ABSTRACT
Objective:To study the mechanism of the effect of Jiajian-Zhujing Decoction on the expression of VEGF on ARPE-19 cells after AKT transfection. Methods:To prepare the serum and blank serum of Jiajian-Zhujing Decoction and divide ARPE-19 cells into the normal group, model group, blank serum group, medicated serum group, Conbercept group and combined group. Except normal group, this research established AKT transfected cell model. Then cultured the normal group and model group with conventional method, and the blank serum group was cultured with 10% blank serum, the medicated serum group was cultured with 10% medicated serum, the Conbercept group was cultured with 20 μg/ml Conbercept, the combined group was cultured with 10% medicated serum and 20 μg/m Conbercept. The proliferation of ARPE-19 cells in each group was detect by the CCK-8 method. The levels of AKT, mTOR and VEGF mRNA were detected by real-time quantitative PCR. Western blot was used to detect the expression of AKT, mTOR and VEGF. Results:After being cultured for 24, 48 and 72 hours, compared with the model group, the cell proliferation rate in blank serum group, medicated serum group, Conbercept group and combined group significantly decreased ( P<0.05). Compared with the model group, the expression of AKT mRNA (24 h: 3.10 ± 0.48, 1.97 ± 0.14, 1.26 ± 0.24 vs. 4.77 ± 0.68; 48 h: 3.52 ± 0.82, 2.62 ± 0.77, 1.10 ± 0.19 vs. 6.12 ± 1.21), mTOR mRNA (24 h: 3.02 ± 0.26, 2.45 ± 0.75, 1.13 ± 0.15 vs. 4.48 ± 0.80; 48 h: 1.29 ± 0.30, 1.30 ± 0.57, 0.65 ± 0.19 vs. 2.54 ± 0.62), VEGF mRNA (24 h: 3.33 ± 0.62, 2.18 ± 0.20, 1.55 ± 0.28 vs. 5.53 ± 1.02; 48 h: 2.35 ± 0.54, 1.23 ± 0.28, 0.93 ± 0.25 vs. 3.59 ± 0.40), AKT protion (24 h: 0.45 ± 0.09, 0.25 ± 0.05, 0.14 ± 0.04 vs. 0.62 ± 0.04; 48 h: 0.36 ± 0.06, 0.23 ± 0.04, 0.14 ± 0.03 vs. 0.54 ± 0.08), mTOR protion (24 h: 0.35 ± 0.05, 0.24 ± 0.02, 0.18 ± 0.02 vs. 0.52 ± 0.09; 48 h: 0.23 ± 0.04, 0.29 ± 0.04, 0.14 ± 0.03 vs. 0.40 ± 0.10), VEGF protion (24 h: 0.14 ± 0.03, 0.33 ± 0.04, 0.24 ± 0.03 vs. 0.54 ± 0.10; 48 h: 0.24 ± 0.03, 0.17 ± 0.02, 0.11 ± 0.02 vs. 0.42 ± 0.10) significantly decreased ( P<0.05), and the combined group was significantly lower than that of the Conbercept group ( P<0.05). Conclusions:AKT transfection can promote the proliferation of ARPE-19 cells, and Jiajian-Zhujing Decoction can significantly inhibit this proliferation. Jiajian-Zhujing Decoction may inhibit the activity of AKT/mTOR signaling pathway to reduce the expression of VEGF.
ABSTRACT
Objective To evaluate the effect of dexmedetomidine on Toll-like receptor 4 ( TLR4)/myeloid differentiation factor 88 ( MyD88)/nuclear factor Kappa B ( NF-κB) signaling pathway in the pe-ripheral blood mononuclear cells of elderly patients with diabetes mellitus undergoing lower extremity surger-y. Methods Forty elderly patients of both sexes with type 2 diabetes mellitus, aged 65-80 yr, with body mass index of 18. 5-27. 9 kg/m2 , of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with New York Heart Association Ⅰ or Ⅱ, undergoing lower extremity surgery with tourniquets under general anesthesia, were divided into 2 groups ( n=20 each) by using a random number table method: control group ( group C) and dexmedetomidine group ( group D) . Combined intravenous-inhalational anesthesia was applied. Dexmedetomidine was infused over 15 min in a dose of 1μg/kg after induction of anesthesia, fol-lowed by a continuous infusion of 0. 5 μg·kg-1 ·h-1 until the end of surgery in group D, while the equal volume of normal saline was given instead of dexmedetomidine in group C. Before using the the tourniquet and at 15 min, 1 h and 24 h after loosing the tourniquet, arterial blood samples were collected for determi-nation of the expression of TLR4, NF-κB and MyD88 in peripheral blood mononuclear cells and concentra-tions of tumor necrosis factor-alpha ( TNF-α) , interleukin-1β ( IL-1β) , cardiac troponin I ( cTnI ) and creatine kinase-MB ( CK-MB) in plasma. Results Compared with group C, the expression of TLR4, NF-κB and MyD88 in peripheral blood mononuclear cells was significantly down-regulated, and the concentra-tions of TNF-α, IL-1β, cTnI and CK-MB in plasma were decreased at each time point after loosing the tourniquet in group D ( P＜0. 05) . Conclusion The mechanism by which dexmedetomidine reduces myo-cardial damage may be related to inhibiting TLR4/MyD88/NF-κB signaling pathway and reducing systemic inflammatory responses in elderly patients with diabetes mellitus undergoing lower extremity surgery.
ABSTRACT
Objective To evaluate the efficacy of dexmedetomidine for improvement of postoperative analgesia with sufentanil in pediatric patients with a large area of burn after tangential excision and skin grafting.Methods Forty-two pediatric patients of both sexes with a large area of burn,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,aged 2-10 yr,weighing 13-36 kg,scheduled for elective tangential excision and skin grafting under combined propofol-remifentanil-sevoflurane anesthesia,were randomly divided into 2 groups (n=21 each) using a random number table:sufentanil group (group Suf) and dexmedetomidine plus sufentanil group (group Dex-Suf).The patient-controlled intravenous analgesia (PCIA) was used for postoperative analgesia.PCIA solution contained sufentanil 2 μg/kg and granisetron 100 μg/kg in 100 ml of normal saline in group Suf,and contained dexmedetomidine 2.5 μg/kg,sufentanil 1.5 μg/kg,and granisetron 100 μg/kg in 100 ml of normal saline in group Dex-Suf.The PCA pump was set up with a 0.5 ml bolus dose,a 15 min lockout interval and background infusion at a rate of 2 ml/h after a loading dose of sufentanil 0.1 μg/kg.When Faces Pain Scale score＞2,sufentanil 0.1 μg/kg was injected intravenously as rescue analgesic.The consumption of sufentanil was recorded within 48 h after operation.Ramsay sedation scores at static and dynamic (during dressing changes) conditions were assessed after operation.The parents' satisfaction,requirement for rescue analgesics and incidence of adverse reactions such as agitation,nausea and vomiting were recorded after operation.Results Compared with group Suf,Ramsay sedation scores at static and dynamic conditions and patients' satisfaction scores were significantly increased,and the consumption of sufentanil,requirement for rescue analgesics and incidence of agitation,nausea and vomiting were significantly decreased after operation in group Dex-Suf (P＜0.05).Conclusion Dexmedetomidine significantly improves postoperative intravenous analgesia with sufentanil in the pediatric patients with a large area of burn after tangential excision and skin grafting,and combination of dexmedetomidine and sufentanil is recommended for this type of pediatric patients.