Sunday, April 28, 2013

Hyperglycaemia is bad

Hyperglycaemia does whatever you want it to. Want to show it increases glycolysis and/or oxidative phosphorylation? No problem. Want to show it decreases both? Equally no problem. Choose your tissue, choose your duration, choose your insulin level, choose your glucose level, choose your tissue culture medium before test, choose... With the correct combination you can show anything.

But certain patterns emerge from lots of papers. In the short term hyperglycaemia increases both glycolysis and oxidative phosphorylation. Acute hyperglycaemia in neurons induces an equally acute hyperpolarisation of the inner mitochondrial membrane (a pre requisite for reverse electron flow through complex I), followed by a burst of free radicals (from reverse electron transport in the face of a low NAD+/NADH ratio?), followed by a collapse of the inner mitochondrial membrane potential (from free radical induced loss of cytochrome c?), soon to be followed by apoptosis, as you might expect

These guys set out the events nicely but suggest the mechanism is unclear. I would be willing to bet on G-3-P dehydrogenase as driving reverse electron flow using the high membrane potential from glycolysis. It seems that, under "mitochondrial preparation" conditions, ignoring reverse electron flow, G-3-P dehydrogenase also spills a reasonable dose of free radicals not only inwards towards the matrix but also outwards to the inter membrane space, in roughly equal amounts. As does complex III of course, but complex III is not specifically driven by a short side branch of hyperglycaemia-induced hyperactive glycolysis. Goodness only knows if this happens in-vivo, but let's accept that it does. Cytochrome c is on the outer surface of the inner mitochondrial membrane and spilling free radicals outwards seems a good way to oxidise the cardiolipin anchors and release one of the most important pro apoptotic proteins we have, cytochrome c.

So acute hyperglycaemic injury, in a cell type where glucose entry is essentially concentration driven, is potentially apoptotic if the injury is severe enough. Lesser but sill significant injury may come from spills of superoxide from complex I on to the mitochondrial DNA, another potentially interesting effect. Research on G-3-P dehydrogenase is still in its infancy and there are no clear cut answer as to how important this scenario might be, but I rather like it. Is it true? Who knows. It's hard to tell.

Exactly how difficult it is to transfer information from "preparations" to any semblance of "in vivo" is reviewed by Martin Brand. I like this chap, he really looks at the limitations of how much we currently know (not much, it appears) plus he came up through Naked Mole Rat research, another positive. Here's his summary of where free radicals might be produced:

Outwards spillage, directly on to cytochrome c, from G-3-P dehydrogenase and complex III...

It's quite clear that hyperglycaemia is not invariably acutely fatal to all neurons on first exposure. It takes years of following the advice of the ADA and AHA to develop diabetic neuropathy or to kill off enough central neurons (around 70%) to get the clinical label of Alzheimers and, while recurrent hyperglycaemia might get us there directly, the indirect effects are much more interesting to a mitochondriac like myself.

Chronic hyperglycaemia is where we have a depressed inner mitochondrial membrane potential, reduced glycolysis and electron transport with subsequent failure to generate superoxide.

Badness too.


Wednesday, April 24, 2013

Axen and Axen (4) Ketogenic insulin resistance. It's all over now...

I have so many posts I want to get finished, all of which are inter-related and all of which need waaaaaay too much work, that I thought I would just throw this one out in the interim. I began with this paper which came as a pdf from Liz. While I was getting the pubmed link to it I noticed the same group had another rather similar paper out which was equally interesting and then the third link down the page was an accidental find which is this one, subject of this post.

I don't know if it's worth going through the figures individually, they are very similar to those from Axen and Axen which produced a series of posts a few years ago, except that the feature of COMPLETE reversal of insulin resistance is, here, presented right up front in Figure 6 and in the abstract too:

That figure for insulin looks a little dubious at 120 minutes but I'll let that go, I guess p was still > than 0.05... Pretty close to full reversal.

It's quite hard to know exactly how much this group understand about their results. They give roughly equal weight to the adverse (sic) effects of a ketogenic diet as they do to the fact it is reversible within a week (or less, they only checked at a week) of re-introducing carbohydrate.

What they seem to lack is the concept that rats fed a very restricted carbohydrate diet MUST be insulin resistant to survive, as happens in starvation. But maybe they are creeping towards some sort of understanding. It's about time. Good.

When people cite Axen and Axen to prove ketogenic diets are going to make you diabetic (there are folks who believe this, or at least wish you to believe it!) you have an answer in Kinzig et al 2010.

BTW, the links which led me here relate to using ketogenic diets to control both pain and inflammation. This is a potentially very useful tool but the beneficial effect does appear to be as rapidly reversible as the physiological insulin resistance... Ketogenic diets are a fix, not a cure (in the short term anyway). But inflammation appears to be a feature of ageing, long term, and if KDs work in "ageing inflammation" all we have to decide is the age at which we should all start on a KD. Unless someone has a method of stopping the ageing process of course....