The effects of antifreeze proteins(AFP)on the quality changes of pork during freezing were investigated in comparison to those of the commercial antifreeze agent alginate(4 mg/mL).Samples with no added AFP were used as a control.The effects of adding different amounts of AFP(0.05,0.1,0.2,and 0.3 mg/mL)on the quality,physicochemical properties of myofibrillar proteins,microstructure,and water distribution of frozen pork were investigated.The results showed that AFP decreased the water loss of frozen pork,which was beneficial to maintaining pork color and microstructure.Addition of AFP at 0.3 mg/mL had the highest cryoprotective effect on the cooking loss,thawing loss and color characteristics of pork during frozen storage,followed by AFP at 0.2 mg/mL and alginate at 4 mg/mL.The shear force and total sulfhydryl content of myofibrillar proteins in the 0.2 and 0.3 mg/mL AFP groups were significantly higher than those of the control group(P＜0.05).Nuclear magnetic resonance imaging(NMRI)discovered that the infiltration of AFP reduced the loss of bound and immobile water in pork.Scanning electron microscopy showed that in the presence of 0.2 mg/mL AFP,the arrangement of muscle fibers was orderly,appearing more compact and denser.Overall,AFP addition had a positive effect on improving the quality of frozen pork and might have the potential of being a safe and efficient cryoprotection method.

Background: Current treatments for osteoarthritis (OA) pain and stiffness have limitations, including adverse effects. Therefore, effective and safe complementary or alternative therapies are needed. Dietary supplement GJ 191, comprising Epimedium, Dioscorea, and Salvia miltiorrhiza extracts, may address this need. Methods: This randomized, double-blind, placebo-controlled study investigated GJ 191 supplementation on knee OA symptoms. Seventy-two adults (40-75 years) with mild to moderate knee OA and mild to moderate knee pain were enrolled. The Knee Injury and Osteoarthritis Outcome Score (KOOS), Pain Visual Analog Scale (VAS), Quality of Life questionnaire, knee joint range of motion, serum C-reactive protein, and rescue medication use were assessed. The Western Ontario McMaster Universities Osteoarthritis Index (WOMAC) pain and stiffness scores were computed using KOOS scores. Results: Decreases in WOMAC pain scores were reported by both GJ 191 and placebo groups after 6 (-1.78 +/- 2.71 and -1.34 +/- 1.93, respectively; p < 0.01) and 12 (-2.31 +/- 2.83 and -1.59 +/- 2.69, respectively; p < 0.01) weeks, with no significant difference between groups. There were decreases in WOMAC stiffness scores for participants supplemented with GJ 191 by 0.53 +/- 1.22 and 0.72 +/- 1.46 (p <= 0.02) after 6 and 12 weeks, respectively, with respective decreases of 0.81 +/- 1.51 and 0.75 +/- 1.85 (p <= 0.03) for those on placebo. Significant improvements in current pain, as assessed by the Pain VAS, and bodily pain were reported by the GJ 191 group after 6 and 12 weeks, while the placebo group only reported significant improvements in these measures after 12 weeks. GJ 191 supplementation was safe and well tolerated. Conclusion: There was no significant difference in pain and stiffness scores between GJ 191 and placebo over the 12 weeks. While both groups reported improvements in WOMAC pain from baseline, improvements in current and bodily pain were experienced sooner with GJ 191 than placebo and were sustained over the study period. GJ 191 supplementation was safe and well tolerated.

ETHNOPHARMACOLOGICAL RELEVANCE:Cardiovascular disease (CVD) and fatigue are two common diseases endangering human life and health that may interact and reinforce one another. Myocardial infarction survivors frequently experience fatigue, and acute myocardial infarction (AMI) is one of the most common cardiovascular diseases that cause fatigue-induced sudden death. Sheng Mai Yin (SMY), a Chinese medicine prescription, is traditionally used for the treatment of diabetes and cardiovascular disease, and has been demonstrated to reduce fatigue and safeguard cardiac function.AIM OF THE STUDY:This study aims to investigate the effects and underlying mechanisms of SMY in treating fatigue and AMI.MATERIALS AND METHODS:The pharmacological mechanisms of SMY in treating fatigue and AMI were predicted by bioinformatics and network pharmacology methods. After administering SMY at high, medium and low doses, the swimming time to exhaustion, hemoglobin level, serological parameters and hypoxia tolerance time were detected in C57BL/6N mice, and the left ventricular ejection fractions (LVEF), left ventricular fractional shortening (LVFS), grasp strength, cardiac histopathology, serological parameters and the expression of PINK1 and Parkin proteins were examined in Wistar rats.RESULTS:371 core targets for SMY and 282 disease targets for fatigue and AMI were obtained using bioinformatics and network pharmacology methods. Enrichment analysis of target genes revealed that SMY might interfere with fatigue and AMI through biological processes such as mitochondrial autophagy, apoptosis, and oxidative stress. For in vivo experiments, SMY showed significant anti-fatigue and hypoxia tolerance effects in mice; It also improved the cardiac function and grasp strength, decreased their cardiac index, myocardial injury and fibrosis degree, and induced serological parameters levels and the expression of PTEN-induced putative kinase 1 (PINK1) and Parkin proteins in myocardium, suggesting that SMY may exert cardioprotective effects in a joint rat model of fatigue and AMI by inhibiting excessive mitochondrial autophagy.CONCLUSION:This study revealed the anti-fatigue, anti-hypoxia and cardioprotective effects of SMY in a joint model of fatigue-AMI, and the pharmacological mechanism may be related to the inhibition of mitochondrial autophagy in cardiomyocytes through the PINK1/Parkin pathway. The discoveries may provide new ideas for the mechanism study of traditional Chinese medicine, especially complex prescriptions, in treating fatigue and AMI.

Previously, we have prepared clam peptides (RBPs) that exhibited potential antihypertension and kidney protective effects in SHR rats by fermentation of Ruditapes philippinarum with Bacillus natto. In this study, we established an adenine-induced chronic kidney disease (CKD) in SD rats and investigated the improvement effects of RBPs on CKD. The results showed that compared with the model group, 8 weeks gavage of RBPs restored the structure of the kidney and reduced serum levels of CKD-related biochemical indicators, including NGAL, CR, Ca, UA, TG, IP, L-FABP, BUN, CHO, and cystatin C (p<0.05). RBPs treatment also reduced serum levels of MDA, IL-1β, TGF-β1, and TNF-α and increased those of CAT and T-AOC (p<0.05). In addition, the results of intestinal microbiota analysis showed that RBPs improved the intestinal flora composition by downregulating the proportion of Firmicutes and Bacteroidetes, decreasing the relative abundance of Romboutsia, Ruminococcaceae_UCG-005, and Christensenellaceae_R-7_group and increasing that of Lactobacillus and Muribaculaceae (p<0.05). The results of this study indicated that RBPs can also be used as bioactive substances in marine drugs or functional foods to alleviate CKD.

Curcumin, from the rhizome of turmeric (Curcuma longa L.), has a wide variety of biological activities. Unfortunately, its poor water-solubility greatly limits its bioavailability. The purpose of this study was to evaluate CUMINUP60®, a novel preparation utilizing a solvent-free, co-grinding method designed to improve curcumin's bioavailability. We performed a single-center crossover experiment to compare the new product with standard 95% curcumin in the blood plasma of twelve healthy adults (10 males, 2 females). Total bioavailability of curcumin and its sulfate and glucuronide conjugates from the test product, measured by their areas under the curve over 12 h (AUC0-T), showed a combined increase of 178-fold over standard curcumin and its conjugates from the reference product. The new product represents a significant improvement for providing greater bioavailability of curcumin, as compared with several other branded preparations. It therefore has broad applications for preparing curcumin as a more effective health ingredient in functional foods, beverages, and nutraceuticals.

目的:研究了水溶性姜黄素制剂CuminUP30在大鼠体内的药动学情况,探究了CuminUP30的抗疲劳和免疫支持的效果.方法:采用单一剂量给药的方法,通过收集不同时间点的血液,进行大鼠体内药动学分析.采用运动疲劳小鼠模型,通过连续灌胃给药 28 d,检测小鼠各运动性能指标.采用免疫功能损伤斑马鱼模型,与受试样品共孵化,检测其免疫细胞和免疫因子水平.结果:结果发现,CuminUP30 相对生物利用度提升 255.03%.CuminUP30 可以显著延长小鼠负重游泳和耐缺氧的时间,显著降低血乳酸含量,提高血液中乳酸脱氢酶的含量,显著降低肌乳酸的分泌,显著提高肌肉腺嘌呤核苷三磷酸(adenosine-triphosphate,ATP)的产生,提高糖原的合成,提高超氧化歧化酶(superoxide dismutase,SOD)的活性.CuminUP30 可以提高免疫受损斑马鱼的吞噬细胞和T细胞水平,降低促炎因子白介素-1β(Interleukin-1 beta,IL-1β)表达,显著提高干扰素γ1(Interferon Gamma1,IFNG1)和重组激活基因 2(recombination activating gene 2,rag2)的表达.结论:CuminUP30 提升了姜黄素的生物利用度,可以缓解因有氧或无氧运动过度,引起的运动疲劳,可以调节机体免疫细胞和免疫因子的水平,提高机体免疫力.

Hyperuricemia is a metabolic disease caused by disturbances in purine metabolism and imbalances in the formation and excretion of uric acid. It is closely related to the gut microbiota, which is considered a therapeutic target. Some lactic acid bacteria with a uric acid lowering effect have the potential to ameliorate hyperuricemia in the host. However, the mechanism of action remains unclear. Here, we screened a strain of Lactobacillus acidophilus F02 that efficiently degrades uric acid precursors (inosine and guanosine) and further investigated its underlying mechanism on the alleviation of hyperuricemia in high fructose-adenine induced mice. Results showed that L. acidophilus F02 administration reduced serum uric acid levels in hyperuricemia mice by inhibiting xanthine oxidase and adenosine deaminase activity, while protecting hepatorenal injury and systemic inflammation by regulating the relative expression of NLRP3 inflammasome. Most importantly, it restored gut microbial homeostasis and the abundance of bacterial taxa (i.e., a balance in the relative abundance of Firmicutes and Actinobacteria and an increase in the abundance of Bacteroides, Ruminococcus and Lactobacillus). Furthermore, the ability of L. acidophilus F02 to lower uric acid and suppress NLRP3 inflammasome expression was confirmed in the HK-2 cell model, as well as the multiple immunomodulatory effects on RAW264.7 cells. Taken together, we speculate that fructose-adenine induced hyperuricemia triggers hepatorenal injury and gut microbiota dysbiosis, which is ameliorated by L. acidophilus F02. Collectively, the results suggest that lowering uric acid synthesis, increasing uric acid excretion and reducing NLRP3-related IL-1β levels are effective strategies to ameliorate hyperuricemia. Such information could inform future studies on the ecology of L. acidophilus and guide the use of this species for dietary applications in the food industry.

Ethnopharmacological relevance:RFAP is a compound extraction complex of four Traditional Chinese Medicine (TCM), including the dry bark of Paeonia lactiflora Pall. (Radix Paeoniae Alba), Gardenia jasminoides J. Ellis (Fructus Gardeniae), Albizia julibrissin Durazz. (Albizia julibrissin Durazz), and Paeonia × suffruticosa Andrews (Peony bark). Not only RFAP but also the individual ingredients have been commonly used for the treatment of depression in the clinic. However, the underlying mechanism of pharmacology is difficult to interpret since its holistic and multidrug nature. Aim of the study:This study aimed to elucidate the potential antidepressant mechanism of RFAP in the treatment of chronic unpredictable mild stress (CUMS) rats' model via the quantitative proteomics approach. Materials and methods:We established the CUMS rats' model and evaluated the efficacy of RFAP using multiple behavior assays, including the sugar preference test, open field test, and forced swimming test. Then label-free quantitative proteomics analyses were performed to evaluate the integrated changes of proteome profiling in control, CUMS, RFAP low dose, and RFAP high dose groups. Finally, we validated the critical changed proteins in the pathways of long-term depression and potentiation via RT-PCR and Western blotting assays. Results:We successfully established the CUMS rats' model. The behavior assays indicated that the rats demonstrated a tendency to behavioral despair after four weeks. Label-free quantitative proteomics showed that 107 proteins were significantly upregulated and 163 proteins were downregulated in the CUMS group compared to the control group. These differentially expressed proteins were involved in long-term potentiation, long-term depression, nervous system development, neuronal synaptic structural constituent of ribosome, ATP metabolic process, learning or memory, and cellular lipid metabolic process. RFAP treatment partially restored the differentially expressed protein profile. The protective effect of RFAP on behavioral assessment were consistent with the results of proteomics. Conclusions:The results indicated that RFAP exerted a synergistic effect on CUMS by regulating long-term inhibition and potentiation-related proteins.

壳寡糖主要以壳聚糖为原料,经生物酶解加工而成,具有分子量小、聚合度低、水溶性好等特点,分子量小于3 000 Da的壳寡糖很容易通过肠道代谢被肠上皮吸收,具有多种生物功能活性.该文就小分子量的壳寡糖在调节肠道菌群、提高免疫力、抗肿瘤、保护肝脏、降尿酸、降血糖等方面的功能活性进行综述,对小于3 000 Da分子量壳寡糖的功能活性进行比较分析,并充分探究不同分子量和聚合度的壳寡糖的作用机制,结合目前壳寡糖的应用现状,分析壳寡糖未来的发展趋势,为壳寡糖在食品等领域的广泛应用提供参考.

AIM:In vivo and in vitro toxicity tests of JointAlive® were studied in animal models to support the safe use of JointAlive® as a drug for knee osteoarthritis treatment.METHODS:The acute toxicity study in Sprague Dawley (SD) rats was conducted at a 20 g/kg bw/day dose of JointAlive®. For 13-week subchronic toxicity tests, SD rats were orally dosed daily with 0.5, 1.5 and 5 g/kg bw/day of JointAlive®. To assess the potential genotoxicity, Ames test, cellular chromosome aberration and mouse micronucleus test in vivo were carried out.RESULTS:Based on a lack of notable findings other than histopathology finding of co-incidental prostate inflammation at the high dose, the "No Observed Adverse Effect Level (NOAEL)" of JointAlive® was concluded as 5 g/kg bw/day in males and females. Results also indicated that JointAlive® has no risk of genotoxicity.CONCLUSIONS:General toxicity and genotoxicity studies empirically demonstrated that JointAlive® poses a low risk of potential health risks, providing safety supports for the application of JointAlive® as a potential drug candidate to treat knee osteoarthritis.

Abstract Under the influence of the COVID-19, people's awareness of physical health and immunity has increased significantly. Chitooligosaccharide is an oligomer of β-(1, 4)-linked D -glucosamine, furthermore, is one of the most widely studied immunomodulators. Chitooligosaccharide can be prepared from the chitin or chitosan polymers through enzymatically, chemically or physically processes. Chitooligosaccharide and its derivatives have been proven to have a wide range of biological activities including intestinal flora regulation, immunostimulant, anti-tumor, anti-obesity and anti-oxidation effects. This review summarizes the latest research of the preparation methods, biological activities in immunity and safety profiles of Chitooligosaccharide and its derivatives. We recapped the effect mechanisms of Chitooligosaccharide basing on overall immunity. Comparing the effects of Chitooligosaccharide with different molecular weights and degree of aggregation, a reference range for usage has been provided. This may provide a support for the application of Chitooligosaccharide in immune supplements and food. In addition, future research directions are also discussed.

目的 对比间隔12 h与间隔24 h观察动情周期的方法在评价动情周期紊乱程度中的优缺点,并探究间期取材对于雷公藤诱导的卵巢早衰大鼠模型评价的意义.方法 将60只动情周期规律的雌性SD大鼠随机分为空白1、2组,模型1、2组(n=15).空白组灌胃生理盐水,模型组灌胃雷公藤多苷片(TG)混悬液,连续15 d.空白1组、模型1组每日8:00行阴道脱落细胞学检测;空白2组、模型2组间隔12 h行阴道脱落细胞学检测,每周称量体重.干预后第15～17天于间期处死动情周期正常的大鼠,在第15天处死动情周期紊乱的大鼠.测定血清性激素,计算卵巢、子宫指数,观察卵巢病理改变.结果 与空白组比,模型组大鼠动情周期紊乱,间隔24 h观察,正常动情周期数目显著减少、动情期显著缩短、间期显著延长;间隔12 h观察,8:00较20:00观察的结果更能体现两组间的差异性,且综合间隔12 h两次观察的结果在动情周期紊乱水平(I与IV级)上差异具有显著性.于间期取材,TG组血清INHB、E2显著降低,FSH显著升高,全周期取材结果差异无显著性.因血清AMH水平不受动情周期变化而波动,全周期或间期取材的结果均显示TG组血清AMH显著降低.TG组卵巢与子宫指数显著下降,卵泡间结构混乱,总卵泡、原始卵泡、生长卵泡、黄体数显著减少,闭锁与囊状扩张卵泡数显著增多,并出现浆细胞浸润.结论 雷公藤可致大鼠卵巢功能明显衰退,卵泡发育障碍,卵巢储备下降.间隔12 h行阴道脱落细胞学检测,有助于捕捉亚周期阶段细胞学变化,反映真实的动情周期变化,较间隔24 h观察结果更能体现模型组动情周期的紊乱程度,但其可行性有待进一步探究.于间期处死,能反映大鼠基础性激素水平,利于评价雷公藤建立卵巢早衰大鼠模型的成功率.亚周期阶段下对比组间卵巢病理变化,客观地体现雷公藤对卵泡发育及黄体功能的不良影响.

We investigated the blood lipid regulation effects and mechanism of a functional Natto yogurt in a high-fat diet-induced hyperlipidemia mouse model. Natto yogurt was characteristically fermented by Bacillus natto and Lactobacillus plantarum with milk-soy dual protein as substrates. After 5 weeks of Natto yogurt consumption, the body weight, fat, and liver weight of mice were significantly improved, while serum levels of TG, TC, LDL, ALT, TBIL, and TBA were reduced. Natto yogurt significantly decreased the area of liver fat infiltration and the number of lipid droplets. In mechanism, we found that Natto yogurt can inhibit fatty acid synthesis and enhance fatty acid catabolism by regulating the expression of PPARα, PPARγ, CD36 and FAS in the liver. Moreover, Natto yogurt increased the ratio of Bacteroidetes to Firmicutes in the intestine. These results provide a possibility for Natto yogurt as a dual protein functional food to prevent and treat hyperlipidemia and obesity. PRACTICAL APPLICATIONS: Traditional-fermented yogurt promotes nutritional absorption and reduces blood pressure and fat, while Bacillus natto and its fermented food have been proved to play a significant role in improving cardiovascular and cerebrovascular diseases and obesity. Therefore, we developed a new dual protein functional yogurt (Natto yogurt) fermented by B. natto and Lactobacillus plantarum with milk and soy as substrates. We found that Natto yogurt could notably regulate blood lipid by inhibiting the synthesis of fatty acids, accelerating the catabolism of fatty acids, reducing liver damage, and increasing the abundance of beneficial intestinal microorganisms. This study suggested that Natto yogurt could improve hyperlipidemia and obesity as a safe, effective, and healthy functional food.

In this study, bioactive peptides (RBPs) from Ruditapes philippinarum were prepared by fermentation with Bacillus natto and the effect and mechanisms of RBPs on obesity and hyperlipidemia were explored in mice. We found that RBPs significantly reduced body weight, adipose tissue weight, accumulation of hepatic lipids, and serum levels of total cholesterol (CHO), triglyceride (TG), and low-density lipoprotein (LDL). Mechanistic studies showed that RBPs up-regulated the hepatic expression of genes related to lipolysis, such as hormone-sensitive lipase (HSL), phosphorylated AMP-activated protein kinase (p-AMPK), and peroxisome proliferator-activated receptors α (PPARα), and down-regulated the expression of peroxisome proliferator-activated receptors γ (PPARγ) which is related to lipid synthesis. In addition, RBPs could attenuate obesity and hyperlipidemia by regulating disordered gut microbiota composition, such as increasing the abundance of microflora related to the synthesis of short chain fatty acids (SCFAs) (Bacteroidetes, Prevotellaceas_UCG_001, norank_f_Muribaculaceae, and Odoribacter) and controlling those related to intestinal inflammation (reduced abundance of Deferribacteres and increased abundance of Alistipes and ASF356) to exert anti-obesity and lipid-lowering activities. Our findings laid the foundation for the development and utilization of RBPs as a functional food to ameliorate obesity and hyperlipidemia.

In the present study, we prepared pea peptides with high angiotensin-converting enzyme (ACE) inhibitory activity in vitro using an enzymatic hydrolysis of pea protein and compounded them with clam peptides to obtain a pea-clam double peptide. The effects of the two-peptide composite and pea peptides on hypertension and the damage-repair of corresponding organs were studied in spontaneously hypertensive rats (SHRs). We found that both pea peptides and the two-peptide composite significantly reduced the blood pressure upon a single or long-term intragastric administration, with the two-peptide composite being more effective. Mechanistically, we found that the two-peptide composite could regulate the renal renin-angiotensin system (RAS), rebalance gut microbial dysbiosis, decrease renal and myocardial fibrosis, and improve renal and cardiac function and vascular remodeling. Additionally, hippocampal lesions caused by hypertension were also eliminated after two-peptide composite administration. Our research provides a scientific basis for the use of this two-peptide composite as a safe antihypertension ingredient in functional foods.

Osteoporosis is one of the most common diseases in the world which resulted in heavy socioeconomic burden and a public health threat. Glucocorticoid-induced osteoporosis (GIO) is the most common secondary reason of osteoporosis. Therapeutic strategies using traditional Chinese medicine are under investigation for osteoporosis, with efforts to improve efficacy and clarify the mechanism. The combination of Eucommia, Cuscuta, and Drynaria is widely used in traditional Chinese decoction for osteoporosis treatment, but the experimental efficacy and mechanism are still unclear. Administration of E.C.D. extracts (Eucommia, Cuscuta, and Drynaria) in experimental GIO rats resulted in decreased urinal calcium, phosphorus loss, and decreased expression of RANKL, CTX in serum, increased serum calcium, phosphorus, and OPG level. E.C.D. extracts also improved bone density, structural integrity, and biomechanical function in experimental GIO rats. These finding were associated with E.C.D. extracts’ treatment efficacy to GIO in vivo . The balance between osteoclast and osteoblast activity is essential for bone remodeling and bone related disease. The E.C.D. extracts inhibited Raw 264.7 cell differentiation to osteoclast in vitro . On the other hand, it promoted OPG expression of bone marrow mesenchymal stromal cells (MSCs) which can suppress the osteoclast genesis. E.C.D. extracts also increased the Wnt1 and Runx2 expression which are related to osteoblast formation. It also regulated the paracrine effect of MSC to inhibit osteoclast differentiation. The analysis of HPLC and comprehensive pharmacology identified the constituents of E.C.D. extracts and the potential osteoporosis-related targets mediated by E.C.D. extracts. The KEGG enrichment analysis suggested that PI3K/Akt pathway may be involved in the regulation osteoclast genesis by E.C.D. extracts and the result of Western blot of vitro assays proved it. Collectively, these data demonstrate E.C.D. extracts can inhibit osteoclast differentiation to foster experimental osteoporosis both in vivo and in vitro and it may exert the function of inhibiting osteoclast differentiation through PI3K/Akt pathway.

Curcuma longa L. is one of the most recognized Curcuma species (Sharifi-Rad et al., 2020 [3]). Curcumin, the primary polyphenolic compound found in turmeric has been used for a variety of purposes for centuries. CuminUP60® is a curcumin complex composed of Curcuma longa L. rhizome extract and Poloxamer 407. The results of GLP compliant in vitro and in vivo safety studies conducted with CuminUP60® including a bacterial reverse mutation assay, an in vitro mammalian cell chromosome aberration study and an in vivo micronucleus study are reported here. In addition, a GLP compliant, a single dose toxicity study in Sprague-Dawley rats and a 4-week repeat dose study were also conducted. CuminUP60® was shown to not be mutagenic in a number of in vitro and one in vivo study, the results of which are reported here. A single oral dose of 5000 mg CuminUP60® was well tolerated by male and female Sprague-Dawley rats. The no observed adverse effect level (NOAEL) for CuminUP60® in male and female Sprague-Dawley rats in a 4-week repeat dose study was determined to be 1000 mg/kg bw/day.

Background: Alzheimer’s disease (AD) as an age-related, irreversible neurodegenerative disease, characterized by cognitive dysfunction, has become progressively serious with a global rise in life expectancy. As the failure of drug elaboration, considerable research effort has been devoted to developing therapeutic strategies for treating AD. TCM is gaining attention as a potential treatment for AD. Gastrodia elata Blume, Polygala tenuifolia Willd., Cistanche deserticola Ma, Rehmannia lutinosa (Gaertn.)DC., Acorus gramineus Aiton, and Curcuma longa L. (GPCRAC) are all well-known Chinese herbs with neuroprotective benefits and are widely used in traditional Chinese decoction for AD therapy. However, the efficacy and further mechanisms of GPCRAC extracts in AD experimental models are still unclear. The purpose of this study was to investigate the synergistic protective efficacy of GPCRAC extracts (composed of extracts from these six Chinese medicines), and the protein targets mediated by GPCRAC extracts in treating AD. Methods: Scopolamine-induced cognitive impairment mouse model was established to determine the neuroprotective effects of GPCRAC extracts in vivo, as shown by behavioral tests and cerebral cholinergic function assays. To identify the potential molecular mechanism of GPCRAC extracts against AD, label-free quantitative proteomics coupled with tandem mass spectrometry (LC-MS/MS) were performed. The integrated bioinformatics analysis was applied to screen the core differentially expressed proteins in vital canonical pathways. Critical altered proteins were validated by qPCR and Western blotting. Results: Administration of GPCRAC extracts significantly recovered scopolamine-induced cognitive impairment, as evidenced by the improved learning and memory ability, increased Ach content and ChAT activity, as well as decreased AchE activity in the hippocampus of mice. In total, 390 proteins with fold-change>1.2 or <0.83 and p < 0.05 were identified as significant differentially expressed proteins, of which 110 were significantly up-regulated and 25 were significantly down-regulated between control and model group. By mapping the significantly regulated proteins, we identified five hub proteins: PPP2CA, Gsk3β, PP3CC, PRKACA, and BCL-2 that were associated with dopaminergic synapse and apoptosis signaling pathway, respectively. Western blotting and QPCR demonstrate that the expression levels of these core proteins could be significantly improved by the administration of GPCRAC extracts. These pathways and some of the identified proteins are implicated in AD pathogenesis. Conclusion: Administration of GPCRAC extracts was effective on alleviating scopolamine-induced cognitive impairment, which might be through modulation of dopaminergic synapse and apoptosis signaling pathway. Consequently, our quantitative proteome data obtained from scopolamine-treated model mice successfully characterized AD-related biological alterations and proposed novel protein biomarkers for AD.

Phytotherapy offers obvious advantages in the intervention of Coronary Artery Disease (CAD), but it is difficult to clarify the working mechanisms of the medicinal materials it uses. DGS is a natural vasoprotective combination that was screened out in our previous research, yet its potential components and mechanisms are unknown. Therefore, in this study, HPLC-MS and network pharmacology were employed to identify the active components and key signaling pathways of DGS. Transgenic zebrafish and HUVECs cell assays were used to evaluate the effectiveness of DGS. A total of 37 potentially active compounds were identified that interacted with 112 potential targets of CAD. Furthermore, PI3K-Akt, MAPK, relaxin, VEGF, and other signal pathways were determined to be the most promising DGS-mediated pathways. NO kit, ELISA, and Western blot results showed that DGS significantly promoted NO and VEGFA secretion via the upregulation of VEGFR2 expression and the phosphorylation of Akt, Erk1/2, and eNOS to cause angiogenesis and vasodilation. The result of dynamics molecular docking indicated that Salvianolic acid C may be a key active component of DGS in the treatment of CAD. In conclusion, this study has shed light on the network molecular mechanism of DGS for the intervention of CAD using a network pharmacology-driven strategy for the first time to aid in the intervention of CAD.

BackgroundAlzheimer's Disease (AD) as an age-related, irreversible neurodegenerative disease, characterized by cognitive dysfunction, has become progressively serious as a result of a global increase in life expectancy. As more mechanism of AD were discovered, therapeutic strategies using traditional Chinese medicine are under investigation for AD treatment, with efforts to improve efficacy and clarify the mechanism. Gastrodia, elata Blume, Polygala tenuifolia Willd . , Cistanche deserticola Ma , Rehmannia lutinosa (Gaertn.)DC. ,Acorus gramineus Aiton , and Curcuma longa L. are well-known chinese herbs with neuroprotective effects and widely used as a combination in traditional Chinese decoction for AD treatment. The purpose of this study was to investigate the synergistic protective efficacy of the combination (composed of extracts from these six Chinese medicines, CuraUltra), and the protein targets on scopolamine-induced cognitive impairment, using the proteomics analysis.MethodsScopolamine-induced cognitive impairment mouse model was established. Behavioral tests, Nissl Staining, cerebral cholinergic system alterations and neuronal apoptosis characteristics were examined to evaluate the ameliorating effects of CuraUltra on cognitive impairment induced by scopolamine. To identify the potential molecular mechanism responsible for the effect on CuraUltra treatment, label-free quantitative proteomics by tandem mass spectrometry (LC-MS/MS) were performed. Critical altered proteins were validated by qPCR and Western blotting. Molecular docking was finally performed to evaluate the binding potential of chemical components with the target proteins.Results Administration of CuraUltra significantly recovered scopolamine-induced cognitive impairment, as evidenced by the improved learning and memory ability, reduced pathological damage of hippocampus, increased content of Ach, decreased activity of AchE, and ameliorated expression levels of the neuronal apoptosis-related protein in the hippocampus of mice. Using quantitative proteomics technology, we observed 252 differentially expressed proteins. Compared with the model group, after CuraUltra treatments, a remarkedly alteration in PPP3CC , PKA , P38MAPK , RASA4 , DNAJB1 , SNAPIN was also observed. Notably, several significant proteins are in the glutamatergic synapse signaling pathway. Moreover, we confirmed that the PPP3CC appears to decreased as the AD pathological development, and may be positively correlated with the phosphorylation level of PKA , thereby inhibiting the activation of P38MAPK phosphorylation.ConclusionsAdministration of CuraUltra was effective on alleviating scopolamine-induced cognitive impairment, which might be through modulation of glutamatergic synapse. Consequently, our quantitative proteome data obtained from scopolamine-treated model mice successfully characterized AD-related biological alterations and proposed novel protein biomarkers for AD.