[Total triterpenes of Euphorbium alleviates rheumatoid arthritis via Nrf2/HO-1/GPX4 pathway].

This study aims to investigate the effect and mechanism of total triterpenes of Euphorbium in treating rheumatoid arthritis(RA). The rat model of RA was established with Freund's complete adjuvant(FCA). Male rats were randomly assigned into control, model, Tripterygium glycosides(7.5 mg·kg~(-1)), and low-, medium-, and high-dose total triterpenes of Euphorbium(32, 64, and 128 mg·kg~(-1), respectively) groups, with 10 rats in each group. In other groups except the control group, 0.2 mL FCA was injected into the right hind toe. Rats in the intervention groups were administrated with corresponding drugs by gavage, and the control group and the model group with the same volume of 0.5% CMC-Na solution once a day. During the treatment period, the swelling degree of the hind paw was measured and the arthritis was scored until day 30. At the end of drug administration, the pathological changes of the joint tissue were observed by hematoxylin-eosin staining. The content of malondialdehyde(MDA), glutathione(GSH), and Fe~(2+) and the activity of superoxide dismutase(SOD) in the joint tissue were measured by biochemical colorimetry. RT-PCR was performed to determine the mRNA levels of nuclear factor erythroid 2-related factor 2(Nrf2), glutathione peroxidase 4(GPX4), and acyl-CoA synthetase long chain family member 4(ACSL4) in the joint tissue. Western blot was employed to determine the protein levels of Nrf2, Kelch-like ECH-associated protein 1(Keap1), heme oxygenase-1(HO-1), NAD(P)H quinone oxidoreductase 1(NQO1), SOD2, GPX4, and ACSL4 in the joint tissue. The results showed that the treatment with Tripterygium glycosides(7.5 mg·kg~(-1)) and total triterpenes of Euphorbium(32, 64, and 128 mg·kg~(-1)) alleviated the swelling degree of bilateral hind limbs, decreased the arthritis score, reduced joint tissue lesions and the content of MDA and Fe~(2+) in the joint tissue, and increased GSH content and SOD activity. Furthermore, the interventions up-regulated the protein and mRNA levels of Nrf2 and GPX4, down-regulated the protein and mRNA levels of ACSL4, and up-regulated the protein levels of Keap1, NQO1, HO-1, and SOD2 in the Nrf2/HO-1/GPX4. In summary, the total triterpenes of Euphorbium can treat RA by inhibiting lipid peroxidation and abnormal ferroptosis, which may involve the Nrf2/HO-1/GPX4 signaling pathway.

Anti-cataract effects of Dajizhi (Euphorbium) eye drops on selenite-induced cataracts in rat.

OBJECTIVE: To evaluate the effects of Dajizhi (Euphorbium) on selenite-induced cataracts. METHODS: Wistar rat pups were divided into 9 groups. Rats in group 1 were subcutaneously injected with saline, and rats in the other groups were injected with sodium-selenite. Every right eye was treated with 5 µL eye drops 3 times per day, and the left eye received no treatment. The eyes of rats in group 3 were treated with pirenoxine; rats in groups 4, 5, 6, 7, 8 and 9 were respectively treated with Dajizhi (Euphorbium) (25 mg/mL), Dajizhi (Euphorbium) (5 mg/mL), Dajizhi (Euphorbium) methanol extract (25 mg/mL), Dajizhi (Euphorbium) methanol extract (5 mg/mL), euphol (25 mg/mL), euphol (5 mg/mL). Cataracts were observed by a slit lamp before and after treatment. Electroretinograms were acquired at set intervals. The morphological changes of the rat eyes were observed in vitro, and the levels of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in the lenses and aqueous humour were estimated at set intervals. RESULTS: Slit lamp examination showed decreased degrees of cataracts after administration of the different treatments. Morphological comparison showed that Dajizhi (Euphorbium) can reduce the turbidity of the lenses, meaning that Dajizhi (Euphorbium) has the anti-cataract effects. Low-concentration of Dajizhi (Euphorbium), its methanol extract and euphol treatment prevented the b-wave amplitudes of the electroretinograms from falling. Euphorbium treatment significantly restored GSH-Px and SOD levels in the lenses and aqueous humour, especially after 10 and 25 d of administration. Euphorbium may help lenses fight oxidative stress caused by selenite. CONCLUSION: The administration of Dajizhi (Euphorbium) can inhibit selenite-induced cataracts.

[Chemical constituents of triterpenoids from Euphorbia resinifera].

The combination of normal-phase silica gel column chromatography, octadecyl silica(ODS) column chromatography, semi-preparative high performance liquid chromatography(HPLC), etc. was employed to isolate and purify the chemical components from Euphorbia resinifera, and 7 triterpenoids were separated from the ethanol extract of the medicinal materials. Their structures were identified by various spectroscopy methods as cycloartan-1,24-diene-3-one(1), cycloartan-1,24-diene-3-ol(2), 3ß-hydroxy-lanosta-8,24-diene-11-one(3), lnonotusane C(4), eupha-8,24-diene-3ß-ol-7,11-dione(5), eupha-24-methylene-8-ene-3ß-ol-7,11-dione(6), and eupha-8,24-diene-3ß,11ß-diol-7-one(7). Compounds 1 and 2 are new compounds, and compound 3 is obtained from nature for the first time.

Santin inhibits influenza A virus replication through regulating MAPKs and NF-κB pathways.

Influenza A virus (IAV) causes high morbidity and significant mortality worldwide. Given the limitations of existing vaccination and antiviral drugs, it is urgent to develop new anti-influenza drugs. Flavonoids are natural polyphenolic compounds with broad applications to treatments for influenza infection. In this study, we demonstrated that santin, a flavonoid compound, showed anti-influenza activity in MDCK and THP-1 cells. Mechanistic studies revealed that santin depressed the phosphorylation of p38 MAPK, JNK/SAPK, ERK, and NF-κB factor and subsequently attenuated the expression of inflammatory cytokines in IAV-infected cells. Thus, santin is a potential candidate for the future development of anti-IAV drugs.

Asymmetric Synthesis of Rupestonic Acid and Pechueloic Acid.

In this report, the originally proposed rupestonic acid (5) and pechueloic acid (3) were efficiently synthesized. The chiral lactone 13, recycled from the degradation of saponin glycosides, was utilized to prepare the key chiral fragment 11. During the exploration of this convergent assembly strategy, the ring-closing metathesis (RCM), SmI2-prompted intermolecular addition, and [2,3]-Wittig rearrangement proved to be effective transformations for the synthesis of subunits.

Polyisoprenylated benzoylphloroglucinol derivatives from Hypericum scabrum.

Four new polyisoprenylated benzoylphloroglucinol derivatives, hyperscabrones J-M (1-4), were isolated from the air-dried aerial parts of Hypericum scabrum. Their structures were elucidated by spectroscopic methods and were subsequently confirmed by comparing with data of known compounds. The absolute configuration of the bicyclo[3.3.1]nonane-2,4,9-trione core was defined by the experimental and calculated electronic circular dichroism (ECD) spectra. The evaluation of their hepatoprotective activities against paracetamol-induced HepG2 cell damage showed that compounds 2 and 4 exhibited significant hepatoprotection at 10µM.

Polycyclic Polyprenylated Acylphloroglucinol Congeners from Hypericum scabrum.

Twenty polycyclic polyprenylated acylphloroglucinols (PPAPs), including the new compounds hyperscabrones A-I (1-9), were isolated from the air-dried aerial parts of Hypericum scabrum. These compounds comprise seven different structural types. All structures were determined by NMR spectroscopic methods and both experimental and calculated electronic circular dichroism (ECD) spectra. The evaluation of their neuroprotective effects on glutamate-induced toxicity in SK-N-SH cells showed that compounds 4-7 exhibited significant neuroprotection at 10 µM. Additionally, compounds 3, 4, 7, and 9 showed moderate hepatoprotective activities against paracetamol-induced HepG2 cell damage at 10 µM.

[Microbial transformation of glycyrrhetinic acid by Cunninghamella blakesleeana].

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18ß-glycyrrhetinic acid(1), 3-oxo-15ß-hydroxy-18ß-glycyrrhetinic acid (2), 7ß,15α-dihydroxy-18ß-glycyrrhetinic acid (3), 3-oxo-7ß, 15α-dihydroxy-18ß-glycyrrhetinic acid (4), 7ß-hydroxy-18ß-glycyrrhetinic acid(5) and 15α-hydroxy-18ß-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation

Protective effects of acacetin isolated from Ziziphora clinopodioides Lam. (Xintahua) on neonatal rat cardiomyocytes.

BACKGROUND: The total flavonoids from ethanol extract of the aerial part of Ziziphora clinopodioides Lam. (Lamlaceae) (Xintahua) showed protective activities against rat acute myocardial ischemia in rats. This study aims to isolate acacetin, a flavonoid, from the aerial part of Z. clinopodioides, to develop an HPLC method for its detection, and to evaluate its protective effects on neonatal rat cardiomyocytes. METHODS: Sephadex LH-20 silicagel and pillar layer chromatography silica gel were applied for the isolation and purification of acacetin and its structure was elucidated on the basis of (1)H and (13)C NMR spectroscopy. The content of acacetin in Z. clinopodioides collected from three different origins was determined by HPLC. The neonatal rat cardiomyocytes were isolated and cultured in vitro to establish a hypoxia/reoxygenation injury model. The viability of cardiomyocytes was measured by the MTT method. Changes of malondialdehyde (MDA) content in the medium were also determined. RESULTS: The acacetin content in various batches of Z. clinopodioides ranged from 45.50 to 47.41 µg/g. Acacetin of 25, 10, 5 µg/mL significantly decreased the MDA content in a model of hypoxia/reoxygenation injury (P < 0.001, P < 0.001 and P = 0.033, respectively). CONCLUSIONS: Acacetin protects neonatal cardiomyocytes from the damage induced by hypoxia/reoxygenation stress through reduction of lipid peroxidation and enhancement of the antioxidant activity.

[Study on chemical constituents of Hyssopus cuspidatus].

OBJECTIVE: To study the chemical constituents of Hyssopus cuspidatus. METHODS: Compounds were isolated and purified by extraction and different kinds of column chromatography. Their structures were determined on the basis of the physicochemical properties and spectral analysis. RESULTS: Six compounds were identified as caffeic acid methyl ester (1), luteolin 7-O-alpha-L-rhamnopyranosyl (1 --> 6)-beta-D-glucopyranoside (2), luteolin 7-O-beta-D-glucuronide (3), diosmin (4), acacetin 7-O-alpha-L-rhamnopyranosyl(1 --> 6)-beta-D-glucopyranoside (5) and rosmarinic acid (6). CONCLUSION: Compounds 1, 4 and 6 are isolated from this plant for the first time, and compounds 2, 3 and 5 are firstly obtained from Hyssopus genus.

Effect of pomegranate peel polyphenol gel on cutaneous wound healing in alloxan-induced diabetic rats.

BACKGROUND: Pomegranate (punica granatum) belongs to the family Punicaceae, and its peel has been used as a traditional Chinese medicine because of its efficacy in restraining intestine, promoting hemostasis, and killing parasites. Pomegranate peel has been reported to possess wound-healing properties which are mainly attributed to its polyphenol extracts. The purpose of this study was to investigate the effect of pomegranate peel polyphenols (PPP) gel on cutaneous wound healing in diabetic rats. METHODS: Alloxan-induced diabetic rats were given incisional wounds on each side of the mid-back and then treated daily with PPP gel (polyphenol mass fraction = 30%) post-wounding. Rats were sacrificed on days 4, 7, 14, and 21 post-wounding to assess the rates of wound closure, histological characteristics; and to detect the contents of hydroxyproline, production of nitric oxide (NO), and activities of NO synthase (NOS), as well as the expressions of transforming growth factor-ß1 (TGF-ß1), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) in wound tissue. RESULTS: Wound closure was significantly shortened when PPP gel was applied to the wounds of diabetic rats. Histological examination showed the ability of PPP gel to increase fibroblast infiltration, collagen regeneration, vascularization, and epithelialization in the wound area of diabetic rats. In addition, PPP gel-treated diabetic rats showed increased contents of hydroxyproline, production of NO, and activities of NOS and increased expressions of TGF-ß1, VEGF, and EGF in wound tissues. CONCLUSION: PPP gel may be a beneficial method for treating wound disorders associated with diabetes.

(E)-1-[2-Hy-droxy-4,6-bis-(meth-oxy-meth-oxy)phen-yl]-3-[3-meth-oxy-4-(meth-oxy-meth-oxy)phen-yl]prop-2-en-1-one.

The title compound, C(22)H(26)O(9), crystallizes with two independent mol-ecules in the asymmetric unit in which the dihedral angles between the two benzene rings are 21.4 (2) and 5.1 (2)°. An intra-molecular O-H⋯O hydrogen bond occurs in each mol-ecule. Inter-molecular C-H⋯O hydrogen bonds stabilize the crystal structure.