Natural product/Terpenoid; MMP2/9

Morroniside is a glycoside that has been reported in Gentiana straminea, Tripterospermum japonicum, and other organisms. Previous studies have reported the protective effect of morroniside, an active extract of Cornus officinalis, against neuronal apoptosis, oxidative stress and inflammation [1].

In an I/R injury model, monogside can lower MMP2 and MMP9 expression. In a dose-dependent manner, monoside therapy markedly decreased I/R-related neuronal death. The findings demonstrated that whereas Bcl-2 was dramatically down-regulated in the model group, active caspase-3 and Bax were significantly up-regulated when compared to the control group. Following monoside therapy, there was a considerable up-regulation of Bcl-2 expression and a dose-dependent significant down-regulation of active caspase-3 and Bax [1]. In the liver of type 2 diabetic db/db mice, monogside has a mitigating effect on alterations brought on by diabetes, including oxidative stress, inflammation, and apoptosis [2].

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The aim of the present study was to investigate the effect of morroniside against matrix metalloproteinase (MMP)2/9 and focal cerebral ischemia/reperfusion (I/R) injury in rats. A rat model of focal cerebral I/R injury rats was established and rats were administered with 30, 90 or 270 mg/kg/day morroniside for 7 days. The expression of MMP2/9 and neuronal apoptosis were assessed. In addition, the expression of active caspase-3, B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) were measured. The results revealed that MMP2 and MMP9 expression was upregulated and the percentage of apoptotic neurons was increased in rats with focal cerebral I/R injury compared with the control. However, treatment with morroniside significantly inhibited I/R-induced MMP2/9 expression and neuron apoptosis compared with the untreated I/R injury group. Morroniside administration also decreased the expression of active caspase-3 and increased the Bcl-2/Bax ratio compared with untreated rats with focal cerebral I/R injury. The inhibitory effect of morroniside on MMP2/9 expression and neuron apoptosis was dose dependent. In summary, the results of the present study suggest that morroniside is able to protect against cerebral I/R injury in the brain and may have potential as a therapeutic treatment for patients who have suffered a stroke.[1]

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The present study was conducted to examine whether morroniside has an ameliorative effect on diabetes-induced alterations such as oxidative stress, inflammation, and apoptosis in the liver of type 2 diabetic db/db mice. Morroniside (20 or 100 mg/kg body weight/d, per os (p.o.)) was administered every day for 8 weeks to db/db mice, and its effect was compared with vehicle-treated db/db and m/m mice. The administration of morroniside decreased the elevated serum glucose concentration in db/db mice, and reduced the increased oxidative biomarkers including the generation of reactive oxygen species and lipid peroxidation in the liver. The db/db mice exhibited the up-regulation of nicotinamide adenine dinucleotide phosphate oxidase subunits, NF-E2-related factor 2 (Nrf2), heme oxygenase-1, nuclear factor-kappa B, cyclooxygenase-2, inducible nitric oxide synthase, monocyte chemotactic protein-1, and intracellular adhesion molecule-1 levels in the liver; however, morroniside treatment significantly reduced those expressions. Moreover, the augmented expressions of apoptosis-related proteins, Bax and cytochrome c, were down-regulated by morroniside administration. Hematoxylin-eosin staining showed that the increased hepatocellular damage in the liver of db/db mice improved on morroniside administration. Taking these into consideration, our findings support the therapeutic evidence for morroniside ameliorating the development of diabetic hepatic complications via regulating oxidative stress, inflammation, and apoptosis. [2]

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Morroniside attenuated the progression of OA in mice, resulting in substantially reduced osteophyte formation and subchondral sclerosis and lower OARSI scores. Specifically, morroniside significantly promoted cartilage matrix synthesis by increasing collagen type II expression and suppressing chondrocyte pyroptosis. Morroniside administration led to inhibition of matrix metalloproteinase-13 (MMP13), Caspase-1 and nod-like receptor protein-3 (NLRP3) expression in DMM mice and IL-1β-stimulated chondrocytes. In addition, morroniside attenuated the progression of OA by enhancing chondrocyte proliferation and inhibiting chondrocyte apoptosis. Morroniside also attenuated the progression of OA by inhibiting nuclear factor-κB (NF-κB) signaling. Conclusion: Morroniside was protective against cartilage matrix degradation and reduced DMM-induced chondrocyte pyroptosis and apoptosis by the inhibition of NF-κB signaling [3].

Apoptotic cell analysis [1]
Apoptotic cells in brain tissues were detected using a TUNEL assay. Briefly, ischemic penumbra cortex area was resected and the frozen sections (4-µm-thick) were fixed using 4% polyoxymethylene at room temperature for 20 min. and and a colorimetric TUNEL kit was used according to the manufacturer's protocol for in situ apoptosis detection. The sections were washed by PBS and then incubated with prepared TUNEL solution for 1 h at 37°C in a dark chamber. A 50 µl DAB solution (2.5 µl DAB, 0.5 µl 30% H2O2 and 47 µl PBS) was added for nuclear staining at 37°C for 20 min. The TUNEL-positive cells were observed and counted under a light microscope at a magnification of ×200. Apoptotic cells were stained brown and the mean was calculated from 10 independent fields.

Rats were randomly assigned into five groups (n=10 in each). Rats in the control group underwent sham surgery. All other rats underwent suture-occluded surgery as described by Longa et al, with a 0.26 mm nylon monofilament inserted through the right common carotid artery and were divided into groups as follows: The cerebral I/R injury model group (model), no treatment; low dose group, 30 mg/kg/day morroniside by gavage; moderate dose group, 90 mg/kg/day morroniside by gavage; high dose group, 270 mg/kg/day morroniside by gavage. Rats in the control and model groups received an equal volume of normal saline. Longa's five-grade scale methods were used to score neurological deficit following surgery and rats with a score of 0 or 4 were excluded from the current study. [1]

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Aim of the study: The study aims to assess the therapeutic effects of morroniside on cartilage degeneration using a mouse model of OA. Material and methods: 8-week-old male C57BL/6J mice were randomly divided into 4 groups: Sham, destabilization of the medial meniscus (DMM)-treated with vehicle, DMM-treated with low dose morroniside and DMM-treated with high dose morroniside. Histological staining, immunostaining, and TUNEL staining were conducted to detect changes in tissue morphology, expression of key molecules in chondrocytes, and chondrocyte apoptosis, respectively. Osteophyte formation, meniscus calcification, and subchondral sclerosis were quantitated using micro-CT. The expression of chondrocyte markers was also analyzed by Western blot in primary chondrocytes derived from mice treated with morroniside [3].

[1]. Morroniside protects against cerebral ischemia/reperfusion injury by inhibiting neuron apoptosis and MMP2/9 expression. Exp Ther Med. 2018 Sep;16(3):2229-2234.

[2]. Evaluation of morroniside, iridoid glycoside from Corni Fructus, on diabetes-induced alterations such as oxidative stress, inflammation, and apoptosis in the liver of type 2 diabetic db/db mice. Biol Pharm Bull. 2011;34(10):1559-65.

[3]. Morroniside attenuates apoptosis and pyroptosis of chondrocytes and ameliorates osteoarthritic development by inhibiting NF-κB signaling. J Ethnopharmacol. 2021 Feb 10;266:113447.

Morroniside is a glycoside.
Morroniside has been reported in Gentiana straminea, Tripterospermum japonicum, and other organisms with data available.

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In conclusion, the present study revealed that protective effect of morroniside in rats with cerebral I/R injury. Morroniside inhibits the I/R-induced upregulation of MMP2/9 and neuron apoptosis, suggesting that morroniside may promote behavioral deficit repair following cerebral I/R injury. However, the underlying mechanism responsible for these effects remains to be elucidated.[1]

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Ethnopharmacological relevance: Corni Fructus (CF), the red fruit of Cornus officinalis Siebold & Zucc, has been used both as food and medicinal herb in traditional Chinese medicine (TCM). Our previous studies showed that Yougui pills and Bushenhuoxue formula, both TCM prescriptions containing Corni Fructus (CF), have protective effects on osteoarthritis (OA). However, the underlying detailed components in both TCM prescriptions that play therapeutic roles have not been fully defined. Morroniside is a major iridoid glycoside and one of the quality control metrics of CF, but the effects of morroniside on OA remain largely elusive.[3]

C17H26O11

406.3817

406.147

25406-64-8

11228693

White to off-white solid powder

1.5±0.1 g/cm3

635.6±55.0 °C at 760 mmHg

227.0±25.0 °C

0.0±4.2 mmHg at 25°C

1.597

-3.16

5

11

5

28

596

10

C[C@H]1[C@@H]2[C@H](C[C@@H](O1)O)C(=CO[C@H]2O[C@H]3[C@@H]([C@H]([C@@H]([C@H](O3)CO)O)O)O)C(=O)OC

YTZSBJLNMIQROD-SFBCHFHNSA-N

InChI=1S/C17H26O11/c1-6-11-7(3-10(19)26-6)8(15(23)24-2)5-25-16(11)28-17-14(22)13(21)12(20)9(4-18)27-17/h5-7,9-14,16-22H,3-4H2,1-2H3/t6-,7+,9+,10+,11+,12+,13-,14+,16-,17-/m0/s1

methyl (1S,3R,4aS,8S,8aS)-3-hydroxy-1-methyl-8-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,3,4,4a,8,8a-hexahydropyrano[3,4-c]pyran-5-carboxylate

Morroniside; 25406-64-8; CHEBI:80852; (7Beta-Hydroxy)-Morroniside; methyl (1S,3R,4aS,8S,8aS)-3-hydroxy-1-methyl-8-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-1,3,4,4a,8,8a-hexahydropyrano[3,4-c]pyran-5-carboxylate; Methyl (1S,3R,4aS,8S,8aS)-3-hydroxy-1-methyl-8-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-4,4a,8,8a-tetrahydro-1H,3H-pyrano[3,4-c]pyran-5-carboxylate; CHEMBL1209803; DTXSID60948257;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~246.08 mM)
H2O : ~50 mg/mL (~123.04 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.12 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (5.12 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (5.12 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 100 mg/mL (246.08 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)