Decoding active compounds and molecular targets of herbal medicine by high-throughput metabolomics technology: A systematic review.

Clinical experiences of herbal medicine (HM) have been used to treat a variety of human intractable diseases. As the treatment of diseases using HM is characterized by multi-components and multi-targets, it is difficult to determine the bio-active components, explore the molecular targets and reveal the mechanisms of action. Metabolomics is frequently used to characterize the effect of external disturbances on organisms because of its unique advantages on detecting changes in endogenous small-molecule metabolites. Its systematicity and integrity are consistent with the effective characteristics of HM. After HM intervention, metabolomics can accurately capture and describe the behavior of endogenous metabolites under the disturbance of functional compounds, which will be used to decode the bioactive ingredients of HM and expound the molecular targets. Metabolomics can provide an approach for explaining HM, addressing unclear clinical efficacy and undefined mechanisms of action. In this review, the metabolomics strategy and its applications in HM are systematically introduced, which offers valuable insights for metabolomics methods to characterizing the pharmacological effects and molecular targets of HM.

Baicalin, silver titanate, Bletilla striata polysaccharide and carboxymethyl chitosan in a porous sponge dressing for burn wound healing.

OBJECTIVE: This study tests the efficacy of Bletilla striata polysaccharide (BSP), carboxymethyl chitosan (CMC), baicalin (BA) and silver titanate (ST) in a wound dressings to fight infection, promote healing and provide superior biocompatibility. METHODS: The antibacterial activity of BA and ST was evaluated in vitro using the inhibition zone method. BA/ST/BSP/CMC porous sponge dressings were prepared and characterized. The biocompatibility of BA/ST/BSP/CMC was assessed using the cell counting kit-8 assay. The therapeutic effect of BA/ST/BSP/CMC was further investigated using the dorsal skin burn model in Sprague-Dawley rats. RESULTS: The wound dressing had good antibacterial activity against Escherichia coli and Staphylococcus aureus through BA and ST, while the combination of BSP and CMC played an important role in promoting wound healing. The BA/ST/BSP/CMC porous sponge dressings were prepared using a freeze-drying method with the concentrations of BA and ST at 20 and 0.83 mg/mL, respectively, and the optimal ratio of 5% BSP to 4% CMC was 1:3. The average porosity, water absorption and air permeability of BA/ST/BSP/CMC porous sponge dressings were measured to be 90.43%, 746.1% and 66.60%, respectively. After treatment for 3 and 7 days, the healing rates of the BA/ST/BSP/CMC group and BA/BSP/CMC group were significantly higher than those of the normal saline (NS) group and silver sulfadiazine (SSD) group (P < 0.05). Interleukin-1ß expression in the BA/ST/BSP/CMC group at 1 and 3 days was significantly lower than that in the other three groups (P < 0.05). After being treated for 3 days, vascular endothelial growth factor expression in the BA/BSP/CMC group and BA/ST/BSP/CMC group was significantly higher than that in the NS group and SSD group (P < 0.05). Inspection of histological sections showed that the BA/ST/BSP/CMC group and BA/BSP/CMC group began to develop scabbing and peeling of damaged skin after 3 days of treatment, indicating accelerated healing relative to the NS group and SSD group. CONCLUSION: The optimized concentration of BA/ST/BSP/CMC dressing was as follows: 6 mg BSP, 14.4 mg CMC, 0.5 mg ST and 12 mg BA. The BA/ST/BSP/CMC dressing, containing antibacterial constituents, was non-cytotoxic and effective in accelerating the healing of burn wounds, making it a promising candidate for wound healing. Please cite this article as: Gong YR, Zhang C, Xiang X, Wang ZB, Wang YQ, Su YH, Zhang HQ. Baicalin, silver titanate, Bletilla striata polysaccharide and carboxymethyl chitosan in a porous sponge dressing for burn wound healing. J Integr Med. 2023; 21(5): 487-495.

Mass spectrometry-based metabolomics for discovering active ingredients and exploring action mechanism of herbal medicine.

Natural products derived from herbal medicine are a fruitful source of lead compounds because of their structural diversity and potent bioactivities. However, despite the success of active compounds derived from herbal medicine in drug discovery, some approaches cannot effectively elucidate the overall effect and action mechanism due to their multi-component complexity. Fortunately, mass spectrometry-based metabolomics has been recognized as an effective strategy for revealing the effect and discovering active components, detailed molecular mechanisms, and multiple targets of natural products. Rapid identification of lead compounds and isolation of active components from natural products would facilitate new drug development. In this context, mass spectrometry-based metabolomics has established an integrated pharmacology framework for the discovery of bioactivity-correlated constituents, target identification, and the action mechanism of herbal medicine and natural products. High-throughput functional metabolomics techniques could be used to identify natural product structure, biological activity, efficacy mechanisms, and their mode of action on biological processes, assisting bioactive lead discovery, quality control, and accelerating discovery of novel drugs. These techniques are increasingly being developed in the era of big data and use scientific language to clarify the detailed action mechanism of herbal medicine. In this paper, the analytical characteristics and application fields of several commonly used mass spectrometers are introduced, and the application of mass spectrometry in the metabolomics of traditional Chinese medicines in recent years and its active components as well as mechanism of action are also discussed.

Dietary supplementation with metformin improves testis function and semen quality and increases antioxidants and autophagy capacity in goats.

ATP is essential for mammalian sperm to maintain fertilizing capacity. Metformin (Met) can activate 5'-AMP-activated protein kinase (AMPK) to maintain energy homeostasis. Thus, the aim of the present study was to investigate whether Met can improve testis function, semen quality, antioxidant and autophagy capacity through AMPK mediation of energy metabolism in goats. Twelve adult goats were randomly divided into three dietary treatments. All goats were fed a basal diet for 3 weeks and then assigned to a Met supplementation diet containing 0, 150, or 300 mg/kg for 8 weeks. The results showed that sperm viability, sperm membranal functional integrity, and acrosome integrity increased (P < 0.05) relative to the other treatments in the 300 mg/kg Met group. Growth hormone (GH) and insulin-like growth factor (IGF-1) in the 300 mg/kg Met group significantly decreased (P < 0.05) relative to the control group. Estrogen levels (E2) in the 300 mg/kg Met group remarkably improved (P < 0.05) compared with the control group. The activities of the antioxidant enzymes catalase (CAT), glutathione peroxidase (GSH-px), and superoxide dismutase (SOD) significantly increased (P < 0.05) in the 300 mg/kg Met group relative to the control group. A significant increase in AMPK and p-AMPK protein expression in the 300 mg/kg Met group was observed relative to the control group (P < 0.05). Belicin-1 and LC3II/I protein expression was significantly increased by adding Met to the diet (P < 0.05) and reached a maximum in the 300 mg/kg Met group. In addition, differentially expressed genes (DEGs) of goat testis were confirmed by RNA-seq. GO enrichment analysis revealed that DEGs were enriched in testicular metabolism and sperm development-related functional pathways. Overall, the results indicate that Met may play an important role in the regulation of testis function, semen quality, antioxidant, and autophagy capacity. These findings will help elucidate the role of Met in goat testis development.

l-Argine regulates the proliferation, apoptosis and endocrine activity by alleviating oxidative stress in sheep endometrial epithelial cells.

l-arginine (L-Arg) is a semiessential amino acid that plays crucial roles in the reproductive performance of animals. This research aimed to investigate the effect of supplementing L-Arg on endometrial epithelial cells (EECs) of Hu sheep. In vivo, female Hu sheep were randomly divided into three groups: control group (n = 5), nutrient-restricted group (n = 5), and L-Arg supplemented nutrient-restricted group (n = 5). Then, the effect of L-Arg on ovine endometrial growth and antioxidant capacity was assessed. We found that L-Arg supplementation promoted the growth of endometrial ductal gland invaginations (DGI), and alleviated oxidative stress in nutrient-restricted sheep. In order to investigate its mechanism, a H2O2-induced EECs oxidative stress model was established, and roles of L-Arg in EECs oxidation resistance, proliferation, apoptosis and endocrine activity were studied in vitro. Our results showed that L-Arg markedly decreased the release of reactive oxygen species (ROS) and malonaldehyde (MDA), and enhanced the expression and activity of certain antioxidant enzymes in EECs challenged by the H2O2 (p < 0.05). Supplementation of L-Arg significantly reduced the effect of 200 µM H2O2 on the viability of EECs (p < 0.05). In addition, EECs treated with L-Arg significantly alleviated the G0/G1-phase cell cycle arrest, apoptosis, and the inhibition of endometrial growth factors expression caused by H2O2 (p < 0.05). Overall, the results demonstrate that L-Arg performs crucial roles in maintaining the proliferation of ovine EECs, endocrine activity and inhibiting apoptosis through reducing oxidative stress. This study offers a theoretical basis for using L-Arg to improve sheep the uterine function.

[Chronic heart failure due to Qi deficiency and blood stasis and intervention mechanism of Compound Renshen Buqi Granules:a proteomics-based study].

Compound Renshen Buqi Granules have been widely used to treat chronic heart failure(CHF) due to Qi deficiency and blood stasis, but the mechanism of action remains unclear. This paper explored the pathogenesis of CHF due to Qi deficiency and blood stasis and the intervention mechanism of Compound Renshen Buqi Granules based on quantitative proteomics for uncovering the biological basis. SD rats were divided into the normal control(N) group, normal+Compound Renshen Buqi Granules(ND) group, model(M) group, model+Compound Renshen Buqi Granules(D) group, and positive control(Y) group. The rat model of CHF due to Qi deficiency and blood stasis was established by ligation of the left anterior descending(LAD) coronary artery and chronic sleep deprivation. The rats in the ND group and D group were provided with Compound Renshen Buqi Granules, while those in the Y group received valsartan. Six weeks later, the serum was sampled and the data-dependent acquisition(DDA) was employed for the non-targeted quantitative proteomics analysis of the differences in protein expression among groups, followed by the targeted analysis of differentially expressed proteins(DEPs) generated by data-independent acquisition(DIA). Compared with the N group, the rats in the M group pre-sented with decreased body weight, grip strength, and pulse amplitude and increased RGB value on the tongue surface. The pathomorphological examination revealed inflammatory cell infiltration, cell degeneration and necrosis, tissue fibrosis, etc. After the intervention with Compound Renshen Buqi Granules, multiple indicators were reversed. As demonstrated by proteomics results, there were 144 and 111 DEPs found in the M group and ND group in comparison with the N group. Compared with the M group, 107 and 194 DEPs were found in the D group and the Y group, respectively. Compared with the ND group, 119 DEPs were detected in the D group. As illustrated by DIA-based verification, the quantitative results of six proteins in each group were consistent with those by DDA. The syndrome indicators and pathomorphological examination results demonstrated that the protein expression profile of rats with CHF due to Qi deficiency and blood stasis changed obviously. However, Compound Renshen Buqi Granules were able to reverse the differential expression of immune proteins to regulate CHF of Qi deficiency and blood stasis syndrome, which has provided clues for figuring out the pathogenesis of CHF due to Qi deficiency and blood stasis and the intervention mechanism of Compound Renshen Buqi Granules.

γ-Linolenic Acid Prevents Lipid Metabolism Disorder in Palmitic Acid-Treated Alpha Mouse Liver-12 Cells by Balancing Autophagy and Apoptosis via the LKB1-AMPK-mTOR Pathway.

Excessive fat deposition is the main character in nonalcoholic fatty liver disease (NAFLD), while γ-linolenic acid (GLA) is a polyunsaturated fatty acid that can reduce lipid deposition. This study investigated the effect and regulatory mechanism of GLA (100 µM) on lipid metabolism in alpha mouse liver 12 (AML-12) cells treated by 400 µM palmitic acid (PA). GLA reduced lipid content and increased fatty acid ß oxidation, as indicated by decreasing triglyceride and cholesterol contents and increasing mRNA and protein expressions of CPT1α and PPARα. GLA relieved oxidative stress caused by PA, upregulated mRNA levels of superoxide dismutase and glutathione peroxidase, and decreased reactive oxygen species content. GLA reduced apoptosis, as indicated by decreases in the BAX/BCL2 expression level and apoptosis percentage. GLA activated autophagy, autophagosome-lysosome fusion, and LKB1-AMPK-mTOR signaling and upregulated mRNA and protein expressions of Beclin-1, autophagy-related 5, and liver kinase B1 (LKB1). These effects of GLA on lipid metabolism disorders of PA-treated hepatocytes were reversed by autophagy inhibitor 3MA and AMPK inhibitor compound C, confirming our conclusions. Overall, GLA can protect AML-12 cells from lipid metabolism disorder caused by PA via balancing autophagy and apoptosis mediated by the LKB1-AMPK-mTOR pathway. Consequently, GLA, as a dietary supplement, can help to prevent and treat NAFLD by regulating lipid metabolism and autophagy.

Spirulina supplementation improves lipid metabolism and autophagic activities in the liver and muscle of Hu lambs fed a high-energy diet.

The current study aimed to examine the effects of dietary spirulina supplementation in high-energy (HE) diets on fatty acid metabolism in sheep, and preliminarily explored the potential mechanisms underlying the associated autophagy-mediated regulation of lipid metabolism. In a 2 × 3 factorial design, including six treatment combinations of two metabolisable energy diets (10 and 11 MJ/kg DM), three spirulina supplementation levels (0, 1%, and 3%) were used. Serum alanineaminotransferase (ALT) (p = 0.003) and aspartatetransaminase (AST) (p = 0.002) activities increased, whereas total PUFA content (p < 0.001) decreased in the liver of lambs fed a HE diet. With the addition of spirulina, serum ALT (p = 0.037) and AST (p = 0.014) activities decreased, whereas EPA (p = 0.004), GLA (p = 0.019), n-6 PUFA (p = 0.005), and total PUFA contents (p = 0.019) increased. Moreover, the crude protein content in the Longissimus thoracis et lumborum (LTL) increased (p = 0.013), the expression of PPARα and PPARγ was up-regulated, while ELOVL2 was down-regulated in liver and LTL (p < 0.05). Spirulina supplementation increased mRNA expression levels of autophagy-associated genes, including that of Beclin-1, AMPK, and ULK1 (p < 0.05). In conclusion, spirulina supplementation in a HE diet exerted a protective effect on the liver, increased PUFA content, and modulated expression levels of autophagy-related genes in growing lambs.

Roles of vitamin D and its receptor in the proliferation and apoptosis of luteinised granulosa cells in the goat.

The objective of this study was to investigate the dose-dependent effect of 1α,25-(OH)2VD3 (Vit D3) on invitro proliferation of goat luteinised granulosa cells (LGCs) and to determine the underlying mechanisms of its action by overexpressing and silencing vitamin D receptor (VDR) in LGCs. Results showed that VDR was prominently localised in GCs and theca cells (TCs) and its expression increased with follicle diameter, but was lower in atretic follicles than in healthy follicles. The proliferation rate of LGCs was significantly higher in the Vit D3-treated groups than in the control group, with the highest proliferation rate observed in the 10nM group; this was accompanied by changes in the expression of cell cycle-related genes. These data indicate that Vit D3 affects LGC proliferation in a dose-dependent manner. Contrary to the VDR knockdown effects, its overexpression upregulated and downregulated cell cycle- and apoptosis-related genes respectively; moreover, supplementation with 10nM of Vit D3 significantly enhanced these effects. These results suggest that changes in VDR expression patterns in LGCs may be associated with follicular development by regulation of cell proliferation and apoptosis. These findings will enhance the understanding of the roles of Vit D3 and VDR in goat ovarian follicular development.

Safety of Shujinjianyao pill in clinical real world: A prospective, observational, multicenter, large-sample study Protocol.

BACKGROUND: Low back pain is a common health problem worldwide, which also is a leading cause of long-term disability and has an important effect on the global economy and society. Usually, conservative therapies are used to treat low back pain. As a kind of Chinese patent medicine, Shujinjianyao pill (SJJYP) has a great curative effect on low back pain. However, its safety has not been studied yet. Therefore, we carried out this clinical trial to observe the safety of SJJYP in the real world. METHODS: First, participants need to meet the medication standards according to inclusion and exclusion criteria. Then, participants are conducted safety examination before taking SJJYP. After qualified screening, participants can be enrolled into the group. Second, all enrolled participants will receive SJJJYP for a period of 4 weeks. During the observation period, participants need to return to the hospital for a subsequent visit after 2 weeks of medication, and come to the hospital for safety check after 4 weeks of medication. Third, telephone follow-up is used to investigate any participants' physical discomfort after 6 to 8 weeks (2-4 weeks after medication withdrawal). After all these steps are completed, clinical observation is finished. If any adverse events occur during this process, we will record them in time. When serious adverse events occur, we will use nested case-control study to explore the causes and mechanisms. DISCUSSION: This study will obtain the safety results of SJJYP in clinical real world, which will offer a scientific basis for clinicians in the treatment of low back pain, and also provide a methodological basis for the safety study of other medicines. TRIAL REGISTRATION: ClinicalTrial.gov registration number is NCT03598153. This study was approved by the ethics committee of Wangjing hospital, China Academy of Chinese Medical Sciences (WJEC-KT-2018-012-P002).

[Dynamic scalp acupuncture combined with PNF therapy for upper limb motor impairment in ischemic stroke spastic hemiplegia].

OBJECTIVE: On the basic therapy, to assess the clinical effects of dynamic scalp acupuncture, scalp acupuncture combined with proprioceptive neuromuscular facilitation (PNF) therapy and simple PNF therapy for upper limb motor impairment in ischemic post-stroke spastic hemiplegia. METHODS: A total of 90 cases were randomly assigned into a PNF group, a dynamic scalp acupuncture group and a scalp acupuncture group, 30 cases in each group. Basic therapy and PNF therapy were applied in the three groups. PNF therapy was used during scalp acupuncture in the dynamic scalp acupuncture group. PNF therapy was applied after scalp acupuncture in the scalp acupuncture group. The points were the upper 1/5 and middle 2/5 of Dingnieqianxiexian (MS 6) and Dingniehouxiexian (MS 7) at the lesion side, the hemiparalysis contralateral side. The treatment was given for 6 months, once a day and 1 month as a course. The modified Ashworth scale (MAS), the Fugl-Meyer motor assessment (FMA) and Barthel index (BI) were observed before treatment and 2 weeks, 1 month, 3 months, and 6 months after treatment. RESULTS: The MAS 1 month, 3 months and 6 months after treatment were improved compared with those before treatment in the three groups ( all P<0.05), and the MAS results in the dynamic scalp acupuncture group were better than those in the PNF and scalp acupuncture group (all P<0.05). The FMA and BI scores 1 month, 3 months and 6 months after treatment were higher than those before treatment (all P<0.05). The FMA Scores in the 3 time points and after treatment in the dynamic scalp acupuncture group were higher than those in the other two groups (all P<0.05). CONCLUSION: PNF therapy during scalp acupuncture can relieve the spasmodic condition of patients with upper limb motor impairment in ischemic post-stroke spasmodic hemiplegia, and improve the limb function and life activity, which is better than PNF therapy after scalp acupuncture and simple PNF therapy.

No-tillage effects on N and P exports across a rice-planted watershed.

No tillage (NT) can be used as a management tool to alleviate the negative effects of agricultural practices on the environment by reducing the runoff volume and nutrient exports. The main objective of this research was to quantify the effect of NT on nitrogen (N) and phosphorus (P) exports across a rice-planted watershed using the soil and water assessment tool (SWAT) model. Results show that total N and P runoff exports from rice fields across the watershed ranged from 7.2 to 22.8 kg N/ha and 0.56 to 6.80 kg P/ha, respectively, over five rice-growing seasons under conventional tillage (CT) practice. The adoption of NT reduced the runoff volume, and the total N and total P exports by 25.9, 8.5, and 7.8 %, respectively, compared with the total exports under CT practice in the same study area. Rice yields were reduced by 0.7-1.9 % within the first 4 years after the adoption of NT, but began to rise in the fifth year. These results suggest that a long-term period of NT practice is necessary to reduce N and P exports without comprising the rice yield on rice-planted watersheds. In addition, the benefits of implementing NT practice alone were limited, and other practices, such as water and nutrient management, should be combined with NT practice.

Nutrient removal efficiency in a rice-straw denitrifying bioreactor.

Rice straw was used as a carbon source in a denitrifying bioreactor, for the removal of nutrients from agricultural drainage. Nutrient removal efficiency was evaluated by: (a) nutrient loading rates (low, medium, and high); (b) hydraulic retention time, and (c) comparison with another carbon source (woodchip). The results show that concentrations of nitrate nitrogen (NO3(-)-N), ammonia nitrogen (NH4(+)-N), total nitrogen (TN), and orthophosphate phosphorus (PO4(3-)-P) in the rice-straw bioreactor effluents were reduced by 53%, 25%, 40%, and 35%, respectively, compared with influents at the medium nutrient loading rate (NO3(-)-N: 10-15 mg N L(-1), NH4(+)-N: 10-15 mg N L(-1), PO4(3-)-P: 1.0-1.5 mg P L(-1)) and long hydraulic retention time (HRT, 24h), with a corresponding denitrification rate (DR) of 0.40 mg N L(-1)h(-1). Moreover, the rice-straw bioreactor showed significantly higher (p<0.05) nutrient removal efficiency than the woodchip bioreactor at the medium nutrient loading rate and 24h HRT.

Effect of salvianic acid A on lipid peroxidation and membrane permeability in mitochondria.

Salvia miltorrhiza Bunge is a traditional Chinese medicine and has long been used for treating liver and heart diseases in China. Salvianic acid A is the main active component of Salvia miltorrhiza Bunge. In the present study, the ability of salvianic acid A in scavenging free radicals, inhibiting lipid peroxidation and mitochondrial membrane permeability transition, as well as respiration and protein thiol oxidation in rat liver mitochondria, was evaluated. The results show that salvianic acid A scavenges superoxide anions in a dose-dependent manner (IC50 52 microg/ml). Salvianic acid A could scavenge lipid free radicals and inhibit lipid peroxidation as effectively as Vitamin E. Salvianic acid A also inhibited the mitochondrial membrane permeability transition assessed as the extent of mitochondrial swelling. Salvianic acid A inhibited the oxidation of mitochondrial protein thiols involved in the mitochondrial membrane permeability transitions. We conclude that salvianic acid A is able to reduce lipid peroxidation in the mitochondrial membrane by scavenging free radicals, and inhibit mitochondrial membrane permeability transition by reducing protein thiol oxidation. These data indicated the pharmacological potential of salvianic acid A against pathological processes related to oxidative stress.