ã€Key words】 Ropivacaine Abstract: Objective: To observe the changes of peripheral blood immune-related factors CD3, CD4, CD8, CD20, IL-2, INF-γ in patients with lumbar disc herniation under epidural anesthesia , To explore the effect of ropivacaine on lymphatic immunity, so as to guide the perioperative anesthetic medication and management. Methods: Collect 30 patients with ropivacaine undergoing epidural anesthesia for lumbar disc herniation nucleus pulposectomy 1 h before anesthesia, 24 h after surgery and 48 h after surgery. CD3 was measured by flow cytometry at different phases , CD4, CD8, CD20, IL-2, INF-γ content. Results: There was no significant change in CD3 after operation, the content of CD4, CD20, IL-2, INF-γ increased, and the content of CD8 decreased. Conclusion: Patients undergoing ropivacaine anesthesia have increased T lymphocytes and B lymphocytes, increased cytokine secretion, and no significant immunosuppression. Keywords: Ropivacaine; Immunization; Anesthesia The Effects of Ropivacaine on the Content of CD3 CD4 CD8 CD20 IL-2 INF-γ suffer epidural anesthesia with ropivacaine. Venous blood samples were collected to measure the contents of CD3, CD4, CD8, CD20, IL-2, and INF-γ. Result: After operation, the contents of CD4, CD20, IL-2 and INF -γ were increased, that of CD3 was constant and that of CD8 was decreased. Conclusion: Ropivacatine has not marked influence on lymphocyte immunity. Key words: Ropivacaine; Immunity; Anesthesia
Ropivacaine is a newer long-acting amide local anesthetic [1], currently used mainly for epidural anesthesia and surgical analgesia, with a wide range of applications. Current research has shown that anesthetic drugs and pain stress have certain effects on cellular immunity, humoral immunity and cytokines of surgical patients [2], and the status of immune function of patients during perioperative period on infection, tumor spread and prognosis of postoperative patients has great significance. This study analyzed the content of immune-related factors CD3, CD4, CD8, CD20, IL-2, INF-γ in peripheral blood of patients undergoing epidural anesthesia with ropivacaine for lumbar disc herniation before and after operation Changes, provide a reference for the better use of ropivacaine.
1 Materials and methods
1.1 General information: 30 patients with nucleus pulposus extraction underwent lumbar disc herniation, ASA I to II, 22 males and 8 females, aged 18 to 48 years.
1.2 Anesthesia: The patient is punctured by epidural at T12L1 or L1 ～ 2. After successful, the catheter is placed 3 ～ 4cm to the head and injected with 0.75% ropivacaine. The anesthesia level is controlled at T10 ～ 5. ECG and SpO2 are monitored during operation And BP. The patient was infused with Ringer's solution during the operation without using hemostatic drugs, plasma replacement or blood transfusion to control the volume and speed of infusion to maintain hemodynamic stability.
1.3 Flow cytometry: draw 2ml of venous blood 1h before anesthesia (T1), 24h (T2) and 48h (T3) after operation, add to EDTA-2Na anticoagulation tube, and perform flow cytometry within 24h . Adopt FACScan flow cytometer of American BD company for detection, the flow cytometry reagent used is provided by BD company. CD3, CD4, CD8 and CD20 were measured by taking 200 μl of anticoagulated blood, adding 20 μl of the corresponding fluorescently labeled monoclonal antibody, placing it for 30 min, adding 1 ml of erythrocyte lysate, and centrifuging (2000 r / min) 3 min after it became transparent , 4 min), discard the supernatant, wash with fluorescent washing solution (PBS + 0.1% NaN3 sodium azide + 5% calf serum) or Hanks solution 1ml twice, suspend 0.5 ml washing solution, put it on the machine and measure. The method of measuring IL-2 and INF-γ is that cytokines are secreted only after lymphocyte activation and are quickly secreted out of the cell. Therefore, the stimulant PMA is used to activate lymphocytes in vitro, and then the Monensin protein transport inhibitor is used to prevent the cytokine from being secreted out of the cell, so that the cytokine is retained in the cell to a certain amount, and then the flow cytometry intracellular antigen is used for detection .
1.4 Result statistics: The measurement data is expressed as mean ± standard deviation, the count data should be tested by X2 test, and P <0.05 means that there is a significant difference.
2 Results Postoperative patients had no significant changes in CD3, CD4, CD20, IL-2, INF-γ content increased, CD8 content decreased. The specific statistical results are shown in Tables 1 and 2. Table 1 Changes in the content of CD3, CD4, and CD8. Table 2 Changes in the content of CD20, IL-2, and INF-γ. Table 3. Discussion During the operation, the patient's immune status is affected by many factors, including emotional anxiety, pain, anesthesia, and surgical stress Factors such as infection will affect the patient's immune status, trigger the distribution of different components of lymphocytes in the peripheral blood circulation, and change the secretion of intracellular factors. Current studies have shown that anesthesia has a certain immunosuppressive effect, affecting the patient's postoperative recovery [3, 4]. T lymphocytes are an important component of the body's immunity. The interaction between T cells and other cells and cytokines together perform immune recognition, activation, and killing. Th cells can stimulate B cells to produce antibodies and assist other T cells to complete immune responses. Ts cells have the function of immunosuppression. Th and Ts cells coordinate with each other to ensure normal immune function in the body. B cells mediate the humoral immune response in the body. B lymphocytes interact with T lymphocytes and are activated with the help of Th2 cells. The cells proliferate, mature and produce antibodies, and participate in the humoral immune response [5]. Interleukin 2 (IL-2) is an important T cell growth factor in the body, is the key to protect the body's normal immune function, and has a variety of biological functions. For example, it can maintain the proliferation of active T cells, induce LAK cells, promote the secretion of antibodies by B cells, promote the production of interferon by NK cells, and can increase the number and function of TH cells to enhance the immune function of the body. With other cytokines, such as: sIL2R, IL-6, IL-1, IL-4, TNF, etc. form a dynamic immune regulatory network to maintain the body's normal immune regulatory function. INF-γ is an important negative regulator of growth, which also has antiviral replication, inhibits tumor cell proliferation, regulates the function of the lymphatic system, and enhances its own immunity. INF-γ is the main macrophage-activating factor (macrophage-activating factor, MAF), promote macrophage phagocytosis and inflammation, and directly promote T and B cell differentiation and CTL maturation, stimulate B cells to secrete antibodies, thereby enhancing the body's immune function. The function of inducing macrophages to produce NO. Under the appropriate stimulation of TNF and lipopolysaccharide, INF-γ can be used as a suitable inducer of NO production, which is of great significance for inhibiting tumor tissue proliferation and cardiovascular remodeling. At the same time, INF-γ plays a role in maintaining the steady state of the body's autoimmunity. In this study, postoperative CD4 and CD20 increased significantly, while CD8 showed a downward trend, the concentration of immunosuppressive Ts cells decreased, Th cells increased, and B cells increased, indicating that the patients' T cells and B cells responded significantly Enhancement, and also observed a gradual upward trend of cytokines IL-2 and INF-γ, which is consistent with the change of CD4, indicating that more Th0 cells differentiate into Th1 cells in patients, and Th1 cells secrete a large amount of IL-2 and INF-γ enhance cellular immunity in vivo. This study shows that ropivacaine has no significant inhibitory effect on patients' T lymphocytes and B lymphocytes.
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