This is due to the different physicochemical properties of flavonoids. Furthermore, the function of these flavonoid compounds can be maximized from the initial process of separating them from the plant of origin known as extraction. This study compared the effectiveness and efficiency of using microwave extraction and maceration of Gedi leaves (Abelmoschus manihot L.). Then, the extract was encapsulated to protect the unstable flavonoids. There were two methods to encapsulate the extract in this study. They were the coprecipitation and kneading methods. The result shows that microwave extraction assisted has higher product in less solvent and was time-consuming. The coprecipitation also shows better performance in encapsulating the extract. The calculation of the efficiency of encapsulation was done for 3 different volumes of both methods. The addition of 1 mL extract by coprecipitation showed approximately 92.12%. It was also supported by the data of FTIR and UV VIS.

Topical Administration of Red Gedi Leaves (Abelmoschus Manihot L. Medik) Gel Extract Increases Neovascularization, Number of Fibroblasts and Re-Epithelialization on Wounds in Diabetic Rats

Flavonoids (quercetin), saponins, tannins, alkaloids and phenols (gallic acid) in red gedi leaves (Abelmoschus manihot L. Medik) can accelerate wound healing which is often found in diabetes mellitus. This was an experimental randomized post-test only control group design using 30 male Wistar rats with diabetes mellitus that were divided into 6 groups. On the 4th and 14th days after treatment, skin tissue samples were taken, histology preparations were made using Hematoxilin-Eosin (HE) staining to examine neovascularization, number of fibroblasts and re-epithelialization. Comparative analysis showed that the average neovascularization on the 4th day of the treatment I group was higher than treatment group II and the lowest was in the control group. Meanwhile, on 14th day, treatment group I had the highest neovascularization rate (p&lt;0.001). The average of fibroblasts in treatment group I on the 4th day higher than treatment group II and the lowest in the control group. Meanwhile, on 14th day, treatment group I had the highest fibroblast average (p&lt;0.001). The mean re-epithelialization on 4th day was highest in treatment group I and lowest in the control group. Meanwhile, on the 14th day, treatment group I had the highest re-epithelialization average (p&lt;0.001). In conclusion, administration of red gedi leaf ethanol gel extract with concentrations of 12.5% and 25% increased neovascularization, number of fibroblast cells and re-epithelialization in wound healing of male Wistar rats with diabetes mellitus, with a concentration of 12.5% being better for all parameters.

Abelmoschus Manihot ameliorates the levels of circulating metabolites in diabetic nephropathy by modulating gut microbiota in non-obese diabetes mice.

Huangkui capsule (HKC), a traditional Chinese medicine, has been used for medication of kidney diseases, including diabetic nephropathy (DN). The current study aimed to evaluate the effects of HKC in the modulation of gut microbiota and the amelioration of metabolite levels by using non-obese diabetes (NOD) mice with DN. The microbiota from three parts of intestines (duodenum, ileum and colon) in NOD mice with and without HKC treatment were analysed using 16S rDNA sequencing techniques. Untargeted metabolomics in plasma of NOD mice were analysed with liquid mass spectrometry. Results showed that HKC administration ameliorated DN in NOD mice and the flora in duodenum were more sensitive to HKC intervention, while the flora in colon had more effects on metabolism. The bacterial genera such as Faecalitalea and Muribaculum significantly increased and negatively correlated with most of the altered metabolites after HKC treatment, while Phyllobacterium, Weissella and Akkermansia showed an opposite trend. The plasma metabolites, mainly including amino acids and fatty acids such as methionine sulfoxide, BCAAs and cis-7-Hexadecenoic acid, exhibited a distinct return to normal after HKC treatment. The current study thereby provides experimental evidence suggesting that HKC may modulate gut microbiota and subsequently ameliorate the metabolite levels in DN.