CBR1 are expressed primarily on presynaptic peripheral and central nerve terminals

The 2-AG basal level is ∼1,000 times greater than AEA in the brain. The enzyme acylphosphatidylethanolamine-phospholipase D is involved in the formation of AEA, and the enzyme diacylglycerol lipase is involved in 2-AG formation. Once synthesized and released, endocannabinoids are removed from the extracellular space through an endocannabinoid membrane transporter, subsequently AEA is hydrolyzed by the enzyme fatty acid amide hydrolase , and 2-AG is degraded by cytosolic monoacylglycerol lipase. There are two G protein-coupled ECB receptors. CBR1 receptors are highly expressed on presynaptic neurons in the brain, spinal cord, and dorsal root ganglion. CBR2 receptors are primarily expressed in immune cells. AEA maintains basal endocannabinoid tone and has a high selectivity for the CBR1 receptor over the peripheral CBR2 receptor. The CBR1 receptor is the most abundant G protein-coupled receptor in the brain and one of the most abundant in both the peripheral and central nervous system.CBR1 is the central receptor responsible for the behavioral and psychotropic effects of the “high” caused by THC. The CBR1 receptor is also present in multiple immune cells, making it important when considering anti-inflammatory properties of endocannabinoids. The CBR2 receptor is largely present on peripheral immune cells and participates in regulation of the immune system. The principal endogenous ligand for the CBR2 receptor is 2-AG. In the brain, stimulation of CBR2 receptors does not produce cannabis-like effects. The anti-inflammatory effects of exogenous cannabinoids are mediated by the endocannabinoid system,cannabis grow facility layout likely through CB2Rs in the periphery that have immunomodulatory functions. 

In neural and non-neural systems, in response to tissue injury or excessive nociception, the ECB system generally suppresses inflammation, suppresses sensitization, and suppresses pain ; however, ECB activity on the nociception system can be complicated, with ECB antinociceptive or pronociceptive depending on the site of expression and the underlying physiological brain state. When considering ECB pain biomarkers, it is important to consider that the ECB system interacts significantly with inflammation mechanisms as well as opioid mechanisms. The ECB system regulates inflammation at multiple levels and generally inhibits inflammation. In preclinical and laboratory investigation, it has been shown that the ECB system inhibits pro- inflammatory cytokines while increasing production of antiinflammatory cytokines. The ECB system also inhibits immune cell activation, immune cell proliferation and migration, and can increase immune cell apoptosis via multiple mechanisms. CBR1 receptors are 10 times more concentrated than mu-opioid receptors in the brain, and cannabinoid receptors co-localize with opioid receptors in many regions involved in pain circuitry including the dorsal horn of the spinal cord and in the supra-spinal periaqueductal gray and rostroventral medulla. ECB and opioid pain biomarkers potentially can be identified via endogenous opioid function which can be assumed when comparing pain sensitivity in the presence of opioid blocking treatment, such as naloxone vs. placebo. Higher endogenous opioid function is associated with decreased benefit from opioid treatments such as morphine. Exercise induced analgesia involves both opioid and endocannabinoid mechanisms. Exercise induced increases of endocannabinoid ligands such as AEA are blocked by naltrexone. This indicates that opioids are involved in the increase of endocannabinoid ligands following exercise. Intrathecal morphine decreases circulating levels of endocannabinoids demonstrating how the opioid and endocannabinoid systems are linked. 

Lower endogenous opioid function is associated with greater analgesia from pain treatment with opiates. One study demonstrated that low endocannabinoid activity is also associated with greater analgesia from pain treatment with opiates. Pathogenic alterations in the distribution of microbial species within the gut is associated with neuropathic pain in a variety of clinical conditions. One study found that reductions in the diversity and increases in the ratios of two microbial species may contribute to HIV associated neuropathic pain. This may be particularly relevant in the context of the endocannabinoid system, as the endocannabinoid system regulates homeostasis of multiple organ systems, including the gut. Because dysregulation of the gut-brain axis can result in chronic inflammation and neuroinflammation, endocannabinoids have anti-oxidant, and anti-inflammatory properties relevant to modulation of inflammation that occurs along the gut-brain axis. MicroRNA modulated inflammation has a major role in the induction and maintenance of neuropathic pain. inflammationregulating microRNA profiles in patients with peripheral neuropathies have been characterized. In patients with polyneuropathies of different etiologies the expression of miR-21-5p, miR-146a, and miR-155 were upregulated. In painful neuropathies, tissues from skin biopsies from the lower leg, where neuropathic changes are most common, had reduced miR-146a and miR-155 expression compared to the thigh; furthermore, peripheral neuropathies are associated with aberrant microRNA expression in the sural nerve and in the skin. In sural nerve biopsies of patients with peripheral neuropathies miR-132-3p expression was more than doubled in white blood cells of neuropathy patients compared to healthy controls as well as in painful compared with non-painful neuropathy. MiRNA’s have been found to be upregulated in polyneuropathies, and miR-21 is increased in painful neuropathies. Cortisol is a proposed stress-related pain biomarker. DHEA and DHEAS are neurosteroids that modulate inhibitory GABA receptors and excitatory NMDA receptors, producing complex neuronal effects. In animal studies, DHEA and DHEAS levels have been proposed as a biomarker for pain. In multi-variable regression analysis, gender, age, and pain perception in the shoulder and upper limbs were significantly related to serum DHEAS. 

In another study plasma DHEAS levels were lower compared with persons with chronic neck pain compared with controls with no pain. One study found that the odds of having depressive symptoms increased with higher cortisol/DHEA-S ratios among people living with HIV on treatment,indoor grow shelves suggesting altered neuroactive steroid metabolism may contribute to the pathophysiological mechanisms of depression in people living with HIV. A study of male war veterans found that reductions in DHEA levels were associated with muscle soreness and were positively associated with chest pain. Self-reported back pain measures in female war veterans were inversely correlated with DHEA and DHEA-S ; those reporting moderate to severe low back pain demonstrated significantly lower DHEA-S levels compared to those with no or mild lower back pain. Two reviews on mechanisms and imaging biomarkers for diabetic neuropathic pain review that diabetic peripheral neuropathy and associated pain have structural and functional central nervous system changes in the spinal cord, subcortex, and cortex. Diabetic peripheral neuropathy has been associated with changes in the thalamus. A decreased thalamic NAA/creatinine ratio is suggestive of thalamic neuronal dysfunction , and thalamic microvascular perfusion changes have also been observed. Smaller spinal cord crosssectional area has been observed in those with diabetic peripheral neuropathy. In fact, in diabetic peripheral neuropathy, diffusion tensor imaging techniques found posterior column damage in the cervical spinal cord. Diabetic neuropathic pain is related to decreased NAA in the thalamus , increased thalamic vascularity , and spinal cord posterior column damage. Diabetic neuropathic pain is associated with increased regional brain gray matter volume loss localized to brain regions involved in somatosensory perception ; furthermore, in diabetic neuropathic pain, increased total gray matter atrophy is associated with impaired ability to walk. Diabetic neuropathic pain has been shown to be related to aberrant default mode functional connectivity , decreased functional connectivity between the thalamus and cortex , and decreased functional connectivity in attention networks. Altered fMRI activation responses to experimental heat pain in limbic and striatal brain circuits are related to the duration of diabetic neuropathic pain. Diabetic neuropathic pain is related to a double dissociation such that neuropathic pain intensity is more associated with thalamus-insular cortex functional connectivity and nerve deficits are more related to thalamus-somatosensory cortex functional connectivity. Diabetic neuropathic pain is also associated with decreased functional connectivity between the thalamus and amygdala , decreased gray matter volumes and decreased white matter connectivity in pain processing and pain modulation brain regions , decreased somatosensory cortical thickness related to cortical function dysfunction , increased activity in the anterior cingulate cortex , as well as ventrolateral periaqueductal gray functional connectivity is altered and correlates with magnitude of spontaneous pain and allodynic pain. 

Structural brain imaging has revealed changes to the brain associated with HIV peripheral neuropathy. Total cortical volume is smaller with HIV distal neuropathic pain. In fact, in HIV distal neuropathic pain the posterior cingulate cortex is the cortical region that was found to be smaller. In another sample of people living with HIV, subjective symptoms of HIV peripheral neuropathy were associated with smaller precuneus volumes which overlap with the posterior cingulate cortex. Smaller brain volumes for HIV distal neuropathic pain are consistent with a general pattern that brain volumes are reduced for a variety of chronic pain conditions. Interestingly, the midbrain, thalamus and posterior cingulate cortex volumes are all reduced in HIV distal neuropathic paresthesia. It has been suggested that brain atrophy associated with HIV distal neuropathic paresthesia may precede brain atrophy associated with HIV distal neuropathic pain. More recent multi-modal brain imaging research has revealed structural brain changes associated with HIV peripheral neuropathy. HIV peripheral neuropathy is correlated with decreased white matter integrity running from the midbrain to the somatosensory cortex. HIV peripheral neuropathy severity is also associated with decreased generalized fractional anisotropy along the tracts of the external capsule in both hemispheres, appearing to lead along the lateral thalamus to sensorimotor cortex. A similar correlation is found in the superior bilateral cingulum. These results indicate ascending deafferentation in HIV peripheral neuropathy extends further downstream from damaged peripheral nerves than seen previously, into the cortex. HIV-associated distal neuropathic pain is associated with decreased fMRI resting state functional connectivity in the default mode network and increased functional connectivity in the salience network. Decreased connectivity between the medial prefrontal cortex and posterior cingulate cortex and stronger connectivity between the ACC and thalamus is associated with HIV distal neuropathic pain. In the setting of experimental heat pain, significant interaction has been found within the right anterior insula during expectation of experimental pain offset in that a group with HIV distal neuropathic pain compared group without HIV distal neuropathic pain exhibited increased insula activation in the feet compared to the hand. These findings are consistent with abnormal processing of expectation of experimental pain offset or abnormal pain relief mechanisms potentially due to increased negative expectation regarding the experience of chronic endogenous neuropathic pain. Anterior cingulate cerebral perfusion and gray matter density correlate with chemotherapy-induced peripheral neuropathy symptoms including pain. Patients with chemotherapyinduced peripheral neuropathy symptoms demonstrated greater activation during painful stimulation in the precuneus compared to healthy controls and exhibited hypo-activation of the right superior frontal gyrus compared to healthy controls. Painful stimuli delivered chemotherapy induced peripheral neuropathy symptoms patients evoke differential activation of distinct cortical regions, reflecting a unique pattern of central pain processing compared with healthy controls providing a tool for monitoring cerebral changes during anti-cancer and analgesic treatment. A population of mixed small-fiber peripheral neuropathy was used to investigate how dysfunction of skin nerves led to abnormal recruitment of pain-related brain regions, suggesting that the brain may be affected in SFN. Greater volume reduction in pain-processing regions, particularly the bilateral anterior cingulate cortices was associated with greater depletion of intraepidermal nerve fibers. There was significant reduction in functional connectivity from the anterior cingulate cortex to the insula pain-processing cortex that is linearly correlated with the severity of intraepidermal nerve fiber depletion. Similarly, another population of mixed small-fiber peripheral neuropathy the degree of skin nerve degeneration was associated with the reduction of connectivity between the thalamus and pain-related areas. Despite altered white matter connectivity, there was no change in white matter integrity assessed with fractional anisotropy. These findings indicate that alterations in structural connectivity may serve as a biomarker of maladaptive brain plasticity that contributes to neuropathic pain after peripheral nerve degeneration. A population of Charcot-Marie-Tooth patients had abnormal diffusion tensor imaging findings indicative of significant cerebral white matter abnormalities. diffusion tensor imaging abnormalities were correlated with clinical disability, suggesting that there is comorbidity of central nervous system damage with peripheral neuropathy in Charcot-Marie-Tooth patients. A population of patients with hereditary neuropathy with liability to pressure palsies were compared to a population of normal controls and the fractional anisotropy values of the patients were significantly lower in bilateral frontal, orbitofrontal, and temporal areas of white matter. Patient populations of paresthesia-dominant and pain-dominant patient groups were compared and contralesional cortical thickness were correlated with pain severity.