Mechanism of catgut embedding at back- points for nonalcoholic steatohepatitis based on IKK/IKB/NF-κB signaling pathway / 中国针灸

OBJECTIVE@#To explore the mechanism of catgut embedding at back- points on nonalcoholic steatohepatitis (NASH) in rats based on IKK/IKB/NF-κB signaling pathway and downstream inflammatory factors.@*METHODS@#Eighty SPF SD rats were selected, among them 10 rats were selected divided into a normal group (group A), and the remaining 70 rats were fed with high-fat diet to establish NASH model. At the end of 12 weeks, 10 rats were randomly selected to verify whether the model establishment was successful. Then the remaining 60 rats were randomly divided into a model group (group B), a catgut embedding at back- points group (group C), a catgut embedding at abdominal points group (group D), an acupuncture at back- points group (group E), a sham catgut embedding group (group F) and a western medication group (group G), 10 rats in each group. The rats in the group C were treated with catgut embedding at "Ganshu" (BL 18), "Pishu" (BL 20), "Weishu" (BL 21) and "Shenshu" (BL 23); the rats in the group D were treated with catgut embedding at "Daheng" (SP 15), "Fujie" (SP 14), "Huaroumen" (ST 24) and "Tianshu" (ST 25); the rats in the group E were treated with acupuncture at the same acupoints as the group C; the rats in the group F were treated with catgut embedding at back- points but the needle did not enter subcutaneous tissue gamma; the rats in the group G were treated with intragastric administration of vitamin E capsule. All the treatment was given for 4 weeks. The rats in the group A were fed with normal diet until the end of 16 weeks without any intervention. The rats in the group B continued to be fed with high-fat diet until the end of 16 weeks. After the intervention, the liver index was calculated; the liver histomorphology was observed by HE staining; the liver function [alanine aminotransferase (ALT), gamma glutamyl transferase (γ-GGT), alkaline phosphatase (ALP)] and blood lipid [serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL)] were measured by serum biochemistry. The serum levels of TNF-α, IL-6 and IL-1βwere detected by ELISA, and the expressions of IKK-α, NF-κBp65, IL-6, IL-1β and TNF-α proteins in liver tissue were detected by Western blot. The temperature of the conception vessel and the governor vessel was measured by infrared thermography.@*RESULTS@#Compared with the group A, the obvious steatosis and inflammatory cell infiltration were observed in the group B, and the body weight, liver wet-weight and liver index were all increased (0.05), while the temperature of the governor vessel in the group C was superior to that in the group D (<0.05).@*CONCLUSION@#The catgut embedding at back- points might inhibit the activation of IKK/IKB/NF-κB signaling pathway to interrupt the inflammatory cascade, and reduce the "second hit" of inflammatory factors on liver, which could slow down NASH progress and prevent and treat NASH.

Anti-inflammatory effect of insulin and its mechanism: An exploratory study / 中国糖尿病杂志

Objective To discuss the anti-inflammatory effect and its possible mechanism of different insulin concentration on IκB, NF-κB and TNF-α mRNA in monocytes stimulated by lipopolysaccharide and to detect the key molecule P-Akt in PI3K/ Akt signaling pathway at the cellular level. Methods The peripheral blood (10 ml)is collected from 30 type 2 diabetics (T2DM). Then, the mononuclear cells are centrifugally separated based on density gradient and divided into five different groups:the control (Con)group; the low concentration insulin 100 mU/L(L-Ins)group; the combination of low concentration insulin 100 mU/L and LY-294002 10 μmol/L(L-Ins+LY) group; the high concentration insulin 1000 mU/L (H-Ins) group, The combination of high concentration insulin 1000 mU/L and LY- 294002 10 μmol/L(H-Ins+LY) group. Two subgroups were set in each category. After incubating for 24 hours, the lipopolysaccharide (100 ng/ml)was added and the mononuclear cell culture of peripheral blood was continued. One hour later, the activities of P-Akt, IκBα and NF-κB of one subgroup were detected using western blot; two hours later, monocytes of the other subgroup were collected and the level of TNF-a mRNA was detected using real-time PCR Results Compared with Con group, the expressions of P-Akt and IkB were higher in L-Ins group and H-Ins group (P<0. 05), the expressions of NF-κB and TNF-a mRNA were lower in L-Ins group and H-Ins group (P<0. 05). Compared with L-Ins group, the expressions of P-Akt and IκB were higher in H-Ins group (P<0. 05), but the expressions of NF-κB and TNF-a mRNA were lower (P< 0. 05). Compared with L-Ins + LY group (H-Ins + LY group), the expressions of IκB and P-Akt were higher(P<0. 05)and the expressions of NF-κB and TNF-a mRNA were lower in L-Ins group (H-Ins group)(P<0. 05). Compared with Con group, no significant variations were shown in the expressions of P-Akt, IkB and NF-κB in L-Ins+LY group and H-Ins+LY group (P>0. 05) except for the high expressions of TNF-a mRNA (P<0. 05). There is no significant difference between L-Ins+LY group and H-Ins+LY group (P>0. 05). Conclusion The direct anti-inflammatory effect of insulin is verified in a dose dependent manner. Insulin may regulate the synthesis and secretion of inflammatory cytokines by activating PI3K/Akt pathway, increasing IkB and affecting the state of NF-κB. Insulin may increase the synthesis and secretion of inflammatory cytokines through other pathways when the PI3K/Akt pathway is blocked.

Protective effect of Longxue Tongluo capsule on oxidized low-density lipoprotein damage to human umbilical vein endothelial cells / 中国中药杂志

To observe the protective effect of Longxue Tongluo capsule (LTC) on human umbilical vein endothelial cells (EAhy.926 cells) injury induced by oxidized low-density lipoprotein (ox-LDL, 100 mg·L⁻¹). The effect of the cell viability of LTCin alleviating OX-LDL-induced endothelial cell injury was determined by MTT and LDH assay. The effect of LTC on lactic dehydrogenase (LDH), nitric oxide (NO), super oxide dlsmutase (SOD) and malondialdehyde (MDA) levels were detected by corresponding assay kits according to manufacturer's instruction. The effect of LTC on the protein expressions of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), p65, p-p65, IKB and p-IKB were detected by Western blot. The results showed that compared with the normal control group, the activity of EAhy.926 cells was significantly decreased, LDH leakage (<0.01) increased, NO content and SOD activity significantly decreased (<0.01, <0.05), and the expressions of ICAM-1, VCAM-1, p-p65/p65 and p-IKB(<0.05)increased.This study demonstrated that LTC had no significant effect on the growth of normal cells. The treatment with LTC significantly promoted the proliferation of vascular endothelial cells damagedby ox-LDL, decreased MDA content and LDH release, andincreased the activity of SOD and NO content. Meanwhile, ox-LDL significantly increased the expressions of ICAM-1, VCAM-1, p-p65/p65, p-IKB/IKB in Eahy.926 cells; these effects were suppressed by LTC at 1, 2 mg·L⁻¹. In conclusion, LTC has a significant protective effect on human umbilical vein endothelial cells caused by ox-LDL. This study suggested that LTC has a certain therapeutic effect on AS.

Loss of IκB kinase β promotes myofibroblast transformation and senescence through activation of the ROS-TGFβ autocrine loop

Using forward and reverse genetics and global gene expression analyses, we explored the crosstalk between the IκB kinase β (IKKβ) and the transforming growth factor β (TGFβ) signaling pathways. We show that in vitro ablation of Ikkβ in fibroblasts led to progressive ROS accumulation and TGFβ activation, and ultimately accelerated cell migration, fibroblast-myofibroblast transformation and senescence. Mechanistically, the basal IKKβ activity was required for anti-oxidant gene expression and redox homeostasis. Lacking this activity, IKKβ-null cells showed ROS accumulation and activation of stress-sensitive transcription factor AP-1/c-Jun. AP-1/c-Jun activation led to up-regulation of the Tgfβ2 promoter, which in turn further potentiated intracellular ROS through the induction of NADPH oxidase (NOX). These data suggest that by blocking the autocrine amplification of a ROS-TGFβ loop IKKβ plays a crucial role in the prevention of fibroblast-myofibroblast transformation and senescence.