Antihypercholesterolemic and Antioxidant Effects of Blumea balsamifera L. Leaf Extracts to Maintain Luteinizing Hormone Secretion in Rats Induced by High-Cholesterol Diets

BACKGROUND: The discovery of herbal ingredients for antihypercholesterolemic and to improve male reproductive function is very necessary due to high-cholesterol diet factors. Blumea balsamifera leaf extract (BBLE) is known to be able to increase the number and diameter of Leydig cells in rats given high-fat feed. This study was to conducted to determine the levels of total cholesterol, body weight, plasma malondialdehyde (MDA), Superoxide Dismutase (SOD), Luteinizing Hormone (LH) in high-cholesterol-fed male rats provided with BBLE.METHODS: This research utilized a randomized post-test only control group. Ethanol was used to extract the BBLE, which was then evaporated. For 21 days, 16 Wistar rats were given a high-cholesterol diet. To determine the effect of BBLE on the high-cholesterol diet, the samples were divided into two groups (control and BBLE group) on day 22. The treatments lasted 30 days. SOD, plasma MDA, LH, and total cholesterol were measured.RESULTS: The results showed that the SOD and LH parameters were significantly higher in the treatment of BBLE compared to the control group (p<0.05). The parameters of total cholesterol levels, bodyweight, and MDA of rats given BBLE were significantly lower than those of the control group (p<0.05).CONCLUSION: Our findings highlight that BBLE has antihypercholesterolemic and antioxidant effects. The BBLE also has potential to be used as a therapy to maintain male reproductive function because it has a positive effect on the hypothalamic-pituitary axis through increasing LH secretion.KEYWORDS: Malondialdehyde, Superoxide dismutase, Luteinizing hormone, Blumea balsamifera extract, High-cholesterol diet

NLRP3 Inflammasome Activation Controls Vascular Smooth Muscle Cells Phenotypic Switch in Atherosclerosis

(1) Background: Monocytes and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome orchestrate lipid-driven amplification of vascular inflammation promoting the disruption of the fibrous cap. The components of the NLRP3 inflammasome are expressed in macrophages and foam cells within human carotid atherosclerotic plaques and VSMCs in hypertension. Whether monocytes and NLRP3 inflammasome activation are direct triggers of VSMC phenotypic switch and plaque disruption need to be investigated. (2) Methods: The direct effect of oxLDL-activated monocytes in VSMCs co-cultured system was demonstrated via flow cytometry, qPCR, ELISA, caspase 1, and pyroptosis assay. Aortic roots of VSMCs lineage tracing mice fed normal or high cholesterol diet and human atherosclerotic plaques were used for immunofluorescence quantification of NLRP3 inflammasome activation/VSMCs phenotypic switch. (3) Results: OxLDL-activated monocytes reduced α-SMA, SM22α, Oct-4, and upregulation of KLF-4 and macrophage markers MAC2, F4/80 and CD68 expression as well as caspase 1 activation, IL-1β secretion, and pyroptosis in VSMCs. Increased caspase 1 and IL-1β in phenotypically modified VSMCs was detected in the aortic roots of VSMCs lineage tracing mice fed high cholesterol diet and in human atherosclerotic plaques from carotid artery disease patients who experienced a stroke. (4) Conclusions: Taken together, these results provide evidence that monocyte promote VSMC phenotypic switch through VSMC NLRP3 inflammasome activation with a likely detrimental role in atherosclerotic plaque stability in human atherosclerosis.

The possible alleviating effect of garlic supplement on the neural retina in a rat model of hypercholesterolemia: a histological and immunohistochemical study

The purpose of this work was to prove that oxidative stress is the main mechanism responsible for retinal neurodegenerative changes, subsequent apoptosis, and inflammatory cytokine release in rats fed with a high cholesterol diet (HCD) and determine the role of garlic in alleviating these changes. Forty rats were equally divided into four groups: control, garlic-treated (positive control), HCD, and HCD + garlic-treated (HCD + G). By the end of the experiment (24 weeks) blood samples were collected for assessment of serum lipid profile, oxidative stress parameters, and plasma levels of IL-6 and TNF-α. Both eyes of the rats were enucleated; one was used for light microscopic examination and the other for electron microscopic examination. There was a significant increase in the levels of serum lipids, oxidative stress parameters, IL-6 and TNF-α, and area of expression of caspase-3 in the HCD group compared to both the control and HCD + G groups. Histological examination revealed degenerative changes in all layers of the neural retina in the HCD group. Garlic administration resulted in a significant improvement in the biochemical, immunohistochemical, and histological characteristics of hypercholesterolemic rats. These findings support the hypotheses that garlic has strong antioxidant, anti-apoptotic, and anti-inflammatory properties. Garlic ameliorates the neurodegenerative changes in the neural retina of hypercholesteremic rats.

ANTIHYPERLIPIDEMIC EFFECT OF THE ETHANOL EXTRACT FRACTION FROM MULBERRY (MORUS AUSTRALIS POIR.) LEAVES ON RATS INDUCED HIGH FAT-DIET (HFD) AND PROPYLTHIOURACIL (PTU)

Objectives: To evaluate the effect of extract and its fraction (n-hexane, ethyl acetate, and water) of the mulberry leaf (Morus australis Poir.) in reducing triglyceride and total cholesterol levels on HFD and PTU induced rats. Methods: In this study, mulberry leaves were collected from East Nusa Tenggara-Indonesia, extracted using maceration method, and fractionated with various solvents. The thin layer chromatography with silica gel 60 F254 was utilized to identify the active compound on both extract and fraction. Furthermore, using a quasi-experimental with control group design, this study employed 40 healthy male Wistar rats and divided into 8 groups, namely: normal and negative control, positive controls (simvastatin and gemfibrozil), ethanolic extract, and fractions (n-hexane, ethyl acetate, and water). All groups were fed with a high cholesterol diet and 0.01% PTU, except for the normal group. During the study, the blood was drawn at day 0, 28, 35, and 42. The triglyceride level was observed using Glycerophosphate-Oxidase Phenol Aminoantipyrine (GPO-PAP) method, while the total cholesterol was measured by Cholesterol Oxidase Phenol Aminoantipyrine (CHOD-PAP). Results: On the 42nd day, ethyl fraction of mulberry leaf (Morus australis Poir.) showed the most significant result of reducing the level of triglyceride and total cholesterol compared to the high-fat diet and PTU induced rat on day 0: triglyceride of 104.89±2.70 mg/dl vs 64.76±1.97 mg/dl and total cholesterol of 96.70±2.45 vs 88.02±1.38 mg/dl. Ethyl acetate fraction was similar to simvastatin in lowering the level of total cholesterol. TLC identification also showed that mulberry leaf contains flavonoids and polyphenols that act as anti-hyperlipidemic. Conclusion: Ethyl acetate fraction of mulberry leaf (Morus australis Poir.) showed best activity on lowering both triglyceride and total cholesterol.

Evaluation of Hypolipidemic and Hepatoprotective Effects of Linum usitatissimum L. Seeds on Hypercholesterolemic Rats

Background: Hyperlipidaemia is a chronic progressive disease, which encompasses various genetic and acquired conditions resulting in inappropriately elevated lipid levels in humans. Several preclinical and clinical studies have demonstrated that dietary supplementation with Linum Usitatissimum have beneficial cardiovascular effects like antihypertensive action, antiatherogenic effects, lowering of cholesterol, anti-inflammatory action and inhibition of arrhythmia. There are few studies on hepatoprotective effects of flax seeds in hyperlipidemic animal models. Aim and objectives: Hence, this study was undertaken to evaluate the hypolipidemic and hepatoprotective effect of Linum seeds. Methods: The study was conducted for the period of two months in Department of Pharmacology KIMS, Bhubaneswar. Total 36 Albino wistar rats were taken and divided into 6 groups containing 6 rats each. Group II was hypercholesterolemic (HC) control, Group III was treatment control and Group IV to VI rats were given different doses of extract. Blood samples were taken at the end of two months. Results: All the values are expressed as Mean±SD (standard deviation). Significant decrease in lipid parameters and liver enzymes is seen at the end of two months in all 3 groups receiving the extract, where the highest dose of extract used, is near to the standard drug. Conclusion: From this study, it can be concluded that the addition of the flaxseeds to the diet may alleviate the rise in circulating cholesterol levels induced by the high cholesterol diet through its content of alfa-linolenic acid (ALA) and lignins. The hepatoprotective role of flaxseed in hypercholesterolaemia has also been demonstrated in this this study.

Anti-Galectin-2 antibody treatment reduces atherosclerotic plaque size and alters macrophage polarity

Galectins have numerous cellular functions in immunity and inflammation. Short-term galectin-2 blockade in ischaemia-induced arteriogenesis shifts macrophages to an anti-inflammatory phenotype and improves perfusion. Galectin-2 may also affect other macrophage related cardiovascular diseases. This study aims to elucidate the effects of Galectin-2 inhibition in atherosclerosis. ApoE -/- mice were given a high cholesterol diet (HCD) for 12 weeks. After six weeks of HCD, intermediate atherosclerotic plaques were present. To study the effects of anti-Gal2 nanobody treatment on the progression of existing atherosclerosis, treatment with two llama derived anti-Gal2 nanobodies (clones- 2H8 and 2C10), or vehicle was given for the remaining 6 weeks. Galectin-2 inhibition reduced the progression of existing atherosclerosis. Atherosclerotic plaque area in the aortic root was decreased, especially so in mice treated with 2C10 nanobodies. This clone reduced atherosclerosis severity as reflected by a decrease in fibrous cap atheromas in addition to decreases in plaque size. The number of plaque resident macrophages was unchanged, however, there was a significant increase in the fraction of CD206+ macrophages. 2C10 treatment also increased plaque α-smooth muscle content, and Gal-2 may have a role in modulating the inflammatory status of smooth muscle cells. Remarkably, both treatments reduced serum cholesterol concentrations including reductions in VLDL, LDL, and HDL whilst triglyceride concentrations were unchanged. Prolonged treatment with anti-Gal-2 nanobodies reduced plaque size, slowed plaque progression, and modified the phenotype of plaque macrophages toward an anti-inflammatory profile. These results hold promise for future macrophage modulating therapeutic interventions that promote arteriogenesis and reduce atherosclerosis.

Mangrove (Rhizophora sp) Fruit Extract Inhibits Tumor Growth Factor (TGF)-β1 Expression in High Cholesterol Diet-Fed Rats

Background: Atherosclerosis is a major finding in cardiovascular disease. One of the pro-fibrotic cytokines that play an important role in the atherosclerosis process is Tumor Growth Factor (TGF)-β1, where the presence of high TGF-β1 secretion due to hypercholesterolemia will trigger excessive collagen matrix formation. Objective: To analyze Rhizophora sp fruit extract effect on TGF-β1 expression in high cholesterol diet-fed rats. Material and Methods: Eighteen 10-week-old rats weighing 150-200 g were used in this study. These animals were grouped into 3 groups, each consisting of 6 rats. Group A (normal control) is a group of rats that received a normal diet. Group B (atherogenic control) received a diet that induces atherosclerosis (atherogenic diet). This diet contains 2% cholesterol, 5% goat fat, 0.2% cholic acid and standard diet up to 100%. Atherogenic diet was given for 3 days, and on the first day this group also received vitamin D3 700,000 IU/kg. Group C (treated), apart from receiving an atherogenic diet, was also given Rhizophora sp fruit extract 500 mg/kg body weight. The Rhizophora sp fruit extract effect on TGF-β1 expression was evaluated by immunohistochemical procedure. The area of ​​the expression is calculated using the ImageJ. Results: The results of this study indicate that the expression of TGF-β1 is higher in the group receiving the atherogenic diet than the normal control group (17.3 vs. 8.9; P=0.000). Rhizophora sp fruit extract reduced this expression remarkably (17.3 vs. 11.4; P=0.001). Conclusion: Rhizophora sp fruit extract inhibits the the expression of TGF-β1 in high cholesterol diet-fed rats.