Patent Application: US-87277107-A

Abstract:
a method of treating or preventing diseases related to modulation of ppar - α in comprising administering to a human or other mammals in need of such treatment an effective amount of plant material derived from plants of the genera puerila lobata .

Description:
we discovered that treatment of puerarin ( 1 - 100 μm ) in human liver cell line ( hepg2 ) upregulated the peroxisome proliferator - activated receptor ( ppar )- α ( fig2 ), a nuclear transcription factor that modulates gene expressions in governing lipid metabolism and plays an important role in obesity . in exploring the possible clinical uses of puerarinn we conducted experiments in rats and found that rats that were given puerarin ( 250 mg / kg / day ) orally were weight 13 % less than the control rats in average over 5 months period with no apparent toxicity ( fig3 ). upon extensive search of literature , we believe that we are the first to discover that puerarin upregulates ppar - α in liver cells and prevents weight gain in animal . experimental procedures , materials puerarin was obtained from natural pharmacia international , inc . ( belmont , mass .). polyclonal anti - ppar - α antibody was purchased from santa cruz biotechnology , inc . ( santa cruz , calif .). fetal bovine serum was purchased from hydcone laboratories , inc . ( logan , utah ). dulbecco &# 39 ; s modified eagle &# 39 ; s medium ( dmem ) was purchased from mediatech , inc . ( herdon , va . ), penicillin / streptomycin was purchased from invitrogen corp . ( carlsbad , calif .). cell cultures hepg2 cells was purchased from american type culture collection ( manassas , va .). they were maintained in dmem containing 10 % heat - inactivated fetal bovine serum and 1 % penicillin / streptomycin in tissue culture dishes incubated in 95 % air / 5 % co 2 , at 37 ° c . the media were changed 2 times per week until the cells were confluent . protein immuno - blotting of ppar - α . hepg2 cells were collected in ripa buffer ( 150 mm nacl , 1 % triton x - 100 , 1 % sodium deoxycholate , 0 . 1 % sodium dodecyl sulfate , 50 mm tris - hcl , 2 mm edta , ph 7 . 5 ) with protease inhibitors ( mini - complete protease inhibitor cocktail tablets ( roche diagnostics , mannheim , germany )] after incubated in puerarin ( 1 - 100 μm ) for 24 hours . the protein content of the cell lysate was quantified by using a protein assay kit ( bio - rad lab ,, hercules calif .). protein ( 50 μg ) from the whole cell lysate was loaded in each lane of a polyacrylamide ( 10 %) gel , eletrophoresed and blotted to a polyvinylidene difluoride ( pvdf ) membrane . the membrane was first probed with a rabbit polyclonal antiserum against ppar - 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