**7. The changes in SDH and KDH activity in patients with metabolic syndrome and arterial hypertension**

254 Dehydrogenases

animal.

SDH in the intact restless rabbit is hyperactivated nearly up to the level of calm under ADR influence. Thus, as judged by ES level and the SDH activity, the restless rabbit has no quiescence in the quiet state without external excitations. The same state of continuous internal metabolic excitation is inherent in all patients examined in our laboratory in

At the background of the initially high SDH activity in restless rabbit ADR administration leads to only small stimulation, which only somewhat exceeds the level under the influence of ADR in calm rabbit. The influence of SUC, which was somewhat stronger activator when injected to the calm rabbit, leads to realization of risk of transition to inhibition in restless

The difference in ADR and SUC effects on ES can be explained by different mechanisms of their influence on SDH activity. ADR only activates SDH through protein kinase. The influence of SUC is more complicated. SUC is an allosteric activator of SDH [58]. Simultaneously the rise of SUC level increases OAA formation, which inhibits SDH

These effects are better pronounced in a sample without SUC addition, namely ES, because SUC excess partially abolish OAA inhibition. Therefore in sample ES SDH inhibition is better observed than in sample for SDH measurement, containing 5 mM SUC, because it is not masked by the addition of substrate. Probably, the main part of ES under SUC injection is ESUC, which is accumulated due to OAA inhibition. It should be considered that measurements are made for an hour. Insight into dynamics of the processes can be achieved by a comparison of data for calm rabbit, in which oxidation is slower with those for restless one, in which oxidation is accelerated. As shown for calm rabbit, at the beginning rapid oxidation of SUC occurred, resulted in an increase in OAA, which is evidenced by the initial accumulation of ESUC. The next stage is presented by data for restless rabbit in which stronger inhibition of SDH by OAA is already developed, which is not completely abolished

by SUC addition and therefore ESUC accumulation is higher than in calm animal.

The data presented show great difference of the two CBCh properties of the state of mitochondria in animals with opposite behaviour, which corresponds in magnitude to the

We have also found the pronounced difference in the SDH and KDH activity in rats before maturation, 6 week old with domination of ADR regulation, and just after maturation, 8 week old with increase in ACH regulation. These changes are close to differences between

It was shown also that complicated menopause in rats and women, which is manifested under the decrease in ADR regulation in the organism, is related to the increase in the SDH activity and can be corrected by metabolic regulators, containing SUC - "Amberen" [59].

The data considered support the view of existence of the link between activity of studied

contrast to healthy volunteers. The data will be considered in the next sections.

according to the mechanism of the negative feedback.

pronounced individual peculiarities in behaviour.

DHs and the level of ADR and ACH regulation.

calm and restless rabbits.

The stages of the SDH and KDH activity changes found under stress were observed in patients with metabolic syndrome and arterial hypertension.

Disturbance of normal metabolic flexibility between oxidation of glucose and fatty acids is the basis of many diseases. High dietary fat intake and low physical activity provoke this disorder. Complex of accompanied biochemical changes is called metabolic syndrome. Metabolic syndrome underlies various pathological symptoms, such as obesity, insulin resistance, type 2 diabetes, risk of hypertension, heart diseases and others [1, 60-65].

Metabolic syndrome is caused by increased intensity of life accompanied with persistent sympathetic hyperactivation and deficit of physical activity that leads to PES. That is why we have studied the participation of mitochondria in development of PES. The importance of mitochondria in development of metabolic syndrome is caused by their crucial role in intracellular oxidation. Aerobic fibers of skeletal muscle mitochondria make a major contribution to regulation of metabolic syndrome and support metabolic health of the whole organism.

Data are expressed in mean area (S) of formazan granules in 30 cells. Owing to a great number of objects calculated (200-800) the presented data are statistically significant at the level of 0.001 (99.99%) Inscription above - types of patterns.

SDHtru is calculated as the difference between samples SUC 5 mM - MAL 5mM.

The changes of SDH and SDHisc activity are presented in per cents respective to conditional norm of patient 1 (100%). The low row for SUC 5mM, the upper row for SUC 5 mM + ISC 5 mM.

The changes in the SDH activity in non-severe, non-hospitalized and workable patients with allergy to some food products are given in Figure 9. In this case for more precise determination of the SDH activity we subtracted ES contribution from the data for the sample with SUC addition. This value was termed as a true SDH (SDHtru). It was determined under addition of SUC and also SUC+ISL, modulator of the SDH activity. This provides determination of the functional state of SDH as considered in section 5.5. Calculation of SDHtrue help to find clearly pronounced similarity between some patients, which is masked by ES contribution. According to SDHtru the patients can be joined in subgroups with nearly identical data for members inside one subgroup and clearly different from the other.

Study of Succinate Dehydrogenase and α-Ketoglutarate Dehydrogenase in Mitochondria Inside Glass-Adhered Lymphocytes Under Physiological Conditions – The Two Dehydrogenases as Counterparts of … 257

adaptation. However, the changes in more severe patient, N4 are already beyond the physiological range and are analogous to that for damaging stress in Figure 4. It is also seen that the response to signal concentration of SUC corresponds to the SDH activity, measured

The presented examples show that the loss of full value quiescence, which is manifested in the complete switching SDH off, is a general difference of all examined patients from healthy people. Therefore mitochondria of patients in the relatively quiet state of the

According to the influence of ISL functional state of SDH in patients with hypertension differs from that of patients with allergy. Hidden inhibition of SDH is typical of allergy, while hyperactivation of SDH is more typical of hypertension. This coincides with more pronounced sympathetic hyperactivation in hypertensive patients, demanding intake of

with substrate concentration of SUC, as shown in Figure 5.

**Figure 10.** CBCh patterns of patients hospitalized with hypertonic disease. The data are expressed as the total area of formazan granules in 30 lymphocytes.

disease. The gender and age are indicated between.

significant at the level of 0.001 (99.99%)

AP, upper line - the data just before blood taking, low line - more stable data from History of the

Owing to a great number of objects calculated (200-800) the presented data are statistically

organism have no metabolic quiescence.

adrenoblockers.

The first example from the left is for healthy person. The next one, 1, is taken as reference control to other patients. He used diet containing no allergenic products for more than 7 years and for this period his body weight, AD and other physiological characteristics were greatly normalized. Other patients have some pathological symptoms and used recommended diet for only several months. Among them we have found increase in SDH activity.

For healthy people full absence of SDHtrue - complete quiescence state is observed. Low SDHtrue was determined for patient with normalization of the state. In all others patients different levels of activation or hyperactivation of SDH were found. In the state of hyperactivation without inhibition, ISL addition results in only the tendency to stimulation. Progressive hyperactivation is masked by internal inhibition of OAA. This is revealed by the increase in activity with ISL. For the patients with greater symptoms, presented in the right part of the Figure 9, the increase in the internal inhibition is typical, which is manifested in both, the SDH activity decrease and strong increase in activation by ISL.

Hyperactivation of the SDH activity without inhibition is more typical of patients hospitalized with hypertension. In Figure 10 data are presented on the total SDH activity (SDH tot) without subtraction of ES contribution. It is seen also, as in Figure 9, that for the healthy control very low activity of SDH is typical. The rise in the activity can be considered as hyperactivation without inhibition by both, their greater value and because ISL does not stimulate it but, in contrast, decreases. ISL addition serves as unique test of the functional state of SDH developed by our group. Additionally to the explanations of ISL influence in section 5.5 it should be mentioned that effect of ISL is dose-dependent. It can be seen from the example of patient N2. Addition of 5mM of ISL decreases hyperactivation of SDH per 2/3, while 50μM only per 1/3.

The extent of the deflection of the SDH activity from norm should be quantitatively determined using different concentrations of ISL. In Figure 10 (as in Figures 5 and 7) the KDH activity is also given, as well as the response of mitochondria to the signal concentration of SUC - 5μM. In norm the KDH activity is low and nearly equal to the SDH activity. At hyperactivation of SDH in patients N2 and N3, KDH is also greatly increased, however less than SDH. Such coupled changes in the two DH activities in patients are similar to those, which were found in rats under PES within the range of physiological

from the other.

activity.

2/3, while 50μM only per 1/3.

The changes in the SDH activity in non-severe, non-hospitalized and workable patients with allergy to some food products are given in Figure 9. In this case for more precise determination of the SDH activity we subtracted ES contribution from the data for the sample with SUC addition. This value was termed as a true SDH (SDHtru). It was determined under addition of SUC and also SUC+ISL, modulator of the SDH activity. This provides determination of the functional state of SDH as considered in section 5.5. Calculation of SDHtrue help to find clearly pronounced similarity between some patients, which is masked by ES contribution. According to SDHtru the patients can be joined in subgroups with nearly identical data for members inside one subgroup and clearly different

The first example from the left is for healthy person. The next one, 1, is taken as reference control to other patients. He used diet containing no allergenic products for more than 7 years and for this period his body weight, AD and other physiological characteristics were greatly normalized. Other patients have some pathological symptoms and used recommended diet for only several months. Among them we have found increase in SDH

For healthy people full absence of SDHtrue - complete quiescence state is observed. Low SDHtrue was determined for patient with normalization of the state. In all others patients different levels of activation or hyperactivation of SDH were found. In the state of hyperactivation without inhibition, ISL addition results in only the tendency to stimulation. Progressive hyperactivation is masked by internal inhibition of OAA. This is revealed by the increase in activity with ISL. For the patients with greater symptoms, presented in the right part of the Figure 9, the increase in the internal inhibition is typical, which is manifested in

Hyperactivation of the SDH activity without inhibition is more typical of patients hospitalized with hypertension. In Figure 10 data are presented on the total SDH activity (SDH tot) without subtraction of ES contribution. It is seen also, as in Figure 9, that for the healthy control very low activity of SDH is typical. The rise in the activity can be considered as hyperactivation without inhibition by both, their greater value and because ISL does not stimulate it but, in contrast, decreases. ISL addition serves as unique test of the functional state of SDH developed by our group. Additionally to the explanations of ISL influence in section 5.5 it should be mentioned that effect of ISL is dose-dependent. It can be seen from the example of patient N2. Addition of 5mM of ISL decreases hyperactivation of SDH per

The extent of the deflection of the SDH activity from norm should be quantitatively determined using different concentrations of ISL. In Figure 10 (as in Figures 5 and 7) the KDH activity is also given, as well as the response of mitochondria to the signal concentration of SUC - 5μM. In norm the KDH activity is low and nearly equal to the SDH activity. At hyperactivation of SDH in patients N2 and N3, KDH is also greatly increased, however less than SDH. Such coupled changes in the two DH activities in patients are similar to those, which were found in rats under PES within the range of physiological

both, the SDH activity decrease and strong increase in activation by ISL.

adaptation. However, the changes in more severe patient, N4 are already beyond the physiological range and are analogous to that for damaging stress in Figure 4. It is also seen that the response to signal concentration of SUC corresponds to the SDH activity, measured with substrate concentration of SUC, as shown in Figure 5.

The presented examples show that the loss of full value quiescence, which is manifested in the complete switching SDH off, is a general difference of all examined patients from healthy people. Therefore mitochondria of patients in the relatively quiet state of the organism have no metabolic quiescence.

According to the influence of ISL functional state of SDH in patients with hypertension differs from that of patients with allergy. Hidden inhibition of SDH is typical of allergy, while hyperactivation of SDH is more typical of hypertension. This coincides with more pronounced sympathetic hyperactivation in hypertensive patients, demanding intake of adrenoblockers.

**Figure 10.** CBCh patterns of patients hospitalized with hypertonic disease.

The data are expressed as the total area of formazan granules in 30 lymphocytes.

AP, upper line - the data just before blood taking, low line - more stable data from History of the disease. The gender and age are indicated between.

Owing to a great number of objects calculated (200-800) the presented data are statistically significant at the level of 0.001 (99.99%)
