1.Taurochenodeoxycholic acid mediates cAMP-PKA-CREB signaling pathway.
You-Chao QI ; Guo-Zhen DUAN ; Wei MAO ; Qian LIU ; Yong-Liang ZHANG ; Pei-Feng LI
Chinese Journal of Natural Medicines (English Ed.) 2020;18(12):898-906
Taurochenodeoxycholic acid (TCDCA) is one of the main effective components of bile acid, playing critical roles in apoptosis and immune responses through the TGR5 receptor. In this study, we reveal the interaction between TCDCA and TGR5 receptor in TGR5-knockdown H1299 cells and the regulation of inflammation via the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA)-cAMP response element binding (CREB) signal pathway in NR8383 macrophages. In TGR5-knockdown H1299 cells, TCDCA significantly activated cAMP level via TGR5 receptor, indicating TCDCA can bind to TGR5; in NR8383 macrophages TCDCA increased cAMP content compared to treatment with the adenylate cyclase (AC) inhibitor SQ22536. Moreover, activated cAMP can significantly enhance gene expression and protein levels of its downstream proteins PKA and CREB compared with groups of inhibitors. Additionally, TCDCA decreased tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, IL-8 and IL-12 through nuclear factor kappa light chain enhancer of activated B cells (NF-κB) activity. PKA and CREB are primary regulators of anti-inflammatory and immune response. Our results thus demonstrate TCDCA plays an essential anti-inflammatory role via the signaling pathway of cAMP-PKA-CREB induced by TGR5 receptor.
Animals
;
Cell Line
;
Cyclic AMP/metabolism*
;
Cyclic AMP Response Element-Binding Protein/metabolism*
;
Cyclic AMP-Dependent Protein Kinases/metabolism*
;
Cytokines/metabolism*
;
Humans
;
Inflammation
;
Macrophages
;
Rats
;
Receptors, G-Protein-Coupled/metabolism*
;
Signal Transduction/drug effects*
;
Taurochenodeoxycholic Acid/pharmacology*
2.Forskolin inhibits spontaneous contraction of gastric antral smooth muscle in rats.
Jing-Zhi JIANG ; Qian SUN ; Dong-Yuan XU ; Mo-Han ZHANG ; Li-Hua PIAO ; Ying-Lan CAI ; Zheng JIN
Acta Physiologica Sinica 2013;65(2):224-228
The aim of the present study was to investigate the effects of cyclic adenosine monophosphate (cAMP) on rat gastric antral circular smooth muscle function. Forskolin, a direct activator of adenylyl cyclase (AC), was used to observe the influences of cAMP. Multi-channel physiological recorder was used to record spontaneous contraction activity of gastric antral circular muscle from Wistar rats. And ELISA method was used to detect the change of cAMP production in perfusate. The results showed that forskolin concentration-dependently suppressed the amplitude and frequency of the spontaneous contraction of the gastric antral muscle, and lowered the baseline of contraction movement significantly. Forskolin concentration-dependently increased the production of cAMP in the perfusate, which showed a significant negative correlation with the contraction amplitude of gastric antral ring muscle. The inhibitory effect of forskolin on spontaneous contraction activity of rat gastric antral circular muscle could be blocked by cAMP-dependent protein kinase (PKA) inhibitor H-89. These results suggest forskolin increases cAMP production and then activates PKA pathway, resulting in the inhibition of the spontaneous contraction activity of rat gastric antral circular smooth muscle.
Adenylyl Cyclases
;
metabolism
;
Animals
;
Colforsin
;
pharmacology
;
Cyclic AMP
;
pharmacology
;
Cyclic AMP-Dependent Protein Kinases
;
metabolism
;
Isoquinolines
;
pharmacology
;
Muscle, Smooth
;
drug effects
;
Pyloric Antrum
;
drug effects
;
Rats
;
Rats, Wistar
;
Sulfonamides
;
pharmacology
3.The effects of antisenes oligodeoxynucleotide on the cyclic nucleotide monophosphates in smooth muscle cells of human corpus cavernosum.
Wen-Jun BAI ; Shu-Kun HOU ; Xiao-Feng WANG ; Zheng YAN ; Pei-Ying HE ; Qing-Ping DENG ; Xiao-Peng HU ; Kao-Peng GUAN
National Journal of Andrology 2002;8(2):88-91
OBJECTIVESTo investigate the effects of antisense oligodeoxynucleotide(ASON) on the cyclic nucleotide monophosphates (cNMP) in smooth muscle cells of human corpus cavernosum, and provide experimental groundwork for the gene therapy of erectile dysfunction.
METHODSPDE5 gene ASON(containing exon 1) was transfected into the corpus cavernosum smooth muscle cells with the presence of liposome DOTAP. Another sense oligodeoxynucleotide(SON) and 1% of bovine serum were also transducted into the cells as controls. Two of cNMP, cAMP and cGMP, were probed and measured by ELISA at 1, 2, 4, 6, 10, 24 and 48 h after transfection.
RESULTSAfter transfection, the level of cGMP(1-6 h) in human corpus cavernosum smooth muscle cells was significantly higher than that in controls(P < 0.01).
CONCLUSIONSThe PDE5 gene ASON had been showed to manifest stimulative effect on the cGMP in smooth muscle cells of human corpus cavernosum in vitro, and it provides experimental groundwork for the gene therapy of erectile dysfunction.
3',5'-Cyclic-GMP Phosphodiesterases ; antagonists & inhibitors ; genetics ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; Humans ; Male ; Muscle, Smooth ; drug effects ; metabolism ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Penis ; cytology
4.Comparison of platelet activators used in slide platelet aggregation test.
Xi-Lin OUYANG ; Jing-Han LIU ; Gui-Xiang SUN ; Xi-Jin LI ; Zi-Lin LIN ; Da-Yong GAO
Journal of Experimental Hematology 2004;12(5):680-683
The study was purposed to explore the suitable platelet activators to be used in slide platelet aggregation test. Experiments were as follows: (1) to detect the intensity and time in 15 healthy donors' platelet aggregation tests induced by cationic propyl gallate (c-PG) and the usual platelet activators: ADP, collagen, epinephrine, arachidonic acid and ristocentin, respectively; (2) to detect the time in platelet aggregation tests of 15 healthy donors induced by c-PG and the above usual platelet activators respectively after addition of PGI2, cAMP or EDTA; (3) to detect the time in 15 healthy donors' platelet aggregation tests induced by c-PG after addition of heparin; (4) to detect the intensity and time of platelet aggregation induced by c-PG at the platelet count of (240-15) x 10(9)/L, (5) to detect the time of platelet aggregation induced by c-PG in eight patients each of whom had taken 100 mg aspirin per day for five days. The results showed that (1) c-PG reduced the strongest intensity of platelet aggregation and the time taken was appropriate, (2) c-PG was the most effective activator to reveal the inhibitive effect on platelet by PGI2, cAMP or EDTA, (3) 0.5 - 3 U/ml heparin did not significantly change the platelet aggregation induced by c-PG, (4) 15 healthy donors' platelet aggregation induced by c-PG displayed clearly on the slide until the platelet count below 30 x 10(9)/L, (5) The platelet aggregation time induced by c-PG was significantly prolonged in eight patients who had taken aspirin. In conclusion, compared to the usual platelet activators, c-PG has remarkable potential advantages when used in slide platelet aggregation test.
Cyclic AMP
;
pharmacology
;
Edetic Acid
;
pharmacology
;
Epoprostenol
;
pharmacology
;
Heparin
;
pharmacology
;
Humans
;
Male
;
Platelet Activation
;
drug effects
;
Platelet Aggregation
;
drug effects
;
Propyl Gallate
;
pharmacology
5.L-4F Inhibits Oxidized Low-density Lipoprotein-induced Inflammatory Adipokine Secretion via Cyclic AMP/Protein Kinase A-CCAAT/Enhancer Binding Protein β Signaling Pathway in 3T3-L1 Adipocytes.
Xiang-Zhu XIE ; Xin HUANG ; Shui-Ping ZHAO ; Bi-Lian YU ; Qiao-Qing ZHONG ; Jian CAO
Chinese Medical Journal 2016;129(9):1108-1112
BACKGROUNDAdipocytes behave like a rich source of pro-inflammatory cytokines including monocyte chemoattractant protein-1 (MCP-1). Oxidized low-density lipoprotein (oxLDL) participates in the local chronic inflammatory response, and high-density lipoprotein could counterbalance the proinflammatory function of oxLDL, but the underlying mechanism is not completely understood. This study aimed to evaluate the effect of apolipoprotein A-I mimetic peptide L-4F on the secretion and expression of MCP-1 in fully differentiated 3T3-L1 adipocytes induced by oxLDL and to elucidate the possible mechanisms.
METHODSFully differentiated 3T3-L1 adipocytes were incubated in the medium containing various concentration of L-4F (0-50 μg/ml) with oxLDL (50 μg/ml) stimulated, with/without protein kinase A (PKA) inhibitor H-89 (10 μmol/L) preincubated. The concentrations of MCP-1 in the supernatant, the mRNA expression of MCP-1, the levels of CCAAT/enhancer binding protein α (C/EBPα), and CCAAT/enhancer binding protein β (C/EBPβ) were evaluated. The monocyte chemotaxis assay was performed by micropore filter method using a modified Boyden chamber.
RESULTSOxLDL stimulation induced a significant increase of MCP-1 expression and secretion in 3T3-L1 adipocytes, which were inhibited by L-4F preincubation in a dose-dependent manner. PKA inhibitor H-89 markedly reduced the oxLDL-induced MCP-1 expression, but no further decrease was observed when H-89 was used in combination with L-4F (50 μg/ml) (P > 0.05). OxLDL stimulation showed no significant effect on C/EBPα protein level but increased C/EBPβ protein level in a time-dependent manner. H-89 and L-4F both attenuated C/EBPβ protein level in oxLDL-induced 3T3-L1 adipocytes.
CONCLUSIONSOxLDL induces C/EBPβ protein synthesis in a time-dependent manner and enhances MCP-1 secretion and expression in 3T3-L1 adipocytes. L-4F dose-dependently counterbalances the pro-inflammatory effect of oxLDL, and cyclic AMP/PKA-C/EBPβ signaling pathway may participate in it.
3T3-L1 Cells ; Animals ; CCAAT-Enhancer-Binding Protein-beta ; analysis ; physiology ; Chemokine CCL2 ; genetics ; secretion ; Cyclic AMP ; physiology ; Cyclic AMP-Dependent Protein Kinases ; physiology ; Humans ; Lipoproteins, LDL ; antagonists & inhibitors ; pharmacology ; Mice ; Peptides ; pharmacology ; Signal Transduction ; physiology
6.New Frontiers in Pharmacology.
Yonsei Medical Journal 1979;20(2):87-91
7.Effects of berberine on cyclic GMP and cyclic AMP levels in rabbit corpus cavernosum in vitro.
Yan TAN ; Qiang TANG ; Ben-Rong HU ; Ji-Zhou XIANG
National Journal of Andrology 2005;11(6):406-408
OBJECTIVETo further investigate the action mechanisms of berberine (Ber), and assess the effects of Ber on the in vitro formation of cGMP and cAMP in the isolated rabbit corpus cavernosum.
METHODSIsolated segments of the rabbit corpus cavernosum were exposed to different concentrations of Ber, and, the dosage-dependent accumulations of cGMP and cAMP were determined in the tissue samples by means of 125I radioimmunoassay. Responses of the isolated tissue preparations to Ber were compared with those obtained with the reference compound sildenafil (Sil).
RESULTSBer increased cGMP concentrations directly (P < 0.05). In the presence of sodium nitroprusside (SNP), a stimulatory agent of cGMP, both Ber and Sil increased cGMP with increasing dosage (P < 0.01), the EC, values being 1.32 and 0.67 micromol/L respectively. With the same concentration, neither Ber nor Sil influenced the cAMP level significantly (P > 0.05). In the presence of PGE1, a stimulator of cAMP, Ber and Sil also raised the cAMP level concentration (P < 0.01 ), the EC, values being 4.90 (Ber) and 6.53 (Sil) micromol/L respectively.
CONCLUSIONBer can increase cGMP and cAMP concentrations in the corpus cavernosum smooth muscles, which may contribute to its action of relaxing corpus cavernosum smooth muscles.
Animals ; Berberine ; pharmacology ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Dose-Response Relationship, Drug ; In Vitro Techniques ; Male ; Muscle, Smooth ; drug effects ; metabolism ; Penis ; drug effects ; metabolism ; Rabbits ; Radioimmunoassay
8.Multi-facet expressions of adenylate cyclase isoforms in B16-F10 melanoma cells differentiated by forskolin treatment.
Du Hyong CHO ; Chang Dae BAE ; Yong Sung JUHNN
Experimental & Molecular Medicine 2000;32(4):235-242
The terminal differentiation of malignant melanoma cells is known to be induced by activating cAMP signaling pathway with alpha-MSH or cAMP analogues. However, sustained activation of cAMP signaling system that induces the differentiation of melanoma cells, also induces the desensitization of the pathway at the receptor level. Nevertheless, the adaptation of adenylate cyclase (AC) expression by sustained activation of cAMP signaling system has not been clearly understood. This study was performed to examine whether the sustained activation of cAMP system induce changes in the expression AC isoforms as an adaptation mechanism. Treatment of B16/F10 murine melanoma cells with 100 mM forskolin for 6 days resulted in differentiation, melanin accumulation and increased expression of tyrosine hydroxylase mRNA. In the forskolin-treated melanoma cells, change in expression of various AC isoform at the transcription level was detected by reverse-transcription polymerase chain reaction (RT-PCR). Expression of AC isoform mRNA: ACI, III, VI, VII, and IX increased to the level of 196-392% of the control whereas the level of ACII was decreased by 30%. The cAMP concentration was increased both in basal and alpha-MSH stimulated cells, but the AC activity was decreased in the forskolin treated cells. Thus, these results suggest that sustained activation of cAMP system induces differential expression of AC isoforms, which results in increase of cAMP accumulation.
Adenylate Cyclase/*genetics
;
Animal
;
Cell Differentiation
;
Cyclic AMP/*metabolism
;
Forskolin/*pharmacology
;
Isoenzymes/genetics
;
Melanoma, Experimental/*enzymology/*pathology
;
Mice
;
Signal Transduction
10.Effects of baicalin on contents of PGE2 and cAMP in hypothalamus of fever rats.
Hong-Yan ZHAO ; Fan ZHANG ; Shu-Duo FAN
Chinese Journal of Applied Physiology 2002;18(2):139-141
AIM AND METHODSBoth PGE2 and cAMP are important neural mediator of fever. In order to discuss if PGE2 and cAMP are involved in the antipyretic mechanism of baicalin, fever models of rats were made by i.v. injection of endotoxin (ET). The antipyretic action and effects of baicalin on contents of PGE2 and cAMP in hypothalamus were observed.
RESULTSBaicalin possessed obvious antipyretic effect on fever rats and reversed the effect of ET on contents of PGE2 and cAMP in hypothalamus. Correlation analysis showed that contents of PGE2 and cAMP in hypothalamus were positively correlated with the change of body temperature of rats.
CONCLUSIONBaicalin may exert its antipyretic effect on fever rats by inhibiting increase of contents of PGE2 and cAMP in hypothalamus.
Animals ; Cyclic AMP ; metabolism ; Dinoprostone ; metabolism ; Fever ; metabolism ; Flavonoids ; pharmacology ; Hypothalamus ; drug effects ; metabolism ; Male ; Rats ; Rats, Wistar