Effects of Gelsemium elegans Combined with Mussaenda pubescens on Transporter BCRP and Drug-metabolizing Enzyme CYP3A11 / 中国实验方剂学杂志

ObjectiveTo explore the effect of Gelsemium elegans combined with Mussaenda pubescens on efflux transporter breast cancer resistance protein （BCRP） and cytochrome P450 3A11 （CYP3A11） and their attenuation mechanism， and to investigate whether the nuclear receptors were involved in such regulation by intervening it with nuclear receptor activators. MethodC57BL/6 mice were divided into the blank group， G. elegans （GE， 0.25 g·kg-1）group， GE + M. pubescens （MP） （0.25 g·kg-1+10 g·kg-1） group， GE + pregnane X receptor （PXR） activator （rifampicin）（GE + Rif，0.25 g·kg-1+50 mg·kg-1） group， GE + MP + Rif （0.25 g·kg-1+10 g·kg-1+50 mg·kg-1） group， GE + constitutive androstane receptor （CAR） activator （1，4-Bis ［2-（3，5-Dichloropyridyloxy）］ benzene， TCPOBOP）（GE + TCP， 0.25 g·kg-1+0.5 mg·kg-1） group， and GE + MP + TCP （0.25 g·kg-1+10 g·kg-1+0.5 mg·kg-1） group. The medication lasted for 14 successive days. One hour after the last administration， the mice were sacrificed by cervical dislocation and the liver tissue was harvested. The left liver tissue was stained with hematoxylin- eosin （HE） for observing the pathological changes. The right liver tissue was used for BCRP and CYP3A11 mRNA and protein expression detection by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultThe survival rates of mice in the GE + Rif group， GE group， and GE + MP group were 25% （the lowest）， 40%， and 80%， respectively， and no death was observed in the other groups. Compared with the obvious lesions in the liver cells of the GE group， the pathological changes in liver cells of the GE + MP group were alleviated， while those in the GE + Rif group were worsened. Compared with the GE + Rif group， the GE + MP + Rif group exhibited relieved pathological changes in liver cells. Both the GE + TCP group and the GE + MP + TCP group showed mild liver lesions. The comparison with the GE + MP group revealed that the pathological changes in the GE + MP + TCP group were slightly relieved. Compared with the blank group， the expression of BCRP protein and mRNA in GE group were significantly decreased （P<0.05，P<0.01）.The expression of CYP3A11 protein in GE group were significantly decreased （P<0.01）. Compared with the GE group， the GE + MP group displayed remarkably up-regulated BCRP protein and mRNA expression （P<0.05，P<0.01） and CYP3A11 protein expression （P<0.05）， but slightly up-regulated CYP3A11 mRNA expression. Compared with the GE group， the GE + Rif group exhibited down-regulated BCRP protein expression （P < 0.05）. The protein and mRNA expression levels of BCRP were lower in the GE + MP + Rif group than in the GE + MP group （P<0.05，P<0.01）. The PXR activator rifampicin regulated BCRP before and after the combination of G. elegans with M. pubescens. The CYP3A11 protein and mRNA expression levels in the GE + TCP group were higher than those in the GE group （P<0.05，P<0.01）. Compared with the GE + MP group， the GE + MP + TCP group showed up-regulated CYP3A11 protein and mRNA expression （P<0.05，P<0.01）. CAR activator TCPOBOP also had a regulatory effect on CYP3A11 before and after the compatibility of G. elegans with M. pubescens. ConclusionThe attenuated toxin after the combination of G. elegans with M. pubescens is closely related to the efflux transporter BCRP and the drug-metabolizing enzyme CYP3A11.

Huayu Jiedu Prescription Alleviates Gut Microbiota and Fecal Metabolites in Mice with Endometriosis / 中国实验方剂学杂志

Objective:To investigate the effect of Huayu Jiedu prescription （HYJDP） on gut microbiota and fecal metabolites in mice with endometriosis. Method:Normal female C57BL/6J mice were divided into normal control group （CO）， endometriosis group （EM） and Chinese medicine Huayu Jiedu decocotion group （CM）. CO and EM groups received normal saline and CM group received HYJDP by intragastric administration. Untargeted metabolomics method was used to detect metabolites in fecal supernatant of mice， and receiver operating characteristic （ROC） analysis was used to screen the differential metabolites， 16S rRNA high-throughput sequencing was used to detect the gut microbiota， and Spearman correlation coefficient was used to represent the degree of correlation between differential metabolites and intestinal flora. Lipopolysaccharides （LPSs） in intestinal wall tissue， serum and peritoneal lavage fluid were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of Vimentin and E-cadherin in ectopic lesions was detected by immunohistochemistry. Result:HYJDP alleviated the disorders of fecal metabolites and gut microbiota in EMS mice， especially with the recovered levels of homoveratric acid， melilotoside C and physapubescin in fecal supernatant. In the comparison of these three factors between EM group and CO group as well as between EM group and CM group， the variable important in projection （VIP） value was both above 2， and AUC in ROC analysis was both >0.9. As compared with EM group， HYJDP restored the abundance of species such as <italic>Lachnospiraceae_NK4A136_group</italic>， <italic>Lactobacillus</italic> and <italic>Blautia </italic>（<italic>P</italic><0.05）. In addition， the level of LPS in peritoneal fluid supernatant of EM group was significantly higher than that of CO group （<italic>P</italic><0.05） and CM group （<italic>P</italic><0.05）. The protein expression of vimentin and E-cadherin in endometriosis decreased significantly （<italic>P</italic><0.05）. Conclusion:HYJDP which can improve the intestinal environment and reduce the level of LPS in mice with endometriosis， is an effective drug for the treatment of endometriosis.

Study on chemical constituents of Abies chensiensis / 中草药

Objective: To study the chemical constituents of the sterns and leaves parts of Abies chensiensis. Methods: The isolation and purification were carried out by HP-20 macroporous resin, Sephadex LH-20, silica gel column chromatography, semi-preparative HPLC. Their structures were elucidated on the basis of physicochemical properties and spectroscopic data. Results: Twenty-four compounds were isolated and elucidated from the sterns and leaves parts of A. chensiensis, their structures were identified as 7,14,24-mariesatrien-26,23-olide-3α,23-diol (1), (23R)-3α-hydroxy-9,19-cyclo-9β-lanost-24-en-26,23-olide (2), 7-oxocallitrisic acid (3), 23-hydroxy-3-oxomariesia-8 (9),14,24-trien-26,23-olide (4), neoabiestrine E (5), 3-oxo-9β-lanosta-7,24-dien-26,23R-olide (6), 7,14,22Z,24-mariesatetraen-26,23-olide-3-one (7), (+)-rel-3α-hydroxy-23-oxocycloart-25 (27)-en-26-oic acid (8), cycloart-25-en- 3β,24-diol (9), neoabiestrine H (10), 3-oxo-24,25,26,27-tetranolanost-8-en-23-oic acid (11), abiesadine C (12), 13β-epidioxy-8 (14)-abieten-18-oic acid (13), dehydroabietic acid (14), 15-hydroxydehydroabietic acid (15), methyl 18-methoxydehydroabietate (16), 7β-hydroxy dehydroabietic acid (17), dehydroabietan-18-ol (18), centdaroic acid (19), abietic acid (20), 6,8,11,13-abi-etatrien-18-oic acid (21), abiesadine B (22), abiesadine N (23), 12β-hydroxyabietic acid (24). Conclusion: All compounds are isolated from the plants of A. chensiensis for the first time, and compound 11 is isolated from the Abies genus for the first time.

Study on chemical constituents of Ligularia veitchiana / 中草药

Objective: To study the chemical constituents of the aerial parts of Ligularia veitchiana. Methods: The isolation and purification were carried out by macroporous resin, silica gel column chromatography, Sephadex LH-20, and semi-preparative HPLC. Their structures were elucidated on the basis of physicochemical properties and spectroscopic data. Results: A total of 16 compounds were isolated and elucidated from the aerial parts of Ligularia veitchiana, their structures were identified as: 2-hydroxplatyphyllide (1), ligudentatin A (2), 2-metyoxy-platyphillide (3), 7-hydroxy-11-hydroperoxybisabol-2,9E-diene (4), bisabola-2,11-diene-7α,10ζ-diol (5), 8β,10β-dihydroxyeremophilenolide (6), 3β(ax)-hydroxyeremophillenolide (7), 6β-hydroxy- 8α-ethoxyeremophil-7(11)-en-12,8β-olide (8), 3β,28-dibydroxylup-20(29)-ene (9), oleanolic acid (10), 3β-hydroxyolean-12-en-27- oic acid (11), ursolic acid (12), coniferyl ferulate (13), 4-O-Geranyl-sinapyl alcohol (14), 4-O-geranyl-coniferyl alcohol (15), auraptene (16). Conclusion: All compounds are isolated from Ligularia veitchiana for the first time. Compounds 4, 5, 7, 8, 11 are isolated from the genus Ligularia for the first time.

Effects 'of β3 adrenoceptors on the contractility of rat thoracic aorta smooth muscle and the mechanism / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the effect of β₃adrenoceptors (β₃-AR) activation on rat thoracic aorta smooth muscle contractility and the possible related mechanism.</p><p><b>METHODS</b>The endothelium removed thoracic aorta was pre-contracted with 30 mmol/L KCl physiological saline solution (PSS). Then the tension of the thoracic aorta was recorded in presence of BRL37344 (BRL) to determine the action of β₃-AR. The tension of the thoracic aorta was also recorded in the presence of Propranolol (PRA), SR59230A (SR), L-NNA, H-89 and Iberiotoxin (IBTX) respectively to reveal the underling mechanism of β₃-AR activation on rat vascular smooth muscle. Immunohistochemistry was adopted to confirm the existence and the distribution of β₃-AR in rat thoracic aorta.</p><p><b>RESULTS</b>The results showed that: (1) The thoracic aorta was relaxed by β₃-AR activation, with a relaxation percentage of (10.59 ± 0.79). (2) β₃-AR was expressed in both endothelial and smooth muscle layer in thoracic aorta sections of rats. (3) PRA did not block the effect of BRL on the thoracic aorta. The relaxation actions of BRL could be antagonized by pre-incubating the thoracic aorta with SR. (4) L-NNA (a NOS inhibitor) and H-89 (a PKA inhibitor) reversed the relaxation effect of BRL on vascular smooth muscle. (5) The effect of BRL was decreased after application of Ibriotoxin (IBTX), a large conductance calcium dependent potassium channel blocker.</p><p><b>CONCLUSION</b>The results confirmed that activation of β₃-AR led to relaxation of thoracic aorta smooth muscle. The relaxation action of β₃-AR on smooth muscle of rat thoracic aorta was related to activation of NOS and PKA signaling pathway. Large conductance Ca²⁺-K⁺ channels were involved in the relaxation action of β₃-AR activation on rat thoracic aorta smooth muscle.</p>

Research on inhibition mechanism of polypeptide extract from scorpion venom combined with Rapamycin on angiogenesis of H22 hepatoma / 中草药

Objective: To study the inhibition of polypeptide extract from scorpion venom (PESV) combined with Rapamycin (RAPA) on angiogenesis of H22 hepatoma and its mechanism. Methods: The H22 carcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups, i.e., control group, PESV group (20 mg/kg), RAPA group (2.5 mg/kg), and PESV + RAPA group, 10 mice in each group. The intervention was lasted 14 d. The tumor volume was measured once every other day, the tumor volume growth curve was drawn, and the tumor inhibitory rate was calculated. The protein expression levels of mammal target of RAPA (mTOR), hypoxia-inducible factor-1α (HIF-1α), and vascular endothelial growth factor-A (VEGF-A) were detected by immunohistochemical assay. Microvessel was signed by factor VIII and then detected by immunohistochemical assay. Results: The growth speed of H22 hepatoma transplantation tumor was obviously inhibited in the PESV group, RAPA group, and PESV + RAPA group compared with control group after 14 d intervention (P 22 hepatoma, the mechanism maybe related to the inhibition on the protein expression of mTOR, HIF-1α, and VEGF-A.

Mechanism of Polypeptide Extract from Scorpion Venom Combined Rapamycin in Enhancing Autophagy of H22 Hepatoma Cells: an Experimental Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe enhanced effects of polypeptide extract from scorpion venom (PESV) combined Rapamycin on autophagy of H22 hepatoma cells in mice and to explore its possible mechanism.</p><p><b>METHODS</b>The H22 hepatocarcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups, i.e., the control group,the high dose PESV group, the low dose PESV group, and the combination group (high dose PESV + Rapamycin), 10 in each group. Mice in high and dose PESV groups were administered with 20 mg/kg and 10 mg/kg PESV respectively by gastrogavage. Mice in the combination group were administered with 2 mg/kg rapamycin and 20 mg/kg PESV by gastrogavage. The intervention lasted for 14 successive days. The tumor volume was measured once every other day, the tumor growth curve was drawn, and then the tumor inhibitory rate calculated. Pathological changes of the tumor tissue were observed by HE staining. Protein expression levels of mammal target of rapamycin (mTOR), UNC-51-like kinase-1 (ULK1), microtubule-associated protein1 light chain3 (MAPILC3A), and Beclin1 were detected by immunohistochemical assay.</p><p><b>RESULTS</b>The growth of H22 hepatoma transplantation tumor was inhibited in high and low dose PESV groups and the combination group (P < 0.05). And there was statistical difference in tumor weight and tumor volume between the combination group and high and low dose PESV groups (P < 0.05). There was no statistical difference in tumor weight or tumor volume between the high dose PESV group and the low dose PESV group (P > 0.05). lmmunohistochemical assay showed that the protein expression of mTOR was higher, but protein expressions of ULK1, MAP1LC3A, Beclin1 were lower in the control group than in the rest 3 groups (P < 0.05, P < 0.01). Compared with the high dose PESV group, protein expressions of ULK1, MAP1LC3A, and Beclin1 were obviously lower (P < 0.05).</p><p><b>CONCLUSION</b>PESV combined Rapamycin might inhibit the development of H22 hepatoma transplantation tumor in mice possibly through inhibiting the activity of mTOR, enhancing expressions of ULK1, MAP1LC3A, and Beclin1.</p>

Formulation optimization of ginkgo flavonoids matrix tablets by orthogonal design / 中国中药杂志

Sustained-release tablet has become one of the hottest research spots in the area of sustained release preparations with its unique advantages. At present, a series of shortcomings were exited in the ordinary ginkgo preparations, which were used for the treatment of cardiovascular and cerebrovascular diseases. In order to avoid these shortcomings, ginkgo flavonoids matrix tablets were prepared in this paper. Furthermore, the amount and varieties of matrix material, adhesives and fillers were investigated. Meanwhile, the formulation was optimized by using the method of orthogonal design, and Zero-order, First-order, Higuchi, Ritger-peppas equation were used for the model fitting and mechanism discussing of drug release.

Expression of high mobility group protein-B1 in mice with hyperoxia-induced bronchopulmonary dysplasia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of hyperoxia exposure on high mobility group protein-B1 (HMGB1) expression in neonatal mice and the role of HMGB1 in the pathogenesis of bronchopulmonary dysplasia (BPD).</p><p><b>METHODS</b>C57BL/6 mice were randomly exposed to 60% O2 or air 1 day after birth. BPD was induced by 60% O2 exposure. The pulmonary tissue samples were harvested 3, 7 and 14 days after exposure. The pathologic changes of pulmonary tissues were observed by hematoxylin and eosin staining, Masson staining and radical alveolar count. The expression of HMGB1 protein in lungs was detected by immunofluorescence. The expression of HMGB1 mRNA was detected by real-time fluorescent quantitative PCR.</p><p><b>RESULTS</b>In the BPD group, the lungs developed decreased alceolar septation, swollen alveolar epithelium, stroma edema, interstitial fibrosis and developmental lag when compared with the control group. These changes became more obvious with more prolonged hyperoxia exposure. The expression of HMGB1 protein and mRNA 7 and 14 days after exposure increased significantly in the BPD group compared with that in the control group.</p><p><b>CONCLUSIONS</b>Hyperoxia exposure results in an increase in lung HMGB1 expression. The increased HMGB1 expression may be associated with the development of BPD.</p>