PMID- 1649742
OWN - NLM
STAT- MEDLINE
DCOM- 19910828
LR  - 20131121
IS  - 0013-7227 (Print)
IS  - 0013-7227 (Linking)
VI  - 129
IP  - 2
DP  - 1991 Aug
TI  - Ovarian actions of tumor necrosis factor-alpha (TNF alpha): pleiotropic effects 
      of TNF alpha on differentiated functions of untransformed swine granulosa cells.
PG  - 641-8
AB  - We have examined interactions between tumor necrosis factor-alpha (TNF alpha), a 
      product of the immune system, and ovarian cells using serum-free monolayer 
      cultures of untransformed swine granulosa cells. Recombinant human TNF alpha, a 
      potent cytoactive product of activated macrophages, bound specifically and with 
      high affinity to intact granulosa cells. Binding sites had an apparent Kd of 0.17 
      nM (95% confidence interval, 0.065-0.31), and a binding capacity of 80 
      nmol/micrograms DNA (95% confidence interval, 52-110). The binding capacity of 
      granulosa cells for TNF alpha (but not the binding affinity) was increased 
      approximately 2-fold by treatment with FSH and insulin. The biological effects of 
      TNF alpha on pig granulosa cells were expressed after 48 and 96 h in culture. At 
      the latter time, TNF alpha significantly suppressed insulin- and insulin- plus 
      FSH-stimulated progesterone accumulation, with respective ID50 values of 0.08 +/- 
      0.008 and 0.06 +/- 0.014 nM, but did not affect basal progesterone accumulation 
      or DNA content. TNF alpha also significantly attenuated the stimulatory effect of 
      combined treatment with FSH and insulin on cAMP generation during 48-96 h of 
      culture. TNF alpha inhibited the stimulatory effects of forskolin, cholera toxin, 
      and the cAMP analog 8-bromo-cAMP on progesterone accumulation, indicating 
      multiple sites of action of this immune modulator. Inhibition of progestin 
      biosynthesis was observed even in the presence of 25-hydroxycholesterol, a 
      soluble oxygenated sterol substrate for the cholesterol side-chain cleavage 
      reaction, and was accompanied by decreased concentrations of specific cellular 
      mRNA encoding cholesterol side-chain cleavage enzyme. There were no changes in 
      the amounts of a constitutively expressed enzyme, phosphoglyceraldehyde 
      dehydrogenase. Inhibitory actions of TNF alpha were specific to de novo steroid 
      hormone biosynthesis, since nanomolar concentrations of this cytokine stimulated 
      accumulation of prostaglandin E2 and prostaglandin F2 alpha basally and during 
      treatment with FSH, cholera toxin, or 8-bromo-cAMP. In contrast, prostaglandin 
      accumulation was not enhanced by interferon-gamma or interleukin-2. In summary, 
      untransformed porcine granulosa cells exhibit specific, high affinity, low 
      capacity saturable binding sites for TNF alpha, and the number of such binding 
      sites can be regulated by combined treatment with insulin and FSH. Granulosa 
      cells are susceptible to the inhibitory actions of TNF alpha on FSH- and 
      insulin-supported progesterone biosynthesis and cAMP accumulation. One important 
      locus of TNF alpha action is blockade of hormonally stimulated increases in 
      specific mRNA encoding the cholesterol side-chain cleavage cytochrome P450 
      enzyme.(ABSTRACT TRUNCATED AT 400 WORDS)
FAU - Veldhuis, J D
AU  - Veldhuis JD
AD  - Department of Internal Medicine, University of Virginia Health Sciences Center, 
      Charlottesville 22908.
FAU - Garmey, J C
AU  - Garmey JC
FAU - Urban, R J
AU  - Urban RJ
FAU - Demers, L M
AU  - Demers LM
FAU - Aggarwal, B B
AU  - Aggarwal BB
LA  - eng
GR  - 3-MO1-RR-00847-1491/RR/NCRR NIH HHS/United States
GR  - DK-38942/DK/NIDDK NIH HHS/United States
GR  - KO4-HD-00634/HD/NICHD NIH HHS/United States
GR  - etc.
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Endocrinology
JT  - Endocrinology
JID - 0375040
RN  - 0 (Insulin)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 4G7DS2Q64Y (Progesterone)
RN  - 9002-68-0 (Follicle Stimulating Hormone)
RN  - B7IN85G1HY (Dinoprost)
RN  - E0399OZS9N (Cyclic AMP)
RN  - EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme)
RN  - K7Q1JQR04M (Dinoprostone)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Cholesterol Side-Chain Cleavage Enzyme/genetics/metabolism
MH  - Cyclic AMP/metabolism
MH  - Dinoprost/metabolism
MH  - Dinoprostone/metabolism
MH  - Female
MH  - Follicle Stimulating Hormone/pharmacology
MH  - Granulosa Cells/drug effects/*metabolism
MH  - Insulin/pharmacology
MH  - Nucleic Acid Hybridization
MH  - Progesterone/biosynthesis
MH  - RNA, Messenger/metabolism
MH  - Recombinant Proteins/pharmacology
MH  - Swine
MH  - Tumor Necrosis Factor-alpha/*physiology
EDAT- 1991/08/01 00:00
MHDA- 1991/08/01 00:01
CRDT- 1991/08/01 00:00
PHST- 1991/08/01 00:00 [pubmed]
PHST- 1991/08/01 00:01 [medline]
PHST- 1991/08/01 00:00 [entrez]
AID - 10.1210/endo-129-2-641 [doi]
PST - ppublish
SO  - Endocrinology. 1991 Aug;129(2):641-8. doi: 10.1210/endo-129-2-641.