Influence of acute blood loss on the dynamics of pro-inflammatory cytokine content in the context of traumatic brain injury, blunt abdominal trauma, and skeletal trauma

Aim: To determine the dynamics of the content of pro-inflammatory cytokines, Tumor Necrosis Factor-α (TNF-α) and Interleukin-1β (IL-1β), in blood serum under conditions of traumatic brain injury (TBI), blunt abdominal trauma (BAT), and skeletal trauma (ST), complicated by acute blood loss, after 3, 7, 14, 21, and 28 days of the post-traumatic period. Materials and Methods: Under thiopental sodium anesthesia, male Wistar line rats were subjected to modeled TBI, BAT, and ST. In separate groups, acute blood loss of the amount 1,5% of body weight was additionally modeled. The content of TNF-α and IL-1β in blood serum was determined after 3, 7, 14, 21, and 28 days of the post-traumatic period. Results: Experimental traumas of various localizations were accompanied by a pronounced systemic pro-inflammatory reaction, manifested by a significant increase in serum levels of pro-inflammatory cytokines TNF-α and IL-1β. The maximum of disturbances was observed in the early post-traumatic period, followed by a decrease, which only reached the control level in the case of TBI and ST. In conditions of BAT, the indicators were significantly higher compared to other localized traumas. When the modeled mechanical traumas of various localizations were complicated by acute blood loss of the amount 1,5% of body weight, the disturbances in the content of TNF-α and IL-1β in the blood serum became greater. The TNF-α content in all experimental groups increased up to the 7-th day, remained at the same level up to day 14, and then decreased. The IL-1β content in blood serum reached its maximum as early as 1 day of the posttraumatic period and then decreased, but both indicators did not reach the control level by the 28-th day. In conditions of BAT, the indicator was significantly higher after 3-21 days of the post-traumatic period compared to other experimental groups. Conclusions: Mechanical traumas of various localizations significantly differ in the nature of the manifestations of the systemic inflammatory reaction and react differently when complicated by acute blood loss. The greatest increase in the serum content of TNF-α and IL-1β is noted in conditions of blunt abdominal trauma.

Aim: To determine the features of disturbances in the antioxidant-prooxidant balance of the liver in conditions of mechanical trauma of various localizations complicated by acute blood loss. Materials and Methods: Traumatic brain injury(TBI), blunt abdominal trauma (BAT), and skeletal trauma (ST) were modeled in mature male Wistar line rats under conditions of thiopental sodium anesthesia. Additionally, acute blood loss in the amount of 1,5 % of body weight was modeled in the traumatized rats. After 3, 7, 14, 21, and 28 days of the post-traumatic period, catalase activity and malondialdehyde content were determined in the liver homogenate extract, and the antioxidant-prooxidant index (API) was calculated based on their ratio. Results: Modeling mechanical traumas of different localizations, compared to the control, caused a shift in the antioxidant-prooxidant balance in the liver towards the predominance of prooxidant mechanisms. In the case of traumatic brain injury, the result was statistically significant starting from the 7-th day of the post-traumatic period, and for blunt abdominal trauma and skeletal trauma - starting from the 3-rd day. The greatest decrease in the antioxidant-prooxidant index at all times of the post-traumatic period was observed with blunt abdominal trauma. Complication of the modeled mechanical traumas with acute blood loss was accompanied by a deepening of metabolic disorders in the liver. In the case of traumatic brain injury, the liver API reached its minimum after 14 days of the post-traumatic period; for traumas of other localizations - already after 3 days. The amplitude of impairments at all times of the post-traumatic period was greater in the group of rats with blunt abdominal trauma. Conclusions: Modeling mechanical traumas of different localizations compared to the control is accompanied by a shift in the liver's antioxidant-prooxidant balance towards the prevalence of prooxidant mechanisms, which is intensified by additional acute blood loss in the amount of 1,5 % of body weight. The largest disorders in the index are noted with blunt abdominal trauma.

The aim of this study was to determine whether anodal and cathodal high-voltage monophasic pulsed currents (HVMPC) provided to the area of a pressure injury (PI) change the blood level of cytokines (interleukin [IL]-1β, IL-10, and tumor necrosis factor [TNF]-α) and growth factors (insulin-like growth factor [IGF]-1 and transforming growth factor [TGF]-β1) in patients with neurological injuries and whether the level of circulatory cytokines and growth factors correlates with PI healing progression. This study was part of a randomized clinical trial on the effects of HVMPC on PI healing. All patients with neurological injuries (spinal cord injury, ischemic stroke, and blunt trauma to the head) and a stage 2, stage 3, or stage 4 PI of at least 4 weeks' duration hospitalized in one rehabilitation center were eligible to participate if older than 18 years of age and willing to consent to donating blood samples. Exclusion criteria included local contraindications to electrical stimulation (cancer, electronic implants, osteomyelitis, tunneling, necrotic wounds), PIs requiring surgical intervention, patients with poorly controlled diabetes mellitus (HbA1C > 7%), critical wound infection, and/or allergies to standard wound treatment. Participants were randomly assigned to 1 of 3 groups: anodal (AG) or cathodal (CG) HVMPC treatment (154 μs; 100 Hz; 360 µC/sec; 1.08 C/day) or a placebo (PG, sham) applied for 50 minutes a day, 5 days per week, for 8 weeks. TNF-α, IL-1β, IL-10, TGF-β1, and IGF-1 levels in blood serum were assessed using the immunoenzyme method (ELISA) and by chemiluminescence, respectively, at baseline and week 4. Wound surface area measurements were obtained at baseline and week 4 and analyzed using a digitizer connected to a personal computer. Statistical analyses were performed using the maximum-likelihood chi-squared test, the analysis of variance Kruskal-Wallis test, the Kruskal-Wallis post-hoc test, and Spearman's rank order correlation; the level of significance was set at P ≤.05. Among the 43 participants, 15 were randomized to AG (mean age 53.87 ± 13.30 years), 13 to CG (mean age 51.08 ± 20.43 years), and 15 to PG treatment (mean age 51.20 ± 14.47 years). Most PIs were located in the sacral region (12, 74.42%) and were stage 3 (11, 67.44%). Wound surface area baseline size ranged from 1.00 cm2 to 58.04 cm2. At baseline, none of the variables were significantly different. After 4 weeks, the concentration of IL-10 decreased in all groups (AG: 9.8%, CG: 38.54%, PG: 27.42%), but the decrease was smaller in the AG than CG group (P = .0046). The ratio of pro-inflammatory IL-10 to anti-inflammatory TNF-α increased 27.29% in the AG and decreased 26.79% in the CG and 18.56% in the PG groups. Differences between AG and CG and AG and PG were significant (AG compared to CG, P = .0009; AG compared to PG, P = .0054). Other percentage changes in cytokine and growth factor concentration were not statistically significant between groups. In the AG, the decrease of TNF-α and IL-1β concentrations correlated positively with the decrease of PI size (P <.05). Anodal HVMPC elevates IL-10/TNF-α in blood serum. The decrease of TNF-α and IL-1β concentrations in blood serum correlates with a decrease of PI wound area. More research is needed to determine whether the changes induced by anodal HVMPC improve PI healing and to determine whether and how different electrical currents affect the activity of biological agents responsible for specific wound healing phases, both within wounds and in patients' blood. In clinical practice, anodal HVMPC should be used to increase the ratio of anti-inflammatory IL-10 to pro-inflammatory TNF-α , which may promote healing.

Effects of Bai-Hu decoction on fever induced by lipopolysaccharide

Serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFalpha) concentrations were measured in subjects during two-hour glucose loading in order to investigate the effects of glucose on serum IL-6 and TNFalpha concentrations. Twenty-six female subjects (mean age 60 +/- 10 years) had normal glucose tolerance (NGT) and nineteen female subjects (mean age: 63 +/- 9 years) had impaired glucose tolerance (IGT) according to WHO criteria. Serum IL-6 and TNFalpha concentrations were measured by chemiluminescent immunometric assay. Subjects with IGT have higher fasting serum TNFalpha levels than subjects with NGT (p < 0.01). Serum IL-6 and TNFalpha concentrations were elevated during glucose loading (for each comparison, p < 0.01). The increase in serum TNFalpha concentrations in IGT was greater than in NGT (p < 0.01). Serum IL-6 and TNFalpha concentration significantly correlated with insulin and glucose in IGT group (for each comparison, p < 0.01). The correlation between serum glucose and cytokines concentrations was significant in IGT (for each comparison, p < 0.01). There was also a positive correlation between serum IL-6 and TNFalpha in NGT and IGT (for each comparison, p < 0.01). In conclusion, hyperglycemia is associated with increased circulating cytokine concentrations and fasting TNFalpha concentrations seem to be more associated with IGT than IL-6.

185P - Evaluation of the effect of cholesorption on the concentration of TNF-α in the serum of the blood in tumours of the biliopancreatoduodenal zone

To study the effect of early use of sodium valproate on neuroinflammation after traumatic brain injury (TBI). A total of 45 children who visited in Xuzhou Children's Hospital Affiliated to Xuzhou Medical University from August 2021 to August 2022 were enrolled in this prospective study, among whom 15 healthy children served as the healthy control group, and 30 children with TBI were divided into a sodium valproate treatment group and a conventional treatment group using a random number table (n=15 each). The children in the sodium valproate treatment group were given sodium valproate in addition to conventional treatment, and those in the conventional group were given an equal volume of 5% glucose solution in addition to conventional treatment. The serum concentrations of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3), high-mobility group box 1 (HMGB1), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were measured in the healthy control group on the day of physical examination and in the children with TBI on days 1, 3, and 5 after admission. Glasgow Outcome Scale-Extended (GOS-E) score was evaluated for the children with TBI 2 months after discharge. Compared with the healthy control group, the children with TBI had significantly higher serum concentrations of NLRP3, HMGB1, TNF-α, and IL-1β on day 1 after admission (P<0.017). The concentration of NLRP3 on day 5 after admission was significantly higher than that on days 1 and 3 after admission in the children with TBI (P<0.017). On days 3 and 5 after admission, the sodium valproate treatment group had a significantly lower concentration of NLRP3 than the conventional treatment group (P<0.05). For the conventional treatment group, there was no significant difference in the concentration of HMGB1 on days 1, 3, and 5 after admission (P>0.017), while for the sodium valproate treatment group, the concentration of HMGB1 on day 5 after admission was significantly lower than that on days 1 and 3 after admission (P<0.017). On day 5 after admission, the sodium valproate treatment group had a significantly lower concentration of HMGB1 than the conventional treatment group (P<0.05). For the children with TBI, the concentration of TNF-α on day 1 after admission was significantly lower than that on days 3 and 5 after admission (P<0.017). On days 3 and 5 after admission, the sodium valproate treatment group had a significantly lower concentration of TNF-α than the conventional treatment group (P<0.05). The concentration of IL-1β on day 3 after admission was significantly lower than that on days 1 and 5 after admission (P<0.017) in the children with TBI. On days 3 and 5 after admission, the sodium valproate treatment group had a significantly lower concentration of IL-1β than the conventional treatment group (P<0.05). The GOS-E score was significantly higher in the sodium valproate treatment group than that in the conventional treatment group 2 months after discharge (P<0.05). Early use of sodium valproate can reduce the release of neuroinflammatory factors and improve the prognosis of children with TBI.

Acupuncture is one of the most useful tools in complimentary medicine, and has demonstrated potential value for treating chronic renal failure (CRF). However, the underlying mechanisms for its therapeutic effect remain unknown. In the present study, the effects of acupuncture on renal interstitial fibrosis (RIF) were explored in a rabbit model of CRF. Rabbits were assigned to the following five groups: sham, model, losartan potassium (Posi), acupuncture (Acup) and acupuncture+inhibitor (Acup+Inhib) groups. The CRF rabbits were administered a drug or/and acupuncture on Shenshu, Mingmen and Pishu. The body weights, urine protein, serum creatinine (SCr) and blood urea nitrogen (BUN) levels of the rabbits were measured. Transforming growth factor β (TGF-β), integrin-linked kinase (ILK) and Smad3 expression were detected by qRT-PCR. Tumor necrosis factor-α (TNF-α) and endothelial nitric oxide synthase (eNOS) expression were analyzed by western blot methods. The concentrations of TGF-β, IL-8, TNF-α and IL-1β in blood serum were detected using ELISA kits. In addition, pathological characteristics of the rabbit tissues were evaluated by H&E and Masson's trichrome staining methods, and TGF-β expression was detected by immunohistochemistry (IHC) assays. Results showing decreased body weights and increased urine protein, SCr and BUN levels confirmed that the CRF model had been successfully constructed. It was also found that acupuncture significantly reduced the levels of TNF-α, Smad3, ILK and TGF-β expression, dramatically decreased the concentrations of TGF-β, IL-8, TNF-α and IL-1β in blood serum, and significantly increased eNOS expression in the CRF model rabbits by affecting the TGF-β/Smad signaling pathway. In addition, it was demonstrated that acupuncture could relieve RIF by affecting the TGF-β/Smad pathway. These observations indicate that acupuncture may be useful for treating CRF, and suggest the TGF-β/Smad pathway as a target for CRF therapy.

The aim of the study. To investigate the relationship between the content of TNF-α in the blood of patients with diabetes and the development of diabetic neuropathy. Materials and Methods. Type 2 DM2 patients without complications and patients with diabetic neuropathy (DN) under inpatient treatment in RSSPMC of endocrinology of HM RUz participated in the study. Biochemical analysis and TNF-α cytokine were determined in blood. Results and discussion. The results of determining the content of TNF-α in the blood serum of the studied groups using ELISA showed that in patients with diabetes there is a significant increase in the content of TNF-α (3.05±0.44 pg/ml) relative to the values in the control group. In the group of patients with DN, an increase in the concentration of TNF-α in the blood serum was observed, which amounted to 5.2±0.44 pg/ml. A study of the concentration of TNF-α in patients without complications of diabetes and in patients with DN mild degree did not reveal statistically significant differences. In patients with DN mild degree, the average concentration of TNF-α was 3.62±0.89 pg/ml, and in DN severe degree -7.13±1.14 pg/ml, i.e. increased almost 2 times. Conclusion. It was found that the content of TNF-α in the serum of patients with diabetic neuropathy is in direct correlation with the degree of severity of the disease. Funding: by Mininnovation.uz and WorldBank ФЗ-2020092959 (Ф-ОТ-2021-153) This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.

To observe the effect of moxibustion on serum inflammatory factors, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor Kappa B (NF-κB) in colon tissue of rats with diarrhea-predominant irritable bowel syndrome (IBS-D)，so as to explore the mechanism of moxibustion in the treatment of IBS-D. Healthy male SD rats were randomly divided into blank group, model group and moxibustion group, with 8 rats in each group. The IBS-D model was established by chronic restraint combined with gavage of Senna-leaf solution. The rats of the moxibustion group was treated with moxibustion at "Tianshu"(ST25)and "Shangjuxu"(ST37)for 30 min， once a day for 7 d. After the intervention, the rate of loose stools and the minimum threshold volume of abdominal withdrawal reflex (AWR) induced by colorectal distension were observed. The contents of serum inflammatory factors IL-1β, TNF-α and IL-6 were detected by ELISA. The pathological changes of rat colon were observed by H.E. staining. The average optical density of TLR4 and NF-κB (p65) in colon tissue was detected by immunohistochemistry. The relative expressions of TLR4, MyD88 and NF-κB (p65) mRNAs in colon tissues were detected by real-time fluorescence quantitative PCR, and the expression levels of TLR4, MyD88 and NF-κB (p65) proteins in colon tissues were detected by Western blot. There was a mild degree of inflammation in colon in rats of the model group. Compared with the blank group, the minimum volume threshold of AWR decreased significantly (P< 0.01), and the rate of loose stools, the contents of IL-1β, IL-6 and TNF-α in serum as well as the expressions of TLR4, MyD88, NF-κB (p65) mRNAs and protein in colon tissues were significantly increased in the model group (P<0.01). Compared with the model group, the inflammation was alleviated in colonic mucosa, the minimum volume threshold of AWR increased significantly (P<0.01), and rate of loose stools decreased significantly (P<0.01) in the moxibustion group, the contents of IL-1β, TNF-α and IL-6 in serum and the expression levels of TLR4, MyD88, NF-κB (p65) mRNAs and proteins in colon tissue decreased significantly in the moxibustion group (P<0.01). Moxibustion ST25 and ST37 can improve diarrhea symptom and visceral hypersensitivity in IBS-D rats, which may be related with its effects in inhibiting TLR4/MyD88/NF-κB signaling pathway and reducing the expression of inflammatory factors.

Objective To evaluate the effects of pretreatment with exogenous insulin-like growth factor-1 (IGF-1) on lung injury in rats undergoing cardiopulmonary bypass(CPB). Methods Seventy-two SPF healthy male Sprague-Dawley rats, weighing 350-500 g, were divided into 4 groups (n=18 each) using a random number table: sham operation group (group S), CPB group, CPB plus left lung ischemia-reperfusion group (group LI) and IGF-1 group.The chest was only opened, and the rats underwent no CPB in group S. Only the CPB model was established in group CPB.The model of left lung ischemia-reperfusion injury was established based on the CPB model in group LI.The model of CPB and left lung ischemia-reperfusion injury was established, and IGF-1 30 μg/kg was intravenously injected at 10 min before clamping the hilum of lung and immediately after opening the hilum of lung in group IGF-1.Six rats were selected before operation (T1), 10 min after opening the left hilum (T2) and at the end of operation (T3), and blood samples were collected from the femoral artery for blood gas analysis.The oxygenation index (OI) and respiratory index (RI) were calculated.Serum was obtained from blood, and the concentrations of interleukin-6 (IL-6), IL-1β and tumor necrosis factor-alpha (TNF-α) in serum were measured using enzyme-linked immunosorbent assay.The left upper lung tissues were removed for examination of the pathological changes which were scored with a light microscope. Results Compared with the baseline at T1, OI was significantly decreased, and RI, concentrations of IL-6, IL-1β and TNF-α in serum and pathological scores of lung tissues were increased at T3 in CPB, LI and IGF-1 groups (P<0.05). Compared with group S, OI was significantly decreased, and RI, concentrations of IL-6, IL-1β and TNF-α in serum and pathological scores of lung tissues were increased at T3 in CPB, LI and IGF-1 groups (P<0.05). Compared with group CPB, OI was significantly decreased, and RI, concentrations of IL-6, IL-1β and TNF-α in serum and pathological scores of lung tissues were increased at T3 in group LI (P<0.05). Compared with group LI, OI was significantly increased, RI, concentrations of IL-6 and IL-1β in serum and pathological scores of lung tissues were decreased at T3, and the concentration of serum TNF-α was increased at T3 in group IGF-1 (P<0.05). Conclusion IGF-1 pretreatment can reduce lung injury in rats undergoing CPB, and the mechanism is related to inhibiting inflammatory responses. Key words: Insulin-like growth factor Ⅰ; Cardiopulmonary bypass; Respiratory distress syndrome, adult

To explore the effect of occupational stress on serum tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-2 and IL-4. A cross-sectional epidemiological study was conducted in 200 workers from the refrigerator assembly line in Henan province in China. Psychosocial work conditions were measured by using the job demand-control model, the effort-reward imbalance model questionnaires and occupational stress measurement scale. Serum TNF-α, IL-1β, IL-2, and IL-4 concentrations were measured by radioimmunoassay or immunoradiometric assay method respectively. Serum TNF-α concentration was statistically significantly different between workers with higher affective balance level and control groups [(1.947 ± 0.173) and (2.029 ± 0.240) fmol/ml] (P < 0.05). Serum IL-1β concentration was statistically significantly different between workers with higher effort level and control groups [(0.133 ± 0.034) and (0.118 ± 0.031) ng/ml] (P < 0.05). Serum IL-2 concentration was statistically significantly different between workers with higher role ambiguity level [(1.658 ± 0.376) and (1.491 ± 0.033) ng/ml] and control groups (P < 0.05), as well as between workers with higher role conflict level and control groups [(1.774 ± 0.311) and (1.589 ± 0.380) ng/ml] (P < 0.05), between workers with higher daily life stress level and control groups [(1.759 ± 0.361) and (1.606 ± 0.381) ng/ml] (P < 0.05). Serum IL-4 concentration was statistically significantly different between workers with higher reward level and control groups [(1.449 ± 0.025) and (1.466 ± 0.041) pg/ml] (P < 0.05). Stepwise regression analysis indicated that affective balance was the predictor of serum TNF-α (R(2) = 0.029). Effort and mental health were the predictors of serum IL-1β (R(2) was 0.029 and 0.055, respectively). Role conflict, daily life stress and role ambiguity were the predictors of serum IL-2 (R(2) was 0.040, 0.078 and 0.104, respectively). Reward was the predictor of serum IL-4 (R(2) = 0.030). Unhealthy psychological stress factor might be induce a marked increase in the concentrations of serum TNF-α, IL-1β, IL-2, as well as IL-4.

Introduction. In the structure of injuries from gunshot wounds dominate the limb injuries that are often complicated by massive bleeding from the main vessels. The only means of rescue on the battlefield is a timely application of a tourniquet. This ensures complete exsanguination of the limb. The duration of exsanguination should not exceed 2 hours. It is proved that in conditions of acute blood loss complicated by ischemia-reperfusion of the limb, there are prerequisites for the development of dysfunction of internal organs, which in turn exacerbates dysmetabolic and functional disorders, stimulates endotoxicosis and may be complicated by multiple organ failure. However, the patterns of formation of the syndrome of endogenous intoxication in the conditions of ischemia-reperfusion of the limb, acute blood loss and their combinations have not been studied. There are no data on the effect of Carbacetam on these processes. It is known that Carbacetam has proven itself to reduce the manifestations of dysfunction of internal organs in the conditions of ischemic-reperfusion syndrome of the limb.The objective of research: To establish the dynamics of endogenous intoxication in the conditions of ischemia-reperfusion of the limb, acute blood loss and their combination. To evaluate the effectiveness of Carbacetam in the correction of identified abnormalities.Materials and methods. The experimental studies were conducted on 108 white nonlinear male rats weighing 200-220 g, which were housed in standard vivarium conditions. All animals were divided into five groups: control and four experimental. Under thiopental-sodium anesthesia in the first experimental group was simulated ischemia-reperfusion of the limb, in the second – acute blood loss, and in the third – these injuries were combined. In the fourth experimental group, animals with acute blood loss and ischemia-reperfusion of the limb were intraperitoneally administered Carbacetam at a dose of 5 mg per kilogram of animal weight. After 1 and 2 hours, as well as after 1, 7 and 14 days in the blood serum was determined the content of middle-mass molecules (MMM254, MMM280).The results and discussion. It was found that ischemia-reperfusion of the limb in comparison with the control group is accompanied by a significant increase of the MMM254 fraction in the blood serum content. From the first hour of the experiment with a maximum after 1 day and normalization of the index up to 14 days. The content in the blood serum of the MMM280 fraction became significantly higher, from the control only after 1 day with subsequent normalization in the following terms of the experiment. After simulation of acute blood loss, both studied fractions of MMM increased from 1 hour of the experiment, reached a maximum after 1 day and did not return to the control level after 14 days. For the first time it was found that two-hour ischemia-reperfusion of the limb significantly aggravates the course of acute blood loss, which revealed a significantly higher content in the serum of fractions MMM254 and MMM280 compared to other experimental groups in almost all periods of the experiment. Due to the use of Carbacetam, the content of both studied fractions of MMM in the serum compared with animals without correction was significantly reduced after 7 and 14 days of the experiment. Although the studied parameters did not return to the level of the Carbacetam control group, it can be considered a pathogenetically justified means of reducing the level of endotoxicosis in the conditions of ischemia-reperfusion of the limb and acute blood loss.Conclusions. Simulation of limb ischemia-reperfusion is accompanied by accumulation of MMM254-280 fractions with a maximum after 1 day of the experiment and normalization up to 14 days. Acute blood loss causes an increase in the serum content of fractions MMM254-280 from 1 hour of the experiment, which up to 14 days do not reach the level of control. The detected disorders are significantly exacerbated after a combination of acute blood loss and ischemia-reperfusion of the limb. The use of Carbacetam in comparison with animals without correction is accompanied by a significant decrease in the content of MMM254-280 fractions in the serum after 7 and 14 days of the experiment.

To observe the effects of electroacupuncture (EA) penetration needling on Toll-like receptors 4/myeloid differentiation factor 88/nuclear factor-kappa B (TLR4/MyD88/NF-κB) signaling pathway in rat synovium and the serum-related inflammatory factors, so as to explore the mechanism of EA penetration needling on synovial inflammation in rats with knee osteoarthritis (KOA). SD male rats were randomly divided into sham-operation group, model group, EA+penetration needling group, and conventional EA group, with 16 rats in each group. The rats model was prepared by anterior cruciate ligment transection and these rats were forced to exercise for 8 weeks after operation. After successful modeling, in the EA+penetration needling group, the needles were inserted at "Dubi" (ST35) "Neixiyan" (EX-LE4), and at "Xuehai"(SP10) "Liangqiu"(ST34) on the right hind limb, towards each other, 5-8 mm in depth, respectively. In the conventional EA group, the needles were inserted at ST35 and EX-LE4 on the right hind limb, obliquely, at 30° angle to the skin, 3-5 mm in depth; and were inserted at SP10 and ST34 on the right hind limb perpendicularly, 3-5 mm in depth. In these two groups, electric stimulation was operated with dense-disperse wave, 2 Hz/10 Hz in frequency and 0.5-1.5 mA in intensity, retained for 20 min in each treatment. The treatment was given once daily, 10 days as 1 course of treatment, and 2 courses were required at the interval of 2 days. After the intervention, the knee joint effusion was observed by musculoskeletal ultrasound; the contents of IL-1β, IL-6 and TNF-α in serum were determined by ELISA; the morphological changes in the synovium were observed after H.E. staining; the positive expression of NF-κB p65 in the synovial membrane was detected by immunohistochemical method; the expression levels of TLR4, MyD88, TRAF-6 and NF-κB p65 proteins in the synovial membrane were determined by Western blot. Compared with the sham-operation group, in the model group, the knee joint effusion was obviously increased, the synovial lining cells were distributed irregularly, the cells were disarranged, the pannus was formed largely, and a great number of the inflammatory cells were infiltrated; the contents of serum IL-1β, IL-6 and TNF-α, the positive expression of NF-κB p65, the protein expression levels of TLR4, MyD88, TRAF-6 and NF-κB p65 in the synovial tissue were increased (P<0.05). Compared with the model group, the knee joint effusion was reduced, the synovial lining cells were proliferated, a small number of the inflammatory cells were infiltrated, and the pannus was formed lightly; the contents of serum IL-1β, IL-6 and TNF-α, the positive expression of NF-κB p65, the protein expression levels of TLR4, MyD88, TRAF-6 and NF-κB p65 in the synovial tissue were lower (P<0.05) in the EA+penetration needling group and the conventional EA group. In the conventional EA group, the knee joint effusion was increased, the synovial lining cells were proliferated, the inflammatory cells were infiltrated largely, and the pannus was formed increasingly; the contents of serum IL-1β, IL-6 and TNF-α, and the protein expression levels of TLR4, MyD88 and NF-κB p65 in the synovial tissue were increased when compared with the EA+penetration needling group (P<0.05). The EA+penetration needling can significantly relieve the synovial inflammatory reaction and the knee joint effusion in KOA rats. The mechanism is probably related to down-regulating the downstream inflammatory cascade through inhibiting the transduction of TLR4/MyD88/NF-κB signaling pathway.

Relevance. Activation of innate and acquired immunity, accompanied by increased production of classical (interleukin (IL) IL-1β, IL-6, tumor necrosis factor-α (TNF-α) and interferon-γ (INF-γ)) proinflammatory cytokines in synovial fluid and blood serum, plays an important role in the pathogenesis of rheumatoid arthritis (RA).Objective. To determine the concentration of IL-1β, IL-6, TNF-α and INF-γ in RA patients in the advanced stage of the disease, to evaluate the relationship between them, clinical indices of disease activity, the presence of rheumatoid factor (RF), and antibodies to cyclic citrullinated peptide (ACCP).Material and methods. We examined 154 patients with RA (41 men and 113) who were middle-aged (56.0 (50.0; 64.0) years), disease duration (9.4 (3.0; 13.0) years), seropositive 129 (83.8 %) for IgM RF and/or 106 (68.8 %) ACCP with moderate to high (DAS28-ESR — 5.40 (4.65; 6.00)) disease activity. The concentration of IL-1β, IL-6, TNF-α and INF-γ in serum determined by multiplex technology.Results. The concentration of IL-1β was not significantly different between patients with RA and controls. The values of IL-6 and INF-γ were significantly higher, and TNF-α — significantly lower than in donors. IL-6 hyperproduction was detected most frequently (51.6 %), whereas elevated levels of INF-γ (38.96 %), IL-1β (26.62 %) and TNF-α (23.38 %) were less common. Significant positive correlations were observed between the concentrations of all cytokines and their high levels. The strongest correlations were characteristic for IL-1β, TNF-α and INF-γ. No statistically significant differences in cytokine levels were observed between patients with RA who were positive or negative for IgM-RF and ACCP. The concentration of IL-6 alone significantly positively correlated with the values of the indices (DAS28-ESR, CDAI, SDAI) of RA clinical activity (p&lt;0.05).Conclusions. There are differences in the levels and frequencies of proinflammatory cytokines among patients with advanced-stage RA. In the presence of a close relationship between them, there are certain differences in their associations with clinical and laboratory indicators of disease activity.