Alkaloid uptake pathways in renal tubular epithelial cells from different processed products of Phellodendri chinensis Cortex.

This study aimed to investigate the absorption of alkaloids from Phellodendri chinensis Cortex (PC) by human renal tubular epithelial cells (HK-2). Cellular uptake and affinity ultrafiltration assays were employed to determine the alkaloid uptake pathway in HK-2 cells. Stemming from the hypothesis that salt-water processed PC introduces these alkaloids into the kidney at a cellular level, this research focused on different processed products of PC that are tailored for renal targeting. Utilizing the UPLC-QqQ-MS method, we quantified variations in the uptake capacity of phellodendrine, magnoflorine, jatrorrhizine, berberrubine, and berberine from raw Phellodendri chinensis Cortex (RPC), salt-water processed Phellodendri chinensis Cortex (SPC), and wine processed Phellodendri chinensis Cortex (WPC) in HK-2 cells. This study also tracked the concentration changes of these five alkaloids in HK-2 cells during the administration phase. Further, we evaluated the influence of two inhibitors on the absorption of these five alkaloids from PC and its processed products into HK-2 cells: the organic anion transporters (OATs) inhibitor-probenecid (PRO), and the organic cationic transporters (OCTs) inhibitor-tetraethylammonium chloride (TEAC). A pivotal component of this research was an investigation into the effects of PC and its processed products on the expression levels of OCT2, OAT1, and OAT3 proteins in HK-2 cells, facilitated by Western blot analysis. Finally, we appraised the binding affinity of PC's alkaloids to OCT2, OAT1, and OAT3 proteins using an ultrafiltration centrifugation technique. The uptake of different processed products of PC by HK-2 cells showed the following trend: SPC group > RPC group > WPC group. When considering inhibitor uptake in HK-2 cells, the group treated with PRO (an OATs inhibitor) demonstrated a higher uptake than the group treated with TEAC (an OCTs inhibitor). It was observed that different processed products of PC elevated the expression of OCT2 and OAT1 proteins in HK-2 cells. Specifically, both the SPC and berberrubine groups displayed enhanced expression of these proteins, with a marked increase noted for OCT2. Through affinity ultrafiltration assays, it was determined that the binding affinity of alkaloids from different processed products of PC to OCT2 and OAT1 significantly exceeded that to OAT3. These results indicate that PC-derived alkaloids are absorbed by HK-2 cells, predominantly through transport mechanisms mediated by OCT2 and OAT1, with OCT2 serving as the dominant transporter. The higher intake of alkaloids in SPC group can likely be linked to the amplified activity of kidney uptake transporters.

[Slicing specifications in general rule of processing in Chinese Pharmacopoeia from slicing development of Chinese materia medica processed product and thinking and suggestion on slicing techniques].

Slicing is critical in the processing of Chinese materia medica(CMM) processed product and the specification(thickness) is closely related to the quality of the decoction. On the basis of clarifying the concept and evolution of slicing of CMM processed product by reviewing the Chinese herbal classics of the past dynasties and general rules of local processing standards, this study discussed the development history of slicing specifications in general rules of Chinese Pharmacopoeia(2020 edition), analyzed the current situation and key problems, and proposed the thinking and suggestion on promoting the sound development of slicing of CMM processed product. Since 2000, the slicing thickness of CMM processed product in the general rules of local CMM processed product processing specifications newly revised and issued by 27 provinces, autonomous regions, and municipalities has been consistent with that in the general rules of the Chinese Pharmacopoeia(2020 edition). The standard that the thickness of extremely thin pieces is less than 0.5 mm is rarely retained, and the pieces in 0.5-1 mm thickness have not been found on the market, which is consistent with the provisions of the general rules of the Chinese Pharmacopoeia. This study can provide a historical and modern basis for the rationality of slicing of CMM processed product.

Cornus officinalis prior and post-processing: Regulatory effects on intestinal flora of diabetic nephropathy rats.

Background: Diabetic nephropathy (DN) is one of the most common and serious chronic complications in the clinic. Cornus officinalis has the effects of replenishing qi and nourishing yin, tonifying liver and kidney, and it is one of the main traditional Chinese medicines used clinically to treat diabetes and its complications. However, the effect and mechanism of Cornus officinalis before and after processing on intestinal flora of diabetic nephropathy need to be further elucidated. Methods: SD rats were randomly divided into a blank group (10 rats) and DN groups (70 rats). After 4 weeks of high-sugar and high-fat diet, the DN rat model was established by intraperitoneal injection of streptozotocin. After successful modeling, the rats were randomly divided into DN model group, irbesartan group (1.35 mg·kg-1), Cornus officinalis group (281.25 mg·kg-1), wine Cornus officinalis group (281.25 mg·kg-1), wine honey Cornus officinalis group (281.25 mg·kg-1), auxiliary wine group (10 ml·kg-1), auxiliary wine honey group (10 ml·kg-1). During the observation of the rats' general state, after 6 weeks of continuous administration, the fasting blood glucose of rats in each group was detected, and the kidney index was calculated. The serum creatinine levels, urea nitrogen and 24 h urinary microalbumin were detected by enzyme-linked immunosorbent assay. The expression levels of YKL-40, Wnt4, ß-catenin and TGF-ß 1 mRNA in renal tissue were detected by fluorescence quantitative PCR. Hematoxylin-eosin staining was used to observe the changes in renal pathological injury in each group; GC-MS detected the changes of short chain fatty acid content. Feces were collected for 16 s high-throughput sequencing to analyze the effects of Cornus officinalis on the diversity of intestinal flora in DN before and after processing. Results: Compared with the blank group, the serum creatinine, urea nitrogen, 24 h urinary microalbumin, kidney index and fasting blood glucose in the DN model group were significantly increased (p < 0.05). The renal tissue morphology was disordered and a large number of inflammatory cells were infiltrated. The expression of YKL-40, Wnt4, ß-catenin and TGF-ß 1 mRNA was significantly increased (p < 0.05). Compared with the DN model group, the serum creatinine, urea. Nitrogen, 24 h urine microalbumin, kidney index and fasting blood glucose of rats in each administration group were significantly decreased (p < 0.05), and the general condition and pathological renal damage of DN rats were improved. The effect of wine honey Cornus officinalis was the best, and the expression of YKL-40, Wnt4, ß-catenin and TGF-ß 1 mRNA was significantly decreased (p < 0.05). In each administration group, the improvement of the above indicators in the wine honey Cornus officinalis group was significantly better than that in the raw Cornus officinalis group and wine Cornus officinalis group (p < 0.05), There was no significant difference compared with the irbesartan group (p > 0.05). Each administration group had a significant callback effect on the content of short-chain fatty acids in rat feces, with increased intestinal beneficial bacteria and decreased pathogenic bacteria. Compared with the blank group, the abundance of Firmicutes in the DN model group increased, the abundance of Bacteroidetes decreased, and the ratio showed an upward trend in the DN model group decreased. Each administration group could improve the relative abundance of the above intestinal flora in the model group to varying degrees. Conclusion: The processing of Cornus officinalis may improve the renal injury of DN rats by blocking the activation of Wnt/ß-catenin signaling pathway, regulating the structural composition of intestinal microorganisms, and ultimately playing a role in renal protection.

[Comparison of 12 constituents of Astragali Radix directionally processed with organic acid by UPLC-MS].

To establish an UPLC-MS method for the simultaneous content determination of 4 saponins and 8 flavonoids, in order to analyze the effect of Astragali Radix directionally processed with organic acid on the content of glycosides and aglycones. The separation was carried out on ACQUITY UPLC HSS T_3(2.1 mm×100 mm, 1.8 µm), the mobile phase was eluted with the mixture of 0.1 mol·L~(-1) formic acid water solution and 0.1 mol·L~(-1) formic acid acetonitril in a gradient mode. The detection wavelength was 260 nm, the flow rate was 0.5 mL·min~(-1), the column temperature was 30 ℃, and the injection volume was 2 µL. Mass spectrometry analysis was performed with an electrospray ionization(ESI) source in a positive ion mode. The 12 constituents showed good linear relations within their own ranges(R~2≥0.999 2),with good average recoveries. The results showed no significant change in saponins but both qualitative and quantitative changes in flavonoids after directional processing of Astragali Radix with organic acid. The established method can provide methodological reference for analyzing the effect of Astragali Radix directionally processed with organic acid on glycosides and aglycones.

Determination of orcinol glucoside by LC-MS in Curculigo orchioides and its application to a pharmacokinetic study

ABSTRACT This study was designed to explore the pharmacokinetic regularity of the plasma concentration, tissue distribution and excretion of orcinol glucoside from aqueous extracts of raw and processed Curculigo orchioides Gaertn., Hypoxidaceae. The experiment first used an ultrahigh-performance liquid chromatography-tandem mass spectrometry approach with multiple reaction monitoring and a positive mode to separate orcinol glucoside from naringin to obtain the plasma concentration curves, bar graph of tissue distribution and excretion curves. These results might be beneficial for reasonable clinical application of C. orchioides and for further development of its wine and salt-processing mechanism.

Simultaneous Determination of Five Chromones of Radix Saposhnikoviae Extract in Rat Plasma by UPLC-MS/MS: Application to a Comparative Pharmacokinetic Study in Normal and Febrile Rats.

A rapid and sensitive quantitative analytical method was established for the simultaneous determination of five chromones (prim-O-glucosylcimifugin, cimifugin, 4'-O-ß-D-glucosyl-5-O-methylvisamminol, 5-O-methylvisammiol, and sec-o-glucosylhamaudol) in the plasma of RS-treated rats for the first time using ultra performance liquid chromatography- (UPLC-) tandem mass spectrometry. The Waters Acquity UPLC BEH C18 (50 mm × 2.1 mm, 1.7 µm) was used as the chromatographic column, 0.1% formic acid water and 0.1% formic acid acetonitrile comprised the mobile phases, and all samples were determined under positive ion mode. The results showed that all analytes had good linearity (r > 0.9902), between-day and within-day precisions less than 15%, accuracy between -5.50% and 5.53%, and extraction recovery between 88.26% and 97.65%. Both the matrix effect and stability met the requirements. This method was successfully applied for the comparative pharmacokinetics of five active components of RS in normal and febrile rats. The results showed that the pharmacokinetic behavior of RS extract significantly differed between the two types of rats.

Endothelium corneum gigeriae galli extract inhibits calcium oxalate formation and exerts anti-urolithic effects.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional Chinese Medicine is preferred because of its safety and minimal/reduced side effects. Endothelium Corneum Gigeriae Galli (ECGG) extract, a traditional Chinese drug consisting of the dried gizzard membrane of Gallus gallus domesticus Brisson, was assessed for its effects and mechanism on urolithiasis. AIMS OF STUDY: To evaluate the effects of ECGG extract on calcium oxalate (CaOx) crystal formation in vitro, and assess the anti-urolithic effects of ECGG extract in vivo and explore the underlying mechanism. MATERIALS AND METHODS: In vitro, CaOx crystals were treated with ECGG extract (0.05, 0.2, and 0.8 g/mL), and assessed by scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray powder diffraction and electrical conductivity. Then, a rat model of renal calculi was established by ethylene glycol and ammonium chloride treatment, and ECGG extract (5.0, 10.0 and 20.0 g/kg) was administered orally. After treatment, urine, serum and kidney bioindicators were analyzed, as well as kidney's pathological features. RESULTS: In the presence of ECGG extract, calcium oxalate dihydrate (COD) crystals with typical tetragonal bipyramidal morphology were obtained; meanwhile, the formation of calcium oxalate monohydrate (COM), a major urinary stone component, was inhibited; in addition, the equilibration time of the chemical reaction of Ca2+ and C2O42- ions was delayed in a concentration dependent manner. ECGG extract actually showed anti-urolithic effects; the incidence rates of crystal formation in the kidney in the model, low, middle and high dose groups were 100%, 90%, 70% and 60%, respectively, with a dose-dependent alleviation of kidney stone amounts and kidney damage. Treatment with middle and high ECGG extract doses significantly decreased urine uric acid and oxalic acid amounts, serum creatinine, urea nitrogen and uric acid contents, and kidney tissue oxalic acid and calcium levels, while increasing kidney and urinary magnesium and superoxide dismutase levels (P < 0.05). CONCLUSION: ECGG extract has outstanding anti-urolithic effects, potentially with included bioorganic molecules inducing COD crystal nucleation and growth. Therefore, ECGG extract is a promising drug for preventing and treating urolithiasis.

Metabolomics analysis and rapid identification of changes in chemical ingredients in crude and processed Astragali Radix by UPLC-QTOF-MS combined with novel informatics UNIFI platform.

Astragali Radix, the root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge., is widely used as a tonic decoction pieces in the clinic of traditional Chinese medicine (TCM). Astragali Radix has various processed products with varying pharmacological actions. There is no modern scientific evidence to explain the differences in pharmacological activities and related mechanisms. In the present study, we explore the changes in chemical components in Astragali Radix after processing, by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with novel informatics UNIFI platform and multivariate statistical analysis. Our results showed that the crude and various processed products could be clearly separated in PCA scores plot and 15 significant markers could be used to distinguish crude and various processed products by OPLS-DA in UNIFI platform. In conclusion, the present study provided a basis of chemical components for revealing connotation of different processing techniques on Astragali Radix.

A Crude 1-DNJ Extract from Home Made Bombyx Batryticatus Inhibits Diabetic Cardiomyopathy-Associated Fibrosis in db/db Mice and Reduces Protein N-Glycosylation Levels.

The traditional Chinese drug Bombyx Batryticatus (BB), which is also named the white stiff silkworm, has been widely used in Chinese clinics for thousands of years. It is famous for its antispasmodic and blood circulation-promoting effects. Cardiomyocyte hypertrophy, interstitial cell hyperplasia, and myocardial fibrosis are closely related to the N-glycosylation of key proteins. To examine the alterations of N-glycosylation that occur in diabetic myocardium during the early stage of the disease, and to clarify the therapeutic effect of 1-Deoxynojirimycin (1-DNJ) extracted from BB, we used the db/db (diabetic) mouse model and an approach based on hydrophilic chromatography solid-phase extraction integrated with an liquid Chromatograph Mass Spectrometer (LC-MS) identification strategy to perform a site-specific N-glycosylation analysis of left ventricular cardiomyocyte proteins. Advanced glycation end products (AGEs), hydroxyproline, connective tissue growth factor (CTGF), and other serum biochemical indicators were measured with enzyme-linked immunosorbent assays (ELISA). In addition, the α-1,6-fucosylation of N-glycans was profiled with lens culinaris agglutinin (LCA) lectin blots and fluorescein isothiocyanate (FITC)-labelled lectin affinity histochemistry. The results indicated that 1-DNJ administration obviously downregulated myocardium protein N-glycosylation in db/db mice. The expression levels of serum indicators and fibrosis-related cytokines were reduced significantly by 1-DNJ in a dose-dependent manner. The glycan α-1,6-fucosylation level of the db/db mouse myocardium was elevated, and the intervention effect of 1-DNJ administration on N-glycan α-1,6-fucosylation was significant. To verify this result, the well-known transforming growth factor-ß (TGF-ß)/Smad2/3 pathway was selected, and core α-1,6-fucosylated TGF-ß receptor II (TGFR-ßII) was analysed semi-quantitatively with western blotting. The result supported the conclusions obtained from LCA lectin affinity histochemistry and lectin blot analysis. The expression level of α-1,6-fucosyltransferase (FUT8) mRNA was also detected, and the results showed that 1-DNJ administration did not cause obvious inhibitory effects on FUT8 expression. Therefore, the mechanism of 1-DNJ for relieving diabetic cardiomyopathy (DCM)-associated fibrosis can be concluded as the inhibition of N-acetylglucosamine (N-GlcNAc) formation and the reduction of substrate concentration.

[Determination of four coumarin constituents before and after Angelicae Dahuricae Radix stewed with yellow rice wine and research on its mutual transformation mechanism].

To determine the contents of oxypeucedanin, oxypeucedanin hydrate, byakangelicol and byak-angelicin both before and after Angelicae Dahuricae Radix was stewed with yellow rice wine by high-performance liquid chromatography, and study the mutual transformation mechanisms of oxypeucedanin into oxypeucedanin hydrate, as well as byakangelicol into byak-angelicin. The research results indicated that the contents of oxypeucedanin and byakangelicol were decreased, but the contents of oxypeucedanin hydrate and byak-angelicin were increased after Angelicae Dahuricae Radix was processed with yellow rice wine. The contents' changes of these chemical compounds were due to the ring opening reaction of epoxy compounds, such as oxypeucedanin and byakangelicol under the weak acidity and heating conditions of yellow rice wine. This research could provide a scientific basis for the processing mechanism of Angelicae Dahuricae Radix with yellow rice wine stewing.

[Study on Variation of Gastroprotective Effect of Atractylodes lancea Rhizome Before and After Processing].

Objective: To compare the gastroprotective effect between the crude and bran-processed Atractylodes lancea rhizome in the rat model of gastric ulcer. Methods: The model of gastric ulcer was imitated by acetic acid in rats. The levels of epidermal growth factor( EGF) and trefoil factor 2( TFF2) in serum and stomach were determined by the double-antibody sandwich enzyme-linked immunosorbent assay( ELISA),and the mRNA expressions of EGF and TFF2 in stomach were analyzed by quantitative real-time reverse transcription polymerase chain reaction( RT-PCR). The protein expressions of EGF and TFF2 in stomach were examined by immunohistochemistry. Results: The productions of EGF,TFF2 and the mRNA expressions of EGF and TFF2 were notably up-regulated by both crude Atractylodes lancea rhizome and bran-processed Atractylodes lancea rhizome. Compared with the crude Atractylodes lancea rhizome,the processed Atractylodes lancea rhizome was more effective. Conclusion: The gastroprotective effect of Atractylodes lancea rhizome was enhanced after stir-frying with wheat bran in the rat model of gastric ulcer induced by acetic acid.

[Effect of Morinda officinalis Root and Its Salt-Processing Product on Energy Metabolism in Rats with Yang Deficiency].

Objective: To compare the effects of Morinda officinalis root and its salt-processing prodduct on rats with kidney yang deficiency from the viewpoint of energetic metabolism. Methods: The rat models for the kidney yang deficiency were established through the perfusion of methimazole. The models were treated with different dosages of Morinda officinalis root without woods and its salt-processing products extracts. The activity of liver glycogen,muscle glycogen,free fatty acid( NEFA),lipoprotein lipase( LPL),hepatic lipase( HL),liver and skeletal muscle Na+,K+ATPase,Ca2 +,Mg2 +ATPase,succinodehydrogenase( SDH) were measured by colormetric methods,and the coefficient of body fat were observed by weighing method. Results: Morinda officinalis root without woods and its saltprocessing products could improve the energetic metabolism of kidney yang deficiency of rats. The effects of salt-processing products were better than Morinda officinalis root without woods in the same dosage. Conclusion: The effect of invigorating the kidney and strengthening yang of Morinda officinalis were correlated with energetic metabolism. The correct processing-products of Morinda officinalis root should be chosen with the suitable measurements in clinic.

[Comparasion of Antis Egg-case Before and After Preparing in Reproducing Kidney-Yang Deficiency Model Rats].

Ojective: To compare the effects of antis egg-case before and after preparing in reproducing kidney-Yang deficiency model rats,and to explore the best medicinal part of antis egg-case. Methods: Divide the rats into nine groups randomly: antis egg-case group, salt fried antis egg-case group, steamed antis egg-case group, eggs from antis egg-case group, eggs from salt fried antis egg-case group, eggs from steamed antis egg-case group, egg shell from antis egg-case group, egg shell from salt fried antis egg-case group, and egg shell from the steamed antis egg-case group. Make kidney-Yang deficient model by giving the rats hydrocortisone. Two weeks later, give the rats treatment for four weeks, compare the effects of antis egg-case by determining the contents of NO,TSH,T3,T4,EPI,NE,17-OH,CORT,T,E2,coefficient of thyroid, body temperature, coefficient of adrenal and kidney, weight and water intake. Results: Antis egg-case before and after preparing increased the contents of T3,T4,EPI,NE and T, increased weight, body temperature, coefficient of thyroid and adrenal, but reduced the contents of NO and E2,the coefficient of kidney and water intake were also decreased. Conclusion: The medicinal materials came from different medicinal parts of antis egg-case before and after preparing can treat the symptoms of kidney-Yang in different degrees. The effects of antis egg-case to treat kidney-Yang deficient has been strengthened after preparing, salt fried antis egg-case and eggs from salt fried antis egg-case played the more important role.

[Investigation of potential toxic factors for fleece-flower root： from perspective of processing methods evolution].

In recent years, the rapid growth of reports on fleece-flower root-caused liver damages has drawn wide attention of both at home and abroad, however, there were rare literature on toxicology of fleece-flower root in ancient Chinese medicine. But why there are so many reports on toxicology of fleece-flower root now compared with the ancient literature? As a typical tonic medicine, the clinical utility of fleece-flower root was largely limited by its standardization and reliability of processing methods in ancient Chinese medicine. The ancient processing methods of fleece-flower root emphasized nine times of steaming and nine times of drying, while the modern processes have been simplified into one time of steaming. Whether the differences between ancient and modern processing methods are the potential cause of the increased events of fleece-flower root-caused liver damages. We will make deep analysis and provide new clues and perspectives for the research on its toxicity. This article, therefore, would discuss the affecting factors and key problems in toxicity attenuation of fleece-flower root on the basis of sorting out the processing methods of fleece-flower root in ancient medical books and modern standards, in order to provide the reference for establishing specification for toxicity attenuation of fleece-flower root.

[Comparison of anti-inflammatory activity between crude Atractylodes lancea and their processed products by stir-baking with bran in rat models of gastric ulcer].

To compare the anti-inflammatory activity of the crude Atractylodes lancea (AL) and AL processed products by stir-baking with bran in rat models of gastric ulcer, and preliminarily explore the anti-ulcer mechanisms of AL, the model of gastric ulcer was imitated by local acetic acid injection into gastric mucosa in rats by surgery according to the modified Okabe method. All rats were randomly divided into the following 10 groups： sham-operation group, model group, omeprazole group, Sanjiu Weitai granule group, crude AL low dose group, crude AL middle dose group, crude AL high dose group, processed AL low dose group, processed AL middle dose group, and processed AL high dose group. Rats were administered via intragastric (ig) two times each day, for 10 consecutive days. Blood was collected from the abdominal aorta, serum was separated, and the ulcer tissues were taken. The levels of inflammatory factors interleukin 6, 8 (IL-6, 8), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) in serum and gastric tissues were determined by enzyme-linked immunosorbent assay (ELISA), and the mRNA expressions of TNF-α and IL-8 in gastric tissues were detected by quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of TNF-α and IL-8 in gastric tissues were detected by immunohistochemistry. Compared with sham-operation group, the levels of TNF-α, IL-8, IL-6, PGE2 as well as the mRNA expressions and protein expressions of TNF-α, IL-8 in gastric tissues were significantly higher in model group. The above levels were reduced in different degrees in all treatment groups. Compared with the crude AL, same dose of processed AL was more effective in decreasing the levels of TNF-α, IL-8, IL-6, PGE2 in serum and gastric tissues and down-regulating the mRNA expressions of TNF-α and IL-8 in gastric tissues, with significant difference in middle dose groups and high dose groups. The results showed that AL had potent anti-inflammatory effects in rat models of gastric ulcer induced by acetic acid, and the processed AL had more obvious effect. The anti-ulcer action of AL could be attributed partly to down-regulating the levels of TNF-α, IL-8, IL-6 and PGE2.

[Study on protective effect of Sijunzi decoction (red/white ginseng) on H2O2-induced H9c2 cardiomyocytes apoptosis].

In this study, efforts were made to screen out the drug concentration of Sijunzi decoction (red ginseng) for in vitro intervention of H9c2 cardiomyocytes, select high, medium and low groups for subsequent experiments, establish the H2O2-induced myocardial cell apoptosis to investigate the protective effect of Sijunzi decoction (white/red ginseng), provide reference ginseng ingredients in Sijunzi decoction used to treat ischemic heart disease and reflect its curative effect, and observe its impacts on SOD, MAD, LDH and other indexes to preliminarily define the action mechanism. According to the results, red ginseng in Sijunzi decoction showed a better protective effect on H2O2-induced myocardial cell injury than that of white ginseng. Both of them could enhance SOD activity and reduce MDA production and LDH release, so as to significantly reduce the amount of apoptotic myocardial cells and play protective role.

[Determination of Five Active Components in Morinda officinalis from Different Habitats by HPLC].

OBJECTIVE: To establish a sensitive and specific high performance liquid chromatography (HPLC) method for simultaneous determination of alizarin anthraquinones (1-methoxy-2-hydroxy anthraquinone,1,2-dimethoxy-3-hydroxy anthraquinone, rubiadin-1- methylether, 1,3-dihydroxy-2-methoxy anthraquinone, rubiadin) in Morinda officinalis from different habitats. METHODS: The analysis was carried out on an Ecosil C18 column (250 mm x 4.6 mm, 5 µm) and detected with an UV detector at the wavelength of 277 nm. Gradient elution was carried out with acetonitrile-0.2% phosphoric acid at the flow rate of 0. 8 mL/min. The column temperature was set at 30 °C. RESULTS: The calibration curves were linear in the range of 0.2856-34.27 g/mL for 1-methoxy-2-hydroxy anthraquinone (r = 0.9999), 0.3268-39.22 g/mL for 1,2-dimethoxy-3-hydroxy anthraquinone (r = 0.9999), 0.3450-41.40 µg/mL for rubiadin-1- methylether (r = 0.9999), 0.1248-14. 98 µg/mL for 1,3-dihydroxy-2-methoxy anthraquinone (r = 0.9999) and 0.0508-6.096 µg/ mL for rubiadin (r = 0.9994), respectively. The average recoveries of the five components were 99.4%, 100.2%, 101.4%, 97.2%, 103.2%, respectively. The content of active components of samples from 10 different hatitats were in the range of 0.0025-0.0722 mg/ g, 0.0016-0.0658 mg/g, 0.0022-0.0684 mg/g, 0.0182-0.3965 mg/g and 0. 0014-0.0179 mg/g. CONCLUSION: The established method is accurate, reliable, and can be used for the simultaneous determination of the five components in Morinda officinalis, which provides a scientific basis for the quality evaluation of Morinda officinalis.

[Application of Cocktail Probe Drugs for Detecting Influences of Raw and Processed Phellodendri Cortex on Cytochrome P450 Isoforms].

OBJECTIVE: To research and compare the influences of raw and processed Phellodendri Cortex on the cytochrome P450 four isoforms by Cocktail probe drugs, and to explore the processing principle of Phellodendri Cortex. METHODS: SD rats were randomly divided into raw group,processed with rice-wine group, processed with salt-water group and blank control group, which were given raw decoction, processed with rice-wine decoction, processed with salt-water decoction (3.24 g/kg) and normal saline respectively for one week, then given the mixture of four probe drugs on the 8th day, and soon after the blood samples were obtained through the orbits at a series of time-points. HPLC method was used to determine the concentrations of probe drugs in rat plasma, and pharmacokinetic parameters were estimated by DAS3.0. The effect of raw and processed Phellodendri Cortex on cytochrome P450 were judged indirectly by the pharmacokinetic parameters. RESULTS: Compared with the blank control group, the t½ significantly increased of theophylline in raw and processed with salt-water group. The CL/F significantly decreased and AUC(0-t) AUC(0-∞). significantly increased of theophylline in raw and processed with rice-wine groups. The t(½) AUC(0-∞) and AUC(0-∞) significantly decreased and CL/F significantly increased of dapsone in raw, processed with rice-wine and processed with salt-water group. The AUC(0-t) significantly increased of chlorzoxazone in raw and processed with salt-water group. The t(½), AUC(0-∞). and AUC(0-t) significantly decreased and CL/F significantly increased of chlorzoxazone in processed with rice-wine group. The AUC(0-t), significantly decreased of tolbutamide in raw, processed with rice-wine and processed with salt-water groups. CONCLUSION: The raw Phellodendri Cortex can inhibit CYP1A2, induce CYP3 A4 and also is need to make a further research work on CYP2C9 and CYP2E1. Meanwhile, it also can change the activities of cytochrome P450 after processed with rice-wine and salt-water. The Phellodendri Cortex processed with rice-wine can reduce the inhibitory effect of CYP1A2 and enhance induction of CYP3A4, it provides reference and basis to make an interpretation about Phellodendri Cortex processed with rice-wine.

[Effects of Processing on Phellodendri Chinensis Cortex Based on Material and Energy Metabolism].

OBJECTIVE: To compare the pharmacodynamic indicators of Phellodendri Chinensis Cortex with its processed products, in order to study the processing mechanism. METHODS: The effects of raw Phellodendri Chinensis Cortex and its processed products on 17 kinds of indicators in rats were compared, such as the biological characteristics (weight and rectal temperature), material metabolism (lactic acid, acetone acid and triglyceride), energy metabolism (Na+ - K+ -ATPase, Ca2+ -Mg2+ -ATPase, LDH, SDH and liver glyco- gen), thyroid function axis (T3, T4, TSH and TRH), cyclic nucleotide ( cAMP, cGMP and cAMP/cGMP value). RESULTS: Phellodendri Chinensis Cortex and its salt processing product changed the indicators including weight, rectal temperature, material metabolism, energy metabolism, thyroid function shaft and cyclic nucleotide in rats. The effect of salt processing product was better than that of the raw one, but wine processing product had little effects. CONCLUSION: The "cold" influence of Philodendri Chinensis Cortex is enhanced after pro- cessing with salt,but induced after processing with wine.

[Effect of Different Processed Products and Extracts of Morinda officinalis Root on Adjuvant-Induced Arthritis in Rats].

OBJECTIVE: To study the therapeutic effects of different processed products and extracts of Morinda officinalis root in rat adjuvant-induced arthritis (AA) model, and to explore the possible mechanism. METHODS: AA model in SD rats was established, then the arthritic rats were randomly divided into seven groups such as model, Tripterygium Glycosides Tablet (TGT), morinda with xylem, morinda, salt-steamed morinda, licorice-processed morinda, morinda EtOAc extract, morinda n-BuOH extract, H2O residue, total polysaccharides and control groups. The paw edema and arthritis indexes were observed, and the contents of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6, IL-2 and INF-γ in AA rats serum were detected by ELISA. RESULTS: Morinda processed by salt and n-BuOH extract could reduce paw swelling and levels of TNF-α, IL-1ß, IL-6 and INF-γ in AA rats serum, which followed by morinda EtOAc extract. The differences were significant compared with the model group (P < 0.05 or P < 0.01). CONCLUSION: Salt-steamed morinda and morinda n-BuOH extract have the best curative effect, followed by morinda EtOAc extract. Reducing TNF-α, IL-1ß, IL-6 and INF-γ contents in serum may be the mechanism of drug treatment of rheumatoid arthritis.