Combined Administration of Curcumin and Chondroitin Sulfate Alleviates Cartilage Injury and Inflammation via NF-κB Pathway in Knee Osteoarthritis Rats.

Objective: Osteoarthritis (OA) is a degenerative chronic disease that most often occurs in the knee joint. Studies have shown that some food supplements, such as curcumin and chondroitin sulfate, are effective in treating knee osteoarthritis (KOA) by exhibiting different protective effects. In this study, we further investigated the combined therapeutic effects of curcumin and chondroitin sulfate on cartilage injury in rats with arthritis. Methods: An experimental KOA model was induced by monosodium iodoacetate (MIA) in rats. All rats were randomly divided into five groups: Ctrl (control), model (saline), Cur (20 mg/kg curcumin in saline), CS (100 mg/kg chondroitin sulfate in saline), and CA (20 mg/kg curcumin and 100 mg/kg chondroitin sulfate in saline); drugs were given 2 weeks after MIA injection. The histomorphological changes of cartilage were observed by safranin fast green staining, H&E staining, and micro-CT scanning. Also, the levels of PGE2, TNF-α and IL-1ß in the arthral fluid and serum were determined by the ELISA kits. The activities of SOD, CAT, COMP, MMP-3, and type II collagen were detected by biochemical kits. The expressions of TLR4, p-NF-κB, NF-κB, and COX-2 in cartilage were detected by Western blot. Results: Data show that serum levels of IL-1ß (p < 0.05), SOD (p < 0.0001), and MMP-3 (p < 0.001) were downregulated significantly in the CA group when compared to those in the model group. Meanwhile, obvious repair of cartilage with higher contains collagen II (p < 0.0001) could be observed in the CA group than the ones in Cur or CS group. In addition, significant downregulation of the expression of p-p65/p65 (p < 0.05) was found in the CA group. Conclusion: Our findings showed that combined administration of curcumin and chondroitin sulfate could exert better repair for KOA in rat models. This may hold great promise for discovering potential drugs to treat KOA and may improve treatment options for it.

Evaluation of toxicity studies of flavonoid fraction of Lithocarpus polystachyus Rehd in rodents.

The aim of the study was to evaluate the safety of flavonoid fraction of Lithocarpus polystachyus Rehd (Sweet Tea-F, ST-F) in mice and rats through acute and sub-chronic toxicity studies respectively. For acute toxicity study, a single dose of 5000 mg/kg of ST-F was given orally to healthy KM mice. The mice were observed mortality and toxic symptoms for 24 h, then once a day up to 14 days. In the sub-chronic toxicity study, ST-F was administered orally at doses of 0, 70, 140, 560 mg/kg/day to rats for 26 weeks. Body weight and food intake were recorded weekly. Hematological, biochemical, coagulation and organ parameters were analyzed at the end of 26 weeks administration. Vital organs were evaluated by histopathology. In the acute toxicity study, ST-F caused neither significant toxic symptoms, nor mortality in mice. In sub-chronic toxicity study, daily oral administration of ST-F at the dose of 70 mg/kg resulted in a significant increase (P < 0.05) in the relative body weight at the 10-week, and the same situation brought at the dose of 140 mg/kg/day at the 22-week. Hematological and biochemical showed significant changes (P < 0.01 or P < 0.05) in WBC, GLU, ALP, AST and serum electrolytes levels at the dose of 560 mg/kg/day. The amount of RBC decreased significantly (P < 0.05) while the content of PLT slightly increased (P < 0.05) at the dose of 140 mg/kg/day. In additional, no obvious histological changes were observed in vital organs of ST-F treated animals compared to control group. The ST-F may be exit slight side effects at the dose of 560 mg/kg/day in rats. Thus, the overall results show that the no-observed adverse effect level (NOAEL) of ST-F was considered to be 140 mg/kg for male SD rats.

Effect of Dendrobium officinale on D-galactose-induced aging mice.

OBJECTIVE: To examine the effect of Dendrobium officinale (DO) on D-galactose-induced aging mice. METHODS: Aging mice was induced by D-galactose at 0.125 g/kg for 10 weeks through subcutaneous injection except for the negative control group. After 10 days, according to complete random design, the aging modeling mice were randomized into 4 groups: aging control group (10 ML·kg-1·d-1) of distilled water), positive control group (vitamin B6 and ganodema lucidum tablets with a dose of 1 tablet/kg), DO-1 treatment group (DO juice with a dose of 1 g/kg), DO-2 treatment group (DO Polysaccharide with a dose of 0.32 g/kg), 14 mice in each group. All the animals were orally medicated daily for 9 weeks. Cognitive function assessment was performed using the maze test and step-down test. At the end of experiment, the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC) levels in the serum, the SOD, GSH-Px and nitric oxide (NO) levels in the cerebrum, the SOD and catalase (CAT) levels in the liver, the SOD and NO levels in the heart, and the SOD level in the kidney, were determined using commercial kits. The spleen, liver, heart, cerebrum and kidney were excised for histological study. RESULTS: Compared to aging control group, DO shortened the time of passing through the maze and prolong the step-down latency of aging mice (P <0.05 or P<0.01). DO markedly up-regulated serum levels of SOD, GSH-Px and T-AOC, and restored SOD levels in the heart, liver, kidney and cerebrum to normal status (P<0.05 or P<0.01). DO at the dose of 1 g/kg also signififi cantly improved the degree of spleen lesions (P<0.01). CONCLUSIONS: DO had marked anti-aging effect on D-galactose-induced model of aging. The underlying mechanism could be related to modulation on antioxidation system and immune system. The results indicated that DO could potentially be used as natural drugs or functional foods for preventing aging.

Evaluation of the effects of the water-soluble total flavonoids from Isodon lophanthoides var.gerardianus (Benth.) H. Hara on apoptosis in HepG2 cell: Investigation of the most relevant mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: The water-soluble total flavonoids (WSTF) were extracted from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, a common folk herbal medicine in China, which has been recorded by the "Chinese Pharmacopoeia" in 2015 and used for prevention and clinical treatment of common diseases of liver and gall for many years. OBJECTIVE OF THE STUDY: The aim of this study is to evaluate the effects of WSTF on apoptosis in HepG2 cell and investigate the relevant mechanisms underlying. MATERIALS AND METHODS: Cytotoxicity was evaluated in HepG2 cells (human hepatoma cell lines) using MTT assay. The influence of the WSTF on the intracellular reactive oxygen species (iROS) and the mitochondrial membrane potential were also determinated. We used flow cytometry analysis to detect the effects of WSTF on apoptosis, cell cycle. Then we applied RT-PCR for genetic expression of main effectors and western blot analysis for activation of main effectors involved in the potential apoptosis signaling pathways. RESULTS: WSTF inhibited cell growth in HepG2 cells. Moreover, WSTF stimulates to increase amount of iROS, mitochondrial membrane potential, and the apoptotic relevant factors (cytochrome c, caspase-3) in HepG2 cells. WSTF could significantly induce apoptosis through downregulating apoptosis-antagonizing protein (Bcl-2, Survivin, mcl-1) and upregulating apoptosis-promoting proteins (Bax) and cell cycle G0/G1 arrest in HepG2 cells. CONCLUSION: The results indicate that WSTF induces cell apoptosis through mitochondrial pathway in the HepG2 cells. Therefore, these studies suggest that WSTF could be used as a chemotherapeutic agent to treat hepatoma.

Dendrobium officinale Prevents Early Complications in Streptozotocin-Induced Diabetic Rats.

Background. Dendrobium officinale (DO) Kimura et Migo is a precious Chinese herb that is considered beneficial for health due to its antioxidant and antidiabetes properties, and so on. In this research, we try to determine the preventive effect of DO on the early complications of STZ-induced diabetic rats. Methods. Type 1 diabetic rats were produced with a single intraperitoneal injection of STZ (50 mg/kg). DO (1 g/kg/day) was then orally administered for 5 weeks. Blood glucose, TC, TG, BUN, CREA, and GSH-PX levels were determined, and electroretinographic activity and hypoalgesia were investigated. Pathological sections of the eyes, hearts, aortas, kidneys, and livers were analyzed. Results. Treatment with DO significantly attenuated the serum levels of TC, TG, BUN, and CREA, markedly increased the amplitudes of ERG a- and b-waves and Ops, and reduced the hypoalgesia and histopathological changes of vital organs induced by hyperglycemia. The protective effect of DO in diabetic rats may be associated with its antioxidant activity, as evidenced by the marked increase in the serum level of glutathione peroxidase. However, DO had no significant effect on blood glucose levels and bodyweight of diabetic rats. Conclusions. DO supplementation is an effective treatment to prevent STZ-induced diabetic complications.

New dihydrochalcone and propenamide from Lithocarpus polystachyus.

Two new compounds, (E)-4-(3, 4-dihydroxyphenyl)-N-(1 -hydroxy-2-(4-hydroxyphenyl) ethyl)-2-oxobut-3-enamide (1) and phloretin2-O-beta-apiofuranosyl (1-->06)-beta-D-glucopyranoside (2) were isolated from Lithocarpus polystachyus Rehd. Their structures were determined on the basis of analysis of their 1D and 2D NMR spectroscopic and mass spectral data.

White pepper and piperine have different effects on pharmacokinetics of puerarin in rats.

This study attempted to explore the effects of white pepper and its major component piperine on puerarin administered to rats. Pharmacokinetic parameters of puerarin in rats were determined by oral administration (400 mg/kg) or intravenous injection (40 mg/kg) of puerarin, pretreated with or without white pepper and piperine given orally. Compared to the control group given oral puerarin only, the combined use of piperine (10 or 20 mg/kg) increased the C max of puerarin by 1.30-fold or 1.64-fold and the AUC0-∞ by 133% or 157%, respectively. In contrast, coadministration of white pepper (125 or 250 mg/kg) decreased oral absorption of puerarin to 83% or 74%, respectively. On the other hand, pretreatment with piperine orally did not alter the intravenous pharmacokinetics of puerarin, while the AUC of puerarin after intravenous administration was increased by pretreatment with white pepper. The results indicate that pretreatment with piperine or pepper exerts different effects on pharmacokinetics of puerarin administrated via intragastric and intravenous routes. Therefore, it is suggested that the combined application of piperine or white pepper with puerarin should be carefully monitored for potential diet-drug interactions.

Sweet tea leaves extract improves leptin resistance in diet-induced obese rats.

AIM OF THE STUDY: Dietary obesity is usually characterized by leptin resistance and abnormal lipid metabolism. Lithocarpus polystachyus Rehd.(Sweet Tea) leaf is a kind of Chinese folkloric medicine, and it has been widely used for obesity, diabetes, and hypertension in South China. The present study is aimed at investigating the pharmacological mechanism of the anti-hyperleptinaemia effects of Sweet Tea leaves extract in high fat diet-induced obese rats. MATERIALS AND METHODS: We induced high fat diet obesity for 14 weeks to test the corrective effects of three ST doses (75, 150 and 300 mg/kg per day) for 8 weeks. At the end of the experiment, body weight, fasting blood glucose and serum lipids, superoxide dismutase (SOD), malondialdehyde (MDA), fasting serum insulin and leptin, C-reactive protein, adiponectin and resistin levels were measured, Homeostasis Model Assessment for Insulin Resistance (HOMA-IR) was also calculated. mRNA gene expression of PPARγ (peroxisome proliferator-activated receptor γ) and C/EBPα(CCAAT/enhancer-binding protein α) in epididymal adipose tissue of DIO control and experimental groups were evaluated. RESULTS: Sweet Tea leaves extract could significantly decrease the levels of serum lipids, attenuate body weight gain and lower circulating leptin and insulin levels, ameliorate the state of oxidative stress, raise serum adiponectin, reduce circulating CRP and resistin levels, and depress the expression of PPARγ and C/EBPα in epididymal adipose tissue of obese rats. CONCLUSION: The present findings suggest that ST can effectively attenuate the leptin resistance at least through anti-hyperlipidemic activity and thus has the therapeutic potential in treating hyperlipidemia and hyperleptinaemia related to dietary obesity.

Effect of the flavonoid fraction of Lithocarpus polystachyus Rehd. on spontaneously hypertensive and normotensive rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Lilhocarpus polystachys Rehd. (Sweet Tea, ST) is a folk herbal medicine that has been traditionally used as a natural remedy for hypertension in China, whose mechanism remains unveiled. Flavonoid fraction is considered as the major active components in ST. This study aimed to provide experimental evidence for the anti-hypertension activity of flavonoid fraction of ST (ST-F) and investigate the underlying mechanism. The effect of ST-F on the blood pressure of normotensive rats was also to be determined. MATERIALS AND METHODS: Spontaneously hypertensive rats (SHRs) were treated with ST-F daily for 10 weeks. Blood pressure of SHRs was measured before and biweekly during ST-F treatment. Subsequently, animals were sacrificed either immediately at the end of treatment or 2 weeks after ST-F treatment discontinuance. The activities of plasma rennin (PRA), angiotensin II (Ang-I), endothelin (ET), nitric oxide (NO), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured as well as skin microcirculatory flux. In normotensive rats, blood pressure was determined after six months' treatment of ST-F. RESULTS: ST-F treatment significantly reduced the blood pressure of SHRs along with decreasing plasma levels of PRA and Ang II. ST-F did not show obvious effects on plasma levels of ET, NO or SOD, but it significantly decreased the plasma level of MDA and improved skin microcirculatory flux. Compared to the anti-hypertensive drug enalapril, ST-F showed a modest effect on lowering blood pressure of SHRs without obvious withdrawal reactions. But long-term intake of ST-F did not change the blood pressure in normotensive rats. CONCLUSION: ST-F had an antihypertensive effect on SHRs. The underlying mechanism could be related to modulation on the rennin-angiotensin-aldosterone system (RAAS) and antioxidation system, as well as regulation of skin microcirculation. Compared to its anti-hypertensive effect on SHRs, ST-F did not cause hypotension in normotensive rats. The results indicated that ST-F could potentially be used as natural drugs or functional foods for preventing hypertension.

The hypoglycemic activity of Lithocarpus polystachyus Rehd. leaves in the experimental hyperglycemic rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves of Lithocarpus polystachyus Rehd. are used for the treatment of disorders such as diabetes, hypertension, and epilepsy in folk medicine of South China. The possible antidiabetic effects of the leaves were investigated in experimental type 2 and type 1 diabetic rats. MATERIALS AND METHODS: Type 2 diabetic rats received orally three different extracts of Lithocarpus polystachyus Rehd. leaves for 4 weeks (aqueous extract [ST-1], ethanol extract [ST-2], flavonoid-rich fraction [ST-3]). At the end of the experiment biochemical parameters were tested and livers and pancreases were excised for histological study. After the comparison of the pharmacological test results of the three extracts, the one which showed the best bioactivity was further studied to confirm its antidiabetes effect on both type 2 and type 1 diabetic rats. RESULTS: Compared to ST-1 and ST-2, ST-3 had better effects on regulation of blood glucose, glycosylated serum protein, cholesterol, triglyceride, malondialdehyde, superoxide dismutase and attenuation of liver injury in type 2 diabetic rats (p<0.01 or p<0.05). ST-3 administration for four weeks also significantly reduced the fasting serum insulin and C-peptide level and improved the insulin tolerance (p<0.05). In type 1 diabetic rats, ST-3 supplement for three weeks caused significant reduction in fasting blood glucose, total cholesterol, triglyceride, urea nitrogen, creatinine and liver mass, along with significantly inhibiting the decline of insulin level compared to diabetic control (p<0.05 or p<0.01). CONCLUSION: The flavonoid-rich fraction of Lithocarpus polystachyus Rehd. leaves (ST-3) had better beneficial effect than that of the ethanol or aqueous extract in experimental diabetic rats, which means that the bioactivity of the herbal leaves is probably due to the presence of flavonoids. The results also strongly suggest that the antidiabetic effect of ST-3 was possibly through multiple mechanisms of action including blood lipid and antioxidant mediation. The results indicated that the aqueous flavonoid-rich fraction of Lithocarpus polystachyus Rehd. leaves possessed significant protective activity in type 2 and type 1 diabetes.

Role of the interaction between puerarin and the erythrocyte membrane in puerarin-induced hemolysis.

Adverse drug reactions (ADR), especially intravenous hemolysis, have largely limited the application of puerarin injections in clinics. This study investigated the underlying mechanisms of puerarin-induced hemolysis. Our results show that puerarin induced concentration-dependent and time-dependent hemolysis when human erythrocytes were incubated in saline solution with more than 2mM puerarin for over 2h. However, incubation in PBS or addition of 1mM of lidocaine to the saline solution completely abolished the hemolysis. Providing materials that could start ATP synthesis did not reverse the hemolysis, and puerarin did not affect Na(+)-K(+)-ATPase activity. In addition, puerarin (0.1-2mM) did not cause calcium influx or exhibited pro-oxidant activity in erythrocytes. Puerarin exhibited different influences on the membrane microviscosity of erythrocytes in saline and PBS. Moreover, 1mM lidocaine inhibited 8mM puerarin-induced reduction of membrane microviscosity in saline solution. SDS-PAGE analysis of membrane proteins revealed that 2mM puerarin treatment induced the appearance of several new protein bands but attenuated the expression of protein bands 2.1, 3, 4.1, 4.2 and 5. These results suggest that high concentrations of puerarin-induced hemolysis were associated with the changes of membrane lipids and of the composition of erythrocytes membrane proteins but not with ATP depletion, pro-oxidation and calcium influx. These changes could be related to the intercalation of amphiphilic puerarin at high concentration into the erythrocyte membrane in certain media, resulting in membrane disorganization and, eventually, cytolysis. Hence, in clinics, determining the optimal dose of puerarin is critical to avoid overdosing and ADR.

Ginseng protects rodent hearts from acute myocardial ischemia-reperfusion injury through GR/ER-activated RISK pathway in an endothelial NOS-dependent mechanism.

ETHNOPHARMACOLOGICAL RELEVANCE: Ginseng (Panax ginseng C.A. Meyer) is widely used in Asian communities for treating cardiovascular diseases. However, the mechanism by which it protects the myocardium in ischemia-reperfusion (I/R) injury remains unclear. In this study, we aim to investigate whether a standardized ginseng extract (RSE) protects rodent hearts against I/R injury and if glucocorticoid and/or estrogen receptor-mediated activation of Akt and Erk1/2 (the reperfusion injury salvage kinase pathway, RISK) and subsequent nitric oxide (NO) synthesis signaling are involved in this effect. MATERIALS AND METHODS: Rats or gene-deleted mice were subjected to 30 min ischemia by occluding the left anterior descending coronary artery and 90 min reperfusion. Infarct size, serum level of creatine kinase (CK), lactate dehydrogenase (LDH), and NO, expression and phosphorylation of glucocorticoid receptor (GR), estrogen receptor (ER), phosphatidylinositol-3 kinase (PI3K), Akt, NO synthase (NOS), extracellular signal-regulated kinase (Erk) 1/2, p38, and c-Jun NH2 terminal kinases (JNK) were examined in rat or mice treated with or without RSE in the absence or presence of pharmacological inhibitors. RESULTS: RSE significantly reduced infarct size in a dose-dependent manner and reduced the incidence of arrhythmia, increased serum NO production, reduced serum activities of creatine kinase and lactate dehydrogenase. The infarct size reduction effect of RSE was abolished by RU468 (an inhibitor of GR), tamoxifen (an inhibitor of ER), LY294002 (an inhibitor of PI3K), Akt inhibitor IV (an inhibitor of Akt protein kinase), U0126 (an inhibitor of Erk1/2) and NG-nitro-l-arginine methyl ester hydrochloride (an inhibitor of NOS), but not actinomycin D (an inhibitor of transcription process). RSE also significantly increased the activation of GR/ER, PI3K-Akt-eNOS cascades and Erk1/2 signaling in rat heart. However, RSE did not markedly reduce infarct size in endothelium NOS(-/-) mice. This differs from its effect in inducible NOS(-/-) and wild type mice, suggesting that endothelium NOS is required for the beneficial effect of RSE on the heart. CONCLUSION: Our findings showed for the first time that RSE protects hearts subjected to acute I/R injury and the infarct size reduction effect of RSE is associated with GR and/or ER-mediated Akt and Erk1/2 activation in an endothelium NOS-dependent manner.

[Protective effect of Isodon lophanthoides on acute hepatic injury induced by D-galactosamine in rats].

OBJECTIVE: To study the protective effects of Isodon lophanthoides (ILVG) aqueous extract on acute hepatic injury induced by D-galactosamine (D-Gal) in rats. METHODS: 60 rats were divided into control group, model group, and low, middle, high dosage group, Bifendate group randomly. In test groups, rats received either ILVG aqueous extract (15, 7.5, 3.75 g/kg) or Bifendate (45 mg/kg) by gastric perfusion daily for 7 consecutive days. On the sixth day, D-Gal (550 mg/kg) was given to rats by oral administration. The levels of ALT, AST, ALP, TBA, T-Bil, TP and ALB in serum were analyzed. The weight of body, liver, spleen and thymus of each rat were measured. The hepatic glycogen content was analyzed individually. Liver tissue pathology was observed. RESULTS: ILVG coud decrease the ALT, AST, ALP, TBA and T-Bil in serum, increase TP, ALB and hepatic glycogen content and restrain the enlargement of liver and the shrinkage of thymus, reduce necrosis in pathological observation. CONCLUSION: ILVG aqueous extract possesses the effect on protecting on acute hepatic injury induced by D-Gal in rats, its effect is related to multifarious mechanism.