Sterol O-acyltransferase (SOAT/ACAT) activity is required to form cholesterol crystals in hepatocyte lipid droplets.

OBJECTIVE: Excess cholesterol storage can induce the formation of cholesterol crystals in hepatocyte lipid droplets. Such crystals distinguish metabolic dysfunction associated steatohepatitis (MASH) from simple steatosis and may underlie its pathogenesis by causing cell damage that triggers liver inflammation. The mechanism linking cholesterol excess to its crystallization in lipid droplets is unclear. As cholesteryl esters localize to and accumulate in lipid droplets more readily than unesterified free cholesterol, we investigated whether cholesterol esterification by sterol O-acyltransferase (SOAT), also known as acyl co-A cholesterol acyltransferase (ACAT), is required for hepatocyte lipid droplet crystal formation. METHOD: Cholesterol crystals were measured in cholesterol loaded Hep3B hepatocytes, RAW264.7 macrophages, and mouse liver using polarizing light microscopy. We examined the effect of blocking SOAT activity on crystal formation and compared these results to features of cholesterol metabolism and the progression to intracellular crystal deposits. RESULTS: Cholesterol loading of Hep3B cells caused robust levels of lipid droplet localized crystal formation in a dose- and time-dependent manner. Co-treatment with SOAT inhibitors and genetic ablation of SOAT1 blocked crystal formation. SOAT inhibitor also blocked crystal formation in low density lipoprotein (LDL) treated Hep3B cells, acetylated LDL treated RAW 264.7 macrophages, and in the liver of mice genetically predisposed to hepatic cholesterol overload and in mice with cholesterol enriched diet-induced MASH. CONCLUSION: SOAT1-mediated esterification may underlie cholesterol crystals associated with MASH by concentrating it in lipid droplets. These findings imply that inhibiting hepatocyte SOAT1 may be able to alleviate cholesterol associated MASH. Moreover, that either a lipid droplet localized cholesteryl ester hydrolase is required for cholesterol crystal formation, or the crystals are composed of cholesteryl ester.

Efficacy of using walnuts as statin adjuvants in hypertension management.

BACKGROUND: Due to the widespread unorthodox use of nuts to improve cardiovascular health, this clinical trial was carried out to evaluate the efficacy of walnut as an adjuvant statin in hypertensive subjects. METHOD: A single-blind placebo-controlled randomized clinical trial that lasted for 3 months. Forty-five screened hypertensive subjects on treatment, aged 45-65 years, were randomized into intervention and placebo groups according to their blood pressure defined by the American Heart Association criteria. Fifteen (15) normotensive subjects were also recruited for this study. The participants in the intervention group included daily 7 g of boiled walnut taken as snacks. The study was not controlled for type of diet and frequency of meals in a day. Low-density lipoprotein cholesterol (LDLc) was the primary endpoint for this study. RESULTS: The mean LDLc levels of the intervention groups (84.6 mg/dl and 79.7 mg/dl, respectively) were significantly (p < .005) lower than the placebo (137.6 mg/dl). The high-density lipoprotein cholesterol (HDLc) levels of the intervention groups were significantly higher than the placebo. The mean total cholesterol levels of the intervention groups were significantly lower than the placebo group. The intervention groups recorded a significantly lower systolic and diastolic blood pressure compared to the placebo. The supplementation of walnut significantly decreased the apolipoprotein E (APOE), proprotein convertase subtilisin kexin 9 (PCSK9), and cholesteryl ester transfer protein (CETP) activities relative to the placebo. CONCLUSION: The use of walnut as a statin adjuvant during hypertension treatment reduced LDLc levels within 42.1% and improved HDL levels by 33.6%, and the LDLc decrease related to reduced PCSK9 and APOE activities while the HDLc increase related to reduced CETP activities.

Preclinical Trial of Traditional Plant Remedies for the Treatment of Complications of Gestational Malaria.

Background: Most pregnant women living in high malaria endemic regions of Nigeria use herbal remedies for the management of malaria-in-pregnancy, rather than the commonly prescribed drugs. Remedies common to this area involve a suspension of A. indica (AI) leaves and in some cases, a suspension containing a mixture of AI and D.edulis (PS). Aim: This study examined the therapeutic efficacies of AI, PS, or a combination of AI and PS in a pregnant rat model for exoerythrocytic stages of Plasmodium falciparum parasite. Method: A predetermined sample size of 30 dams was used (for a power level and confidence interval of 95%), and divided equally into six groups made up of non-malarous dams, untreated malarous dams, and malarous dams either treated exclusively with 1 mL of 3000 mg/kg b.w AI, 1000 mg/kg b.w PS, AI + PS (50% v/v), or 25 mg/kg b.w CQ. Result: No maternal mortality was recorded. AI significantly improved maternal weight gain from 32.4 to 82.2 g and placental weight from 0.44 to 0.53 g. In the curative test, AI and AI + PS significantly reduced the average percentage parasitemia (APP) in the pregnant rats from >80% to <20%. No significant difference in the APP was found between the pregnant rats treated with any of CQ or AI during the suppressive test. Results for the prophylactic test of the study groups showed that the APP was significantly reduced from 24.69% to 3.90% when treated with AI and 3.67% when combined with PS. AI + PS reduced diastolic blood pressure from 89.0 to 81.0 mm/Hg and compared with that of the non malarous dams. AI or AI + PS significantly increased the platelet counts (103 µL) from 214.1 to 364.5 and 351.2, respectively. AI and AI + PS improved birth weight from 2.5 to 3.9 g and crown rump length from 2.6 to 4.1 cm. For biomarkers of preeclampsia, combining AI and PS led to the reversal of the altered levels of creatine kinase, lactate dehydrogenase, cardiac troponin, soluble Fms-Like Tyrosine Kinase-1, and placental growth factor. Conclusions: This study validates the use of A. indica for the treatment of gestational malaria due to its antiplasmodial and related therapeutic effects and in combination with pear seeds for the management of malaria-in-pregnancy-induced preeclampsia.

Lettuce-avicularin treatment reverses insulin resistance through stimulation of glycolytic kinases and insulin signaling molecules.

OBJECTIVES: In order to recommend a more effective approach to manage insulin resistance, we monitored the activities of glycolytic kinases, insulin signaling molecules, and incretin hormones and identified the possible targets related to the insulin-sensitizing effects of combined pharmacological and dietary intervention involving avicularin and lettuce. MATERIALS AND METHODS: Insulin resistance was induced in rats with a fructose-rich diet and confirmed from baseline analysis of FBS (>250 mg/dl), insulin (>25 µIU/ml), and HOMA-IR (>10). For 12 weeks, the insulin-resistant rats were treated exclusively with 5000 mg/kg b.w avicularin (DAvi) or by dietary placement on lettuce (DLet) or a combination of both and compared with non-insulin resistant rats. RESULTS: Avicularin reversed alterations in HbA1c and insulin levels. DLet produced no significant effect on the incretins GLP 1 (P=0.909) and GIP (P=0.990), but DAvi slightly stimulated GLP 1 but not GIP. A strong positive correlation was found between improved ß-cell responsiveness and the insulin signaling molecules: Akt2 (r=0.7248), IRS 1 (r=0.5173), and PI3K (r=0.7892). Only the combined avicularin and lettuce reversed the Akt2 levels (P=0.728). The lettuce meal slightly stimulated PI3K but normalized IRS 1 while avicularin treatment slightly stimulated IRS 1 but restored the PI3K levels (P=0.815). The positive correlation between ß-cell responsiveness and hexokinase (r=0.5959), PFK (r=0.6222), and PK (r=0.6591) activities were statistically significant. Alterations in glycolytic kinases were reversed by DLet and in combination with avicularin. CONCLUSION: A combined pharmacological and dietary approach with avicularin and lettuce is required to effectively reverse insulin resistance.

Nutritional modulation of blood pressure and vascular changes during severe menstrual cramps.

OBJECTIVES: This study examined the influence of nutrition on the severity of menstrual pains and associated transient changes in blood pressure (BP) and vascular-health indicators. It has also investigated the influence of nutrition on angiotensin (ANG II) and vascular cell adhesion molecules (VCAM-1). METHODS: A total of 207 university students, aged between 18 and 25 years, were grouped into three groups: a no-dysmenorrhoea (control) group, a moderate dysmenorrhoea (MDys) group, and a severe dysmenorrhoea (SDys) group, using the NRS-11 scale and initial contactin-1 (CNTN-1) levels. The groups were separately fed vegetable, protein, and carbohydrate meals. The meal plan involved three different types of food served three times a day (for breakfast, lunch, and dinner), beginning 48 h before menstruation. RESULTS: We found that 73.9% and 100% of the MDys patients on the protein and carbohydrate diets, respectively, had severe dysmenorrhoea. As many as 69.6% of the SDys patients on vegetable diets experienced no dysmenorrhoea; the BP of 61% of SDys normalised to the standard values of 120/80. The BP of 87% MDys had systolic BP ≥ 130 and ≥90 diastolic BP after carbohydrate meals. On the other hand, 30% of SDys had higher BP after protein meals. With respect to the choice of food, the severity of menstrual pain was positively correlated with ANG II (r = 0.5158) and VCAM-1 (r = 0.5849). ANG-II. Similarly, VCAM-1 were significantly elevated (p < 0.05) in the dysmenorrhoeal participants. The mean VCAM-1 and ANG-II levels of dysmenorrhoeal participants placed on vegetable meals were comparable to the control baseline levels. CONCLUSIONS: This study recommends the intake of a vegetable meal at least 48 h before menstruation as an effective nutritional approach to preventing and managing severe menstrual cramps. This approach can also prevent associated vascular changes. Carbohydrate meals should be avoided at least 48 h before menstruation.

Okra Modulates Regulatory Enzymes and Metabolites of Glucose-Utilizing Pathways in Diabetic Rats.

OBJECTIVE: Using a rat diabetes model, the authors examined how substrates and products of glycolysis and key regulatory enzymes for glycolysis, gluconeogenesis, Kreb's cycle, and glycogen metabolism react to treatment with okra diet therapy, relative to glibenclamide treatment. METHOD: The animal grouping involved normoglycemic rats, untreated diabetic rats, and diabetic rats treated with glibenclamide, 50% w/w okra sauce, exclusive okra sauce diet, or sauce without okra. Alloxan monohydrate was the diabetogenic agent. Insulin and adiponectin were assayed with enzyme-linked immunosorbent assay (ELISA) while the metabolites and enzymes were assed using standard spectrophotometric methods. RESULTS: The exclusive diet therapy significantly (p < 0.05) improved insulin activities after 60 days and reversed the altered adiponectin activities. Glucose-6-phosphate, fructose-6-phosphate, and fructose-1,6-bisphosphate levels were depleted during diabetes, but phosphoenolpyruvate and pyruvate accumulated during the first short phase of diabetes. Rats in the glibenclamide and 100% okra diet groups showed comparable hexokinase, phosphofructokinase, and pyruvate kinase activities relative to the normoglycemic rats, while the gluconeogenic enzymes, glucose-6-phosphatase, and fructose-1,6-bisphosphatase remained altered. The authors observed that extended treatment with glibenclamide restored the activities of all the Kreb's cycle enzymes, while succinate dehydrogenase and α-ketoglutarate dehydrogenase were nonresponsive to the okra diet therapy relative to their control levels. The glycogen stores were normalized by the exclusive diet therapy, but glycogen synthase and phosphorylase activities were unresponsive. CONCLUSIONS: Okra diet has shown insulin-sensitizing potentials with prolonged intake during diabetes as well as the potential to reverse alterations in the major carbohydrate-metabolizing enzyme.

Potentiation of incretin hormones and modulation of metabolic enzymes as possible mechanisms behind the insulin sensitizing effects of cabbage-metformin treatment.

In our study, we treated high fructose diet-induced insulin resistance in rats with any of metformin, cabbage (80%w/w) or combined metformin and cabbage (MetCabb), and observed the activities of glycolysis and gluconeogenesis regulatory enzymes, incretin hormones and other hormones affecting glucose homeostasis. Comparisons were made with normoglycemic noninsulin resistance rats (control) and insulin-resistant untreated rats (INres). Baseline analysis showing elevated fasting blood sugar (>250 mg/dl), insulin (>25 µIU/ml) and HOMA-IR (>10) satisfied the criteria for recruitment into the insulin-resistant groups. Treatment lasted for 12 weeks. HOMA-IR values significantly (P < 0.05) decreased from 24.7 to 5.5 and 10.6 respectively with MetCabb treatment. MetCabb normalized HOMA-IR values and mean ß-cell responsiveness of the INres. Cabbage and metCabb normalized the leptin levels relative to control. The mean fasting blood sugar, insulin, and c-peptide levels with MetCabb treatment reverted to control levels. We found a strong positive linear correlation between the glucagon levels (r = 0.9145) and increasing HOMA-IR values while both incretin hormones; GLP-1 and GIP negatively regressed (r = -0.8084 and -0.8488). MetCab treatment produced comparable values of GLP-1 and GIP to the control. A strong positive correlation was found between the HOMA-IR values and the PEPCK (r = 0.9065), F-1,6-BPase (r = 0.7951), and G-6-Pase (r = 0.7893). The hexokinase (r = -0.807), PFK (r = -0.9151), and PK (r = -0.7448) levels regressed as HOMA-IR values increased. The glycolytic and gluconeogenic enzymes except PEPCK reverted to control levels with MetCabb treatment. Combination of metformin and cabbage was more effective than individual treatments.

The Relationship between Pain and Vascular Function Biomarkers in Dysmenorrheal University Students.

Our aim was to establish if the secretion of contactin 1 (CNTN-1), a widely researched pain biomarker correlates with the severity of dysmenorrhea and circulating levels of vascular cell adhesion molecule 1 (VCAM-1) and angiotensin II (ANG-II). This study was a longitudinal randomized clinical study that involved 95 female students between 17-25 years. The control participant group were students who, without medications, had not experienced dysmenorrhea, while the inclusion criteria were primary dysmenorrhea without medications. Data was collected using demographic questionnaires that also contained the Numeric Rating Scale (NRS-11), while blood samples were collected for analysis of CNTN-1, VCAM-1 and ANG-II by ELISA. The participants' mean BMI's across the four pain strata were between 16.60-38.43 kg/m2 and in addition to age and menarche, showed no correlation to either the NRS-11 scale (r=-0.01214) or their CNTN-1 levels (r=0.009622). The severe dysmenorrhea group showed statistically higher (p<0.0001) and positive correlation to systolic (r=0.7304) and diastolic (0.6588) blood pressures. The contactin 1 levels (7.00-55.70 ng/mL) increased with higher menstrual pain and as the pain increased, so did the mean VCAM-1 and ANG-II levels (p<0.0001). A positive linear correlation (r=0.9691) was observed between the NRS-11 scale of the participants and their CNTN-1 activities while the CNTN-1 levels positively correlated with their VCAM-1 (r=0.9334) and ANG-II (r=0.8746) secretion. In summary, the severity of dysmenorrheal pain elevates the contactin 1 levels which affects their vascular health and blood pressure.