Safety evaluation of Tibetan medicine Qishiwei Zhenzhu Pills based on serum pharmacochemistry and network pharmacology / 中国中药杂志

To explore the mechanism of the active ingredients of Qishiwei Zhenzhu Pills in inhibiting the hepatorenal toxicity of the zogta component based on serum pharmacochemistry and network pharmacology, thereby providing references for the clinical safety application of Qishiwei Zhenzhu Pills. The small molecular compounds in the serum containing Qishiwei Zhenzhu Pills of mice were identified by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Then, by comprehensively using Traditional Chinese Medicines Systems Pharmacology(TCMSP), High-throughput Experiment-and Reference-guided Database(HERB), PubChem, GeneCards, SuperPred, and other databases, the active compounds in the serum containing Qishiwei Zhenzhu Pills were retrieved and their action targets were predicted. The predicted targets were compared with the targets of liver and kidney injury related to mercury toxicity retrieved from the database, and the action targets of Qishiwei Zhenzhu Pills to inhibit the potential mercury toxicity of zogta were screened out. Cytoscape was used to construct the active ingredient in Qishiwei Zhenzhu Pills-containing serum-action target network, and STRING database was used to construct the protein-protein interaction(PPI) network of intersection targets. The Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out on the target genes by the DAVID database. The active ingredient-target-pathway network was constructed, and the key ingredients and targets were screened out for molecular docking verification. The results showed that 44 active compounds were identified from the serum containing Qishiwei Zhenzhu Pills, including 13 possible prototype drug ingredients, and 70 potential targets for mercury toxicity in liver and kidney were identified. Through PPI network topology analysis, 12 key target genes(HSP90AA1, MAPK3, STAT3, EGFR, MAPK1, APP, MMP9, NOS3, PRKCA, TLR4, PTGS2, and PARP1) and 6 subnetworks were obtained. Through GO and KEGG analysis of 4 subnetworks containing key target genes, the interaction network diagram of active ingredient-action target-key pathway was constructed and verified by molecular docking. It was found that taurodeoxycholic acid, N-acetyl-L-leucine, D-pantothenic acid hemicalcium, and other active ingredients may regulate biological functions and pathways related to metabolism, immunity, inflammation, and oxidative stress by acting on major targets such as MAPK1, STAT3, and TLR4, so as to inhibit the potential mercury toxicity of zogta in Qishiwei Zhenzhu Pills. In conclusion, the active ingredients of Qishiwei Zhenzhu Pills may have a certain detoxification effect, thus inhibiting the potential mercury toxicity of zogta and playing a role of reducing toxicity and enhancing effect.

Progress on Hypoglycemic Effect and Mechanism of Momordica charantia / 中国实验方剂学杂志

Momordica charantia has been a traditional Chinese medicine （TCM） and food since ancient times. The discussions on its nature， taste and efficacy in ancient books of TCM are almost the same. With a high nutritional value， M. charantia is rich in a variety of vitamins and minerals， and has been widely used in the production of a wide range of dietary supplements and functional foods. At the same time， M. charantia is one of the most deeply studied natural medicines in traditional alternative medicine， with a wide range of pharmacological effects， especially in the treatment of metabolic diseases. Clinical trials have confirmed that M. charantia has a hypoglycemic effect， and could reduce blood lipids and weight loss， so as to improve metabolism in a comprehensive manner. According to the study on the mechanism of M. charantia in the treatment of diabetes， M. charantia could reduce blood sugar by improving islet β-cell function， improving insulin resistance， inhibiting intestinal glucose absorption and resisting inflammation and oxidative stress. However， at present， there is a lack of unified standards for the hypoglycemic effects and various mechanisms of action of M. charantia， and the safety has not been fully confirmed. Further studies shall be conducted to investigate the hypoglycemic effect and mechanisms of M. charantia， explore active components of M. charantia， define the pharmacodynamics material basis， extract monomer compounds with a clear structure and confirm its effectiveness and safety， which is helpful to develop and utilize the homologous value of medicine and food of M. charantia and further apply it in clinic. The application of the hypoglycemic effect of M. charantia in clinic has important economic benefits and a social significance.

Clinical Observation on Orthopedics No.1 Prescription Combined with Celecoxib in Treating Knee Osteoarthritis with Middle Stage of Cold-dampness Syndrome / 中国实验方剂学杂志

Objective:To study the clinical efficacy of orthopedics No.1 prescription combined with celecoxib in the treatment of knee osteoarthritis （KOA） with middle stage of cold-dampness syndrome and investigate its effect on serum cytokines levels. Method:The 72 patients were randomly divided into control group and observation group， with 36 cases each. Patients in both groups were given basic treatment with oral celecoxib capsules （0.2 g/ time， 1 time/day）. On the basis of western medicine treatment， patients in observation group were treated with orthopedics No.1 prescription decoction-free granules by fumigation， 1 bag/time， 1 time/day， 5 times/week. Both groups received treatment for 4 weeks. The visual analog pain score （VAS）， American knee society knee score （KSS）， serum interleukin-1<italic>β </italic>（IL-1<italic>β</italic>）， tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>）， and transforming growth factor-<italic>β</italic>₁（TGF-<italic>β</italic>₁） levels were observed before and after treatment， and their clinical efficacy was evaluated. Result:After treatment， VAS score significantly decreased in both groups （<italic>P</italic><0.01）， and KSS score significantly increased （<italic>P</italic><0.01）， with better clinical effect in observation group. After treatment， serum IL-1<italic>β</italic> and TNF-<italic>α</italic> levels decreased significantly in both groups （<italic>P</italic><0.01）， and the levels in observation group were lower than those in control group after treatment （<italic>P</italic><0.05）. TGF<italic>-β</italic>₁content was significantly higher than that before treatment in two groups （<italic>P</italic><0.01）. Conclusion:Orthopedics No.1 prescription combined with celecoxib for the treatment of KOA with middle stage of cold-dampness syndrome can effectively relieve the clinical symptoms of patients with KOA， improve joint function， improve quality of life， reduce the contents of inflammatory factors IL-1<italic>β</italic> and TNF-<italic>α</italic> in serum， and increase the expression of TGF-<italic>β</italic>₁ level.

Effect of Zishenwan on Pyroptosis and Epithelial-Mesenchymal Transition of Renal Tubular Epithelial Cells in Diabetic Nephropathy Mice / 中国实验方剂学杂志

Objective:To study the efficacy and mechanism of Zishenwan （ZSW） against pyroptosis and epithelial-mesenchymal transition （EMT） of renal tubular epithelial cells in diabetic nephropathy （DN） mice， so as to provide evidence for the treatment of DN with ZSW. Method:The <italic>db/db</italic> mice with spontaneous diabetes were randomly divided into the model group， dapagliflozin （1.0 mg·kg^-1） group， and high-， medium-， and low-dose （6.0， 3.0， 1.5 g·kg^-1） ZSW groups. The non-diabetic <italic>db/m</italic> mice were classified into the normal group. The ones in the model and normal groups were given an equal volume of deionized water by gavage， while those in the other groups were intervened with the corresponding drugs for 12 weeks. The fasting blood glucose （FBG） level was tested at tail vein once every two weeks. The levels of urine albumin-creatinine ratio （ACR）， <italic>β</italic>-N-acetyl-D-glucosaminidase （NAG）， and cystatin C （CysC） were detected once every four weeks. After 12 weeks of administration， the blood sampled from eyeballs was used for measuring the blood urea nitrogen （BUN） and serum creatinine （SCr）. The pathological changes in renal tissues were observed by light microscopy and transmission electron microscopy. The expression of EMT markers in the renal tubular epithelium was analyzed by immunohistochemistry （IHC）. The in situ terminal end-labeling （TUNEL） staining was conducted to analyze the nuclear damage of renal tubular epithelial cells. The protein and mRNA expression levels of EMT markers， nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） inflammasome and pyroptosis-related inflammatory cytokines in renal tissues were separately assayed by Western blot and Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. Result:Compared with the normal group， the model group displayed significantly increased FBG， BUN， serum SCr， ACR， NAG， and CysC （<italic>P</italic><0.01）， impaired renal tissues， altered EMT marker expression intensities and levels （<italic>P</italic><0.01）， and elevated TUNEL-positive rate and protein and mRNA expression levels of pyroptosis-related inflammatory cytokines and NLRP3 inflammasome （<italic>P</italic><0.01）. Compared with the model group， ZSW and dapagliflozin significantly decreased the levels of FBG， BUN， serum SCr， ACR， NAG， and CysC （<italic>P</italic><0.01）， relieved the pathological injuries in renal tissues， changed the EMT marker expression intensities （<italic>P</italic><0.01） and protein and mRNA expression levels （<italic>P</italic><0.05， <italic>P</italic><0.01）， and down-regulated the TUNEL-positive rate （<italic>P</italic><0.01） of renal tubular epithelial cells as well as the protein and mRNA expression levels of pyroptosis-related inflammatory cytokines （<italic>P</italic><0.01） and NLRP3 inflammasome （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:ZSW alleviates DN possibly by inhibiting pyroptosis and EMT in renal tubular epithelial cells.

Research Progress on Medicinal Effect of Flower of Edgeworthia gardneri / 中国实验方剂学杂志

Tibetan medicine Edgeworthia gardneri is the flower bud of Edgeworthia gardneri, mainly distributed in the eastern Tibet as well as the northwest and west parts of Yunnan. It is called one of the " Eighteen treasures of Qinghai-Tibet" and has been used in many Tibetan secret recipes for a long time. Nowadays, the local people often drink E. gardneri Meisn tea for blood glucose regulation and health care. In this study, the related papers were searched in CNKI, Wanfang, Weipu, PubMed and other databases by using the keywords " Lv luo hua" and " flower of E. gardneri Meisn" . Then the status quo of the flower of E. gardneri was summarized from three aspects: provenance, chemical compositions and extraction method, as well as efficacy and function. It is found that E. gardneri mainly contains coumarins, flavonoids, polysaccharides and polyphenols. The extraction methods of E. gardneri include ultrasonic extraction, microwave extraction, supercritical CO2 extraction, and bio-enzyme-assisted extraction method, et al. The extraction of flavonoids and polyphenols is more common. In terms of medicinal efficacy, it has many medicinal effects such as regulating blood sugar, lowering blood fat, anti-oxidation, enhancing immunity and anti-tumor. The flower of E. gardneri has broad pharmacological effect, attracting more and more scholars to research it. However, studies on its medicinal effectiveness both at home and abroad are mainly focusing on it' s crude extractions or extracts, with no in-depth studies on action mechanism. This review aims to summarize the provenance, main chemical compositions and extraction methods, efficacy and effect of the flower of E. gardneri, especially focusing on the research on the medicinal efficacy and mechanism of the flower of E. gardneri, and provide a reference for the further research of the flower of E. gardneri.

Chemical Constituents of Ethyl Acetate Extract from Ajuga ovalifolia var. calantha / 中国实验方剂学杂志

Objective:To investigate the chemical constituents from the ethyl acetate extract of 95% ethanol extract from Ajuga ovalifolia var. calantha. Method:A. ovalifolia var. calantha was percolated with 95% ethanol，and the percolate was concentrated under reduced pressure to obtain the extract. The extract was then dispersed with water and extracted with petroleum ether to obtain the petroleum ether extract fraction and water layer. Then ethyl acetate was used to extract the water layer to obtain the ethyl acetate extract fraction. The Compounds from the ethyl acetate extract fraction were isolated and purified by silica gel，Sephadex LH-20，MCI column，ODS column chromatography and semi-preparative high performance liquid chromatography. Their structures were elucidated by interpretation of NMR，ESI-MS and other spectral evidence. Result:17 Compounds were isolated and elucidated as bakkenolide-E（1），loliotide（2），isololiotide（3），（E）-linalool-1-oic acid（4），umbelliferone（5），phillygenin（6），1-（4-hydroxyphenyl）-ethanol（7），（2E）-8-hydroxy-2，6-dimethyl-2-octenoic acid（8），1H-indole-3-carboxylic acid（9），ajugarin I（10），ajugalactone（11），luteolin（12），20-hydroxyecdysone-2-acetate（13），benzyl-4'-hydroxy-benzoyl-3'-O-β-D-glucopyranoside（14），harpagide（15），6-deoxy-8-acetylharpagide（16），and acteoside（17）. Conclusion:Compounds 1，3-4，6-9，14 were isolated from the plants of Ajuga for the first time，and compounds 2，5，10，13，15-16 were isolated from this plant for the first time.

Metabolic transformation and attenuation characteristics of endogenous alkaloid toxic components of Tibetan medicinal herbs:a review / 中国药理学与毒理学杂志

The active ingredients in some Tibetan medicinal herbs are toxic components as well,and we need to have a clear understanding of their mechanism and metabolic pathways in use. The endogenous toxic components of highly toxic Tibetan herbal medicines are mainly alkaloids, such as aconitum alkaloids, methyllycaconitine, tropane alkaloids, brucine, strychnine, papaverine and swainso-nine. The majority of endogenous toxic alkaloids in Tibetan medicine herbs exist in roots, fruits and seeds of plants, exerting neurotoxicity or cardiotoxicity as highly toxic inherent chemicals. Most alka-loids are metabolized in phaseⅠvia de-alkylation, hydroxylation, hydrolysis and other reactions, as well as in phaseⅡvia glucuronic acid and sulfonic acid conjugation. They form various metabolites with high polarities and reduced toxicities so as to be easily excreted. The closeness between the therapeutic dose and toxic dose of alkaloids components in Tibetan medicinal herbs leads to their attenuated prep-aration via frying, dairy, highland barley wine soaking, or in combination with Terminalia Chebula to decrease toxicity, as is cited classic books on in Tebitan medicine. Focused on twelve alkaloids of five classes including aconitine, tropane and brucine, we have reviewed the characteristics of their metabo-lism and transformation, as well as their toxicity attenuation and safety evaluation.

Effects of Heyutai Fuzhu Jiangtang Tablets Combined with Metformin on Insulin Resistance in Skeletal Muscle of Diabetic Rats / 中国中医药信息杂志

Objective To observe effects of Heyutai Fuzhu Jiangtang Tablets combined with metformin in insulin resistance (IR); To discuss its mechanism of action. Methods 6–7 week old male ZDF (fa/fa) rats were randomly divided into model group,metformin group,Heyutai Fuzhu Jiangtang Tablets group(Jiangtang Tablets group),and metformin combined with Heyutai Fuzhu Jiangtang Tablets group.ZDF(fa/+)rats were chosen as normal group.Each medication group was given relevant medicine for gavage for 6 weeks. Body weight, FBG, TG, TC, FFA, FINS, HOMA-IR, OGTT and HE staining were tested. HE staining was used to observe the pathological changes of skeletal muscle. RT-PCR and Western blot were used to detect skeletal muscle corresponding gene and protein expression. Results Compared with Jiangtang Tablets group and metformin group, TC, FFA, FBG, and HOMA-IR in metformin combined with Heyutai Fuzhu Jiangtang Tablets group decreased significantly (P<0.05, P<0.01). Blood glucose level and AUC significantly decreased at each time point in OGTT. HE staining of skeletal muscle fibers arranged in order; nucleus increased and internal movement was not significant, without obvious infiltration of inflammatory cells. Expressions of skeletal muscle InsR, Akt, and Glut4 mRNA expression increased (P<0.05, P<0.01). Expressions of skeletal muscle p-InsR, p-Akt, and Glut4 protein expression increased (P<0.05, P<0.01). Conclusion Heyutai Fuzhu Jiangtang Tablets combined with metformin can improve IR in type 2 diabetic rats, and the effect is better than single-application.

Effects of Heyutai Fuzhu Jiangtang Tablets Combined with Metformin on Insulin Resistance in Skeletal Muscle of Diabetic Rats / 中国中医药信息杂志

Objective To observe effects of Heyutai Fuzhu Jiangtang Tablets combined with metformin in insulin resistance (IR); To discuss its mechanism of action. Methods 6–7 week old male ZDF (fa/fa) rats were randomly divided into model group,metformin group,Heyutai Fuzhu Jiangtang Tablets group(Jiangtang Tablets group),and metformin combined with Heyutai Fuzhu Jiangtang Tablets group.ZDF(fa/+)rats were chosen as normal group.Each medication group was given relevant medicine for gavage for 6 weeks. Body weight, FBG, TG, TC, FFA, FINS, HOMA-IR, OGTT and HE staining were tested. HE staining was used to observe the pathological changes of skeletal muscle. RT-PCR and Western blot were used to detect skeletal muscle corresponding gene and protein expression. Results Compared with Jiangtang Tablets group and metformin group, TC, FFA, FBG, and HOMA-IR in metformin combined with Heyutai Fuzhu Jiangtang Tablets group decreased significantly (P<0.05, P<0.01). Blood glucose level and AUC significantly decreased at each time point in OGTT. HE staining of skeletal muscle fibers arranged in order; nucleus increased and internal movement was not significant, without obvious infiltration of inflammatory cells. Expressions of skeletal muscle InsR, Akt, and Glut4 mRNA expression increased (P<0.05, P<0.01). Expressions of skeletal muscle p-InsR, p-Akt, and Glut4 protein expression increased (P<0.05, P<0.01). Conclusion Heyutai Fuzhu Jiangtang Tablets combined with metformin can improve IR in type 2 diabetic rats, and the effect is better than single-application.