1. Exploring mechanism of hypolipidemic effect of total Ligustrum robustum (Roxb. ) Blume on hyperlipidemic golden hamsters based on intestinal flora
Chen-Xi XU ; Rui-Le PAN ; Meng-Chen DONG ; Zhi-Hong YANG ; Xiao-Ya LI ; Wen JIN ; Run-Mei YANG
Chinese Pharmacological Bulletin 2024;40(3):476-483
Aim To evaluate the hypolipidemic effect of the total phenylpropanoid glycosides extracted from Ligustrum robustum (Roxb.) Blume (LRTPG) on hyperlipidemic golden hamsters and explore its regulatory effect on intestinal flora. Methods Sixty hamsters were randomly divided into a control group, a model group, a positive drug group, LRTPG-L group, LRTPG-M group, and LRTPG-H group. After the successful induction of the model by high-fat diet, the animals were continuously administered for four weeks, and their blood lipids and liver lipids were detected. The formed feces from the colorectal region of the hamsters in the control group, model group and LRTPG-H group were collected for 16S rDNA sequencing. Results LRTPG reduced serum TG, TC, LDL-C and liver TG, TC concentrations significantly in hyperlipidemic hamsters. The results of the intestinal microbiota sequencing showed that compared to the control group, LRTPG significantly decreased the relative abundance of the phylum Firmicutes and increased the relative abundance of the phylum Bacteroidetes and Verrucomicrobia (P < 0.01) at the phylum level. At the family level, LRTPG significantly increased the relative abundance of Christensenellaceae, Peptococcaceae, and Verrucomicrobiaceae (P < 0.05 or P < 0.01). At the genus level, LRTPG significantly increased the relative abundance of Oscillospira, Oscillibacter, Flavonifractor and Akkermansiaceae (P < 0.05 or P < 0.01). These changes in the flora were beneficial to the hypolipidemic effect of LRTPG. Conclusion LRTPG may exert its hypolipidemic effect by improving the intestinal flora disorder caused by a high-fat diet in golden hamsters.
2.“Liquid seal to detoxification – drying and puffing” of two stage processing technology design and pharmacodynamic study of aconite (Aconiti Lateralis Radix Praeparata) processed by microwave
Ya-nan HE ; Xin YANG ; Jing WU ; Yu-sen HOU ; Qi HU ; Run-chun XU ; Qin-wan HUANG ; Ming YANG ; Ding-kun ZHANG
Acta Pharmaceutica Sinica 2023;58(5):1328-1337
Establish a production line with controllable process and high intelligence, contribute to improve the quality and production efficiency of aconite processed by microwave, and promote the transformation and application of aconite processed by microwave. According to the principle of aconite detoxification and the characteristics of industrial microwave equipment, an industrial production line of aconite processed by microwave was established with diester alkaloids and monoester alkaloids as indicators, and pilot production was carried out. At the same time, the content of active constituents and efficacy were compared with that of the main processed products, such as Shengfupian, Baifupian and Heishunpian. The results showed that the industrial production of aconite processed by microwave can be divided into two stages: "Liquid seal to detoxification - drying and puffing". The content of monoester alkaloids in 10 batches of aconite processed by microwave was 0.071%-0.166% and the content of diester alkaloids was 0.004%-0.016%, which met the relevant requirements of the Chinese Pharmacopoeia in 2020. Compared with Heishunpian and Baifupian, the retention rate of the effective components of aconite processed by microwave was higher. Pharmacological experiments showed that aconite processed by microwave not only retained the anti-inflammatory and analgesic activities of Heishunpian and Baifupian, but also significantly increased the levels of leukocytes and lymphocytes in mice with liver cancer chemotherapy, enhanced the CD4/CD8 ratio in spleen cells of mice (
3.Astragalin Inhibits Apoptosis of Cerebral Cortex Neurons in APP/PS1 Mice
Shu-han WANG ; Cui-zhu YANG ; Run-heng ZHANG ; Jia-hong LIN ; Ya-qi YANG ; Jing LIU ; Guo-ying LI ; Yu-xin MA
Journal of Sun Yat-sen University(Medical Sciences) 2023;44(6):983-990
ObjectiveTo investigate the effect of Astragalin (AST) on apoptosis of cerebral cortex neurons in APP/PS1 transgenic mice. MethodsEighteen six-month-old male APP/PS1 transgenic mice were randomly divided into APP/PS1 group, APP/PS1+ 40 mg/kg AST group and APP/PS1+ 20 mg/kg Donepezil (DNP) group, with six mice in each group. At the same time, six male C57BL/6 mice were selected as the normal control group. After intraperitoneal injection of AST once a day and continuous administration for one month, we used Tunel staining to detect the apoptosis of neurons in the cerebral cortex of APP/PS1 mice; immunofluorescent staining to examine the expression of apoptosis-related proteins Bax, Bcl-2, Caspase9 and Cleaved-Caspase3 in the cerebral cortex neurons of APP/PS1 mice; Western blot method to evaluate the changes of the expression of Bax, Bcl-2, Caspase9 and Caspase3. ResultsTunel staining showed that 40 mg/kg AST and 20 mg/kg DNP both reduced the apoptosis of neurons in the cerebral cortex of APP/PS1 mice, AST with more significant inhibition effect. Immunofluorescent staining revealed that 40 mg/kg AST and 20 mg/kg DNP both inhibited the expression of Bax, Caspase9, and Cleaved-Caspase3, and icreased the expression of Bcl-2 in the cerebral cortex neurons of APP/PS1 mice. Western blot results further confirmed that 40 mg/kg AST and 20 mg/kg DNP both down-regulated the expression of Bax (P < 0.05, P < 0.05), Caspase9 (P < 0.005, P < 0.05) and Caspase3 (P < 0.0001, P < 0.0001) , and up-regulated the expresstion of Bcl-2 (P < 0.05, P < 0.05) in the cerebral cortex neurons of APP/PS1 mice. ConclusionsAST can inhibit the apoptosis of cerebral cortex neurons in APP/PS1 mice.
4.Human menstrual blood-derived stem cells alleviate autoimmune hepatitis via JNK/MAPK signaling pathway in vivo and in vitro.
Fen ZHANG ; Lanlan XIAO ; Ya YANG ; Menghao ZHOU ; Yalei ZHAO ; Zhongyang XIE ; Xiaoxi OUYANG ; Feiyang JI ; Shima TANG ; Lanjuan LI
Frontiers of Medicine 2023;17(3):534-548
Autoimmune hepatitis (AIH) is a severe globally distributed liver disease that could occur at any age. Human menstrual blood-derived stem cells (MenSCs) have shown therapeutic effect in acute lung injury and liver failure. However, their role in the curative effect of AIH remains unclear. Here, a classic AIH mouse model was constructed through intravenous injection with concanavalin A (Con A). MenSCs were intravenously injected while Con A injection in the treatment groups. The results showed that the mortality by Con A injection was significantly decreased by MenSCs treatment and liver function tests and histological analysis were also ameliorated. The results of phosphoproteomic analysis and RNA-seq revealed that MenSCs improved AIH, mainly by apoptosis and c-Jun N-terminal kinase/mitogen-activated protein signaling pathways. Apoptosis analysis demonstrated that the protein expression of cleaved caspase 3 was increased by Con A injection and reduced by MenSCs transplantation, consistent with the TUNEL staining results. An AML12 co-culture system and JNK inhibitor (SP600125) were used to verify the JNK/MAPK and apoptosis signaling pathways. These findings suggested that MenSCs could be a promising strategy for AIH.
Mice
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Animals
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Humans
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Hepatitis, Autoimmune/pathology*
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Signal Transduction
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Disease Models, Animal
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Stem Cells
5.Establishment and application of a rapid quality inspection method for Indigo Naturalis based on quantitative portrayal of water testing process
Xue-mei LIU ; Ya-nan HE ; Fang WANG ; Ming YANG ; Hai ZHANG ; Xiang-bo YANG ; Li HAN ; Run-chun XU ; Ding-kun ZHANG
Acta Pharmaceutica Sinica 2022;57(11):3411-3418
A new rapid, quality control method based on quantitative water tests has been established for the quality evaluation of Indigo Naturalis. The Turbiscan stability index (TSI) of 26 batches of Indigo Naturalis was measured by a stability analyzer. The parameters, including the method by which the ingredients are added, their particle size, amount, and the testing temperature, were systematically optimized and the methodological indexes such as repeatability and stability were determined. The content of indigo and indirubin in 26 batches of Indigo Naturalis was determined by high performance liquid chromatography and the total ash was measured. The correlation analysis between the active ingredients, total ash content and TSI value of Indigo Naturalis was determined by SPSS 26.0 and Origin 2021. This research shows that the best way to prepare samples for testing is to add 0.2 g of Indigo Naturalis powder which has passed through a No. 7 sieve but failed to pass through a No. 9 sieve to a glass bottle containing 20 mL pure water by a funnel and scan at 25 ℃ with a stability analyzer. Consistency analysis showed that the content ranking of indigo and indirubin is opposite to the TSI value, and the content ranking of total ash is generally consistent with the TSI value. Correlation analysis showed that the correlation coefficients of indigo and indirubin content and TSI value were -0.850 and -0.801, respectively, and
6.Comparison of odor and quality of Galli Gigerii Endothelium Corneum derived from domestic chickens and broilers.
Wei HUANG ; Lu-Meng CHEN ; Hao-Zhou HUANG ; Ya-Nan HE ; Xiao-Ming BAO ; Jing YANG ; Jun-Zhi LIN ; Peng TAN ; Li HAN ; Run-Chun XU ; Ding-Kun ZHANG
China Journal of Chinese Materia Medica 2022;47(20):5434-5442
Galli Gigerii Endothelium Corneum(GGEC) is commonly used for the clinical treatment of indigestion, vomiting, diarrhea, and infantile malnutrition with accumulation. In recent decades, omnivorous domestic chickens, the original source of GGEC, has been replaced by broilers, which may lead to significant changes in the quality of the yielding GGEC. Through subjective and objective sensory evaluation, biological evaluation, and chemical analysis, this study compared the odor and quality between GGEC derived from domestic chickens and that from broilers. The odor intensity between them was compared by odor profile analysis and it was found that the fishy odor of GGEC derived from domestic chickens was significantly weaker than that of GGEC from broilers. Headspace-solid phase microextraction-gas chromatography-triple quadrupole tandem mass spectrometry(HS-SPME/GC-QQQ-MS/MS) suggested that the overall odor-causing chemicals were consistent with the fishy odor-causing chemicals. According to the odor activity va-lue and the orthogonal partial least squares discriminant analysis(OPLS-DA) result, dimethyl trisulfide, 2-methoxy-3-isobutylpyrazine, and 2-methylisoborneol were responsible for the fishy odor(OAV≥1) and the content of fishy odor-causing chemicals in GGEC derived from broilers was 1.12-2.13 folds that in GGEC from domestic chickens. The average pepsin potency in GGEC derived from broilers was 15.679 U·mg~(-1), and the corresponding figure for the medicinal from domestic chickens was 26.529 U·mg~(-1). The results of pre-column derivatization reverse-phase high-performance liquid chromatography(RP-HPLC) assay showed that the content of total amino acids and digestion-promoting amino acids in domestic chickens-derived GGEC was 1.12 times and 1.15 times that in GGEC from broilers, and the bitter amino acid content was 1.21 times folds that of the latter. In conclusion, GGEC derived from domestic chickens had weaker fishy odor, stronger enzyme activity, higher content of digestion-promoting amino acids, and stronger bitter taste than GGEC from broilers. This study lays a scientific basis for studying the quality variation of GGEC and provides a method for identifying high-quality GGEC. Therefore, it is of great significance for the development and cultivation of GGEC as both food and medicine and breeding of corresponding varieties.
Animals
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Odorants/analysis*
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Chickens
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Gas Chromatography-Mass Spectrometry/methods*
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Tandem Mass Spectrometry
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Solid Phase Microextraction
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Amino Acids
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Endothelium/chemistry*
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Volatile Organic Compounds/analysis*
7.Challenges and strategies of high-quality development of Indigo Naturalis industry.
Ming YANG ; Ding-Kun ZHANG ; Ya-Nan HE ; Li HAN ; Fang WANG ; Run-Chun XU
China Journal of Chinese Materia Medica 2021;46(13):3171-3179
Traditional Chinese medicine( TCM) processing is a traditional pharmaceutical technology unique to China,which is an important means to ensure the safety and effectiveness of clinical medication. As China' s intangible cultural heritage,it contains a wealth of wisdom. With the evolution and optimization of processing methods,Indigo Naturalis,an ancient dye,has gradually become an effective medicine for the treatment of high fever in children,colitis,psoriasis,and leukemia. However,the quality of Indigo Naturalis pieces is difficult to be fundamentally improved due to the unique traditional processing method,complex technology,unclear principle,and outdated equipment. After spending 20 years in exploring the inheritance,innovation,and transformation of ancient lawtheory-principle-technology-equipment-quality control in Indigo Naturalis processing,our research group has basically expounded the processing principle and realized the modern expression and industrial transformation of traditional technology. As China enters a new era,the TCM industry has begun to undergo the high-quality transformation. It is urgent to carry out new excavations and improve the processing,quality,and clinical application of Indigo Naturalis pieces,to better inherit and innovate traditional processing technologies and meet people's demand for high-quality TCM health services.
Child
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Drugs, Chinese Herbal
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Humans
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Indigo Carmine
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Indigofera
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Medicine, Chinese Traditional
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Psoriasis
8.Microbial community structure and transformation of indoles in soaking and fermentation of Indigo Naturalis.
Ya-Nan HE ; Le-le MA ; Yi WU ; Lu-Meng CHEN ; Ding-Kun ZHANG ; Xiang-Bo YANG ; Run-Chun XU ; Fang WANG ; Li HAN ; Ming YANG
China Journal of Chinese Materia Medica 2021;46(13):3180-3187
The soaking and fermentation of Baphicacanthus cusia( Nees),the important intermediate link of Indigo Naturalis processing,facilitates the synthesis of indigo and indirubin precursors and the dissolution of endogenous enzymes and other effective components,while the role of microorganisms in the fermentation is ignored. The present study investigated the changes of microbial community structure in Indigo Naturalis processing based on 16 S amplicon sequencing and bioinformatics. Meanwhile,the contents of indigo,indirubin,isatin,tryptanthrin,indole glycoside,etc. were determined to explore the correlation between the microorganisms and the alterations of the main components. As demonstrated by the results,the microbial diversity decreased gradually with the fermentation,which bottomed out after the addition of lime. Proteobacteria,Bacteroidetes,and Firmicutes were the main dominant communities in the fermentation. The relative abundance of Proteobacteria declined gradually with the prolongation of fermentation time,and to the lowest level after the addition of lime. The relative abundance of Firmicutes increased,and that of Bacteroidetes decreased first and then increased. The contents of effective substances in Indigo Naturalis also showed different variation tendencies. As fermentation went on,indole glycoside decreased gradually; indigo first increased and then decreased; indirubin and isatin first decreased and then increased; tryptanthrin gradually increased. Those changes were presumedly related to the roles of microorganisms in the synthesis of different components. This study preliminarily clarified the important role of microorganisms in the soaking and fermentation and provided a scientific basis for the control of Indigo Naturalis processing and the preparation of high-quality Indigo Naturalis.
Fermentation
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Indigo Carmine
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Indigofera
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Indoles
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Microbiota
9. Influence of fat and muscle on sit-ups in female college students
Bin-Bin HUANG ; Ze-Rong WANG ; Ya-Zhu YANG ; Cheng TANG ; Hua-Qian WU ; Run CHEN ; Qiao-Li LI ; Li-Qian HUANG ; Wen-Bo SHU ; Peng LIU
Acta Anatomica Sinica 2021;52(5):822-826
Objective To explore the influence of muscle and fat distribution on sit-up ability of female college students. Methods A total of 1560 female students majoring in non-physical education in a Guangxi University were randomly selected. Body composition was measured by the bioresistance antibody component meter. According to the National Physical Health Test Standard, the height, weight and sit-ups of female college students were measured. The students were divided into four groups according to the score of sit-ups:<60 points, 60-69 points, 70-79 points, ≥80 points. SPSS 22.0 statistical software was used to analyze the data. Results There were statistically significant differences in body fat rate, total fat mass, trunk fat mass, trunk fat rate, visceral fat area, visceral fat mass and subcutaneous fat mass in different sit-ups (F value were 3.414, 2.914, 2.881, 3.347, 3.064, 3.235, 2.766, P<0.05). That was, the fat parameters in the 60-69 group were the highest and ≥80 the lowest. However, there was no significant difference between total muscle mass and trunk muscle mass in each score segment. After correcting for age, height, and weight factors, except for visceral fat area and visceral fat mass, the correlation between other fat parameters and sit-up score were negatively correlated, which were statistically significant (r<0, P<0.05). There was a positive correlation between each muscle parameter and sit-up performance (r>0, P<0.05). Compared with other parameters, the correlation coefficient between trunk muscle mass and sit-ups was the largest. Conclusion The distribution of fat and muscle has different effects on sit-up ability. Female college students' sit-up ability is greatly affected by subcutaneous fat and trunk muscle, and has nothing to do with visceral fat.
10. Effects of ginsenoside Re on intestinal mucosa of mice with severe acute pancreatitis and its regulation mechanism
Run LONG ; Ya-Pei XU ; Zhu-Feng YANG ; Jing YANG ; Yan-Hong LI
Acta Anatomica Sinica 2021;52(4):628-634
Objective To explore the effect and mechanism of ginsenoside Re on the intestinal mucosa of severe acute pancreatitis (SAP) mice by regulating the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/ STAT3) pathway. Methods Totally 48 mice were divided into : control group, SAP group, SAP + ginsenoside Re group and SAP + ginsenoside Re + LY2784544 group (n = 12). The mice were intraperitoneally injected with caerulein solution (after fasting for 12 hours, 100 μg/kg, 6 times, injection interval 60 minutes) to establish SAP models. Mice in the SAP+ ginsenoside Re group were intraperitoneally injected with ginsenoside Re (4 mg/kg, 1 time a day for 7 consecutive days). Intraperitoneal injection of 12.7 mg/kg LY2784544 was used to inhibit the JAK2/STAT3 pathway. Pancreatic and intestinal mucosal injury were detected in each group. The wet to dry weight ratio of pancreas, serum amylase and inflammatory factor levels were detected in each group. The intestinal mucosal barrier function was analyzed by detecting the levels of D-lactic acid and fluorescein isothiocyanate dextran (FITC-D). The damage of pancreatic tissue and intestinal mucosa tissue was observed by HE staining. Western blotting was used to detect the levels of apoptotic proteins Bax and Bcl-2 in the intestinal mucosa. The JAK2/STAT3 pathway expression levels in pancreatic tissue and intestinal mucosa tissue were detected by Realtime PCR and Western blotting. Results The pancreatic index, serum amylase, interieukin 6 (IL-6) and tumor necrosis factor a (TNF-a), FITC-D, D-lactic acid, Bax protein levels, JAK2 and STAT3 mRNA and protein levels in the SAP group were significantly higher than those in the control group, while Bcl-2 protein was significantly lower than that in the control group (P<0.05). The Bcl-2 protein of SAP+ginsenoside Re group was significantly higher than that of SAP group, and other indexes were significantly lower than those in SAP group (P<0.05). The Bcl-2 protein of SAP+ginsenoside Re+ LY2784544 group was significantly higher than that of SAP + ginsenoside Re group, and other indexes were significantly lower than those in the SAP+ginsenoside Re group (P<0.05). Conclusion Ginsenoside Re may reduce the pancreatic injury in SAP model mice by inhibiting the JAK/STAT pathway to alleviate the inflammatory response, and may protect the small intestinal mucosal barrier by alleviating pancreatitis and inhibiting the intestinal mucosal JAK/STAT pathway to inhibit cell apoptosis.

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