Mogroside regulates the oxidative stress response of retinal pigment epithelial cells induced by H 2O 2 through silent information regulator of transcription 1/nuclear factor erythroid-2-related actor 2 signaling pathway / 中华眼底病杂志

Objective:To observe and preliminarily explore the effect of mogroside on oxidative stress of retinal pigment epitheliaum (RPE) cells induced by hydrogen peroxide (H 2O 2) and its possible mechanism. Methods:A experimental study. The RPE cells were divided into control group, H 2O 2 group, silent information regulator of transcription 1 (SIRT1) inhibitor EX527 group (EX527 group), mogroside group, mogroside+EX527 group. Methyl thiazolete trazolium method was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis rate. 2' ,7'-dichlorodihydrofluorescein diacetate fluorescent probe method, xanthine method and enzyme-linked immunosorbent assay method were used to detect the level of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells respectively. Real-time quantitative polymerase chain reaction and Western blot were used to detect relative expressions of SIRT1, nuclear factor erythroid-2-related actor 2 (Nrf2), heme oxygenase-1 (HO-1) mRNA and protein in cells. One-way ANOVA was used for comparison among groups. The pairwise comparison between groups was tested by the least significant difference t test. Results:Compared with the control group, the H 2O 2 group cell survival rate decreased, the apoptosis rate increased, the ROS level in the cells increased, the SOD activity decreased, the MDA content increased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein decreased ( P<0.05). Compared with H 2O 2 group, the cell survival rate decreased, apoptosis rate increased, the cell ROS level increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein expression decreased in EX527 group ( P<0.05); the cell survival rate increased, apoptosis rate decreased, ROS level decreased, SOD activity increased, MDA content decreased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein increased in mogroside group ( P<0.05). Compared with the mogrosides group, the cell survival rate decreased, the apoptosis rate increased, the level of ROS increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein decreased in mogrosides+EX527 group ( P<0.05). Conclusions:Mogrosides can alleviate the oxidative stress response of visual RPE cells induced by H 2O 2, promote cell proliferation, and reduce cell apoptosis. Mogrosides may exert antioxidant effects by activating the SIRT1/Nrf2 signaling pathway.

Transcriptomic and proteomic screening of key pathways of mogroside V relieving OVA-induced mouse asthma / 中国药理学通报

Aim To discuss the potential key mechanism of mogroside V in relieving pulmonary inflammation in asthmatic mice based on transcriptomics and proteomics. Methods Ovalbumin(OVA)was chosen to induce female BALB/C mouse asthma model, and the mice were treated with mogroside V to observe the pathological changes of lung tissues. Lung tissues in groups of natural control, ovalbumin control and mogroside V control were chosen for transcriptomic and proteomic analysis, and differential genes and proteins were screened for tendency analysis, followed by KEGG enrichment analysis for the potential genes and proteins. Results The results of lung morphological observation and HE revealed that mogroside V attenuated the OVA-induced pulmonary inflammation. Differential genes and proteins were selected from RNA-seq and DIA analysis. In the analysis of omics 454 genes increased in comparison between groups of natural control with ovalbumin control and decreased in comparison between groups of mogroside V control with ovalbumin control in 1 122 potential genes, and 111 genes were of opposite features. A total of 238 proteins increased in comparison between groups of natural control with ovalbumin control and decreased in comparison between groups of mogroside V control with ovalbumin control in 497 potential proteins, and 91 proteins were of opposite features. The PI3K/Akt signaling pathway was enriched from KEGG and tendency analysis of transcriptomics and proteomics. The key factors of Igha, Ighg1, PI3K and Akt increased in ovalbumin control group and decreased in mogroside V control group by the validation of molecualr biology experiments. Conclusions Transcriptomic and proteomic analysis exhibits that mogroside V relieves asthma in mice through inhibiting the activation of key factors including Lgha, Lgh1, PI3K and Akt, depressing the signaling pathway, attenuating pulmonary inflammation to reach the goal of moistening lung and relieve cough, which provides a reference for drug development of asthma.

Mogroside v stimulates osteoblast proliferation and differentiation by promoting lncRNA TUG1 expression / 中国组织工程研究

BACKGROUND: Osteoporosis is a balance disorder between bone formation of osteoblasts and bone resorption of osteoclasts during bone remodeling. Strict control of bone remodeling at the cellular level is important to maintain bone homeostasis and avoid osteoporosis. Previous studies have shown that 1.25×10-2 g/L mogroside V can promote osteoblast proliferation and differentiation, and its mechanism may be related to LncRNA TUG1. OBJECTIVE: To investigate the role of LncRNA TUG1 in the promotion of osteoblast proliferation and differentiation by mogroside V. METHODS: Osteoblasts from neonatal Sprague-Dawley rats were extracted by two-step enzymatic digestion. The cells were divided into two groups and treated with 0 and 1.25×10-2 g/L Mogroside V. The LncRNA was detected after 2 days of culture. LncRNA TUG1 silencing virus was designed and synthetized. The newly extracted osteoblasts were divided into normal cell control group, mogroside V intervention group, mogroside V+negative virus group, TUG1 silent group, and mogroside V+TUG1 silent group. The proliferation of osteoblasts was observed by FDA fluorescence staining at 2, 4, and 6 days after processing according to the above grouping conditions. After 6 days of treatment on osteoblasts, the effect of TUG1 on osteoblast proliferation and differentiation was studied by alkaline phosphatase staining, alizarin red staining and qRT-PCR. RESULTS AND CONCLUSION: LncRNA detection showed that 1.25×10-2 g/L Mogroside V significantly promoted the expression of LncRNA TUG1 in osteoblasts. FDA fluorescent staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on osteoblast proliferation. After 6 days of treatment, alkaline phosphatase staining and alizarin red staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on mineralization of osteoblasts. The results of qRT-PCR showed that Runx2, BSP, OCN and COL1A1 genes were highly expressed in the mogroside V intervention group, but their expression was inhibited in the mogroside V+TUG1 silent group. Overall findings indicate that mogroside V stimulates the proliferation and differentiation of osteoblasts by promoting the expression of LncRNA TUG1.

Synthetic biology for the synthesis of mogroside V - a review / 生物工程学报

Mogroside V, a component with high content and sweetness in mogrosides, has many pharmacological activities such as relieving cough, reducing sputum, anti-cancer, anti-oxidation, regulating blood sugar, making it a natural nonsugar sweetener with therapeutic functions, and showing a broad market prospect. However, the limited resources and high extraction costs have restricted its widespread use. The rapid development of synthetic biology has provided a new idea for the production of plant natural products. The low-cost and large-scale production will be realized through the construction of a microbial cell factory for mogroside V. Here, we briefly introduce the structure and pharmacological activity of mogroside V, and review progress in applying synthetic biology for its synthesis, and also discuss the challenges faced by the current research, to provide a reference for further studies on the biosynthesis of mogroside V.

Regio-selective glycosylation of mogroside IIIE by glycosyltransferase / 中国药科大学学报

@#Mogrosides, the main sweet components isolated from Siraitia grosvenorii, are a family of cucurbitane-type tetracyclic triterpenoid saponins. Given that the high sweetness, low calorie and excellent taste, mogrosides have become the important resource for the development of natural non-nutritive sweeteners. As reported, 11α-hydroxyl group in the structural skeleton of mogrosides was closely related to sweetness and taste, but it had not been confirmed experimentally. In this work, we used mogroside IIIE as a model compound, which was 300 times sweeter than 5% sucrose and tasted better, and modified its 11α-hydroxyl group through glycosyltransferase to elucidate the relations between structure and sweetness of mogroside compounds. The glycosyltransferase HXSW-GT-2 was obtained to regio-selectively glycosylate the 11α-hydroxyl group of mogroside IIIE through the screening of glycosyltransferase library. And then, the soluble expression of HXSW-GT-02 in Escherichia coli was efficiently achieved by optimizing the induction conditions. Subsequently, the yield of glycosylated mogroside IIIE(MG-IIIE-Glu)was increased to > 85% through optimizing reaction pH, temperature, UDP-G dosage and biocatalyst loading. The product MG-IIIE-Glu was bio-prepared at a 0. 5 L scale and the final purity was 97. 8%. A “mouth feel” test showed that MG-IIIE-Glu had no sweetness and displayed obvious bitterness through the comparison with mogroside IIIE and 5% sucrose. In conclusion, the function of the 11α-hydroxyl group of mogrosides in sweetness and taste was preliminarily elucidated which would be beneficial for the structural modification and development of mogroside sweeteners.

Enzymatic synthesis of mogroside ⅢE / 中国药科大学学报

The aim was to develop the simple preparation method of mogroside ⅢE,and to lay the foundation for the development of the mogroside sweeteners. In the present study,the glycosidase CPU-GH17,which can regio-selectively biosynthesize mogroside ⅢE from mogroside V,was screened from the established library of glycosi-dases. Then,the soluble expression condition of CPU-GH17 in E. coli was exploited by investigating isopropyl β-D-thiogalactoside (IPTG)concentration,culture temperature and induction time,and 0. 4 mmol/L IPTG,15 °C and 12 h was used as optimal condition. The result showed that mogroside V could be completely converted into mogroside ⅢE under the conditions of pH 6. 0,45 °C,3 U/mL enzyme loading,5 mg/mL substrate concentration for 20 h. In conclusion,a biosynthetic system for the regio-selective preparation of mogroside ⅢE by recombinant CPU-GH17 was successfully established and verified at a preparative scale.

Study on the Relieving Cough and Eliminating Phlegm Effects of Stemoninine Combined with Mogroside Ⅴ on Mice / 中国药房

OBJECTIVE:To study the relieving cough and eliminating phlegm effects of stemoninine combined with mogrosideⅤ on mice,and select its optimal ratio. METHODS:70 mice were selected in each experiment and randomly divided into 7 groups,namely solvent group(normal saline), codeine(15 mg/kg)or ambroxol(30 mg/kg)group(positive control),stemoni-nine group(30 mg/kg),mogroside Ⅴ group(30 mg/kg),stemoninine-mogroside Ⅴ combination group(30 mg/kg)with mass ra-tio of 2:1,1:1,1:2,ig,once a day. Ammonia-induced cough experiment(positive drug was codeine)and tracheal phenol red ex-cretion experiment(positive drug was ambroxol)were respectively conducted. Using cough latent period,cough times and volume of tracheal phenol red excretion as indexes,the antitussive and expectorant effects of drug in each group were compared. RE-SULTS:Compared with solvent group,cough latent period was obviously shortened,cough times was obviously reduced,volume of tracheal phenol red excretion was obviously increased in each administration group(P<0.05 or P<0.01). Compared with mogro-side Ⅴ group,cough latent period was obviously shortened,cough times was obviously reduced in stemoninine-mogroside Ⅴgroup with different mass ratios(P<0.05),and 2:1,1:1 groups showed the best effects. Compared with stemoninine group,vol-ume of tracheal phenol red excretion was obviously increased in stemoninine-mogroside Ⅴ group with different mass ratios (P<0.05),and 1:1,1:2 groups showed the best effects. CONCLUSIONS:The combination of temoninine and mogroside Ⅴ shows synergistic effects on relieving cough and eliminating phlegm,stemoninine-mogrosideⅤmass ratio of 2:1,1:1 can be used as pre-ferred combination of reference.

Influence of diferent dehydrations on active components in fructus momordicae / 国际中医中药杂志

Objective To compare the different dehydrations of microwave-vacuurn drying and high temperature drying on contents of the major effective in fructus momordicae.Methods Using two different drying methods for processing of fructus momordicae, the mogroside Ⅴ, vitamin C in dried products which was determined by high performance liquid chromatography (HPLC) was used to a main indicator for comparing different drying methods.Results The contents of mogroside Ⅴ in fructus momordicae which processed by microwave-vacuum drying, microwave-vacuum drying after ripening treatment and high temperature drying were 1.14％, 1.19％ and 0.60％;the contents of vitamin C were 0.21％, 0.30％ and 0.000 06％.Conclusion The processing method of microwave-vacuum drying to keep active components is obviously better than which of high temperature drying.

New natural saponins from fruits of Momordica grosvenori / 中草药

Objective To study the chemical constituents in the fruits of Momordica grosvenori. Methods Isolation and purification of the constituents were carried out on column chromatography. Their structures were identified by NMR and MS spectral analyses. Results Ten compounds were isolated and elucidated as kaempferol (Ⅰ), kaempferol 7-O-?-L-rhamnopyranoside (Ⅱ), kaempferitrin (Ⅲ), mogroside ⅡE (Ⅳ), mogroside Ⅲ (Ⅴ), kaempferol 3-O-?-L-rhamnopyranoside-7-O-[?-D-glucopyranosyl (1-2)-?-L-rhamnopyranoside] (Ⅵ), mogroside ⅣA (Ⅶ), grosmomoside Ⅰ (Ⅷ), mogroside Ⅴ (Ⅸ), and mogroside ⅡA1 (Ⅹ). Conclusion Compounds Ⅶ and Ⅹ are new natural products, and Ⅰ is isolated from the fruits of M. grosvenori for the first time.

Protective Effect of Mogrosides on Experimental Liver Injury in Mice / 中国药房

OBJECTIVE:To study the protective effect of Mogrosides on experimental liver injury in mice. METHODS: Acute hepatic injury model was induced by CCl4 in mice, and the immunological hepatic injury model was induced in mice by bacillus calmette-guerin (BCG) and lipopolysaccharide (LPS). The activities of ALT and AST in serum, SOD activity in liver tissue and content of MDA in the liver tissue were investigated; and the histological changes of liver were observed. RESULTS: Mogroside could decrease the serum activities of ALT and AST in mice with acute hepatic injury or immunological hepatic injury, increase the activity of SOD but decrease MDA content in liver tissue of mice with immunological hepatic injury, and it could lessen the histological changes of liver. CONCLUSION: Mogroside showed protective effect on mice with acute hepatic injury or immunological hepatic injury. The mechanism might be attributed to its action against lipid peroxidation.

Effect of Mogroside extracts on blood glucose,blood lipid and antioxidation of hyperglycemic mice induced by Alloxan / 中国药理学通报

Aim To study antidiabetic effect and mechanism of Mogroside extracts(MGC) on hyperglycemic mice induced by Alloxan.Methods Alloxan-induced diabetic mice were fed with MGC at dose of 50,100?300,500 mg?kg~(-1) bw respectively for 30 d.The blood glucose levels,the blood lipid levels and antioxidation levels were assayed after the last fedding.Results ①MGC at dose of 100?300?500 mg?kg~(-1) bw significantly lowered the fasting blood glucose and the hypoglycemic effect of MGC at dose of 100 mg?kg~(-1) bw was the best ②MGC decreased the content of TC,TG and enhanced the content of HDL-C of alloxan-induced diabetic mice,so it could restore the blood lipid levels of diabetic mice(P0.01).Conclusion Mogroside extracts(MGC) have obvious glucose-lowering effect on hyperglycemic mice,its mechanism may be related with improving antioxidation level and restoring the blood lipid levels of hyperglycemic mice.