Hepatoprotective activity of depsidone enriched Cladonia rangiferina extract against alcohol-induced hepatotoxicity targeting cytochrome P450 2E1 induced oxidative damage.

Alcoholic liver disease (ALD) is a broad-spectrum disorder, covering fatty liver, cirrhosis, alcoholic hepatitis and in extreme untreated condition hepatocellular carcinoma (HCC) may also develop. Cladonia rangiferina (CR) is a class of lichen having a broad spectrum of pharmacological activity. It is used like traditional natural sources in ancient times in India, China, Sri Lanka, etc. Folkloric record about CR has reported their use as an antimicrobial, antitumor, antioxidant, anti-inflammatory activities, etc. Hence, the present study was requested to ascertain the effect of the ethanolic extract of Cladonia rangiferina (CRE) on alcohol-induced hepatotoxicity. The animals were evaluated for the estimation of the liver in vivo biochemical antioxidant parameters. The liver tissues were further evaluated histopathologically and western blotting examination for localization of apoptotic gene expression that plays a pivotal role in hepatotoxicity. The results of this study reveal that CRE proves to be helpful in the treatment of alcohol-induced hepatotoxicity and oxidative stress. Results of different markers have shown that among all, CRE has demonstrated the best hepatoprotective activity. These observations say about the importance of the components of the extract. The ameliorative action of CRE in alcoholic liver damage may exist due to antioxidant, anti-inflammatory, and anti-apoptotic activities.

Oxidative free radicals scavenging activity (in vitro and in vivo assay) of standardized fractions from the seeds of Argyreia speciosa (Ghav-patta) a traditional Indian medicine.

TRADITIONAL PERTINENCE: Argyreia speciosa Sweet (Linn.), belongs to the family convolvulaceae, a traditional Indian medicinal herb, has been used to treat acute/chronic ulcers, gonorrhea, rheumatoid arthritis and several nervous disorders having a long history. AIM OF THE STUDY: A broad spectrum approach of this work was to find out the antioxidant activity of Argyreia speciosa seeds, in vitro and in vivo antioxidant assay were performed. MATERIAL AND METHODS: Total phenolic content (TPC), reducing power (RP), antioxidant activity (AOA), O 2 · - (superoxide anion), DPPH (1,1-diphenyl-2-picrylhydrazyl) and ˙OH (hydroxyl) radicals scavenging activities, GSH (glutathione), CAT (catalase), SOD (superoxide dismutase) and LPO (lipid peroxidase) are the major parameters which were studied for determining in vitro and in vivo antioxidant property of seed extract & their six fractions obtained from A. speciosa. Carbon tetrachloride (CCl4) induced rat model was used to determine in vivo antioxidant assay of extract and its fractions. RESULTS: Butanol fraction (AS-BF) showed strong antioxidant property and protected oxidative DNA damage. AS-BF was found best as compared to all other fraction for determining antioxidant property of seeds with the reduction in lipid peroxide formation and increment in GSH, CAT and SOD. AS-BF showed the presence of phenolic compounds viz. gallic acid, chlorogenic acid, and ellagic acid. CONCLUSION: From these results, it was proved that A. speciosa seeds prevent tissue damage due to oxidative stress with strong antioxidant activity.

Wound healing potential of flowers extracts of Woodfordia fruticosa Kurz.

Wound healing or repair is the body's natural process of regenerating dermal and epidermal tissue. Woodfordia fruticosa Kurz (Family: Lythraceae) is used traditionally in wound healing by the tribals of Chhattisgarh district. However, there is a paucity of scientific data in support. In this study, we evaluated antimicrobial activity of petroleum ether, chloroform, ethanolic and aqueous extracts against a diverse range of gram +ve and gram -ve bacteria along with pathogenic fungi. The wound healing activity of ethanolic extract was also evaluated at dose levels of 250 and 500 mg/kg body wt in rats by excision, incision and dead space wound healing models along with histopathology of wound area of skin. The ethanolic extract showed potent wound healing activity, as evident from the increase in the wound contraction and breaking strength in dose-dependent manner. Treatment with ethanolic extract (250 and 500 mg/kg body wt) showed significant dose-dependently decrease in epithelization period and scar area. Hydroxyproline, hexuronic acid and hexosamine contents, the important constituents of extracellular matrix of healing were also correlated with the observed healing pattern. During early wound healing phase, pro-inflammatory cytokines TNF-alpha, IL-6 and anti-inflammatory cytokine IL-10 levels were found to be upregulated by the ethanolic extract treatment. The ethanolic extract exhibited a strong and broad spectrum antimicrobial activity, as compared to other extracts. It showed very low Minimum inhibitory concentration (MIC) values and inhibited the growth of E. coli, Staphylococcus aureus and Candida albicans in concentration of 2.5 microg/disc. Thus, the results of the present study demonstrated the strong wound healing potential and antimicrobial activities of W. fruticosa, flowers, supporting the folklore use of the plant by the tribal people of Chhattisgarh district.

A review on ethnobotany, phytochemistry and pharmacology of Fumaria indica (Fumitory).

Fumaria indica (Hausskn.) Pugsley (Fumariaceae), known as "Fumitory", is an annual herb found as a common weed all over the plains of India and Pakistan. The whole plant is widely used in traditional and folkloric systems of medicine. In traditional systems of medicine, the plant is reputed for its anthelmintic, diuretic, diaphoretic, laxative, cholagogue, stomachic and sedative activities and is used to purify blood and in liver obstruction in ethnopharmacology. The whole plant is ascribed to possess medicinal virtues in Ayurvedic and Unani systems of medicine and is also used in preparation of important Ayurvedic medicinal preparations and polyherbal liver formulations. The review reveals that phytochemical constituents of wide range have been separated from the plants and it possesses important pharmacological activities like smooth muscle relaxant, spasmogenic and spasmolytic, analgesic, anti-inflammatory, neuropharmacological and antibacterial activities. The separation of hepatoprotective and antifungal constituents from this plant was also reported newly. This review highlights the traditional, ethnobotanical, phytochemical, pharmacological information available on Fumaria indica, which might be helpful for scientists and researchers to find out new chemical entities responsible for its claimed traditional uses.

Morpho-anatomical and physicochemical studies of Fumaria indica (Hausskn.) Pugsley.

OBJECTIVE: To study morpho-anatomical characters and physicochemical analysis of Fumaria indica (F. indica) (Hausskn.) Pugsley, (Fumariaceae), an important medicinal plant used extensively for treating a variety of ailments in various system of indigenous medicine. METHODS: Evaluation of the different parts of the plant was carried out to determine the morpho-anatomical, physicochemical, phytochemical and HPTLC fingerprinting profile of F. indica and other WHO recommended methods were performed for standardization. RESULTS: Morpho-anatomical studies showed compound and pinnatifid leaf, 4 to 6 cm in length, linear and oblong in shape and anomocytic arrangement of stomata, thin walled parenchymatous cells, scattered, sclerenchymatous, capped vascular bundles and radiating medullary rays. Physicochemical studies showed foreign matter 0.2%, loss on drying 6.8%, total ash 16.77%, alcohol and water soluble extractives 8.92% and 20.26%, respectively, sugar 17.75%, starch 22.97% and tannins 2.37%. Phytochemical evaluation revealed the presence of carbohydrate, alkaloids, flavonoids, saponins, tannins and sterol. Thin layer chromatography was carried out with different solvents and the best solvent system was chloroform and methanol in 80:20 ratio and revealed 12 spots with different Rf value under UV light 366λ. CONCLUSIONS: The results of the study can serve as a valuable source of information and provide suitable standards for identification of this plant material for future investigations and applications.

Genetic ablation of luteinizing hormone receptor improves the amyloid pathology in a mouse model of Alzheimer disease.

Amyloid-beta peptide (Abeta) plays an essential pathophysiologic role in Alzheimer disease, and elevation of luteinizing hormone (LH) levels during aging has been implicated in its pathogenesis. To assess the effect of LH receptor deficiency on Abeta accumulation, we generated a bigenic mouse model, APPsw(+)/Lhr(-/-), which expresses human amyloid precursor protein (APPsw) in the background of LH receptor (Lhr) knockout. Genetic ablation of Lhr resulted in a significant decrease in the number of Abeta plaques and protein content in the hippocampus and cerebral cortex in both male and female mice. Accordingly, several Abeta deposition-related neuropathologic features and functionally relevant molecules were markedly improved, including decreased astrogliosis, reductions of elevated phosphorylated tau, c-fos, alpha7-nicotinic acetylcholine receptor, and restoration of the altered neuropeptide Y receptors Y1 and Y2. Diminution of Abeta accumulation in the absence of LH receptor supports the contention that dysregulation of LH may impact the pathogenesis of Alzheimer disease. The APPsw(+)/Lhr(-/-) mouse may be a useful tool for advancing understanding of the role of LH-mediated events in Alzheimer disease and a model in which to test therapeutic interventions.

Study of anti-inflammatory, analgesic and antipyretic activities of seeds of Hyoscyamus niger and isolation of a new coumarinolignan.

A chemical and biological validation of the traditional use of Hyoscyamus niger seeds as anti-inflammatory drug has been established. The methanolic extract of seeds of H. niger (MHN) was evaluated for its analgesic, anti-inflammatory and antipyretic activities in experimental animal models at different doses. MHN produced significant increase in hot plate reaction time, while decreasing writhing response in a dose-dependent manner indicating its analgesic activity. It was also effective in both acute and chronic inflammation evaluated through carrageenin-induced paw oedema and cotton pellet granuloma methods. In addition to its analgesic and anti-inflammatory activity, it also exhibited antipyretic activity in yeast-induced pyrexia model. Furthermore, the bioactive MHN under chemical investigation showed the presence of coumarinolignans as major chemical constituent and yielded a new coumarinolignan, cleomiscosin A methyl ether (1) along with four known coumarinolignans, cleomiscosin A (2), cleomiscosin B (3), cleomiscosin A-9'-acetate (4) and cleomiscosin B-9'-acetate (5). The structure elucidation of 1 was done by spectroscopic data interpretation and comparative HPLC analysis. Cleomiscosin A, but not its isomer cleomiscosin B, reduced dry and wet weight of cotton pellet granuloma in mice. This suggests that cleomiscosin A is an important constituent of MHN responsible for anti-inflammatory activity.

Gastroprotective effect of standardized extract of Ficus glomerata fruit on experimental gastric ulcers in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditionally fruits of Ficus glomerata Roxb (Family: Moraceae) are used to treat anemia and gastrointestinal disorders. AIM: Gastroprotective effect of 50% ethanolic extract of F. glomerata fruit (FGE) was studied in different gastric ulcer models in rats. METHODS: FGE (50, 100 and 200 mg/kg body weight) was administered orally, twice daily for 5 days for prevention from pylorus ligation (PL), ethanol (EtOH) and cold restraint stress (CRS)-induced ulcers. Estimation of H+K+ATPase activity and gastric wall mucous were performed in EtOH-induced ulcer and antioxidant enzyme activities in supernatant mitochondrial fraction of CRS-induced ulcers. RESULTS: FGE showed dose dependent inhibition of ulcer index in pylorus ligation, ethanol and cold restraint stress-induced ulcers. FGE prevents the oxidative damage of gastric mucosa by blocking lipid peroxidation and by significant decrease in superoxide dismutase, H+K+ATPase and increase in catalase activity. High performance thin layer chromatography (HPTLC) analysis showed the presence of 0.57% and 0.36% w/w of gallic acid and ellagic acid in FGE. CONCLUSIONS: Our results show that F. glomerata possess significant gastroprotective activity which might be due to gastric defence factors and phenolics might be the main constituents responsible for this activity.

Protective effect of (+)-catechin against gastric mucosal injury induced by ischaemia-reperfusion in rats.

Ischaemia and reperfusion are known to induce gastric lesions, predominantly due to excessive formation of reactive oxygen metabolites, adhesion of neutrophils to endothelial cells, microvascular dysfunction, gastric acid secretion, endogenous histamine and gastrin release. We have studied the effect of (+)-catechin on a gastric ulcer model involving damage to gastric injury by ischaemia- reperfusion (I/R) in rats. (+)-Catechin 50 mg kg(-1)administered orally, once daily for three days after the initiation of I/R injury showed a significant (P<0.001) anti-ulcer activity against mucosal dam- age. However, (+)-catechin significantly decreased the lipid peroxidation and increased the level of catalase in the I/R condition. Elevated levels of alkaline phosphatase in the I/R group was significantly lowered (P<0.01) by (+)-catechin. The amount of H(+)K(+)ATPase was significantly decreased (P<0.001) in (+)-catechin-treated as compared with I/R rats. (+)-Catechin significantly decreased elevated plasma histamine (P<0.05) and corticosterone (P<0.05). The results suggested that (+)-catechin protected gastric mucosa against ischaemia-reperfusion-induced gastric ulcers by its antioxidant activity and mucus protection.

Pregnancy-promoting actions of HCG in human myometrium and fetal membranes.

Human chorionic gonadotropin (HCG) plays a major role in early human development through a series of well recognized pregnancy-promoting actions that are exerted in the first trimester, including maternal recognition of pregnancy, enhancement of embryo implantation and survival, stimulation of trophoblast growth and differentiation, and prolongation of the functional life of the corpus luteum. Recent research indicates that HCG can exert significant pregnancy-promoting actions also in the remainder of pregnancy through its effect on the myometrium and on fetal membranes. In the myometrium, HCG promotes the inhibition of smooth muscle cell contractility through several mechanisms, including inhibition of gap junction formation, reduction of intracellular calcium concentration, increase in the expression of progesterone receptor, and an increase in the expression of phosphodiesterase 5 (PDE5), an enzyme controlling the intracellular levels of cGMP. This effect appears to be specific for PDE5 since it has not been found for other hormones potentially involved in pregnancy such as estrogen, progesterone and thyroid hormone. In fetal membranes, HCG can modulate expression of the inducible isoform of nitric oxide synthase (iNOS), as well as specific immunoregulatory cytokines such as the high mobility group box 1 (HMGB1) protein. This accumulating evidence suggests that HCG has a wide spread pregnancy-promoting actions that are exerted in various reproductive and gestational tissues.

Evaluation of anti-inflammatory activity of Cissampelos pareira root in rats.

In the present study, 50% ethanolic extract of Cissampelos pareira roots (CPE) in acute, subacute and chronic models of inflammation was assessed in rats. Per os (p.o.) administration of CPE (200, 400 mg/kg) exhibited significant anti-inflammatory activity. In acute inflammation as produced by carrageenin 59.55% and 64.04%, by histamine 15.38% and 30.77%, by 5-hydroxytryptamine 17.78% and 31.11% and by prostaglandin E(2)-induced hind paw edema 19.23% and 30.77% protection was observed. While in subacute anti-inflammatory models using formaldehyde-induced hind paw edema (after 1.5 h) 38.36% and 47.95% and in chronic anti-inflammatory model using cotton pellet granuloma 15.02% and 19.19% protection from inflammation was observed. CPE did not show any sign of toxicity and mortality up to a dose level of 1000 mg/kg, p.o. in rats. Both acute as well as chronic administration of CPE (100, 200 and 400 mg/kg, p.o.) did not produce any gastric lesion in rats. These data indicate that CPE possesses significant anti-inflammatory activity without ulcerogenic activity suggesting its potential as an anti-inflammatory agent for use in the treatment of various inflammatory diseases.

Action of Hygrophila auriculata against streptozotocin-induced oxidative stress.

Hygrophila auriculata (K. Schum.) Heine (Family: Acanthaceae) is a wild herb widely used in 'Ayurveda' as 'Rasayana' drug for treatment of various disorders. Treatment of diabetic rats with aerial parts of Hygrophila auriculata extract (HAEt, 100 and 250 mg/kg body weight) for 3 weeks showed significant reduction in blood glucose, thiobarbituric acid reactive substances (TBARS) and hydroperoxide in both liver and kidney. The treatment with HAEt significantly increased the glutathione (GSH), glutathione peroxidase (GPx), glutathione S-transferase (GST) and catalase (CAT) in the drug-treated group, which is comparable to the control group. HAEt and glibenclamide-treated rats also showed decreased lipid peroxidation that is associated with increased activity of superoxide dismutase (SOD) and catalase. The ability of HAEt on tissue lipid peroxidation and antioxidant status in diabetic animals has not been studied before. The result of this study thus shows that HAEt possesses significant antidiabetic activity along with potent antioxidant potential in diabetic conditions.

Bone medullary arterioles from ovariectomized rats have smaller baseline diameters but normal eNOS expression and NO-mediated dilation.

This study was designed to test the hypothesis that endogenous estrogens decrease the expression of endothelial nitric oxide synthase (eNOS) in resistance-size bone arterioles, thereby reducing endothelium-dependent vasodilator function. Sexually mature female rats were ovariectomized to reduce endogenous estrogens. Age-matched female rats served as controls. Seven to ten days after ovariectomy, bone marrow tissue was collected from the femoral canal. Immuno-histochemistry was performed to detect expression of estrogen receptors, alpha and beta and eNOS. eNOS protein content in medullary bone arterioles was compared using Western blot analysis. Endothelial cell function was assessed by quantitating the dilation of isolated, pressurized bone arterioles in response to acetylcholine. The results indicate that the endothelium of bone arterioles from ovariectomized and control rats express ER-alpha, ER-beta and eNOS. eNOS protein content in the two groups of arterioles did not differ. However, the baseline diameter of arterioles from ovariectomized rats (63+/-4 microm) was significantly smaller than the diameter of arterioles from control rats (75+/-3 microm, p<0.05). The two groups of arterioles dilated equally in response to acetylcholine. L-NAME, an inhibitor of eNOS, almost completely abolished the dilator responses to acetylcholine, but not to sodium nitroprusside. L-Arginine restored acetylcholine-induced dilation after L-NAME treatment. Thus, arteriole dilation to acetylcholine appears to be mediated almost exclusively by NO. The smaller diameter of arterioles from ovariectomized rats suggests that endogenous estrogens exert a significant dilator influence on bone arterioles. However, the dilator influence does not appear to be mediated by an increase in eNOS expression or enhanced NO-dependent vasodilation. These results indicate that estrogens do not decrease eNOS expression or diminish NO-mediated dilation of bone medullary arterioles.

Orthotopic transplantation of LH receptor knockout and wild-type ovaries.

Luteinizing hormone (LH) receptor knockout animals have an ovarian failure due to an arrest in folliculogenesis at the antral stage. As a result, the animals have an infertility phenotype. The present study was undertaken to determine whether this phenotype could be reversed by orthotopic transplantation of wild-type ovaries. The results revealed that transplanting wild-type ovaries into null animals did not result in resumption of estrus cycles. Although the number of different types of follicles increased, none progressed to ovulation. The serum hormone profiles improved, reflecting the ovarian changes. The wild-type animals with null ovaries also failed to cycle and their ovaries and serum hormone levels were more like null animals with their own ovaries. Although the lack of rescue of null ovaries placed into wild-type animals was predicted, the failure of wild-type ovaries placed in null animals was not, which could be due to chronic exposure of transplanted tissue to high circulating LH levels and also possibly due to altered internal milieu in null animals. These findings may have implications for potential future considerations of grafting normal donor ovaries into women who have an ovarian failure resulting from inactivating LH receptor mutations.

Dependence of uterine cyclooxygenase2 expression on luteinizing hormone signaling.

Several previous studies have demonstrated that uterine Cox2 (also known as Ptgs2) is required for implantation. Luteinizing hormone (LH) released from anterior pituitary gland and human chorionic gonadotropin released from placenta (hCG) can upregulate the uterine Cox2 gene expression. The Lhcgr knockout (herein designated LHRKO) animals have implantation failure even after estradiol and progesterone therapy. These findings led us to investigate the dependence of uterine Cox2 gene expression on LH signaling in LHRKO animals. The results revealed that, while Cox1 (also known as Ptgs1) mRNA levels were similar, Cox2 mRNA levels were lower in uterus of null animals than in wild-type siblings. Treatment with hCG did not increase Cox2 mRNA levels in null endometrial stromal or myometrial smooth-muscle cells unless gene therapy was performed to introduce native LHCGR. The Cox1 mRNA levels, on the other hand, did not change regardless of the introduction of native or activated Lhcgr or hCG treatment. The Cox2 mRNA increase paralleled the cAMP raise, suggesting that LH uses the cAMP second messenger system. Treating the wild-type uterine cells with hCG resulted in a Cox2 but not Cox1 mRNA increase. This increase became exaggerated when additional native LHCGR were introduced by gene therapy. In conclusion, deletion and reinsertion of Lhcgr further support that uterine Cox2 gene expression is dependent on LH signaling.

Targeted disruption of LH receptor gene revealed the importance of uterine LH signaling.

Numerous previous studies have demonstrated that LH and hCG can directly regulate several uterine functions. We investigated in the present study, whether uterine phenotype in LH receptor knockout animals resulted also from the absence of direct actions of LH in the uterus. The phenotype consisted of marked growth reduction of uterus, decreased thickness of endometrial and myometrial layers, number of endometrial glands, height of luminal epithelium and vascular space. Analysis of uterine gene expression by mouse genome U74Av2 Affymetrix genechips revealed a differential expression of 155 genes by more than 3-fold (range 3-53-fold) between null and wild-type animals. Of these, 89 genes decreased and 66 increased in uterus of null animals. Semi-quantitative RT-PCR confirmed the differential expression of several selected genes. The decreased genes can be clustered into 18 functional families and the increased into 15 functional families. Semi-quantitative RT-PCR, Western blotting and immunocytochemistry demonstrated a decreased expression of ERbeta, PR-A, PR-B and AR in uterus of null animals as compared with wild-type siblings. Twenty-one-day estradiol and progesterone replacement therapy did not normalize the decrease in the number of endometrial glands and several genes that either decreased or increased in expression. The partial success of therapy suggests that direct LH actions could be required to completely normalize the uterus. In summary, findings on the knockout model reaffirm that LH and hCG control uterine functions directly as well as indirectly through increasing ovarian synthesis of steroid hormones and both actions are required for normal uterine biology.

Anti-inflammatory and anti-nociceptive activity of the water decoction Desmodium gangeticum.

The water decoction of root and aerial parts of Desmodium gangeticum (Leguminosae) was examined for anti-inflammatory and anti-nociceptive activity in experimental animals. The root and aerial decoction in doses of 5, 10 and 20 mg /kg caused a dose-dependent inhibition of swelling caused by carrageenin equivalent to 14.58-51.02% protection and 14.43-38.67%, respectively, in cotton pellet granuloma. There was a significant increase in analgesio-meter-induced force equivalent to 6.56-67.66% protection and 22.18-73.83% protection in acetic acid-induced writhing. The result establishes the traditional use of water decoction of Desmodium gangeticum codified in Indian System of Medicine.

Antiulcer activity of Utleria salicifolia rhizome extract.

The effect of 50% ethanolic extract of Utleria salicifolia (USE) was assessed in different acute and chronic gastric ulcer models in rats. USE, 50-200 mg/kg administered orally, twice daily for 5 days showed dose-dependent ulcer protective effect in pylorus ligation (14.48-51.03% protection, P < 0.5 to P < 0.01), aspirin (28.80-56.52% protection, P < 0.5 to P < 0.001), ethanol (13.22-60.74% protection, P < 0.5 to P < 0.001), cold-restraint stress (21.22-77.14% protection, P < 0.05 to P < 0.001), and acetic acid (20.0-84.37% protection, P < 0.5 to P < 0.001)-induced acute and chronic ulcers. USE also significantly (P < 0.001) reduced the ulcer incidence (50 and 10%) and severity (67.83 and 91.34% protection) of duodenal ulcer, induced by cysteamine. Besides USE offered protection (53.52 and 60.58%) against ethanol-induced depletion of gastric wall mucus. However, USE reduced the ulcer index with significant decrease in plasma corticosterone (25.53 and 39.52% protection, P < 0.1 and P < 0.05), lipid peroxidation (18.75 and 47.92% protection, P < 0.01 and P < 0.001), superoxide dismutase (15.80 and 26.61% protection, P < 0.05 and P < 0.001) and increased in catalase (28.42 and 71.0% protection, P < 0.05 and P < 0.001) activity, respectively. Preliminary phytochemical screening of the USE gave the positive test for steroids, alkaloids, terpenoids, saponins and tannins. The HPTLC studies in the toluene: ethyl acetate: formic acid and the densitometric scanning at 254 nm gave three major spots with area corresponding to 28.16, 17.17, and 13.79% at 0.69, 0.78, and 0.88 R(f) values, respectively. The results indicate that USE possesses antiulcer activity.

Functional luteinizing hormone/chorionic gonadotropin receptors in human adrenal cortical H295R cells.

Previous studies have suggested that activation of normal human adrenal and adrenal tumor luteinizing hormone (LH)/chorionic gonadotropin (hCG) receptors results in an increased secretion of steroid hormones. Since it is not feasible to test this suggestion on normal human adrenal cells, we used human adrenal cortical carcinoma H295R cells, which are similar in some respects to normal adrenal cortical cells. These cells contained LH/hCG receptor transcripts and receptor protein that can bind (125)I-hCG in a hormone-specific manner. Culturing the cells with highly purified hCG resulted in a time- and dose-dependent significant increase in dehydroepiandrosterone sulfate (DHEAS) secretion as compared with the controls. The DHEAS response was hormone as well as steroid specific. Since hCG treatment did not increase DHEA secretion, we suspected that the hCG might increase DHEA sulfotransferase (ST). Consistent with this possibility, hCG treatment increased steady-state DHEA-ST mRNA levels. The hCG effects require its receptors, as inhibition of their synthesis by treatment with antisense phosphorothioate oligodeoxynucleotides (ODN) made from the LH/hCG receptor sequence resulted in loss of DHEA-ST and DHEAS responses. The findings that 1) hCG treatment increased cAMP levels and activated protein kinase A (PKA), 2) 8-bromo cAMP mimicked hCG, and 3) blocking PKA activation prevented hCG as well as 8-bromo cAMP from increasing both DHEA-ST mRNA and DHEAS levels suggested that cAMP/PKA signaling was involved in the hCG actions. In conclusion, H295R cells contain LH/hCG receptors, which are coupled to increasing DHEAS secretion through upregulating the ST enzyme mRNA level. This action is mediated by the cAMP/PKA signaling pathway. These findings support the concept that adrenal function in normal and pathological conditions could be influenced by LH and hCG.