Difference of lipid-lowering efficacy of "Xinjianqu" before and after fermentation and its mechanism based on LKB1-AMPK pathway and 16S rDNA sequencing technology / 中国中药杂志

On the basis of establishing the prescription of Xinjianqu and clarifying the increase of the lipid-lowering active ingredients of Xinjianqu by fermentation, this paper further compared the differences in the lipid-lowering effects of Xinjianqu before and after fermentation, and studied the mechanism of Xinjianqu in the treatment of hyperlipidemia. Seventy SD rats were randomly divided into seven groups, including normal group, model group, positive drug simvastatin group(0.02 g·kg~(-1)), and low-dose and high-dose Xinjianqu groups before and after fermentation(1.6 g·kg~(-1) and 8 g·kg~(-1)), with ten rats in each group. Rats in each group were given high-fat diet continuously for six weeks to establish the model of hyperlipidemia(HLP). After successful modeling, the rats were given high-fat diet and gavaged by the corresponding drugs for six weeks, once a day, to compare the effects of Xinjianqu on the body mass, liver coefficient, and small intestine propulsion rate of rats with HLP before and after fermentation. The effects of Xinjianqu before and after fermentation on total cholesterol(TC), triacylglyceride(TG), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), alanine aminotransferase(ALT), aspartate aminotransferase(AST), blood urea nitrogen(BUN), creatinine(Cr), motilin(MTL), gastrin(GAS), and the Na~+-K~+-ATPase levels were determined by enzyme-linked immunosorbent assay(ELISA). The effects of Xinjianqu on liver morphology of rats with HLP were investigated by hematoxylin-eosin(HE) staining and oil red O fat staining. The effects of Xinjianqu on the protein expression of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK), phosphorylated AMPK(p-AMPK), liver kinase B1(LKB1), and 3-hydroxy-3-methylglutarate monoacyl coenzyme A reductase(HMGCR) in liver tissues were investigated by immunohistochemistry. The effects of Xinjianqu on the regulation of intestinal flora structure of rats with HLP were studied based on 16S rDNA high-throughput sequencing technology. The results showed that compared with those in the normal group, rats in the model group had significantly higher body mass and liver coefficient(P<0.01), significantly lower small intestine propulsion rate(P<0.01), significantly higher serum levels of TC, TG, LDL-C, ALT, AST, BUN, Cr, and AQP2(P<0.01), and significantly lower serum levels of HDL-C, MTL, GAS, Na~+-K~+-ATP levels(P<0.01). The protein expression of AMPK, p-AMPK, and LKB1 in the livers of rats in the model group was significantly decreased(P<0.01), and that of HMGCR was significantly increased(P<0.01). In addition, the observed＿otus, Shannon, and Chao1 indices were significantly decreased(P<0.05 or P<0.01) in rat fecal flora in the model group. Besides, in the model group, the relative abundance of Firmicutes was reduced, while that of Verrucomicrobia and Proteobacteria was increased, and the relative abundance of beneficial genera such as Ligilactobacillus and Lachnospiraceae＿NK4A136＿group was reduced. Compared with the model group, all Xinjianqu groups regulated the body mass, liver coefficient, and small intestine index of rats with HLP(P<0.05 or P<0.01), reduced the serum levels of TC, TG, LDL-C, ALT, AST, BUN, Cr, and AQP2, increased the serum levels of HDL-C, MTL, GAS, and Na~+-K~+-ATP, improved the liver morphology, and increased the protein expression gray value of AMPK, p-AMPK, and LKB1 in the liver of rats with HLP and decreased that of LKB1. Xinjianqu groups could regulate the intestinal flora structure of rats with HLP, increased observed＿otus, Shannon, Chao1 indices, and increased the relative abundance of Firmicutes, Ligilactobacillus(genus), Lachnospiraceae＿NK4A136＿group(genus). Besides, the high-dose Xinjianqu-fermented group had significant effects on body mass, liver coefficient, small intestine propulsion rate, and serum index levels of rats with HLP(P<0.01), and the effects were better than those of Xinjianqu groups before fermentation. The above results show that Xinjianqu can improve the blood lipid level, liver and kidney function, and gastrointestinal motility of rats with HLP, and the improvement effect of Xinjianqu on hyperlipidemia is significantly enhanced by fermentation. The mechanism may be related to AMPK, p-AMPK, LKB1, and HMGCR protein in the LKB1-AMPK pathway and the regulation of intestinal flora structure.

Interventional effect of asiaticosdide on rats exposed to silica dust / 中华劳动卫生职业病杂志

Objective: To investigate the effect of asiaticoside for fibrosis in lung tissues of rats exposed to silica and to explore its possible mechanism. Methods: 144 SD male rats were randomly divided into control group, model group, positive drug control group, asiaticoside high-dose group, medium-dose group and low-dose group, each group included 24 rats. Rats in the control group were perfused with 1.0 ml of normal saline, and the other groups were given 1.0 ml 50 mg/ml SiO(2) suspension. Gavage of herbal was given from the next day after model establishment, once a day. Rats in the positive drug control group were administration with 30 mg/kg tetrandrine and rats in the low-dose group, medium-dose group and high-dose group were given 20 mg/kg, 40 mg/kg and 60 mg/kg asiaticoside for fibrosis respectively. Rats in the control group and the model group were given 0.9% normal saline. The rats were sacrificed in on the 14th, 28th and 56th day after intragastric administration and collect the lung tissues to detect the content of hydroxyproline, TGF-β(1) and IL-18, observe the pathological changes of the lung tissues by HE and Masson staining and determine the expressions of Col-I, a-SMA, TGF-β in lung tissues by Western Blot. Results: On the 14th day, 28th day and 56th day after model establishment, the lung tissues of rats in the model group showed obvious inflammatory response and accumulation of collagen fibers, and the degree of inflammation and fibrosis increased with time. The intervention of asiaticoside could effectively inhibit the pathological changes of lung tissues. The contents of hydroxyproline, IL-18 and TGF-β1 in lung tissues of model group were higher than those in the control group (P<0.05) , while the level of hydroxyproline, IL-18 and TGF-β1 in asiaticoside groups were significantly decreased, and the difference was statistically signicant (P<0.05) . Compared with the control group, the expression levels of Col-I, TGF-β1and α-SMA in lung tissue of model group were increased (P<0.05) , while the expression level of Col-I, TGF-β1 and α-SMA were decreased after the intervention of asiaticoside, and the difference was statistically signicant (P<0.05) . Conclusion: Asiaticoside can inhibit the increase of Col-I, TGF-β1 and α-SMA content in the SiO(2)-induced lung tissues of rats, reduce the release of TGF-β1 and IL-18 inflammatory factors in lung tissue, and then inhibit the synthesis and deposition of extracellular matrix in rat lung tissue, and improve silicosis fibrosis.

Research progress of NLRP3 inflammasome in pulmonary fibrosis / 中国职业医学

In recent years, the prevalence of pulmonary fibrosis is increasing worldwide. Due to its complex pathogenesis and different clinical manifestations, the treatment options are limited. The nucleotide-binding oligomerization domain like receptor family pyrin domain-containing protein 3(NLRP3) inflammasome plays an important role in pulmonary fibrotic diseases, and its activation and inhibition can directly affect the process of pulmonary fibrosis. The structure of NLRP3 inflammasome consists of a sensor molecule, NLRP3, apoptosis-associated speck-like protein CARD, and caspase-1. There are three signaling pathways that include canonical pathway, selective or noncanonical pathway, and alternative pathway. Excessive activation of NLRP3 inflammasome can accelerate the development of idiopathic pulmonary fibrosis and pulmonary fibrosis induced by silicosis. Therefore, the NLRP3 inflammasome may serve as a therapeutic target of pulmonary fibrosis. NLRP3 inflammasome activators and inhibitors can mediate the activation and inhibition of NLRP3 inflammasome, and then regulate the occurrence and development of pulmonary fibrosis. Inhibiting the activation of NLRP3 inflammasome can improve pulmonary fibrosis. However, the specific mechanism has not been elucidated. The therapeutic strategies for pulmonary fibrosis need further research on the activation and regulation of NLRP3 inflammasome.

Analysis on Mechanism of Baiyaojian in Treating Ulcerative Colitis Based on Intestinal Flora and Intestinal Mucosal Barrier Function / 中国实验方剂学杂志

Objective:To compare the effects of Baiyaojian before and after fermentation on intestinal flora and expression of Occludin and zonula occludens protein-1 （ZO-1） in intestinal mucosa of mice with ulcerative colitis （UC）， and to explore the mechanism of Baiyaojian and Galla Chinensis in the treatment of UC. Method:Totally 50 mice were randomly divided into 5 groups with 10 mice in each group， one group was randomly selected as blank group， and the other 4 groups were treated with dextran sodium sulfate （DSS） to induce UC model. After modeling， mice in the blank group and model group were given normal saline， and treatment groups were given Mesalazine （0.8 g·kg^-1）， Galla Chinensis decoction （1.8 g·kg^-1） and Baiyaojian decoction （2.7 g·kg^-1） by intragastric administration for 7 days. The 16S rRNA sequencing technology was used to detect the changes of intestinal flora in mouse feces. The histopathological changes of colon tissue were observed by hematoxylin-eosin （HE） staining， and the expression of Occludin and ZO-1 in colon tissue of mice were compared by immunohistochemistry. Result:Compared with the blank group， the abundance and diversity of intestinal flora in UC mice were significantly decreased， and the colonic tissue was thickened with congestion and obvious ulcers， and the expression levels of Occludin and ZO-1 were significantly decreased （<italic>P</italic><0.01）. After treatment with Galla Chinensis and Baiyaojian， the abundance and diversity of flora were improved. At the phylum level， relative abundance of Firmicutes， Proteobacteria and Actinobacteria increased significantly （<italic>P</italic><0.01）， while the relative abundance of Bacteroidetes decreased significantly （<italic>P</italic><0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of Proteobacteria and Actinobacteria increased significantly （<italic>P</italic><0.01）， while the relative abundance of Firmicutes increased and the relative abundance of Bacteroidetes decreased， but there was no significant difference. At the genus level， the relative abundance of <italic>Bacteroides</italic>， <italic>Allobaculum </italic>and <italic>Ruminococcus</italic> decreased significantly （<italic>P</italic><0.01）， the relative abundance of <italic>Roseburia</italic>， <italic>Prevotella</italic>， <italic>Oscillospira</italic> and <italic>Paraprevotella</italic> increased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01） in Galla Chinensis group. In Baiyaojian group， the relative abundance of <italic>Bacteroides</italic> and <italic>Allobaculum</italic> decreased significantly （<italic>P</italic><0.05， <italic>P</italic><0.01）， and the relative abundance of <italic>Prevotella</italic>， <italic>Oscillospira</italic>， <italic>Roseburia</italic> and <italic>Ruminococcus</italic> increased significantly （<italic>P</italic><0.01）. Compared with model group， colon tissue of Galla Chinensis group and Baiyaojian group was recovered obviously， congestion was alleviated， only scattered ulcers were seen. The expression of Occludin and ZO-1 increased， and the expression level of Baiyaojian group was higher than that of Galla Chinensis group. Conclusion:The effect of Baiyaojian is better than Galla Chinensis in the treatment of UC. The mechanism may be through regulating the abundance and diversity of intestinal flora， improving the disorder of intestinal flora and increasing the expression of ZO-1 and Occludin and protecting the intestinal mucosal barrier function for alleviating intestinal inflammation.

Dynamic Changes of Trace Elements in Yu Salvia miltiorrhiza in Different Growth Periods / 中国实验方剂学杂志

Objective:To establish an atomic absorption spectrometry to determine the content of eight trace elements of Yu Salvia miltiorrhiza in different growth stages，namely K，Cu，Na，Mg，Fe，Zn，Ca and Mn. Method:Micro-digestion-atomic absorption spectrometry was used to determine the trace elements in the roots of Yu S. miltiorrhiza. The HPLC method was used to determine the content of active constituents of Yu S. miltiorrhiza. Principal component analysis and correlation analysis were used to evaluate the results. Result:The contents of trace elements in Yu S. miltiorrhiza in different growth periods were significantly different. Cu had a significant positive correlation with the growth period，while Zn，Ca and Mn had significant negative correlations with the growth period. The comprehensive score of Yu S. miltiorrhiza in December was the best. The content of index components was negatively correlated with Mn，Zn and Ca，and positively correlated with Cu，Fe and Na. In soil，Mg，Fe，Ca and Mn were correlated with Zn，Ca and Mn，while Mn was negatively correlated with Cu. The content of K and Mg in the crude drug increased gradually with the change of the growth period，and the overall score of annual Yu S. miltiorrhiza was the best. Conclusion:The change of trace elements in Yu S. miltiorrhiza in different periods has certain regularity. Trace elements in soil have impacts on trace elements in medicinal materials. Trace elements in medicinal materials are closely correlated with index components and quality of medicinal materials.

Establishment of Thin Layer Chromatography Identification Method for Different Processed Products of Rehmanniae Radix / 中国实验方剂学杂志

Objective:To establish a thin layer chromatography （TLC） identification method for different processed products of Rehmanniae Radix. Method:Catalpol， D-fructose， sucrose， raffinose， stachyose， melibiose and manninotriose were used as control substances， and the effects of extraction solvents （water， 20% methanol， 50% methanol， 80% methanol）， developing solvents （n-butanol-methanol-chloroform-glacial acetic acid-water， ethyl acetate-pyridine-glacial acetic acid-water， n-butanol-glacial acetic acid-water）， color reagents （aniline-diphenylamine-phosphoric acid solution， ninhydrin solution）， sampling volumes （2， 4， 6 μL） and inspection conditions （sunlight， bottom lamp of sunlight， 365 nm， 254 nm） on TLC were investigated to determine the preparation method of sample solution of Rehmanniae Radix and Rehmanniae Radix Praeparata and the optimal TLC conditions. Result:High performance silica gel G plate was used for TLC， n-butanol-methanol-chloroform-glacial acetic acid-water （13∶5∶5∶1∶2） was used as developing agent， aniline-diphenylamine-phosphoric acid solution was sprayed and heated at 110 ℃ for color development， and then inspected under the bottom lamp of sunlight. The separation and color development of different processed products of Rehmanniae Radix were good with clear spots and good characteristics. Conclusion:The established TLC is simple and easy to operate with obvious qualitative characteristics and intuitive results. It can effectively identify different processed products of Rehmanniae Radix and provide experimental basis for determining the end point of processing of Rehmanniae Radix Praeparata.

Progress of Historical Evolution of Processing，Chemical Compositions and Pharmacological Effects of Foeniculi Fructus / 中国实验方剂学杂志

Foeniculi Fructus is warm and spicy in nature and belongs to the liver, kidney, spleen and stomach meridians with the function of dispersing cold and relieving pain, regulating Qi-flowing and harmonizing stomach, and can be used as medicine and food. This article summarized the historical evolution of processing of Foeniculi Fructus by consulting the relevant books since the Han dynasty, the Chinese Pharmacopoeia as well as the processing norms of provinces and cities, and summarized the chemical compositions and pharmacological effects of Foeniculi Fructus by reviewing literatures at home and abroad, then it found that Foeniculi Fructus mainly contains essential oil, flavonoids, phenols and fatty acids, has certain effects on human visceral system, central nervous system, cardiovascular system, endocrine system, immune system and chemotherapy, including the functions of regulating gastrointestinal function, analgesia, anti-inflammatory, etc. Among them, the main chemical components that play the pharmacodynamic effect are volatile oil, flavonoids and phenols, and the chemical components and pharmacological effects in Foeniculi Fructus before and after processing will have corresponding changes, which can provide reference for the further study of Foeniculi Fructus in processing principle, pharmacodynamic substance basis and pharmacological mechanism.

Research Progress on Chemical Constituents and Pharmacological Actions of Psoralea Fructus / 中国实验方剂学杂志

Psoralea Fructus,a commonly used traditional Chinese medicine in clinical application,is warm, spicy and bitter in nature and belongs to the kidney and spleen meridian. Psoralea Fructus has the function of warming kidney and tonifying yang,absorbing Qi and relieving asthma,warming spleen and stopping diarrhea,and in topical use, it can remove beverage. In recent years,considerable progress has been made in the study of chemical constituents and pharmacological effects of Psoralea Fructus. Nearly one hundred compounds have been isolated from it,including coumarins,flavonoids,and meroterpenes,and various types of compounds such as lipids,glycosides,volatile oils,and trace elements have been also found in Psoralea Fructus. In this article, the original literature was reviewed to summarize the main compound types and structural formula, with detailed reviews on pharmacological studies about its anti-tumor,anti-oxidant,anti-bacterial,anti-inflammatory,anti-depressive,regulating estrogen level,promoting bone growth,liver protection,and neuroprotection effects. In addition, the compounds with the same or similar pharmacological effects were sorted out and summarized to correlate the chemical structure and pharmacology. Psoralea Fructus and its modern preparations are widely used in modern clinical practice. The researches and papers on chemical constituents of Psoralea Fructus at home and abroad and the papers on pharmacological activities in recent years were reviewed,and the chemical types of individual compounds were corrected in this article,providing a reference for further study of Psoralea Fructus in efficacy material basis,quality standards and pharmacological activities.

Transformation and attribution of drug properties in Galla Chinesis fermented Baiyaojian based on cold and heat syndrome rats / 中国中药杂志

To reveal the transformation and attribution of drug properties in Galla Chinesis fermented Baiyaojian by studying the effect of Galla Chinesis and Baiyaojian on cold and heat syndrome rats. Euthyrox was used to induce the hyperthyrosis model,ice water stimulation was used to induce the cold syndrome model,and different concentrations of Galla Chinesis and Baiyaojian water decoction were administrated by gavage for 15 d continuously. Symptom indexes were evaluated,content of pyruvic acid( PA),ATPase activity in liver and contents of DA,T4,cAMP,5-HT,NE,17-OHCS,TRH and TSH in serum were assayed by enzyme linked immunosorbent assay and spectrophotometry. The rectal temperature,water consumption and body weight of heat syndrome rats in model group were increased,cAMP,NE,17-OHCS,TRH and PA were increased,TSH,Na-K ATPase and Ca-Mg ATPase were increased significantly( P<0. 01),while 5-HT was decreased,compared with those of the blank group( P< 0. 05),the contents of T4,DA,NE,TSH,TRH,cAMP and 17-OHCS were decreased significantly( P<0. 01),PA and Ca-Mg ATPase in WG and BG groups were decreased compared with those of the model group( P<0. 05),and the Galla Chinesis content of WG group was lower than that of BG group,while the contents of 5-HT in WG and BG groups were increased,and the Galla Chinesis content of WG group was higher than that of BG group,with no significant difference of viscera index between heat syndrome rats in blank group,model group and drug groups. The rectal temperature,water consumption and body weight of cold syndrome rats in model group were decreased,DA,T4,cAMP,NE,17-OHCS,TRH,TSH,PA,Na-K ATPase and Ca-Mg ATPase of rats in model group were decreased,whereas 5-HT was increased compared with those of the blank group( P<0. 05),the indexes of heart,lung and kidney were significantly higher than those in the blank group( P<0. 05). Both Galla Chinesis and Baiyaojian can significantly alleviate the symptoms of heat syndrome rats caused by levothyroxine sodium. Galla Chinesis has a better effect than Baiyaojian,but cannot alleviate the symptoms of cold syndrome caused by ice water stimulation,suggestting that the decoction of Galla Chinesis and Baiyaojian are both cold,but Galla Chinesis is colder than Baiyaojian. Cold property in Galla Chinesis fermented Baiyaojian can be relieved. In clinical application,the property of " slight cold" is more accurate than " neutral property" for Baiyaojian.

Processing of origin integration method and technology of Rehmanniae Radix / 中草药

Objective To study the processing of origin integration method and technology of Rehmanniae Radix (RR) and prove the feasibility of this new and innovative way. Methods The processing of origin integration technology of RR was selected with appearance and character as indexes, and thinking of appearance traits, content of catalpol, acteoside, rehmaionoside A, D and leonuride as an index, adopting comprehensive index scoring method to select the new processing of origin integration technology of RR. Results The optimized technology of the processing of origin integration of RR was as follows: out 2—6 cm diameters of fresh RR, baked completely at 75 ℃, piling up and sweating for 12 h, cutting 4—5 mm thick slices, dried at 75 ℃ up to 4—5 h, taking out and packing after cooling. Conclusion Adopting the processing of origin integration technology directly processes fresh RR into RR decoction pieces, which is simple as well as convenient, reducing the repeated processing and storage steps and achieving the aim of reducing the processing cost on Chinese herbal medicines and improving the quality of Chinese medicine decoction pieces.

Studies on the changes of 5-hydroxymethyl furfuraldehyde content in Radix Rehmanniae steamed for different time / 中国中药杂志

<p><b>OBJECTIVE</b>To find the changing regularity of 5-Hydroxymethyl furfuraldehyde (5-HMF) content in Radix Rehmanniae steamed for different time.</p><p><b>METHOD</b>HPLC method was used, with C18 column (6 mm x 200 mm, 5 microm), a mixture of methanol-water (15: 85) as the mobile phase. The flow rate was 1.0 mL x min(-1). The UV detection was setat 284 nm and the column temperature was set at 24 degrees C.</p><p><b>RESULT</b>The linear range was from 0.01 to 0.08 microg, r = 0.999 9 (n = 5). The average recovery was 100.2% and RSD was 2.7% (n = 5).</p><p><b>CONCLUSION</b>The 5-HMF content increases as the processing time prolongs in a certain time rang.</p>