Contents: 1. PCOS: etiopathogenesis 2. DOGMA theory – Dysbiosis of Gut Microbiota 3. Evidence for microbiological dysbiosis and a “leaky” gut in PCOS 4. Chronic low-grade inflammation as a cornerstone of PCOS pathology 5. Chronic inflammation as a cause of insulin resistance 6. Probiotics and Prebiotics – potential new treatment for PCOS

1. Novel paradigm in PCOS
etiology: gut dysbiosis
Dr. Arun Sharma
Manager, Medical Affairs
Mankind Pharma Ltd.

2. Contents
• PCOS: etiopathogenesis
• DOGMA theory – Dysbiosis of Gut Microbiota
• Evidence for microbiological dysbiosis and a “leaky” gut in
PCOS
• Chronic low grade inflammation as a cornerstone of PCOS
pathology
• Chronic inflammation as a cause of insulin resistance
• Probiotics and Prebiotics – potential new treatment for
PCOS

3. PCOS: Chronic Metabolic & Endocrine disorder
• Affects 3–26% of reproductive-aged women
• In the long term, PCOS leads to an increased risk of:
• Endometrial cancer
• Diabetes
• Metabolic syndrome
• Cardiovascular disease
Michelmoreetal.,1999;Marchetal.,2010;Lietal.,2013
Legroetal.,1999;Hardimanetal.,2003;DumesicandLobo,2013

4. Clinical features
• Hyperandrogenism
• Hyperinsulinemia
• Anovulation
• Polycystic ovarian morphology
Landay et al., 2009
Lim et al., 2013
2.8-fold higher prevalence of obesity compared to women
without PCOS

5. Exact pathophysiology unknown
• Genetic, neuroendocrine and metabolic causes have been
suggested
• No single pathological process can account for all cases of PCOS
• PCOS is heterogeneous with many patients not exhibiting all
three cardinal features of the PCOS ‘‘triad’’
Diabetes 1989;38(9):1165–74.
N Engl J Med 2005;352(12): 1223–36.
Nat Rev Endocrinol 2011;7(4):219–31.

6. Clinical features central to pathophysiology
• Considerable debate on what exactly constitutes PCOS
• 2 key biochemical features present in the vast majority of
women with PCOS are
• Insulin resistance
• Chronic inflammation

9. Novel microbiological paradigm for the
development of PCOS
• Dysbiosis of gut microbiota can increase the ovarian
production of androgens
• Further interference with normal follicle development
• Triggering a chronic inflammatory response and insulin
resistance
Tremellen and Pearce, 2012
DOGMA theory – Dysbiosis of Gut Microbiota

11. TheDOGMAtheoryforcreation
ofPCOS

12. Key pathophysiological links in the DOGMA theory
• A diet high in saturated fat and refined sugars
• Favors the growth of ‘‘bad’’ Gram negative bacteria within the gut
• Reduces the growth of beneficial ‘‘good’’ bacteria such as
Bifidobacteria and Lactobaccilus
• The cell wall of Gram negative bacteria contains a powerful
immuno-stimulant called lipopolysaccaride (LPS)
• This causes profound activation of the innate immune
system if it is allowed to traverse the gut wall and enter the
systemic circulation Diabetes 2007;56(7):1761–72.
Curr Opin Pharmacol 2009;9(6):737–43.
Sci Transl Med 2009;1(6):6–14.
Am J Physiol Gastrointest Liver Physiol 2010;299(2):G440–8.

13. Key pathophysiological links in the DOGMA theory
• High saturated fat-sugar/low fibre diet, & obesity leads to an
increase in gut mucosal permeability
• This facilitates transfer of LPS from the gut lumen into
circulation, initiating a state of ‘‘metabolic endotoxaemia’’.
• Chronic activation of hepatic and tissue macrophages produces
impaired insulin receptor function and resulting insulin resistance
• Hyper-insulinaemia then drives an increase in the ovaries
production of androgens and halts normal ovulatory processes
producing a ‘‘blockade’’ in follicle development
Endocrinology 1988;123(2):733–9.
Obstet Gynecol 198; 64(Suppl. 3):73S–80S.
Endocrinology 1988;122(5):2034–43.

15. Evidence for microbiological dysbiosis and
a “leaky” gut in PCOS

16. High energy intake, with abundant saturated fats and
refined sugars, is more common in PCOS women than
their normal or overweight ovulatory controls

17. A high-fat/high-sugar diet favours colonization of the
intestine with ‘‘bad’’ Gram negative bacteriaresulting in
a metabolic endotoxaemia induced inflammatory state

18. High fat/sugar
diet
Decreased
bifidobacteria
Increased gut
permeability
Metabolic
endotoxemia
Activation of
immune system

19. Obesity alters gut permeability too!
A positive correlation between waist circumference,
visceral fat content on DEXA assessment and
increases in colonic gut permeability

20. • Adipose tissue may trigger an increase in gut permeability
• Visceral fat deposits have been linked with an increase in production of
the pro-inflammatory cytokine Tumour Necrosis Factor alpha (TNFa)
• Resulting in a reduction in the production of anti-inflammatory protein
adiponectin

21. The increase in gut permeability and resulting
metabolic endotoxaemia would help explain the
observed positive correlation between markers of
immune activation such as CRP and increasing BMI

22. Gutpermeability
Metabolic
endotoxemia
CRP
BMI
Positive correlation between metabolic endotoxemia &
markers of immune activation

23. Colonic mucosal permeability in PCOS
• No study to date has directly measured differences in
colonic mucosal permeability between PCOS patients and
normal controls
• Research suggests that such a difference is likely to exist
• The prevalence of Irritable Bowl Syndrome (IBS) & Chronic
Fatigue Syndrome throw some light on this

24. Irritable Bowel Syndrome (IBS) and Chronic Fatigue
Syndrome (CFS), linked to increased gut permeability and
metabolic endotoxaemia have both been shown to be
significantly more common in PCOS

25. Since both IBS and CFS have been linked with
metabolic endotoxaemia and immune activation,
women with PCOS might experience a similar
‘‘leaky gut’’ syndrome

26. Chronic low grade inflammation as a
cornerstone of PCOS pathology

27. PCOS is a pro-inflammatory state
Women with PCOS exhibit an elevation in circulating CRP that is
independent of obesity. This finding corroborates existing molecular
evidence of the chronic low-grade inflammation that may underpin the
pathogenesis of this disorder
Meta-
analysis
of 31
articles

28. Chronic inflammation as a cause of
insulin resistance

29. Insulin resistance and PCOS: strong association
• Common in both obese and lean women with PCOS
• Affects up to 70% of the PCOS population
• 5 to 10-fold higher prevalence and later conversion to frank diabetes
in the PCOS cohort
• Some studies support a genetic predisposition to insulin resistance
• Mounting evidence sugges that the chronic state of inflammation seen
in PCOS may be responsible for initiating insulin resistance Diabetes 1989;38(9):1165–74.
Hum Reprod Update 2010;16(4):347–63.
N Engl J Med 2005;352(12): 1223–36.

30. Role of pro-inflammatory cytokines
• Even in lean PCOS individuals, the presence of a ‘‘leaky gut’’
would generate an increase in serum TNFα and IL-6
• Both TNFα and IL-6 have been linked to the generation of
insulin resistance
• Gut microbiota would be capable of initiating a state of
insulin resistance if the passage of LPS into the systemic
circulation is permitted by an increase in colonic mucosal
permeability (‘leaky gut’’)
Science 1993;259(5091):87–91.
Science 1996;271(5249): 665–8.

31. The direct administration of LPS into the circulation of mice and humans
has been shown to cause an increase in fasting glucose and insulin
levels, confirming insulin resistance

32. A beneficial adjustment in the balance between ‘‘good’’ and ‘‘bad’’
bowel bacteria using Lactobacillus and Bifidobacterium probiotics
can result in improved insulin sensitivity

33. Summary of the DOGMA theory of PCOS

34. Recap
• A high fat and sugar & low fibre diet, alters the normal
balance of beneficial and harmful bacteria in the gut lumen
• Increased translocation of immuno-stimulatory LPS
molecules from gut lumen into the systemic circulation due
to:
• Poor diet related changes
• Direct inflammatory effect of excess adiposity
• Possible increase in numbers of Gram negative ‘‘bad’’ bacteria
• Leading to a state of metabolic endotoxaemia

35. Recap
• ‘‘Metabolic endotoxaemia’’ activates macrophages in the fat,
liver and muscle
• Leading to release of high levels of TNFα and insulin
resistance
• This state of hyper-insulinaemia interferes with normal
follicle development in the ovary, causing a halt in follicular
development with the generation of multiple small follicles
(typical polycystic morphology on ultrasound), and impaired
ovulation with its associated menstrual irregularity.

36. Probiotics and Prebiotics – potential
new treatment for PCOS

37. Probiotic: WHO Definition (2002)
• Live microorganism which, when administered in adequate
amounts, confers a health benefit to the host
• Criteria needed to be met by a microorganism in order for it
to be considered a probiotic:
1. Must be resistant to gastric acidity and bile acid toxicity
2. Must adhere to the intestinal epithelial cells so that it is not
rapidly lost in the faecal material
3. Must convey a benefit to the host human – implying it is non
pathogenic (safe)
Annu Rev Food Sci Technol 2011;2:373–93

38. Most commonly used probiotic organisms
Bifidobacteria Lactobacillus
Annu Rev Food Sci Technol 2011;2:373–93

39. Prebiotics: a selective food source for beneficial
‘‘good’’ bacteria
• First introduced by Gibson and Roberfroid
• An alternative way of boosting the number of ‘‘good’’
bacteria within the gut, rather than by direct application of
these bacteria (prebiotic approach)
• Defined as ‘‘a selectively fermented ingredient that allows
specific changes, both in the composition and/or activity in
the gastrointestinal microflora that confers benefits upon
host wellbeing and heath’’
Annu Rev Food Sci Technol 2011;2:373–93
Br J Nutr 2010;104(Suppl. 2):S1–S63.

40. Prebiotics meeting European Union’s criteria
• Fructo-oligosaccharides (FOS)
• Inulin (a long chain form of FOS)
• Galactooligosaccharides
• Lactulose
Br J Nutr 2010;104(Suppl. 2):S1–S63.

41. Synbiotics
• Combinational therapy consisting of a prebiotic and a
probiotic
• Target synergy being that the prebiotic will selectively
stimulate the growth and/or activity of both the
endogenous and exogenously applied (prebiotic) beneficial
bacteria
• Advantageous over a pure prebiotic approach
• Host has a diet that has insufficient dietary fibre to
support the optimal growth and function of the colonic
microbiota.
Annu Rev Food Sci Technol 2011;2:373–93

42. Current evidence supporting the use
of probiotics and prebiotics in

43. Significant reduction in the incidence of anovulatory
infertility (primarily PCOS mediated) if the subject consumed
high amounts of whole milk, compared to those subjects
drinking skim or low fat milk
Prebiotic effect of whole milk is possible

44. Potential of prebiotics to drive down insulin levels in
PCOS
• Animal studies have shown that prebiotic and probiotic
supplements can help prevent impaired insulin resistance, a
central driver of PCOS pathology
• These animal studies are backed up by a large trial of 256
pregnant women
• Prebiotic treatment significantly reduced the incidence of
gestational diabetes (36% v 13%, p = 0.003), while also
improving insulin sensitivity in both the pregnancy and 12-
month post natal period
Br J Nutr 2010;104(12):1831–8.
Br J Nutr 2009;101(11):1679–87.
Br J Nutr 2010;103(12):1792–9.

45. Probiotics/prebiotics may treat PCOS via a
weight loss mechanism: 3 theories
Prebiotic fibre increases faecal energy and fat excretion,
effectively reducing energy intake

46. Production of short chain fatty acids by beneficial bacteria increases the colonic
mucosa’s production of the ‘‘satiety’’ hormone Glucagon-Like Peptide-1 (GLP-1)
which has central action on the brain reducing appetite and energy intake
The prebiotic oligosaccharide has also been shown to increase serum GLP-1
levels supporting a role for prebiotics in controlling human energy intake

47. • Research now suggests that changes in bowel flora may even be
directly responsible for obesity by increasing the energy harvest
from the colonic luminal contents.
• Lactobacillus gasseri containing probiotic to cause a significant
reduction in abdominal adiposity and body weight, without the
need for an increase in exercise or change in energy intake

48. Summary: role of prebiotics and
probiotics in PCOS

49. Recap: prebiotics and probiotics
• Improve intestinal barrier function
• Reduce the colonic Gram negative bacterial load
• Reduction in transfer of LPS across the mucosal wall
• Reduction of metabolic endotoxaemia
• Probiotics mediate increase in production of GLP-1 which
reduces energy intake
• GLP-1 also drops adipose tissue mass and a decrease in
inflammation

50. Asymbioticapproachtothe
managementofPCOS
MedicalHypotheses79(2012)104–11