Enriched environment combined with fluoxetine ameliorates depression-like behaviors and hippocampal SYP expression in a rat CUS model.

Effects of enriched environment (EE) combined with fluoxetine in a chronic unpredictable stress (CUS) rat model were examined in our study. Thirty male Sprague-Dawley rats were randomly divided into control group, CUS group, CUS+EE group, CUS+fluoxetine group, and CUS+EE+fluoxetine group (n=six per group). Rats in the CUS group were bred under conditions of CUS and separation for 6 weeks; Control group animals were bred in group cages (three rats per cage) under standard laboratory conditions for 6 weeks; Rats in CUS+EE group, CUS+fluoxetine group, and CUS+EE+fluoxetine groups were bred under the conditions of CUS and separation for 6 weeks and had an intervention of EE, an oral gavage of fluoxetine, and an intervention of EE+oral gavage of fluoxetine, respectively, every day for the final 3 weeks. Every rat underwent a behavioral assessment at the beginning of the 1st week, at the end of the 3rd week and at the end of the 6th week. Behavioral assessments included sucrose water consumption, weight measurement, and an open field test (measuring horizontal moving distance, rearing behavior, and defecation). Finally, the level of synaptophysin expressed in the hippocampus was measured with immunohistochemistry. We found that EE, fluoxetine, and EE+fluoxetine all reversed the depression-like behaviors of CUS rats. The effect of EE+fluoxetine appeared to be superior to EE or fluoxetine alone; the expression level of synaptophysin in CA1, CA3, and DG of the hippocampus was decreased in CUS rats, however, exposure to EE, fluoxetine, and EE+fluoxetine all reversed this decrease.

Synthesis and antitumor activity evaluation of lamiridosin A derivatives.

A series of lamiridosin A derivatives were synthesized through simple procedures. Their antitumor activities were evaluated against EC9706, MGC803, and B16 cell lines in vitro. Several compounds showed potent antitumor activity, especially compound 10, with IC50 value of 2.36 µmol/L against MGC803 cell lines, is more potent than marketed positive drug 5-fluorouridine (5-FU).

[Chemical Constituents from Melissa officinalis Leaves].

OBJECTIVE: To investigate the chemical constituents of Melissa officinalis leaves. METHODS: The chemical constituents were separated by silica gel column chromatography and their structures were determined by spectroscopic experiments. RESULTS: 13 compounds were isolated and identified as protocatechuyl aldehyde(1), serratagenic acid(2), vanillin(3), 2α,3ß-dihydroxy-urs-12-en-28-oic acid(4), ursolic acid(5), oleanolic acid(6), daucosterol(7),2α,3ß,23,29-tetrahydroxyolean-12-en-28-oic acid-29-O-ß-D-gluco- pyranoside(8), luteolin(9) rosmarinic acid(10), luteolin-7-O-ß-D-glucoside (11), ß-stitosterol(12) and palmitic acid(13). CONCLUSION: Compounds 1 ~ 8 are separated from this plant for the first time and compounds 1-4 and 8 are isolated from this genus for the first time.

[Chemical constituents from Punica granatum flowers].

OBJECTIVE: To investigate the chemical constituents in the flowers of Punica granatum. METHODS: The chemical constituents were separated by silica gel column chromatography and their structures were determined by spectroscopic experiments. RESULTS: As a result, twelve compounds were isolated and identified as oleanolic acid (1), ursolic acid (2), palmitic acid (3), tricin (4), catechin (5), rutin (6), apigenin (7), apigenin-7-O-glucoside (8), 2S, 3S, 4S-trihydroxypentanoic acid (9), gallic acid (10), beta-stitosterol (11), and daucosterol (12). CONCLUSION: Compounds 3, 4, 7, 8 and 9 are separated from this plant for the first time, and compounds 3, 4 and 9 are isolated from this genus for the first time.

A new sesquiterpenoid from the rhizomes of Homalomena occulta.

Chemical constituents of EtOAc extract from the rhizomes of traditional Chinese medicine Qian-nian-jian (Homalomena occulta) have been studied, a new sesquiterpenoid, named euadesma-4-ene-1ß,15-diol (1), and four related known compounds, polydactin B (2), oplodiol (3), 1ß,4ß,7α-trihydroxyeudesmane (4), and (-)1ß,4ß,6α-trihydroxy-eudesmane (5), were isolated. Their structures were elucidated using spectroscopic methods including 1D and 2D NMR techniques and mass spectrometry. All the isolates were tested against the human lung adenocarcinoma A549 using MTT assay method. Oplodiol (3) and (-)1ß,4ß,6α-trihydroxy-eudesmane (5) were found to show moderate cytotoxic effects on A549 with IC50 values at 25.5 and 15.0 µg/mL, respectively.

Lactuside B decreases aquaporin-4 and caspase-3 mRNA expression in the hippocampus and striatum following cerebral ischaemia-reperfusion injury in rats.

This study aimed to investigate the effects of lactuside B (LB) on aquaporin-4 (AQP4) and caspase-3 mRNA expression in the hippocampus and the striatum following cerebral ischaemia-reperfusion (I/R) injury in rats. Cerebral I/R injury was established in Sprague-Dawley rats by occluding the middle cerebral artery for 2 h and then inducing reperfusion. Rats in the I/R + LB groups were treated with various doses of LB following reperfusion. Neurological deficit scores and brain water content were obtained to determine the pharmacodynamics of LB. Reverse transcription polymerase chain reaction was performed to determine the expression levels of AQP4 and caspase-3 mRNA in the hippocampus and the striatum. The results of the present study indicate that LB decreased the neurological deficit scores and the brain water content. In the hippocampus, AQP4 and caspase-3 mRNA expression levels were significantly downregulated in the I/R + LB groups at 24 and 72 h following drug administration, compared with those in the I/R group (P<0.05). In the striatum, LB was also shown to significantly reduce AQP4 and caspase-3 mRNA expression levels at 24 and 72 h following drug administration, compared with those in the I/R group (P<0.05). The effects became stronger as the LB dose was increased. The most significant reductions in AQP4 and caspase-3 mRNA expression were noted in the I/R + LB 25 mg/kg and I/R + LB 50 mg/kg groups at 72 h following drug administration. The results of the present study show that LB is capable of significantly downregulating AQP4 and caspase-3 mRNA expression in the hippocampus and striatum following cerebral I/R injury in rats. The mechanism by which LB improved ischaemic brain injury may be associated with changes in AQP4 and caspase-3 mRNA expression in the hippocampus and the striatum.

Two new dammarane-type glycosides from Phlomis umbrosa.

Two new dammarane-type glycosides, phlomisumbroside A (1) and phlomisumbroside B (2), together with 15 known compounds (3-17) were isolated from the leaves of Phlomis umbrosa Turcz. Their structures were established by the spectroscopic methods including 2D NMR techniques.

Oxytocin in the periaqueductal grey regulates nociception in the rat.

Studies have demonstrated that oxytocin (OXT) plays important roles in pain modulation in the central nervous system, and there are OXT receptors in the periaqueductal grey (PAG). The experiment was designed to investigate the effect of OXT in the PAG on antinociception. The results showed that (1) intra-PAG injection of OXT increased the pain threshold, whereas the local administration of the high specific OXT receptor antagonist, desGly-NH(2), d(CH(2))(5)[D-Tyr(2), Thr-sup-4]OVT decreased the pain threshold in a dose-dependent manner; (2) Pain stimulation could elevate OXT concentration in the PAG perfusion liquid. The data suggested that OXT in the PAG was involved in the antinociceptive process through the OXT receptor.

Two new diterpenoids and other constituents from Isodon rubescens.

Two new ent-kaurene diterpenoids, 15α-acetoxyl-6,11α-epoxy-6α-hydroxy-20-oxo-6,7-seco-ent-kaur-16-en-1,7-olide (1), 15α-hydroxy-20-oxo-6,7-seco-ent-kaur-16-en-1,7α(6,11α)-diolide (2), together with ten known compounds (5-14) were isolated from the leaves of Isodon rubescens. Their structures were elucidated mainly by various spectroscopic techniques and finally confirmed by single-crystal X-ray diffraction. Compounds 1, 2, 8 and 12 were evaluated for their cytotoxicities against EC-1, U87, A549, MCF-7 and Hela cell lines.

Oxytocin, but not arginine vasopressin is involving in the antinociceptive role of hypothalamic supraoptic nucleus.

Our previous study has demonstrated that the hypothalamic supraoptic nucleus (SON) plays a role in pain modulation. Oxytocin (OXT) and arginine vasopressin (AVP) are the important hormones synthesized and secreted by the SON. The experiment was designed to investigate which hormone was relating with the antinociceptive role of the SON in the rat. The results showed that (1) microinjection of L-glutamate sodium into the SON increased OXT and AVP concentrations in the SON perfusion liquid, (2) pain stimulation induces OXT, but not AVP release in the SON, and (3) intraventricular injection (pre-treatment) with OXT antiserum could inhibit the pain threshold increase induced by SON injection of L-glutamate sodium, but administration of AVP antiserum did not influence the antinociceptive role of SON stimulation. The data suggested that the antinociceptive role of the SON relates to OXT rather than AVP.

Arginine vasopressin in hypothalamic paraventricular nucleus is transferred to the caudate nucleus to participate in pain modulation.

Arginine vasopressin (AVP), which is synthesized and secreted in the hypothalamic paraventricular nucleus (PVN), is the most important bioactive substance in the pain modulation. Our pervious study had shown that AVP plays an important role in pain modulation in caudate nucleus (CdN). The experiment was designed to investigate the source of AVP in CdN by the nucleus push-pull perfusion and radioimmunoassay. The results showed that: (1) pain stimulation increased the AVP concentration in the CdN perfusion liquid, (2) PVN decreased the effect of pain stimulation which was stronger in both sides than in one side of PVN cauterization; and (3) L-glutamate sodium would excited the PVN neurons by the PVN microinjection that could increase the AVP concentration in the CdN perfusion liquid. The data suggested that AVP in the CdN might come from the PVN in the pain process, i.e., AVP in the PVN might be transferred to the CdN to participate in the pain modulation.

[Effect of lactuside B on the expression of bcl-2 and bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury].

This study is to investigate the effect of the major chemical composition in rhizome of Pterocypsela elata, lactuside B, on expression of bcl-2, bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury. First, middle cerebral artery ischemia-reperfusion injury model was established, and each group was treated with the corresponding medicines. Animals were separately sacrificed at 24 h and 72 h. The brain infarct volumes were detected by TTC dye, bcl-2 and bax mRNA expression was checked by RT-PCR, and the proteins of bcl-2 and bax were explored by two-step immunohistochemistry in cerebral cortex of rats. Lactuside B can reduce brain infarct volume of cerebral cortex of rats, increase the expression of bcl-2 mRNA and decrease that of bax mRNA. Moreover, the ratio of bcl-2 to bax mRNA is higher in 12.5 and 25 mg kg(-1) dose group, respectively, which is significantly different from that of model group (P < 0.05 or P < 0.01). Generally, either 12.5 or 25 mg kg(-1) dose group is better than positive control medicine nimodipine (P < 0.05 or P < 0.01). In addition, the expression of bcl-2 and bax protein is consistent with their gene expression. Infarct volume and the ratio of bcl-2 to bax mRNA expression are significantly different (P < 0.05 or P < 0.01) between 72 h and 24 h group. The results demonstrated that lactuside B could play a good role in resisting cerebral ischemia by upregulating the expression of bcl-2 mRNA and protein and downregulating that of bax mRNA and protein.

Norepinephrine plays an important role in antinociceptive modulation of hypothalamic paraventricular nucleus in the rat.

Our previous study has proven that hypothalamic paraventricular nucleus (PVN) plays a role in antinociception. The effects of studied classical neurotransmitter on PVN antinociceptive modulation were investigated in the rat. The results showed: (1) Pain stimulation increased norepinephrine (NE), but not epinephrine, dopamine (DA), 3,4-dihydroxyphenylacetic acid (DA metabolic product), homovanilic acid (DA metabolic product), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HT metabolic product), acetycholine (Ach), choline (Ach metabolic product), gamma-aminobutyric acid (GABA), and L-glutamate acid concentrations in the PVN perfusion liquid; (2) PVN stimulation with L-glutamate sodium, which excited local neurons only, did not influence the concentrations of the studied classical neurotransmitter and metabolic product in the PVN perfusion liquid; (3) Microinjection of NE, epinephrine, or L-glutamate sodium into the PVN elevated pain threshold, and local administration of GABA decreased pain threshold in a dose-dependent manner, but PVN administration of Ach, DA, or 5-HT did not change pain threshold; (4) Microinjection of phentolamine (alpha-receptor antagonist) or MK801 [NMDA-receptor antagonist] into the PVN reduced pain threshold, and local administration of bicuculline (GABA-receptor antagonist) raised pain threshold, but PVN administration of propranolol (beta-receptor antagonist), atropine (Muscarinic cholinergic receptor antagonist), 6-OH gallamine (Nicotinic cholinergic receptor antagonist), fluperidol (DA-receptor antagonist), or cyproheptadine (5-HT-receptor antagonist) did not alter pain threshold. The data suggested that endogenous NE, not epinephrine, 5-HT, Ach, GABA, and L-glutamate acid played an important role in the PVN antinociceptive modulation.

6ß-Acet-oxy-1α,7ß,11ß,15ß-tetra-hydr-oxy-7α,20-ep-oxy-ent-kaur-16-ene.

The title compound, C(22)H(32)O(7), a natural ent-kaurane diterpenoid also referred to as Maoyecrystal F, was obtained from the medicinal plant Isodon nervosa. There are four rings with the expected cis and trans junctions. Cyclohexane ring A adopts a chair conformation, rings B and C adopt boat conformations, while the five-membered ring has an envelope conformation. The mol-ecules stack along the a axis in the crystal and are linked together by inter-molecular O-H⋯O hydrogen bonds.

1α,6ß,7ß,11α,15ß-Penta-hydr-oxy-7α,20-ep-oxy-ent-kaur-16-ene.

The title compound, C(20)H(30)O(6), a natural ent-kaurane diterpenoid, named nervosanin B, was obtained from the medicinal plant Isodon serra. It is composed of four rings with the expected trans and cis junctions. One of the six-membered rings is in a chair conformation, the other two are in boat conformations and the five-membered ring adopts an evenlope conformation. The mol-ecules stack along the a axis and are linked together by inter-molecular O-H⋯O hydrogen bonds. Two intramolecular O-H⋯O interactions also occur.

6ß,15ß-Diacet-oxy-1ß,7ß,13α-trihydr-oxy-7α,20-ep-oxy-ent-kaur-16-ene.

The title compound, C(24)H(34)O(8), a natural ent-kaurane diterpenoid, is composed of four rings with the expected cis and trans junctions. The crystal structure is stabilized by inter-molecular O-H⋯O hydrogen bonds. In addition, an intra-molecular O-H⋯O hydrogen bond occurs.

15α,20ß-Dihydr-oxy-6ß-meth-oxy-6,7-seco-6,20-ep-oxy-1,7-olide-ent-kaur-16-ene.

The title compound, C(21)H(30)O(6), a natural ent-kaurane diterpenoid, was obtained from the medicinal plant Isodon serra. The five rings in the mol-ecule exhibit the expected cis and trans junctions. The three six-membered rings adopt chair, twist-boat and boat conformations, while two five-membered rings adopt envelope conformations. There are two mol-ecules in the asymmetric unit, related by a non-crystallographic twofold screw axis; the main difference is in the different degrees of distortion of ring B. In the crystal, the mol-ecules are linked by inter-molecular O-H⋯O hydrogen bonds, forming chains along the b axis.

Chemical constituents of Isodon nervosus and their cytotoxicity.

Two new ent-kaurane diterpenoids, 6,20,15alpha-trihydroxy-6,7-seco-1alpha,7-olide-ent-kaur-16-ene (1) and 7beta,12alpha-dihydroxy-6beta,15beta-diacetoxy-7alpha,20-epoxy-ent-kaur-2,16-dien-1-one (2), together with the six known compounds, were isolated from the aerial part of Isodon nervosus. The structures of the new compounds were determined by spectral methods (1D, 2D NMR, and MS). Six compounds were assayed for their cytotoxicity against HL60, SMMC-7721, and HeLa human cell lines. Compounds 5, 7, and 8 showed significant cytotoxicity.

1α,11α,15ß-Triacet-oxy-7ß-hydroxy-7α,20-ep-oxy-ent-kaur-16-en-6-one.

The title compound, C(26)H(34)O(9), a natural ent-kaurane diterpenoid, is composed of four rings with the expected cis and trans junctions. In the crystal structure, the mol-ecules stack along the a axis and are linked together by inter-molecular O-H⋯O hydrogen bonds.

1α,6ß,7ß,14ß,15ß-Penta-hydr-oxy-7α,20-ep-oxy-ent-kaur-16-ene.

The title compound, enmenol, C(20)H(30)O(6), a natural ent-kaurane diterpenoid, comprises five fused rings, four of which are six-membered. Cyclo-hexane ring A adopts a chair conformation, rings B and C adopt boat conformations, while ring D has an envelope conformation, and two intramolecular O-H⋯O interactions occur. In the crystal, inter-molecular O-H⋯O hydrogen bonds generate a two dimensional network.