Bioinformatics Analysis of LDLR Gene Mutation that Shed a New Light on Red Yeast Rice Monacolin K Treatment � Systematic Review

Background: Hypercholesteremia is the major cause of cardiovascular diseases. It results from elevated cholesterol levels in the blood. LDL cholesterol is removed from the circulation by using the LDL receptor. Red mold rice or red yeast rice is produced by fermentation of the Monascus Purpureus yeast on rice. Many researchers suggest that the active component in Red Yeast Rice (monacolin k) serves as a treatment for hypercholesteremic patients. Methods: By using NCBI databases, specifically GenBank to analyze DNA sequence and mRNA sequence of LDLR gene. GenBank file format was helpful to extract an accession number of the gene, number of amino acids, exons, and length of nucleotides. FASTA format was also useful to retrieve the nucleotide sequence and get the function of the protein. BLAST was used to compare the protein product of the LDLR gene between humans and pan paniscus (pygmy chimpanzee). Results: In accession number NC_000019, the number of amino acids in protein product is 44389 bp, and the number of exons found is 18. On the other hand, the gene is located in chromosome 19. The function of LDLR gene is to control the production of LDL receptor where the low-density lipoprotein particles attach to it and are taken into the cell ending up in the lysosome where the protein is degraded and cholesterol is made which will inhibit 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase that controls the production of cholesterol. Finally, many organisms have the same gene like dogs, cows, mice, rats, zebrafish, and frogs. Conclusion: Mutation in the LDLR gene causing high level of cholesterol in the blood especially LDL (Low-density Lipoprotein). Monacolin k that found in red yeast rice (RYR) is safe and natural alternative treatment for hypercholesteremic patients by lowering the cholesterol level in the blood.

Effects of cassia seed,red yeast rice,ginkgo biloba leaf and salvia miltiorrhiza compounds on blood lipid level of rats / 预防医学

Objective @#To learn the effects of cassia seed extract,ginkgo biloba leaf extract,salvia miltiorrhiza extract and red yeast rice compounds on blood lipid level of rats with hyperlipidemia. @*Methods @#The model of hyperlipidemia in SD rats was established. The rats in the low,moderate and high dose groups were given 0.267 g/kg body weight,0.533 g/kg body weight and 1.600 g/kg body weight of cassia seed,red yeast rice,ginkgo biloba leaf and salvia miltiorrhiza compounds by gavage for 30 days. A blank control group and a model control group were also set. The levels of serum total cholesterol（TC）,triglyceride（TG）,high-density lipoprotein cholesterol（HDL-C）and low-density lipoprotein cholesterol（LDL-C）in the five groups were measured before and after the intragastric administration. @*Results @#One week after the model was founded,the levels of serum TC,TG and LDL-C in the model control group were higher than those in the blank control group（P<0.05）. Before the intragastric administration,there were no significant differences in the levels of serum TC,TG,HDL-C and LDL-C between the model control group and each dosage group（P>0.05）. After the intragastric administration,there were no significant differences in the levels of serum HDL-C and LDL-C between the model control group and each dosage group（P>0.05）. The levels of serum TC and TG in the high dose group were significantly lower than those in the model control group（P<0.05）;There was no significant differences in the levels of serum TC and TG between the moderate dose,low dose group and the model control group（P>0.05）. @*Conclusion@# The compounds of cassia seed extract,red yeast rice,ginkgo biloba extract and salvia miltiorrhiza extract with a dosage of 1.600 g/kg body weight can reduce serum TC and TG levels in rats with hyperlipidemia.

Gastrointestinal Regulation of Red Yeast Rice on Mice with Syndrome of Food Retention Due to Spleen Deficiency Before and After Fermentation / 中国实验方剂学杂志

Objective: To investigate gastrointestinal regulation of red yeast rice on spleen deficient dyspepsia mice, and provide reference for clarifying mechanism of invigorating spleen to promote digestion of fermented traditional Chinese medicine. Method: Mice were randomly divided into 5 groups, including the blank group, the model group, the unfermented red yeast rice treatment group (1.17 g·kg-1·d-1), the fermented red yeast rice treatment group (1.17 g·kg-1·d-1), the domperidone group (3.9 mg·kg-1·d-1), ten mice in each group were continuous intragastric administration for 10 days. The effects of red yeast rice before and after fermentation on the general behavior, body weight, food intake and gastrointestinal function of spleen deficient dyspepsia mice were compared. The effects of red yeast rice on the gastrointestinal hormone[motilin (MTL), gastrin (GAS), 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP)] levels in serum of spleen deficient dyspepsia mice were determined by enzyme-linked immunosorption assay (ELISA). The regulation function of red yeast rice on the pathological changes of gastrointestinal tissue of spleen deficient dyspepsia mice was investigated by hematoxylin-eosin (HE) staining. The effect of red yeast rice on gut microbiota of spleen deficient dyspepsia mice was investigated, the primers were designed on bacterial 16S rRNA V3-V4 region sequences and Illumina Miseq platform was used for high-throughput sequencing. Result: Compared with the model group, fermented red yeast rice could recover the body weight and food intake, reduce gastric residual rate (PPEscherichia-Shigella, while increasing the relative abundance of the beneficial bacteria such as Lactobacillus and Bifidobacterium. Conclusion: Fermented red yeast rice can enhance the gastrointestinal function of spleen deficient dyspepsia mice, the mechanism may be related to regulating gastrointestinal hormone level, improving histopathology of gastrointestinal tissue and restoring intestinal flora structure.

Feasibility exploration of discriminating red yeast rice for different applications by secondary metabolite fingerprints / 药学学报

Though red yeast rice (RYR) has been used as medicine for centuries, few study has been reported about its biological activities related to traditional medicinal application and marketed RYR showed poor consistency in quality. In this study, with comprehensive investigation of their production processes and field acquisition samples including those from genuine producing area, an ultra performance liquid chromatographic (UPLC) method was firstly established to discriminate RYR for different applications based on their secondary metabolites fingerprint. It was performed on a CAPCELL CORE AQ column (100 mm×4.6 mm, 2.7 μm), with PDA (range: 200-650 nm, extracted: 237 nm) and ELSD detection. The mobile phase used was water (A) and acetonitrile (B) both containing 0.1% formic acid at gradient elution (0-15 min, 50% B→85% B (linear); 15-16 min, 85 % B→50% B (linear) and maintained until 21 min), with a flow rate of 0.5 mL∙min-1. The method established was fully validated in agreement with guidelines of Chinese Pharmacopeia. Common metabolites were found in RYR for same application and the fingerprints of RYR for food coloring or brewing from various manufacturers had similarities above 0.90. Meanwhile, significant differences were observed among the fingerprints for various applications and discrimination could be achieved by principal component analysis (PCA). Lovastatin was absence in RYRs for food coloring or brewing, and the fingerprint of traditional medicinal RYR was similar to that of RYR for brewing. However, standardization was required for RYR containing lovastatin because of their significant differences from various manufacturers in fingerprints and lovastatin content. The results demonstrated the feasibility to discriminate RYR for different applications by the secondary metabolites fingerprint method established in this study, which provides a scientific basis to investigate the relationship between biological activities of medicinal RYR and their corresponding secondary metabolites, and further aid their quality standardization and improvement.

Potential protective effects of red yeast rice in endothelial function against atherosclerotic cardiovascular disease / 中国天然药物

Atherosclerotic cardiovascular disease (ASCVD) is the deadliest disease in the world, with endothelial injury occurring throughout the course of the disease. Therefore, improvement in endothelial function is of essential importance in the prevention of ASCVD. Red yeast rice (RYR), a healthy traditional Chinese food, has a lipid modulation function and also plays a vital role in the improvement of endothelial reactivity and cardiovascular protection; thus, it is significant in the prevention and treatment of ASCVD. This article reviews the molecular mechanisms of RYR and its related products in the improvement of endothelial function in terms of endothelial reactivity, anti-apoptosis of endothelial progenitor cells, oxidative stress alleviation and anti-inflammation.

Six new monacolin analogs from red yeast rice / 中国天然药物

Six novel monacolin analogs, monacolins V-V (1-6), together with seven known ones (7-13), were isolated from the ethyl acetate extract of red yeast rice. Their structures and absolute configurations were determined by spectroscopic methods, especially 2D NMR (H-HCOSY, HSQC, HMBC, and NOESY/ROESY) and CD spectroscopic analyses as well as chemical derivation. Monacolins V (2) and V (3) represent the first examples of monacolins with 3-hydroxybutyrate substitute. The anti-inflammatory inhibitory activities against the lipopolysaccharide (LPS) induced NO production in BV-2 cells as well as antioxidant activities against rat liver microsomal lipid peroxidation were evaluated.

Trace phenolic compounds from Red Yeast Rice / 中国中药杂志

Trace chemical constituents from the ethyl acetate extract of Red Yeast Rice were investigated. Four phenolic compounds were isolated by various column chromatographies, and their structures were identified on the basis of spectroscopic analysis including UV, MS, IR and NMR. The four compounds were identified as 2-methyl-5-(2'R-methyl-4'-hydroxy-butyl)-cinnamic acid(1), 5-(2'-hydroxy-6'-methyl phenyl)-3-methylfuran-2-carboxylic acid(2), daidzein(3), and genistein(4). Compound 1 was new and 2 was firstly discovered from the genus Monascus, while 3-4 were obtained from Red Yeast Rice for the first time.

Safety Evaluation of Red Koji (red yeast rice) in Healthy Adult Volunteers / 日本補完代替医療学会誌

Objective: Red Koji (red yeast rice), which has been shown to reduce serum LDL cholesterol levels, has potential as a material in the development of functional food. The efficacy and safety of fermented food generally depends on the production method (including bacterial strains, bacterial species, and incubation conditions); therefore, it is important to confirm the safety of materials individually. Design: We conducted a randomized, double-blind placebo-controlled clinical study to investigate the safety of red yeast rice (3P-D21) produced by the solid state fermentation of the Monascus pilosus KP1148 strain. Fifty-one healthy volunteers with normal cholesterol levels and borderline and mild hypercholesterolemia were divided into 2 groups: one group given the test food [500 mg of red yeast rice (2.0 mg as monascin)] for 4 weeks, and the other a placebo food. Safety was evaluated using blood and urine analyses, a physical examination, and medical interviews. Results: No adverse events were associated with the intake of the test food in any examinations. Conclusion: These results show that food containing 500 mg of red yeast rice (2.0 mg as monascin) is safe, and red yeast rice produced by the solid state fermentation of the Monascus pilosus KP1148 strain has potential as a material in the development of functional food.

Textual research on Gutian as place where red yeast rice originated / 中草药

China is regarded as the birthplace of red yeast rice (RYR). However, various opinions concerning the place where RYR originated and time when RYR invented exist. Upon newly-discovered documented files, current opinion saying "RYR was invented before Tang Dynasty" was proven to be invalid. Meanwhile, RYR in Gutian county Fujian province was verified to be manufactured before 28th year of Kaiyuan in the Tang Dynasty (AD 740) with rich historical data. Based on the fact dating back to the farthest times among those records reported nowadays, a conclusion was accordingly deduced that Gutian was the birthplace of RYR.

Red yeast rice prevents atherosclerosis through regulating inflammatory signaling pathways / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of red yeast rice (RYR) on blood lipid levels, aortic atherosclerosis (AS), and plaque stability in apolipoprotein E gene knockout (ApoE-/-) mice.</p><p><b>METHODS</b>Twenty-four ApoE-/- mice were fed with a high-fat diet starting from 6 weeks of age. Mice were randomized into three groups (n = 8 in each group): model group (ApoE-/- group), RYR group (ApoE-/- + RYR group), and simvastatin group (ApoE-/- + simvastatin group). Eight 6-week-old C57BL/6 mice were assigned as the control group and fed with a basic diet. After 36 weeks, plasma lipids and inflflammatory factors were measured. Aortic atherosclerotic lesions by microscope, scanning electron microscope and transmission electron microscope were observed. Plasma levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured with enzyme-linked immunosorbent assay. The level of high sensitivity C-reaction protein (Hs-CRP) was detected by the scattering immunoturbidimetric assay. Protein expression of matrix metalloproteinase-9 (MMP-9) and nuclear factor κB (NF-κB) in aorta were tested by immunohistochemistry.</p><p><b>RESULTS</b>Compared with the model group, treatment with RYR significantly decreased the levels of total cholesterol, triglyceride, low-density lipoprotein cholesterol, lipoprotein (a), and apolipoprotein B100 in ApoE-/- mice (P<0.01). Compared with the model group, treatment with RYR decreased the levels of Hs-CRP, IL-6, and TNF-α (P<0.01). RYR also reduced the protein levels of NF-κB and MMP-9 of the aorta.</p><p><b>CONCLUSIONS</b>RYR has the anti-atherosclerotic and stabilizing unstable plaque effects. The mechanism might be related to the inflflammatory signaling pathways.</p>

No additional cholesterol-lowering effect observed in the combined treatment of red yeast rice and Lactobacillus casei in hyperlipidemic patients: A double-blind randomized controlled clinical trial / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effect of combining red yeast rice and Lactobacillus casei (L. casei) in lowering cholesterol in patients with primary hyperlipidemia, the later has also been shown to remove cholesterol in in vitro studies.</p><p><b>METHODS</b>A double-blind clinical trial was conducted to evaluate the cholesterol-lowering effect of the combination of red yeast rice and L. casei. Sixty patients with primary hyperlipidemia were recruited and randomized equally to either the treatment group (red yeast rice + L. casei) or the control group (red yeast rice + placebo). One red yeast rice capsule and two L. casei capsules were taken twice a day. The treatment lasted for 8 weeks, with an extended follow-up period of 4 weeks. The primary endpoint was a difference of serum low-density lipoprotein cholesterol (LDL-C) level at week 8.</p><p><b>RESULTS</b>At week 8, the LDL-C serum level in both groups was lower than that at baseline, with a decrease of 33.85±26.66 mg/dL in the treatment group and 38.11±30.90 mg/dL in the control group; however, there was no statistical difference between the two groups (P>0.05). The total cholesterol was also lower than the baseline in both groups, yet without a statistical difference between the two groups. The only statistically signifificant difference between the two groups was the average diastolic pressure at week 12, which dropped by 2.67 mm Hg in the treatment group and increased by 4.43 mm Hg in the placebo group (P<0.05). The antihypertensive activity may be associated with L. casei. Red yeast rice can signifificantly reduce LDL-C, total cholesterol and triglyceride.</p><p><b>CONCLUSION</b>The combination of red yeast rice and L. casei did not have an additional effect on lipid profifiles.</p>

Protective effect of red yeast rice capsule containing coenzyme Q10 on osteoporosis in rats induced by ovariectomy combined with D-galactose / 中国药理学通报

Aim To investigate the effects of red yeast rice capsules containing coenzyme Q10 on femur with an animal model of osteoporosis, which was induced by OVX with D-galactose in rats, and the results were compared with those obtained from diethylstilbestrol. Methods Three-month old female Sprague-Dawley rats were randomly divided into sham group ( CON ) , ovariectomized group ( OVX ) , model group ( MOD ) , diethylstilbestrol group( DES) , and red yeast rice cap-sule group( RYR) . After 60 days, the left femurs were collected for Ca, P and hyp measurement, while the right femurs were performed with three-point bending test and micro-CT evaluation, respectively. Results Compared with CON group, MOD group had a signifi-cant increase in body weight, Tb. Sp, SMI and signifi-cant decrease in maximum load, stiffness, maximum strength, break strength, elastic modulus, Ca, P, Hyp contents and indicators of BV/TV, Tb. N, BMD, Conn-Dens. On the other hand, compared with MOD group, RYR group had a lower body weight and all bone biomechanics indexes were increased without sta-tistically significant difference. At the same time, the content of Ca, P and indicators of BV/TV, Tb. N, Tb. Th, BMD, Conn-Dens increased significantly;yet Tb. Sp decreased significantly. In DES group, the results of indicators were consistent with those for RYR group. In addition, compared with DES group, in RYR group body weight decreased significantly;the content of Ca, P and indicators of BV/TV, Tb. N, Tb. Th, Conn-Dens were significantly higher, and Tb. Sp, SMI were significantly lower. Conclusions Significant bone loss and deteriorated mechanical properties of femur can be observed in animal model of osteoporosis induced by OVX combined with D-galactose. Red yeast rice cap-sules containing coenzyme Q10 show effective prevention effects. Furthermore, red yeast rice capsules(0. 5 tab-let·kg-1 ) have better effect on increasing the number of trabecular bone than diethylstilbestrol ( 30 μg · kg-1 ) does.

Clinical study on cholesterol-lowering effect of RYR Cholestin among Americans with moderate hy-percholesterolemia / 上海预防医学

[Objective] To evaluate lipid -lowering efficacy and safety of RYR Cholestin , or Monascus purpureus (Red Yeast) Rice, in Americans with moderate hypercholesterolemia. [Methods] This study was an open-label, self-control, and multi-center clinical trial.A total of 187 subjects were entered into this trial (serum LDL-Cholesterol 3.50~4.92 mmol/L,total cholesterol 5.18~7.25 mmol/L, male:female=116∶71) , of whom 162 completed the study .Subjects were placed on the NCEP Step I Diet throughout the study and RYR Cholestin (2.4 g/day) was administered for 8 weeks following initial 4-week diet control . [ Results] Being on the diet alone for 4 weeks resulted in no significant changes in serum lipids .RYR Cholestin treatment for 8 weeks reduced serum total cholesterol , LDL-Cholesterol and triglycerides by 16 .6%, 24 .0%, and 25 .2%, respectively , and increased HDL-Cholesterol by 14 .3%( all P<0 .001 ) .There were 97 .5% of patients having ≥10% improvement in at least one of lipid risk factors, and 79.0%having ≥20% improvement.Discontinuation of RYR Cholestin intervention for 14 d led to a return of serum lipids to baseline of pre -study .And 29 possible product -related mild adverse re-actions were reported . [ Conclusion] RYR Cholestin is well tolerated and effective in reducing total and LDL-Cholesterol, and triglycerides, as well as in increasing HDL-Cholesterol in hypercholesterolemic patients, but those indicators return to the beginning baseline when the treatment is discontinued .

The Effect of Red-Yeast-Rice Supplement on Serum Lipid Profile and Glucose Control in Subjects with Impaired Fasting Glucose or Impaired Glucose Tolerance

This study was aimed at evaluating the effect of red-yeast-rice supplementation on cholesterol-lowering and glucose control in subjects with impaired fasting glucose (IFT) or impaired glucose tolerance (IGT). We conducted a doubleblind, placebo-controlled study with 3 groups ; placebo, low dose group (red yeast rice 210.0 mg/capsule, 2.52 g/day)and high dose group (red yeast rice 420.0 mg/capsule, 5.04 g/day), which were randomly assigned to subjects with impaired fasting glucose or impaired glucose tolerance. We measured fasting serum concentrations of total-, LDL-, HDL-cholesterol, triglyceride, glucose, insulin, free fatty acid (FFA) and 2 h oral glucose tolerence test (OGTT) before and after the supplementation. Both low dose and high dose groups had significant decrease in LDL cholesterol and atherogenic index (AI) compared with placebo group (p < 0.05). Additionally, total and HDL cholesterol improved significantly in high dose group compared with placebo group (p < 0.05). Fasting serum glucose decreased in test groups and increased in placebo group after intervention. However, it was not significant differences. In subjects which fasting blood glucose is more than 110 mg/dL, fasting glucose had a tendency to decrease in high dose group (p < 0.1) and Hemoglobin A1c (HbA1c) had significant decrease in low dose group (p < 0.05), while insulin and HOMA-IR had a tendency to increase in placebo group after intervention. Mean changes of glucose related parameters (fasting glucose, insulin, HOMA-IR) compared with placebo group did not show significant differences. In conclusion, subjects with impaired fasting glucose or impaired glucose tolerance were significantly improved in serum lipid profile by red yeast rice supplementation without serious side effects. These are more effective in the case of a high dose. The effects of red yeast rice supplementation on glucose control were insignificant.

Effects of Red-Yeast-Rice Supplementation on Bone Mineral Density and Bone Mineral Content in Overiectomized Rats

This study was performed to evaluate the effect of red-yeast-rice on bone metabolism in overiectomized (OVX) rats. Forty female Sprague-Dawley rats (body weight 210 +/- 5 g, 9 weeks old age) were divided into two groups. One group were OVX, and the other group received sham operation (SHAM), and received either control diet (20% casein) or a red-yeast-rice power supplemented diet (0.1%) for 9 weeks. And then each rat group was further divided into control diet (casein 20%) and red-yeast-rice powder supplemented (0.1%) diet group. All rats were fed on experimental diet and deionized water ad libitum for 9 weeks. Bone mineral density (BMD) and bone mineral content (BMC) were measured using PIXImus (GE Lunar Co, Wisconsin, USA) in spine and femur on 5, 9 weeks after feeding. The serum and urine concentrations of Ca and P were determined. Bone formation was measured by serum osteocalcin and alkaline phosphatase (ALP) concentrations. And bone resorption rate was measured by deoxypyridinoline (DPD) crosslinks immunoassay and corrected for creatinine. Serum osteocalcin, growth hormone, IGF-1 and calcitonin were analyzed using radioimmunoassay kits. Urinary Ca and P excretion were not significantly different among the groups. Within the OVX group, the red-yeast-rice group had a lower crosslinks value than the casein group. Therefore the redyeast- rice supplemented groups had a lower bone resorption ratio than the casein group in the ovariectomized rats. And, the red-yeast-rice group had significantly higher IGF-1 hormone than casein group in ovariectomized rats. The redyeast-rice group had higher spine bone mineral content than those of control group within the OVX groups. This study was an important first step in establishing that the observed beneficial effects of red-yeast-rice on bone, and this study also established the need for a study on the long-term effect of this supplement in a human.