[Huazhi Rougan Granules attenuates steatosis in cell model of nonalcoholic fatty liver disease by inducing autophagy].

To investigate the effect of Huazhi Rougan Granules(HZRG) on autophagy in a steatotic hepatocyte model of free fatty acid(FFA)-induced nonalcoholic fatty liver disease(NAFLD) and explore the possible mechanism. FFA solution prepared by mixing palmitic acid(PA) and oleic acid(OA) at the ratio of 1∶2 was used to induce hepatic steatosis in L02 cells after 24 h treatment, and an in vitro NAFLD cell model was established. After termination of incubation, cell counting kit-8(CCK-8) assay was performed to detect the cell viability; Oil red O staining was employed to detect the intracellular lipid accumulation; enzyme-linked immunosorbnent assay(ELISA) was performed to measure the level of triglyceride(TG); to monitor autophagy in L02 cells, transmission electron microscopy(TEM) was used to observe the autophagosomes; LysoBrite Red was used to detect the pH change in lysosome; transfection with mRFP-GFP-LC3 adenovirus was conducted to observe the autophagic flux; Western blot was performed to determine the expression of autophagy marker LC3B-Ⅰ/LC3B-Ⅱ, autophagy substrate p62 and silent information regulator 1(SIRT1)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK) signaling pathway. NAFLD cell model was successfully induced by FFA at 0.2 mmol·L~(-1) PA and 0.4 mmol·L~(-1) OA. HZRG reduced the TG level(P<0.05, P<0.01) and the lipid accumulation of FFA-induced L02 cells, while elevated the number of autophagosomes and autophagolysosomes to generate autophagic flux. It also affected the functions of lysosomes by regulating their pH. Additionally, HZRG up-regulated the expression of LC3B-Ⅱ/LC3B-Ⅰ, SIRT1, p-AMPK and phospho-protein kinase A(p-PKA)(P<0.05, P<0.01), while down-regulated the expression of p62(P<0.01). Furthermore, 3-methyladenine(3-MA) or chloroquine(CQ) treatment obviously inhibited the above effects of HZRG. HZRG prevented FFA-induced steatosis in L02 cells, and its mechanism might be related to promoting autophagy and regulating SIRT1/AMPK signaling pathway.

[Pharmacodynamic mechanism of Tibetan medicine Liurui Capsules on experimental autoimmune uveitis in rats based on network pharmacology and animal experiments].

By integrating network pharmacology and animal experiments, we studied the pharmacodynamic mechanism of the Tibetan medicine Liurui Capsules in the treatment of experimental autoimmune uveitis(EAU). The active ingredients and targets of Liurui Capsules were searched against the Encyclopedia of Traditional Chinese Medicine(ETCM), Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM), and relevant literatures. The EAU-related targets were obtained from Gene Expression Omnibus(GEO), GeneCards, Online Mendelian Inheritance in Man(OMIM), and Therapeutic Target Database(TTD). The common targets shared by Liurui Capsules and EAU were identified, and the protein-protein interaction(PPI) network was established via STRING. Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were conducted via g: Profiler. The rat model of EAU was induced by interphotoreceptor retinoid-binding protein(IRBP) and treated with Liurui Capsules. The inflammatory response of anterior segment and the pathological morphology of retina were observed. The mRNA and protein levels of delta-like ligand 4(DLL4), Notch1, interleukin-17(IL-17), and tumor necrosis factor-alpha(TNF-α) were determined by real-time quantitative PCR(q-PCR) and Western blot, respectively. The network pharmacology analysis predicted 51 common targets of Liurui Capsules and EAU, which were mainly involved in IL-17, TNF, and nuclear factor-kappa B(NF-κB) signaling pathways, as well as liposome receptors and other biological processes. Compared with the control group, the modeling of EAU caused inflammatory changes in the anterior segment and retina and up-regulated mRNA and protein levels of DLL4, Notch1, IL-17, and TNF-α in ocular tissue. Compared with the model group, Liurui Capsules reduced the inflammatory reaction of anterior segment and retina and down-regulated the mRNA and protein levels of DLL4, Notch1, IL-17, and TNF-α. Liurui Capsules can down-regulate the expression of the proteins involved in DLL4/Notch1/IL-17 signaling pathway in ocular tissue and alleviate the ocular inflammation, which may be one of the mechanisms of Liurui Capsules in the treatment of EAU.

[Study on medication regularity of traditional Chinese medicine in treatment of COVID-19 based on data mining].

The coronavirus disease 2019(COVID-19) is developing rapidly and posing great threat to public health. There is no specific medicine available for treating the disease. Luckily, traditional Chinese medicine has played a positive role in the fighting against COVID-19. In this paper, We collected and sorted the prescriptions of modern Chinese medicine for COVID-19 released by national government, different provinces, autonomous regions and municipalities, as well as online databases, such as CNKI, WanFang medical network, and VIP database. These prescriptions were combined with the inheritance of traditional Chinese medicine auxiliary V2.5, and the complex system entropy clustering method was used to determine the association rules and frequency of single drug and drug combination in the prescription. In the end, 96 effective prescriptions were included. Among them, the four properties were mainly concentrated in temperature, cold and level, the five tastes were mainly concentrated in bitter, hot and sweet, and the meridians were mainly concentrated in lung, stomach and spleen. The high-frequency drugs were Glycyrrhizae Radix et Rhizoma, Armeniacae Semen Amarum, Gypsum Fibrosum, etc., and the high-frequency combinations are Gypsum Fibrosum-Armeniacae Semen Amarum, Gypsum Fibrosum-Glycyrrhizae Radix et Rhizoma, Armeniacae Semen Amarum-Glycyrrhizae Radix et Rhizoma, the core combinations are Lepidii Semen-Armeniacae Semen Amarum-Gypsum Fibrosum, Pogostemonis Herba-Zingiberis Rhizoma Recens-Magnoliae Officinalis Cortex, Ophiopogonis Radix-Armeniacae Semen Amarum-Scutellariae Radix and so on. Form new prescriptions Lepidii Semen, Armeniacae Semen Amarum, Gypsum Fibrosum, Pogostemonis Herba, Zingiberis Rhizoma Recens, Magnoliae Officinalis Cortex. Ophiopogonis Radix, Armeniacae Semen Amarum, Scutellariae Radix, Schisandrae Sphenantherae Fructus, Panacis Quinquefolii Radix. From the medicinal properties to high-frequency drugs and new prescriptions, it could be seen that the overall treatment of COVID-19 by traditional Chinese medicine was to strengthen body resistance, eliminate pathogenic factors, and give attention to Qi and Yin.

[Medication rules of Professor Zhang Bing in treatment of skin itching based on data mining].

Skin itching is a subjective sensation that causes the desire to scratch. It is one of the most common clinical symptoms at department of dermatology, even the only complaint of dermatological patients, which seriously affects the quality life of patients. Therefore, based on the software of traditional Chinese medicine inheritance auxiliary platform, association rules and complex system entropy clustering were adopted to collect and analyze Zhang Bing's prescriptions for skin itching, and get the drug use frequency and the relationship between drugs. Based on that, we could conclude the experience for skin itching. A total of 147 prescriptions were collected, 20 drugs with a frequency of 34 or more and 20 high-frequency drug combinations were analyzed, and 14 core combinations and 7 new prescriptions were excavated. The high-frequency drugs included Kochiae Fructus, Dictamni Cortex, Mori Cortex. The high-frequency drug combinations included "Kochiae Fructus-Dictamni Cortex" "Angelicae Dahuricae Radix-Chuanxiong Rhizoma" "Paeoniae Radix Rubra-Paeoniae Radix Alba", and the core combinations included "Schizonepetae Herba-Saposhnikoviae Radix-Cinnamomi Ramulus" "Arctii Fructus-Cicadae Periostracum-Houttuyniae Herba" "Ghrysanthemi Indici Flos-Kochiae Fructus-Dictamni Cortex", and new formulations include "Schizonepetae Herba, Saposhnikoviae Radix, Cinnamomi Ramulus, Clematidis Radix et Rhizoma, Tribuli Fructus, Dictamni Cortex", "Phellodendri Chinensis Coritex, Lonicerae Japonicae Flos, Atractylodis Rhizoma, Ghrysanthemi Indici Flos, Kochiae Fructus, Dictamni Cortex" "Arctii Fructus, Cicadae Periostracum, Houttuyniae Herba, Trichosanthis Fructus". The result of this research shows that Professor Zhang Bing's experience in the treatment of skin itching is mainly to dispelling wind and arresting itching, clearing heat and drying dampness.

[Discussion on safety of Xanthii Fructus and consideration on its rational use].

Xanthii Fructus is a traditional Chinese medicine for the treatment of sinusitis and headache,rich in medicinal materials and is widely used for more than 1 800 years. Modern pharmacological studies have showed that Xanthii Fructus has anti-inflammatory,analgesic,anti-tumor,anti-bacterial,hypoglycemic,anti-allergic,immunomodulatory and other pharmacological effects,which can be commonly used in the treatment of diseases relating to immune abnormalities,such as rheumatoid arthritis,acute and chronic rhinitis,allergic rhinitis,and skin diseases,with a high medicinal value. Toxicological studies have shown that Xanthii Fructus poisoning can cause substantial damage to organs,such as the liver,kidney,and gastrointestinal tract,especially to liver. Because of the coexisting of its efficacy and toxicity,Xanthii Fructus often leads to a series of safety problems in the clinical application process. This study attempts to summarize its characteristics of adverse reactions,analyze the root cause of the toxicity of Xanthii Fructus from such aspects as processing,dose,course of treatment and eating by mistake,discuss the substance of its efficacy/toxicity from chemical compositions,and put forward exploratory thinking about how to promote its clinical rational application from the aspects such as strict processing,reasonable compatibility,medication information,contraindication,strict control of the dose,and course of treatment,so as to promote the safe and reasonable application of Xanthii Fructus.

Relationship between High-Performance Liquid Chromatography Fingerprints and Uric Acid-Lowering Activities of Cichorium intybus L.

This study aimed to explore the spectrum-effect relationships between high-performance liquid chromatography fingerprints and the uric acid-lowering activities of chicory. Chemical fingerprints of chicory samples from ten different sources were determined by high-performance liquid chromatography, and then investigated by similarity analysis and hierarchical clustering analysis. Pharmacodynamics experiments were conducted in animals to obtain the uric acid-lowering activity information of each chicory sample. The spectrum-effect relationships between chemical fingerprints and the uric acid-lowering activities of chicory were established by canonical correlation analysis. The structures of potential effective peaks were identified by liquid chromatography with tandem mass spectrometry. The results showed that a close correlation existed between the spectrum and effect of chicory. Aesculin, chlorogenic acid, chicoric acid, isochlorogenic acid A/B/C and 13,14-seco-stigma5(6),14(15)-diene-3α-ol might be the main effective constituents. This work provides a general model of the combination of high-performance liquid chromatography and uric acid-lowering activities to study the spectrum-effect relationships of chicory, which can be used to discover the principle components responsible for the bioactivity.

[Molecular docking analysis of xanthine oxidase inhibition by constituents of cichory].

Human xanthine oxidase is considered to be a target for therapy of hyperuricemia. Cichorium intybus is a Chinese plant medicine which widely used in Xinjiang against various diseases. In order to screen the inhibitors of xanthine oxidase from C. intybus and to explore main pharmacological actions of cichory a compound collection of C. intybus was built via consulting related references about chemical research on cichory. The three-dimensional crystal structure of xanthine oxidase (PDB code: 1N5X) from Protein Data Bank was downloaded.. Autodock 4.2 was employed to screen the inhibitors of xanthine oxidase from cichory 70 compounds were found to possess quite low binding free energy comparing with TEI (febuxostat). C. intybus contains constituents possessing potential inhibitive activity against xanthine oxidase. It can explain the main pharmacological actions of cichory which can significantly lower the level of serum uric acid.

The spectrum-effect relationship-a rational approach to screening effective compounds, reflecting the internal quality of Chinese herbal medicine.

Since the chromatographic fingerprint was introduced, it has been accepted by many countries to assess the quality and authenticity of Chinese herbal medicine (CHM). However, solely using the chromatographic fingerprint to assay numerous chemicals is not suitable for the assessment of the whole internal quality and pharmacodynamics of CHM. Consequently, it is necessary to develop a rational approach to connecting the chromatographic fingerprint with effective components to assess the internal quality of CHM. For this purpose, a spectrum-effect relationship theory was proposed and accepted as a new method for the assessment of CHM because of its potential use to screen effective components from CHM. In this paper, we systematically reviewed the application of the spectrum-effect relationship theory in the research of CHM, including research mentality, different chromatographic analysis techniques, data processing technologies, and structure determination.

[Mechanism of endogenous carbon monoxide effect on hydrogen sulfide in guinea pigs with established allergic rhinitis].

OBJECTIVE: To study the effect of carbon monoxide (CO) on hydrogen sulfide (H2S) in guinea pigs with allergic rhinitis (AR) through intervention treatment. METHODS: AR model in guinea pigs was established by using ovalbumin. The animals were divided into three groups. Group one was sensitized continuously by ovalbumin, group two was treated with Hemin as induction group, and group three was treated with zinc protoporphyrin (ZnPP) as suppression group. The guinea pigs treated with saline were used as control. The behavior science scores, eotaxin concentration of nasal lavage, IgE in blood serum were recorded, and the plasma concentrations of CO and H2S were determined, then the expression of hemeoxygenase (HO)-1, cystathionine-beta-synthase (CBS) and cystathionine-gamma-lyase (CSE) were measured in nasal mucosa by fluorescent quantitative RT-PCR. RESULTS: The behavior science scores, concentration of eotaxin in nasal lavage, IgE in blood serum and concentration of CO in plasma of sensitized group were higher than those of control (P<0.01), and the expression of HO-1 in nasal mucosa was also higher than control [(7.61+/-2.80)x10(-3) vs (2.32+/-1.14)x10(-3), P<0.05]. All these items were higher when treated with Hemin and lower when treated with ZnPP (P<0.05). The concentration of H2S in plasma was lower than control with significant differences [(14.80+/-1.60) micromol/L vs (18.90+/-1.00) micromol/L, P<0.01], the expression of CSE was also lower than control (P<0.05), and both of them were lower with Hemin induced and higher with ZnPP (P<0.05). The expression of CBS was very low and had no significant differences between groups (P>0.05), so it indicated that the CSE was the key enzyme for endogenous H2S product in nasal mucosa. Moreover the concentration of H2S was negatively correlated with CO (r=-0.702, P<0.001). CONCLUSIONS: Endogenous CO and H2S play a significant role in the pathogenesis of AR, and HO-1 and CSE are the main speed-relate enzymes respectively. The H2S is also influenced by CO.

[Influence of carbon monoxide on the expression of inducible nitric oxide synthase mRNA in guinea pigs with allergic rhinitis.].

OBJECTIVE: To study the impact of carbon monoxide (CO) on expression levels of inducible nitric oxide synthase (iNOS) mRNA in guinea pigs with allergic rhinitis (AR). METHODS: Twenty four guinea pigs were divided randomly into four study groups with 6 guinea pigs in each. The guinea pigs in the first group were treated with saline only (Group 1, the healthy controls). The remaing guinea pigs were sensitized by ovalbumin and thus establishing the AR models. After sensitization, the animals in the second group remained untreated (Group 2, AR control group). The third group was treated with Hemin as the induction group, and the fourth group was treated with Zinc protoporphyrin (ZnPP) as the suppression group. The plasma concentration of carboxyhemoglobin (COHb) was measured, which represents the concentration of CO. The expression levels of Heme oxygenase-1 (HO-1) and NOS mRNAs in nasal mucosa were determined by fluorescent quantitative RT-PCR. RESULTS: AR models were established successfully in all study guinea pigs. The concentrations of COHb (x(-) +/- s) in plasma of the second group (2.27% +/- 1.13%) were significantly (q = 4.10, P < 0.01) higher than those of healthy controls (1.08% +/- 0.24%). The plasma concentration of COHb in the third group (3.17% +/- 0.68%) were also significantly higher (q = 3.12, P < 0.05) than those in the second group. The expression levels of HO-1 and iNOS in nasal mucosa of the second group [(7.80 +/- 1.60) x 10(-3) and (5.81 +/- 0.05) x 10(-3), respectively] were also significantly (q equals 5.52 and 7.21, respectively, P < 0.01) higher than those of controls [(1.96 +/- 0.71) x 10(-3) and (0.97 +/- 0.05) x 10(-3), respectively]. The expression levels of HO-1 and iNOS in the nasal mucosa of the third group [(11.89 +/- 4.78) x 10(-3) and (7.42 +/- 0.70) x 10(-3), respectively] were significantly (q equals 3.86 and 2.22, P < 0.05) higher than those of the second group. The expression levels of HO-1 and iNOS in nasal mucosa of the fourth group [(3.82 +/- 0.98) x 10(-3) and (2.34 +/- 0.04) x 10(-3), respectively] were significantly (q equals 3.76 and 5.18, P < 0.05) lower than those in the second group. CONCLUSIONS: Endogenous carbon monoxide influenced the expression levels of iNOS in nasal mocusa in guinea pigs with AR.