Pharmacokinetics and tissue distribution of four alkaloids in Ermiao Pills and Sanmiao Pills in normal and arthritic model rats / 中国中药杂志

This work aimed to investigate the differences of pharmacokinetics and tissue distribution of four alkaloids in Ermiao Pills and Sanmiao Pills in normal and arthritic model rats. The rat model of arthritis was established by injecting Freund's complete adjuvant, and ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) in the positive ion multiple reaction monitoring(MRM) mode was used for the determination of four alkaloids in plasma and tissues of normal and arthritic rats after administration of Ermiao Pills and Sanmiao Pills, respectively. The differences in pharmacokinetics and tissue distribution of the four active components were compared, and the effect of Achyranthis Bidentatae Radix on the major components of Sanmiao Pills was explored. This study established an UPLC-MS/MS for simultaneous determination of four alkaloids, and the specificity, linearity, accuracy, precision, and stability of this method all met the requirements. Pharmacokinetics study found that as compared with normal rats, the AUC and C_(max) of phellodendrine, magnoflorine, berberine and palmatine in model rats were significantly decreased after administration of Ermiao Pills, the clearance rate CL/F was significantly increased, and the distribution and tissue/plasma concentration ratio of the four alkaloids in the liver, kidney, and joint were significantly reduced. Achyranthis Bidentatae Radix increased the AUC of phellodendrine, berberine, and palmatine, reduced the clearance rate, and significantly increased the distribution of the four alkaloids in the liver, kidney, and joints in arthritic rats. However, it had no significant effect on the pharmacokinetics and tissue distribution of the four alkaloids in normal rats. These results suggest that Achyranthis Bidentatae Radix may play a guiding role in meridian through increasing the tissue distribution of effective components in Sanmiao Pills under arthritis states.

Effect of Tongxie Yaofang with or Without Saposhnikoviae Radix on Water Metabolism and 5-HT Pathway in Irritable Bowel Syndrome Rats / 中国实验方剂学杂志

Objective:To investigate the effect of Saposhnikoviae Radix on Tongxie Yaofang in regulating water metabolism and 5-serotonin（5-HT） signal system in diarrhea type irritable bowel syndrome（IBS-D）rats. Method:The 40 IBS-D SD rats were randomly divided into model group, Tongxie Yaofang wituout Saposhnikoviae Radix group (26 g·kg-1), Tongxie Yaofang decoction group (30 g·kg-1), Tongxie Yaofang with double Saposhnikoviae Radix group (34 g·kg-1),another 10 normal SD rats were selected as the normal group.Except for normal group, the rats in other groups were separated from their mothers and induced by acetic acid to establish D-IBS model. These rats in the each group were administered with corresponding drugs for 14 days. The diarrhea indexs and the water contents of the feces were observed and calculated. The Na+-K+-ATPase activities in the intestinal mucosa were detected by micro method. The contents of 5-HT were detected by enzyme linked immunosorbent assay (ELISA). The activities of monoamine oxidase A (MAO-A) were detected by chemiluminescence method. The expressions of aquaporin 4 (AQP4) in colon were detected by immunohistochemistry. Protein expression levels of HT receptor 3 (5-HT3R), HT receptor 4 (5-HT4R) and serotonin transporter (SERT) in hypothalamus and colon were detected by Western blot. Result:Compared with normal group, the fecal water contents, contents of 5-HT in hypothalamus and colon, activities of MAO-A, expressions of 5-HT3R were significantly increased in model group (P<0.01), and the activities of Na+-K+-ATPase, expressions of AQP4, 5-HT4R and SERT were reduced significantly (P<0.01). Compared with model group, the diarrhea indexs and fecal water contents, contents of 5-HT in hypothalamus and colon, activities of MAO-A, expressions of 5-HT3R were significantly decreased in each experimental group, and the activities of Na+-K+-ATPase, expressions of AQP4,5-HT4R and SERT were significantly increased(P<0.05，P<0.01),however, there was no significant difference in the expression of 5-HT3R protein in Tongxie Yaofang wituout Saposhnikoviae Radix group.Compared with Tongxie Yaofang wituout Saposhnikoviae Radix group,the diarrhea index, fecal water content, 5-HT content, MAO-A enzyme activity, and protein expression of 5-HT3R were significantly decreased in Tongxie Yaofang group and Tongxie Yaofang with double Saposhnikoviae Radix group(P<0.05，P<0.01), and the activity of Na+-K+-ATPase and protein expression of SERT were significantly increased(P<0.05，P<0.01). Conclusion:Saposhnikoviae Radix can enhance the effects of Tongxie Yaofang on improving the water metabolism of rats with IBS-D and regulating the multi-target of 5-HT signaling system, further confirming the synergistic effect of Saposhnikoviae Radix.

Study on Effect and Mechanism of Huayu Jiedu Recipe on Apoptosis of Gastric Cancer Cells / 中国实验方剂学杂志

Objective:To observe the effect of Huayu Jiedu recipe （HYJDR） on apoptosis of gastric cancer cells， explore the mechanism of HYJDR on apoptosis of gastric cancer cells and provide experimental basis for clinical application of HYJDR. Method:Thiazolyl blue tetrazolium bromide （MTT） method was used to detect the effect of HYJDR （5，10，15，20，25，30 g·L-1） on the activity of SGC-7901 cells and the median inhibition concentration （IC50） of HYJDR on SGC-7901 cells. Propidium iodide （PI） and Hoechst double fluorescence staining were used to detect the effect of HYJDR on the apoptosis of gastric cancer SGC-7901 cells. Western blot was used to detect the effect of HYJDR on the expression of apoptosis related proteins in gastric cancer cells. Real-time polymerase chain reaction （Real-time PCR） was used to detect the effect of HYJDR on the expression of apoptosis related protein mRNA in gastric cancer cells. Result:As compared with blank group， HYJDR （>10 g·L-1） can significantly inhibit the proliferation of gastric cancer cells. With the increase of the concentration of HYJDR， the cell viability decreased significantly obviously in a concentration-dependent manner（P<0.05）. HYJDR can significantly promote the apoptosis of gastric cancer cells. With the increase of the concentration of HYJDR， the apoptosis of gastric cancer cells gradually increased. As compared with blank group， HYJDR can obviously inhibit the expression of apoptosis-related B -cell lymphoma-2 （Bcl-2） （P<0.05）， and increase the expression of apoptosis-promoting proteins such as Bcl-2 antagonist of cell death （Bad） and Bcl-2-associated X （Bax）（P<0.05）. Real-time PCR results showed that as compared with blank group， HYJDR（5，10 g·L-1） could effectively inhibit the expression of Bcl-2 protein mRNA in gastric cancer cells（P<0.05）， and all HYJDR groups could promote the expression level of Bad mRNA（P<0.05）. Conclusions:HYJDR can obviously promote the apoptosis of SGC-7901， and its effect is probably achieved by increasing the expression of Bad mRNA and inhibiting the expression of Bcl-2 protein.

Biliary excretion characteristics of berberine, palmatine and jateorhizine in sandwich-cultured rat hepatocytes / 中国药理学通报

Aim To study the biliary excretion characteristics of berberine,palmatine and jateorhizine in rat hepatocytes.Methods Berberine,palmatine and jateorhizine were incubated with the sandwich-cultured rat hepatocytes (SCRH) in standard Ca2+ buffer or Ca2+ free buffer.The accumulation of the three compounds under different conditions were measured by UPLC-MS/MS.The biliary excretion index and biliary clearance were calculated,and the effect of P-gp or Mrp2 inhibitor on the transport of three compounds was also investigated.Results While the incubation time increased,the accumulation of the three compounds also increased.There were obvious differences in accumulation of berberine,palmatine and jateorhizine in incubations treated with standard buffer and calcium-free buffer.The P-gp inhibitors ciclosporin A and verapamil could inhibit the biliary excretion of berberine,palmatine and jateorhizine.However,the Mrp2 inhibitors MK571 and probenecid had no effect on biliary excretion of the three compounds.Conclusions The biliary excretion of berberine,palmatine and jateorhizine is mainly through an active process.They are all the P-gp substrates other than Mrp2 substrates.

Effect of Saposhnikoviae Radix on pharmacokinetics and tissue distributions of active components in Tongxie Yaofang in rats / 中国中药杂志

Tongxie Yaofang (TXYF) is a famous formula that has been used for treating gastrointestinal diseases in traditional Chinese medicine(TCM). Saposhnikoviae Radix is considered as a meridian guiding drug in TXYF and could enhance the effectiveness of prescription. However, the scientific evidence for this effect is still not clear. To reveal the interactions of Saposhnikoviae Radix with other herbs, we conducted this study on the pharmacokinetic profile and tissue distribution of active ingredients of TXYF in rats. The concentrations of four components in blood and tissues were determined by UPLC-MS/MS after oral administration with TXYF. The detection was carried out by electrospray ionization mass spectrometry in the multiple reaction monitoring (MRM) mode. The positive and negative ion switching technique was performed in the same analysis. The results revealed that Saposhnikoviae Radix could enhance Cmax, AUC0-t, and AUC0-∞ of paeoniflorin and hesperidin, and increase the distribution of atractylenolide-I, paeoniflorin and hesperidin in liver, spleen, brain and small intestine. Saposhnikoviae Radix increased the ratio of brain to blood concentrations of atractylenolide-I, paeoniflorin and hesperidin. Meanwhile, it reduced the ratio of lung to blood concentrations of atractylenolide-I and paeoniflorin. Saposhnikoviae Radix, and may enhance the effectiveness of prescriptions by promoting distribution of other herbs in brain.

Uptake Characterizations of Berberine, Palmatine and Jateorhizine in Rat Primary Hepatocytes / 中国药学杂志

OBJECTIVE: To evaluate the uptake characterizations of berberine, palmatine and jateorhizine through hepatic uptake model with rat primary hepatocytes. METHODS: Rat hepatocytes were isolated by Seglen's two-step method and cultured in suspension. The activity of transporters was assessed using rosuvastatin which is known as the substrate of the transporters. The tested drugs were incubated with rat hepatocytes at 4 or 37℃ for various periods of time and the drugs were determined using UPLC-MS/MS method to calculate the uptake amounts. The effect of cultural time and temperature on the hepatic uptake of these compounds were assessed. The kinetic parameters such as Km, Vmax and CLactive/CLuptake were calculated from concentration dependent uptake experiments. The effects of positive inhibitors on the uptake of berberine, palmatine and jateorhizine were also surveyed by pre-incubation of the inhibitors with the rat hepatocytes, followed by co-incubation with berberine, palmatine and jateorhizine. RESULTS: The hepatic uptake of berberine, palmatine and jateorhizine increased along with the increase of concentration at 37℃ while the change of hepatic uptake was very little at 4℃. The value of CLactive/CLuptake was 85.26%, 74.90% and 57.74% for berberine, palmatine and jateorhizine. Ibuprofen, digoxin, glycyrrhizin and prazosin which were the known inhibitors of transporters could remarkably reduce the hepatic uptake of berberine and palmatine, respectively. Glycyrrhizin and prazosin could reduce the uptake of jateorhizine. CONCLUSION: The transport of berberine, palmatine and jateorhizine into hepatocytes is mainly through an active process. Berberine and palmatine are the substrates of Oatp1a1, Oatp1a4, Oatp1b2 and Oct1, and the uptake of jateorhizine might be associated with Oatp1b2 and Oct1.

Impact of sub-chronic aluminium-maltolate exposure on catabolism of amyloid precursor protein in rats / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the impact of sub-chronic Aluminium-maltolate [Al(mal)3] exposure on the catabolism of amyloid precursor protein (APP) in rats.</p><p><b>METHODS</b>Forty adult male Sprague-Dawley (SD) rats were randomly divided into five groups: the control group, the maltolate group (7.56 mg/kg BW), and the Al(mal)3 groups (0.27, 0.54, and 1.08 mg/kg BW, respectively). Control rats were administered with 0.9% normal saline through intraperitoneal (i.p.) injection. Maltolate and Al(mal)3 were administered to the rats also through i.p. injections. Administration was conducted daily for two months. Rat neural behavior was examined using open field tests (OFT). And the protein expressions and their mRNAs transcription related with APP catabolism were studied using enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The expressions of APP, β-site APP cleaving enzyme 1 (BACE1) and presenilin-1 (PS1) proteins and their mRNAs transcription increased gradually with the increase of Al(mal)3 doses (P<0.05). The enzyme activity of BACE1 in the 0.54 and 1.08 mg/kg Al(mal)3 groups increased significantly (P<0.05). The expression of β-amyloid protein (Aβ) 1-40 gradually decreased while the protein expression of Aβ1-42 increased gradually with the increase of Al(mal)3 doses (P<0.05).</p><p><b>CONCLUSION</b>Result from our study suggested that one of the possible mechanisms that Al(mal)3 can cause neurotoxicity is that Al(mal)3 can increase the generation of Aβ1-42 by facilitating the expressions of APP, β-, and γ-secretase.</p>

Effect of aluminum exposure on cognitive function in electrolytic workers and its influential factors / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To clarify the effect of aluminum exposure on the cognitive function in electrolytic workers and the prevalence of mild cognitive impairment (MCI) among them by prevalence survey, and to investigate its influential factors.</p><p><b>METHODS</b>Sixty-six retired workers from the electrolysis workshop of an electrolytic aluminum plant were selected as an aluminum exposure group, while 70 retired workers from a flour mill in the same region were selected as a control group. MCI patients were screened out by Mini-Mental State Examination (MMSE); the blood aluminum level was measured by inductively coupled plasma-mass spectrometry; multivariate statistical analysis was used to investigate the influential factors for MMSE scores and the correlation between blood aluminum level and MCI prevalence.</p><p><b>RESULTS</b>The aluminum exposure group showed a significantly higher blood aluminum level than the control group (25.18 ± 2.65 µg/L vs 9.97 ± 2.83 µg/L, P < 0.01). The total MMSE score of the aluminum exposure group (26.13 ± 2.57) was significantly lower than that of the control group (27.89 ± 1.91) (P < 0.05), particularly the scores on time and place orientation, short-term memory, calculation ability, and language skill (P < 0.05). The detection rate of MCI was significantly higher in the aluminum exposure group (18.2%) than in the control group (5.7%) (P < 0.01). The main influential factors for MMSE scores were gender, age, education level, and blood aluminum level. The logistic regression analysis indicated that the MCI prevalence was significantly correlated with blood aluminum level in the study population (OR = 1.168, P < 0.01).</p><p><b>CONCLUSION</b>Long-term exposure to aluminum can cause cognitive disorders in electrolytic workers and may be one of the risk factors for MCI. Advanced age, male, low education level, and high blood aluminum level may be high-risk factors for cognitive impairment.</p>

Multi-state Markov model in expressing the outcome of mild cognitive impairment among community-based elderly residents / 中华流行病学杂志

Objective To introduce the Multi-state Markov model in studying the outcome prediction of mild cognitive impairment (MCI). Methods Based on the intelligence quotient (IQ)changes that reflecting the trends in cognitive function in the patients under follow-up program, we constructed a four states model and used Multi-state Markov model to analyze the patients. Results Among 600 MCI patients, there were 174(29.0%) males and 426(71.0%) females, with age range of 65-90 years-old (average 69.7±6.6). For univariate analysis, gender, age, education level, marital status, smoking, household income, cerebral hemorrhage, hypertension, high cholesterol, diabetes,LDL-C, SBP and DBP were found to be associated with cognitive function. For multivariate analysis,female, older age, cerebral hemorrhage and higher SBP were shown to be the risk factors for transition from the state of cognitive stability to the state of severe deterioration, and their coefficients were 0.762,0.366,0.885, and 0.069, respectively. Age (0.038) could influence the transition from the state of cognitive stability to slight deterioration. Higher education level was shown to be the protective factor for these transitions (-0.219 and-0.297). Transition intensity from the state of cognitive stability to the state of slight and severe deterioration was 1.2 times that of transition to the state of improving. Transition intensity from state of slight deterioration to cognitive stability was 11.4times that of transition to severe deterioration. Conclusion Multi-state Markov model was an effective tool in dealing with longitudinal data.