Mechanisms of cholesterol reduction by Lactobacillus ...
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Kolida S | Hernandez O | OHara SP| Rastall RA| Contact: Dr Sofia Kolida | R&D Director | OptiBiotix Health PLC |Tel: ++ 44 7585001450| Background Lactobacillus plantarum LP LDL Ò is a probiotic with established cholesterol reducing activity in normal to mildly hypercholesterolemic adults. Probiotics can reduce cholesterol by various mechanisms: • Adsorption to cell surface. • Integration to cell membrane. • Bile Salt Hydrolase (BSH) activity- prevents bile salt reabsorption. • Cholesterol esterase activity-conversion to coprostanol. Mechanisms of cholesterol reduction by Lactobacillus plantarum LP LDL ® Aims Investigate the relevance of the above described mechanisms to the cholesterol lowering activity of LP LDL Ò . Methods To investigate the ability of LP LDL Ò of passively removing cholesterol through adsorption on its cell membrane, pure cultures were carried out in the presence and absence of cholesterol. Samples were analysed using scanning electron microscopy (SEM). Faecal, micro-scale, pH-controlled batch cultures (10ml) containing cholesterol-enriched growth media, were carried out in the presence of LP LDL Ò . Culture supernatants were obtained at 0, 10 and 24h for: • Bile Salt Hydrolase (BSH) activity determination. • Cholesterol and coprostanol concentrations were determined using GC-FID. A double blind, placebo controlled, randomized study was carried out in normal to mildly hypercholesterolemic adults to confirm cholesterol reducing efficacy in vivo. Results Adsorption of cholesterol on LP LDL Ò cell surface: SEM of LP LDL Ò grown in the absence (A) and presence of cholesterol (B) following washing in PBS. A B BSH activity in pH- controlled faecal batch cultures: significant increase in bile salt hydrolysis at 8 and 24h with significantly higher preference for glycolate. 0 50 100 150 200 250 300 350 Control Control LPLDL® Control LPLDL® 0h 8h 24h Activity units/ml Glycholate Glycodeoxycholate Taurocholate Taurodeoxycholate 0 5 10 15 20 25 30 35 Control Control LPLDL® Control LPLDL® 0h 8h 24h % cholesterol removal Cholesterol reducing activity in pH-controlled faecal batch cultures: significant increase in cholesterol reduction at 8 and 24h in cultures where LP LDL Ò was present. Coprosanol formation in pH- controlled faecal batch cultures: Conversion of cholesterol to coprostanol in the presence of LP LDL Ò after 8 and 24h fermentation. Once formed, coprostanol is excreted in faeces in vivo. 0 h 8 h 24 h Impact of LP LDL ® intake on serum lipid profiles in vivo: statistically significant reduction in systolic blood pressure, LDL and total cholesterol and an increase in HDL cholesterol in normal to mildly hypercholesterolemic adults Study Highlights Our findings suggest that LP LDL Ò can mediate cholesterol reduction by three mechanisms: I. Adsorption II. BSH activity III. Enzymatic conversion to coprostanol The ability of LP LDL Ò to reduce cholesterol through several mechanisms contributes to its enhanced efficacy in improving serum lipid profiles in human studies.
Transcript of Mechanisms of cholesterol reduction by Lactobacillus ...
Kolida S | Hernandez O | OHara SP| Rastall RA|
Contact: Dr Sofia Kolida | R&D Director | OptiBiotix Health PLC |Tel: ++ 44 7585001450|
BackgroundLactobacillus plantarum LPLDL
Ò is a probiotic with establishedcholesterol reducing activity in normal to mildly hypercholesterolemicadults. Probiotics can reduce cholesterol by various mechanisms:
• Adsorption to cell surface.• Integration to cell membrane.• Bile Salt Hydrolase (BSH) activity- prevents bile salt reabsorption.
• Cholesterol esterase activity-conversion to coprostanol.
Mechanisms of cholesterol reduction by Lactobacillus plantarum LPLDL®
AimsInvestigate the relevance of the above described mechanisms to thecholesterol lowering activity of LPLDL
Ò.
MethodsTo investigate the ability of LPLDL
Ò of passively removing cholesterolthrough adsorption on its cell membrane, pure cultures were carriedout in the presence and absence of cholesterol. Samples wereanalysed using scanning electron microscopy (SEM).Faecal, micro-scale, pH-controlled batch cultures (10ml) containingcholesterol-enriched growth media, were carried out in the presenceof LPLDL
Ò . Culture supernatants were obtained at 0, 10 and 24h for:• Bile Salt Hydrolase (BSH) activity determination.• Cholesterol and coprostanol concentrations were determinedusing GC-FID.
A double blind, placebo controlled, randomized study was carried outin normal to mildly hypercholesterolemic adults to confirmcholesterol reducing efficacy in vivo.
Results
Adsorption of cholesterol on LPLDLÒ cell surface: SEM of LPLDL
Ògrownin the absence (A) and presence of cholesterol (B) following washingin PBS.
A B
BSH activity in pH-controlled faecal batchcultures: significantincrease in bile salthydrolysis at 8 and 24hwith significantly higherpreference for glycolate.
0
50
100
150
200
250
300
350
Control Control LPLDL® Control LPLDL®
0h 8h 24h
Activ
ity u
nits
/ml
Glycholate Glycodeoxycholate Taurocholate Taurodeoxycholate
0
5
10
15
20
25
30
35
Control Control LPLDL® Control LPLDL®
0h 8h 24h
% c
hole
ster
ol re
mov
al
Cholesterol reducing activity in pH-controlled faecal batchcultures: significant increase in cholesterol reduction at 8 and24h in cultures where LPLDL
Òwas present.
Coprosanol formation in pH-controlled faecal batchcultures: Conversion ofcholesterol to coprostanol inthe presence of LPLDL
Ò after 8and 24h fermentation. Onceformed, coprostanol isexcreted in faeces in vivo.
0 h
8 h
24 h
Impact of LPLDL® intake on serum lipid profiles in vivo:
statistically significant reduction in systolic blood pressure, LDLand total cholesterol and an increase in HDL cholesterol innormal to mildly hypercholesterolemic adults
Study HighlightsOur findings suggest that LPLDL
Ò can mediate cholesterolreduction by three mechanisms:I. Adsorption
II. BSH activityIII. Enzymatic conversion to coprostanolThe ability of LPLDL
Ò to reduce cholesterol through severalmechanisms contributes to its enhanced efficacy in improvingserum lipid profiles in human studies.
