Angelica acutiloba Kitagawa flower induces A549 cell pyroptosis via the NF-κB/NLRP3 pathway for anti-lung cancer effects.

Angelica acutiloba Kitagawa, a traditional medicinal herb of the Umbelliferae family, has been demonstrated to have anticancer activity. In this study, we investigated the anti-lung cancer effects of two compounds extracted from A. acutiloba flowers: kaempferol-3-O-α-L-(4″-E-p-coumaroyl)-rhamnoside (KAE) and platanoside (PLA). MTT, cell colony formation, and cell migration (scratch) assays revealed that both KAE (100 µM) and PLA (50 µM and 100 µM) inhibited the viability, proliferation, and migration of A549 cells. Dichlorodihydrofluorescein diacetate assays showed that KAE and PLA also induced the generation of reactive oxygen species in A549 cells. Morphologically, A549 cells swelled and grew larger under treatment with KAE and PLA, with the most significant changes at 100 µM PLA. Fluorescence staining and measurement of lactate dehydrogenase release showed that the cells underwent pyroptosis with concomitant upregulation of interleukin (IL)-1ß and IL-18. Furthermore, both KAE and PLA induced upregulation of NF-κB, PARP, NLRP3, ASC, cleaved-caspase-1, and GSDMD expression in A549 cells. Subsequent investigations unveiled that these compounds interact with NLRP3, augment NLRP3's binding affinity with ASC, and stimulate the assembly of the inflammasome, thereby inducing pyroptosis. In conclusion, KAE and PLA, two active components of A. acutiloba flower extract, had significant anti-lung cancer activities exerted through regulation of proteins related to the NLRP3 inflammasome pathway.

Apigenin exerts chemopreventive effects on lung injury induced by SiO2 nanoparticles through the activation of Nrf2.

Apigenin (APG) is a flavonoid widely distributed in fruits, vegetables, and herbs, with comprehensive pharmacological effects. In this paper, we report that APG can elicit a protective effect, which is comparable to those induced by gymnoside II/n-BuOH extracts of Bletilla striata, on SiO2-induced lung injury in vitro and in vivo. In vitro experiments showed that APG (25 µM) could restore the SiO2-decreased A549 cell viability and lower the apoptotic rate and the production of intracellular reactive oxygen species (ROS) in A549 cells treated with nm SiO2. Western blot results showed that APG (25 µM) could increase the level of Nuclear factor E2-related factor 2 (Nrf2) and its downstream proteins. In vivo experiments showed that APG (20 mg/kg) could potently alleviate the SiO2-elicited lung injury by enhancing the Nrf2 expression and thereby suppressing Bax/Bcl-2 pathway. The present study suggests that APG can significantly alleviate the SiO2-induced lung injury both in vitro and in vivo through, at least partially, activating Nrf2 expression.

Dietary egg white protein hydrolysate improves orotic acid-induced fatty liver in rats by promoting hepatic phospholipid synthesis and microsomal triglyceride transfer protein expression.

We investigated the effects of egg white protein hydrolysates (EWH) on orotic acid (OA)-induced nonalcoholic fatty liver (NAFL) in rats. Effects of the egg white protein (EWP) and EWH were also compared. Four groups of male Sprague-Dawley rats were separately fed AIN-76-based diets, supplemented with 20% casein for control, or with 1% OA, together with either 20% casein (OA), 20% EWP, or 20% EWH, respectively, for 3 d (developing stage) and 14 d (developed stage). In both feeding periods, animals from the OA group showed higher accumulation hepatic triacylglycerol (TAG) compared with those from the control group. In the 14-d experiment, dietary EWP and EWH significantly reduced the hepatic TAG levels. Intake of EWP reduced liver fat in OA-fed rats by 61%, while EWH reduced it by 92%. In addition, EWH restored the OA-induced high serum-TAG level to that seen in the control group. The 3 d experiment showed that consumption of EWH improved the expression of hepatic MTP, that was reduced by OA, without changing Mttp gene expression. It also increased the hepatic synthesis of PC and PE by enhancing the transcription of Pcyt1 and Pemt genes. Inclusion of EWP and EWH in the diet improves the OA-induced NAFL. EWH reduces the liver TAG better than EWP, and works more rapidly. Dietary EWH ameliorates OA-induced NAFL by promoting the secretion of hepatic TAG.

Inhibition of intestinal lipases alleviates the adverse effects caused by high-fat diet in Nile tilapia.

Intestinal lipases are fat-digesting enzymes, which play vital roles in lipid absorption in the intestine. To study the regulation of intestinal lipase activity in systemic lipid metabolism in fish, especially in the metabolic diseases caused by high-fat diet (HFD) feeding, we inhibited intestinal lipases in Nile tilapia to investigate the physiological consequences. In the present study, Nile tilapia were firstly fed with HFD (12% fat) for 6 weeks to establish a fatty fish model. Afterwards, Orlistat as a potent intestinal lipase inhibitor was added into the HFD for the following 5-week feeding trial, with two dietary doses (Orlistat16 group, 16 mg/kg body weight; Orlistat32 group, 32 mg/kg body weight). After the trial, both doses of Orlistat treatment significantly reduced intestinal lipase activity, fat absorption, hepatic lipid accumulation, and gene expression of lipogenesis, whereas increased gene expression of lipid catabolism. Moreover, intestinal lipase inhibition increased immune enzyme activities, antioxidant capacity, and gene expression of anti-inflammatory cytokines, whereas lowered gene expression of pro-inflammatory cytokines. Besides, Orlistat could also improve the structure of the intestine and increase expression of intestinal tight-coupling protein. Taken together, intestinal lipase inhibition alleviated the adverse effects caused by HFD in Nile tilapia. Thus, intestinal lipases played key roles in absorbing dietary lipid and could be a promising target in regulating systemic lipid metabolism in fish.

Progress in Development of Interventions to Prevent Birth Defects in Diabetic Pregnancies.

Diabetic embryopathy is a diabetic complication, in which maternal hyperglycemia in early pregnancy causes birth defects in newborn infants. Under maternal diabetic conditions, hyperglycemia disturbs intracellular molecular activities and organelles functions. These include protein misfolding in the endoplasmic reticulum (ER), overproduction of reactive oxygen species (ROS) in mitochondria, and high levels of nitric oxide (NO). The resultant ER, oxidative, and nitrosative stresses activate apoptotic machinery to cause cell death in the embryo, ultimately resulting in developmental malformations. Based on the basic research data, efforts have been made to develop interventional strategies to alleviate the stress conditions and to reduce embryonic malformations. One of the challenges in birth defect prevention is to identify effective and safe agents to be used in pregnancy. One approach is to search and characterize naturally occurring phytochemicals, including flavonoids, curcuminoids and stilbenoids, for use in prevention of diabetic embryopathy.

Glycyrrhetinic Acid Maintains Intestinal Homeostasis via HuR.

Glycyrrhetinic acid (GA) is one of the main components of the traditional Chinese medicine of licorice, which can coordinate and promote the effects of other medicines in the traditional prescription. We found that GA could promote the proliferation, decrease the apoptotic rate, and attenuate DFMO-elicited growth arrest and delay in restitution after wounding in IEC-6 cells via HuR. GA failed to promote proliferation and to suppress apoptosis after silencing HuR by siRNA in IEC-6 cells. Furthermore, with the model of small intestinal organoids developed from intestinal crypt stem cells, we found that GA could increase HuR and its downstream ki67 levels to promote intestinal organoid development. In the in vivo assay, GA was shown to maintain the integrity of the intestinal epithelium under the circumstance of 48 h-fasting in rats via raising HuR and its downstream genes such as EGF, EGFR, and MEK. These results suggested that via HuR modulation, GA could promote intestinal epithelium homeostasis, and therefore contribute to the absorption of constituents from other medicines co-existing in the traditional prescription with licorice in the small intestine. Our results provide a new perspective for understanding the effect of licorice on enhancing the therapeutic effect of traditional prescriptions according to the traditional Chinese medicine theory.

Anti-Inflammatory Activity of Chemical Constituents Isolated from the Willow Bracket Medicinal Mushroom Phellinus igniarius (Agaricomycetes).

Nuclear factor-κB (NF-κB) has an important role in immune response and inflammation. Phellinus igniarius (known as Sang huang in traditional Chinese medicine) has various pharmacologic effects. In this study, 19 chemical constituents (1-19) were isolated from the fruiting bodies of Ph. igniarius. Their structures were elucidated based on data from nuclear magnetic resonance spectroscopy and comparison with the literature. Overall, 3 compounds (2, 7, and 11) from this species were reported, to our knowledge for the first time. Two compounds (4 and 15) from the genus Phellinus and 9 compounds (1, 3, 5, 8-10, and 12-14) from the Hymenochaetaceae family were also reported for the first time. An NF-κB luciferase reporter assay of these compounds was carried out in tumor necrosis factor-α-induced HeLa cells-again, to our knowledge for the first time. Among them, compounds 5 and 7 showed moderate inhibition of NF-κB, with fold values of 0.48 ± 0.02 and 0.55 ± 0.09, respectively, in HeLa cells at 100 µmol/L. These results suggest that some of the ingredients from Ph. igniarius could be developed into antiinflammatory agents for use in clinical applications.

One new 1,4-napthoquinone derivative from the roots of Juglans mandshurica.

A new 1,4-napthoquinone derivative, namely (S)-(-)-3-(8-hydroxy-1,4-dioxo-1,4-dihydro-naphthalen-2-yl)-3-(4-hydroxy-3-methoxyphenyl)-propionic acid methyl ester (1), was isolated from the roots of Juglans mandshurica Maxim. The structure was identified based on HR-ESI-MS, 1D and 2D NMR spectroscopic methods.

A natural chalcone induces apoptosis in lung cancer cells: 3D-QSAR, docking and an in vivo/vitro assay.

This study was to study the antitumor effect of lonchocarpin (34) from traditional herbal medicine Pongamia pinnata (L.) Pierre and to reveal the underlying mechanism. The cytotoxic activities of lonchocarpin were evaluated in 10 lung cancer cell lines and it exhibited 97.5% activity at a dose of 100 µM in the H292 cell line. A field-based quantitative structure-activity relationship (3D-QSAR) study of 37 flavonoids from P. pinnata was also performed, and the results obtained showed that the hydrophobic interaction could be the crucial factor for the antitumor activity of lonchocarpin. Molecular docking studies revealed that lonchocarpin bound stably to the BH3-binding groove of the Bcl-2 protein with hydrophobic interactions with ALA146. Also, lonchocarpin significantly reduced cell proliferation via modulating Bax/Caspase-9/Caspase-3 pathway. An apoptotic test using flow cytometry showed that lonchocarpin produced about 41.1% and 47.9% apoptosis after treatment for 24 h and 48 h, respectively. Moreover, lonchocarpin inhibited tumor growth in S180-bearing mice with an inhibition rate of 57.94, 63.40 and 72.51%, respectively at a dose of 25, 50 and 100 mg/kg. These results suggest that lonchocarpin is a potentially useful natural agent for cancer treatment.

Natural potential neuroinflammatory inhibitors from Alhagi sparsifolia Shap.

Neuroinflammation is a key contributor to neuronal damage in neurodegenerative diseases. In our previous work on natural effective neuroinflammatory inhibitors, Alhagi sparsifolia Shap. (Leguminosae), a folk medicine widely distributed in Xinjiang, attracted our attention because of its significant anti-neuroinflammatory effect. Therefore, further investigation of the bioactive material basis was carried out. As a result, 33 major components were characterized and identified by chromatographic and spectral methods, respectively. Furthermore, the anti-neuroinflammatory effects of the extract and purified constituents were evaluated in LPS-induced N9 cells in vitro. The results displayed that compounds 1, 2, 3, 5, 6, 8, 11, 15, 16, 17, 22, 23, 25, 26, 28, 30, 33 could exhibit significant inhibitory activities without obvious cytotoxicities at their effective concentrations. Especially, isorhamnetin (1) (IC50 17.87µM), quercetin (2) (10.22µM), 3',7-dihydroxyl-4'-methoxylisoflavone (5) (17.43µM), 3',7-dihydroxyl-4',6-dimethoxylisoflavone (6) (11.21µM), syringgaresinol (16) (2.68µM), bombasinol A (17) (7.61µM), aurantiamide (23) (14.91µM) and 1,3,3,4-tetramethyl cyclopentene (33) (2.63µM) showed much stronger inhibiting effect than that of the positive control minocycline (19.89µM). Therefore, the effective compositions might be responsible for the significant neuroinflammation inhibitory activities exhibited by the herb. Moreover, compounds 16 and 33 could be good leading compounds for the development of potential therapeutic agents against neurodegenerative diseases.

Determination of a new chromone from Aurantii Fructus Immaturus by DFT/GIAO method.

Investigation of EtOAc fraction from the 95% ethanol extract of Aurantii Fructus Immaturus led to the isolation of one new chromone (1) and seven known flavonoids (2-8). Their structures were elucidated mainly by NMR and HR-ESI-MS, as well as on comparison with the reported NMR data. The final substituent pattern of 1 was defined by comparison of (13)C NMR data calculated by DFT/GIAO with those of experimental NMR data.

Natural therapeutic agents for neurodegenerative diseases from a traditional herbal medicine Pongamia pinnata (L.) Pierre.

Neurodegenerative diseases are associated with neuroinflammation, manifested by over-production of nitric oxide (NO) by microglial cells. Now there still lack effective treatment and prevention for the neurodegenerative diseases. Concerning neuroinflammation mediated by microglia cell, bioactivity-guided phytochemical research of Pongamia pinnata (L.) Pierre was performed in this study. A new chlorinated flavonoid, 2',6'-dichlore-3', 5'-dimethoxy-[2'',3'':7,8]-furanoflavone (1) was identified together with 29 known compounds, including flavonoids (compounds 2-17), isoflavonoids (compounds 18-23), chalcones (compounds 24-25), flavonones (compounds 26-27), triterpenes (28-29) and alkaloid (30) from the effective dichloride methane extract of dry stem of P. pinnata (L.) Pierre. Their structures were elucidated by physicochemical and spectral methods. The anti-neuroinflammatory activities were assayed in BV-2 cells by assessing LPS-induced NO production. Then pongaglabol methyl ether (2), lonchocarpin (24) and glabrachromene II (25) were selected as potential therapeutic agents for neurodegenerative diseases because of their significant anti-neuroinflammatory activities. Furthermore, the characteristics of structure type existing in P. pinnata (L.) Pierre and brief SAR were summarized, respectively.

A new phenyl glycoside from the aerial parts of Equisetum hyemale.

A new phenyl glycoside, 2-(sophorosyl)-1-(4-hydroxyphenyl)ethanone (9), was isolated from the ethanolic extract of the aerial parts of Equisetum hyemale L., together with eight known compounds (1-8). The structures of these compounds were elucidated using a combination of spectroscopic analyses and chemical method. Of these nine compounds, 4 and 7 showed hepatoprotective effects towards tacrine-induced cytotoxicity in Hep 3B cells with EC50 values of 42.7 ± 1.5 and 132.6 ± 2.8 µM, respectively.

[Effects of fuzheng huayu recipe in antagonizing rat's renal interstitial fibrosis].

OBJECTIVE: To observe the effects of Fuzheng Huayu Recipe (FHR) on rat's renal interstitial fibrosis induced by mercuric chloride (HgCl2), and to explore preliminarily its mechanism of action. METHODS: Rats were randomly divided into four groups: the normal group, the model group, the FHR group and the vitamin E group, the latter two were treated respectively by FHR 4.6 g/kg and vitamin E 100 mg/kg. Rats model was established by oral administration of 8 mg/kg HgCl2 for 9 weeks. Serum creatinine (Cr) and urea nitrogen (BUN) content were tested with corresponding test kits; hydroxyproline (Hyp) content in kidney was assayed with hydrochloric acid hydrolysis; renal histologic change was observed with HE, Masson and methenamine silver (PASM) staining; and collagen type I (Col I), as well as protein expressions of fibronectin (FN) and alpha-smooth muscle actin (alpha-SMA) was determined with Western blot. RESULTS: Compared with the model group, the kidney/body weight ratio, serum levels of Cr and BUN, kidney Hyp content, and severity of renal interstitial fibrosis in the two treated groups were significantly lower (P<0.05 or P<0.01), and the improvements were more significant in the FHR group than those in the vitamin E group; Col I and FN protein expression was also weaker in the two treated group (Col, P<0.05; FN, P<0.01); while the expression of alpha-SMA was lower in the FHR group (P<0.01), but it wasn't in the vitamin E group (P>0.05). CONCLUSION: FHR could improve the HgCl2-induced renal function injury in rats, decrease extracellular matrix deposition and restrain renal interstitial fibrosis, the mechanism of action might be related with its inhibitory effect on myofibroblast activation.

[Effect of Qingxin Kaiqiao fang on learning and memory ability and morphology of hippocampal nerve cells in AD mice].

OBJECTIVE: To study the effect of the Qingxin Kaiqiao fang on learning and memory ability and shape hippocampal nerve cells in Alzheimer disease (AD). METHOD: One handred and fifty mice were divid into five groups: blank group, model group, two groups of treatment by Qingxin Kaiqiao fang (14.82, 7.41 g x kg(-1)), piracetan comparison group (0.42 g x kg(-1)). The model group was orally given AlCl3 (200 mg x kg(-1)) every day. For Qingxin Kaiqiaofang and piracetan groups, AlCl3 treatment was given for 6 days at the beginning, followed by giving orally AlCl3 in the morning and drug in the afternoon for 8 weeks. Then, learning and memory aility, the contents of nitric oxide (NO), NO synthase (NOS) and acetylcholinesterase (AchE) in brain, and morphology of hippocampal nerve cells were investigated. RESULT: Learning and memory ability of Qingxin Kaiqiaofang groups was improved, compared with comparison group; the difference was significant (P < 0.05, P < 0.01). The drug could prevent hippocampal nerve cells from damaged obviously. The contents of NO, NOS and AchE in mice of treatment groups were lower than those of comparison group; the difference was significant (P < 0.05, P < 0.01). CONCLUSION: Qingxin Kaiqiaofang can improve learning and memory ability of AD mice, prote chippocampal nerve cells, and treat Alzheimer disease.

[Effect of combined therapy of xiaoke shen'an capsule and western medicine in diabetic nephropathy].

OBJECTIVE: To observe the therapeutic effect of xiaoke shen' an capsule on diabetic nephropathy. METHODS: Ninety patients with diabetic nephropathy were randomly divided into two groups, the patients in the control group (CG) were treated with conventional western medicine, and those in the treated group (TG) were treated with combined therapy of xiaoke shen'an capsule and conventional western medicine. The treatment course of both groups was 8 weeks, and the therapeutic effect related indexes were measured before and after treatment. RESULTS: The curative rate was 85.0% and 73.3% in CG and TG respectively, it was better in TG than that in CG (P < 0.05). Indexes such as fasting blood glucose, glycosylated hemoglobin, blood pressure, quantity of 24h urinary protein, urinary albumin excretion rate, renal function, blood lipids and hemoreheologic parameters were significantly different before and after treatment in TG (P < 0.05 or P < 0.01). As compared with CG, some of these indexes after treatment were superior to those in CG (P < 0.01 or P < 0.05). CONCLUSION: Combined therapy of xiaoke shen'an capsule and conventional western medicine has definite therapeutic effect on diabetic nephropathy.

[Discussion on thoughts and methods for treatment of diabetic nephropathy by TCM according to inflammatory pathogenesis].

Aim of this article was to investigate relationship between inflammatory pathogenesis of diabetic nephropathy and the TCM pathogenetic theory of Shen-Collateral impaired by Toxin, and to illustrate the method for removing toxin, activating collateral and protecting Shen can be an effective treatment for inhibiting the inflammatory pathogenesis of diabetic nephropathy.

[Effects of Fuzheng Huayu recipe on renal toxic interstitial fibrosis in rats].

OBJECTIVE: To investigate the effects of Fuzheng Huayu Recipe on renal interstitial fibrosis in rats induced by the toxic substances. METHODS: Forty Wistar rats were divided into 3 groups, including the normal group, model control and Fuzheng Huayu Recipe treated group. The renal fibrotic model was induced with intraperitoneal injection of dimethylnitrosamine (DMN) for 4 weeks and oral administration of food containing heavy metals such as Hg, etc for 8 weeks. From the 5th week, the rats were orally administered Fuzheng Huayu Recipe for 4 weeks. The renal tissue was stained with HE and periodic acid-silver metheramine, respectively. The serum creatinine and urea nitrogen were assayed with the kits, and the renal hydroxyproline (Hyp) contents in the homogenate of kidney tissue were measured with HCl hydrolysis. RESULTS: In the model group, the renal tubule was swollen, and the tubular epithelial cells were degenerated, necrosed and detached, with the infiltration of inflammatory cells; the tubular interstitium was expanded, characterized with the deposition of large collagens; the tubular basement membrane became thicker and wrinkled; and the renal Hyp content and serum creatinine and urea nitrogen levels increased remarkably as compared to the normal group. After the treatment with Fuzheng Huayu Recipe, the renal tubular inflammation and interstitial collagen deposition were alleviated, and the serum creatinine and urea nitrogen levels decreased significantly as compared to the model group. CONCLUSION: Administration of DMN and heavy metals can induce the renal interstitial fibrosis and impair the renal function. Fuzheng Huayu Recipe can improve the impaired renal functions and reverse the renal interstitial fibrosis.