1.Role of natural killer T cells in renal fibrosis in mice with acute kidney injury
Tao ZHANG ; Benquan LIU ; Hua LIANG ; Jiangju HUANG ; Wenqiang YU ; Hanbing WANG
Chinese Journal of Anesthesiology 2021;41(1):92-95
Objective:To evaluate the role of natural killer T cells in renal fibrosis in mice with acute kidney injury (AKI).Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=6 each) using a random number table method: control group (group C), AKI group (group A), control plus CD1d antibody group (group C-MA), and AKI plus CD1d antibody group (group A-MA). The model of renal fibrosis in mice with AKI was established by intraperitoneal injection of folic acid 250 mg/kg.In group C, homotypic control antibody 20 mg/kg was injected via the tail vein.In group AKI, homotypic control antibody 20 mg/kg was injected via the tail vein at 24 h before establishing the model. In group C-MA, anti-CD1d monoclonal antibody 20 mg/kg was injected via the tail vein.In group A-MA, anti-CD1d monoclonal antibody 20 mg/kg was injected via the tail vein at 24 h before establishing the model.On the 14th day after folic acid injection, blood samples were taken from eyeballs to determine the concentrations of blood urea nitrogen (BUN) and creatinine (Cr) in serum.Then the mice were sacrificed, and the renal tissues were taken for Sirius red staining and HE staining to determine the area of renal fibrosis, and renal injury was scored.The expression of fibronectin (FN), type I collagen (Col-Ⅰ) and alpha-smooth muscle actin (α-SMA) in renal tissues was detected by immunofluorescence method.The expression of interleukin (IL)-4, IL-13, arginase-1 (Arg-1) and found in inflammatory zone 1 (FIZZ1) mRNA in renal tissues was detected by real-time polymerase chain reaction. Results:Compared with group C, the concentrations of BUN and Cr in serum, renal injury score, and area of renal fibrosis were significantly increased, the expression of FN, Col-Ⅰ and α-SMA and IL-4, IL-13, Arg-1 and FIZZ1 mRNA was up-regulated in A and A-MA groups ( P<0.05), and no significant change was found in the above indexes in group C-MA ( P>0.05). Compared with group A, the concentrations of BUN and Cr in serum, renal injury score, and area of renal fibrosis were significantly decreased, the expression of FN, Col-Ⅰ and α-SMA and IL-4, IL-13, Arg-1 and FIZZ1 mRNA was down-regulated in group A-MA ( P<0.05). Conclusion:Activation of natural killer T cells is involved in the process of renal fibrosis in mice with AKI, and the mechanism may be related to promoting the release of Th2 cytokines and M2 polarization of macrophages.
2.Role of JMJD3 in cisplatin-induced renal fibrosis following acute kidney injury in mice
Jiayi NIE ; Hua LIANG ; Benquan LIU ; Ying GAO ; Wenqiang YU ; Hanbing WANG
Chinese Journal of Anesthesiology 2022;42(1):77-81
Objective:To evaluate the role of Jumonji domain-containing 3 (JMJD3) in cisplatin-induced renal fibrosis following acute kidney injury in mice.Methods:Forty-eight healthy C57BL/6 male mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n=12 each) using a random number table method: control group (group CON), control plus JMJD3 inhibitor group (group CON-A), cisplatin group (group CIS), and cisplatin plus JMJD3 inhibitor group (group CIS-A). In group CIS and group CIS-A, cisplatin was intraperitoneally administered on 1st and 14th days, respectively, to develop a renal fibrosis model in mice with acute kidney injury, and the JMJD3 inhibitor GSKJ4 10 mg/kg and equal volume of PBS were intraperitoneally injected on 4th day, respectively, once every 3 days, 6 injections in total.The equal volume of PBS and GSKJ4 10 mg/kg were intraperitoneally injected at the corresponding time points in group CON and group CON-A, respectively.Six mice in each group were selected, and orbital blood samples were collected on 3rd day after the first injection of cisplatin to determine the concentrations of serum creatinine (Cr) and blood urea nitrogen (BUN), then the animals were sacrificed, and kidney tissues were obtained for microscopic examination of pathological changes after HE and PAS staining (with a light microscope), and the damage to kidneys was assessed and scored.Six mice were sacrificed on 28th day after the first injection of cisplatin, and kidney tissues were removed for determination of the area of renal fibrosis ( via Sirius red and Masson staining), expression of fibronectin (Fn), collagen type Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA) (by immunofluorescence), F4/80 + cell and CD3 + cell count (using immunohistochemical method), and expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), CXC chemokine ligand 16 (CXCL16), and monocyte chemoattractant protein1 (MCP-1) mRNA (by real-time polymerase chain reaction). Results:Compared with group CON, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly increased, the expression of Fn, Col Ⅰ and α-SMA was up-regulated, the F4/80 + cell and CD3 + cell count was increased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was up-regulated in group CIS ( P<0.05), and no significant change was found in the parameters mentioned above in group CON-A ( P>0.05). Compared with group CON-A, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly increased, the expression of Fn, Col Ⅰ and α-SMA was up-regulated, the F4/80 + cell and CD3 + cell count was increased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was up-regulated in group CIS-A ( P<0.05). Compared with group CIS, the serum BUN and Cr concentrations, renal injury scores, and area of renal fibrosis were significantly decreased, the expression of Fn, Col Ⅰ and α-SMA was down-regulated, the F4/80 + cell and CD3 + cell count was decreased, and the expression of IL-6, CXCL16, TNF-α and MCP-1 mRNA was down-regulated in group CIS-A ( P<0.05). Conclusions:JMJD3 is involved in the process of renal fibrosis following acute kidney injury in mice, and the mechanism may be related to promotion of inflammatory responses.
3.Role of JMJD3 in drug-associated acute kidney injury in mice
Wenqiang YU ; Benquan LIU ; Hua LIANG ; Jiayi NIE ; Chaoqun ZHONG
Chinese Journal of Anesthesiology 2022;42(3):342-346
Objective:To evaluate the role of histone demethylase (JMJD3) in drug-induced acute kidney injury (AKI) in mice.Methods:Twenty-four male C57BL/6 mice, aged 8-10 weeks, weighing 20-30 g, were divided into 4 groups ( n =6 each) using a random number table method: control group (C group), AKI group, a specific JMJD3 inhibitor GSKJ4+ control group (GSKJ4 group), and GSKJ4-AKI group.Folic acid 250 mg/kg was injected intraperitoneally to develop AKI model.GSKJ4 20 mg/kg was intraperitoneally injected at 1 h before developing AKI model in GSKJ4-AKI group and at the corresponding time point in GSKJ4 group.Blood samples were collected at 72 h after development of AKI model for determination of serum BUN and Cr concentrations.The animals were then sacrificed and renal tissues were collected for microscopic examination of histopathological morphology (using HE and PAS staining) and for determination of cell apoptosis (by TUNEL) and expression of JMJD3, Bax and cleaved caspase-3 (by Western blot), the number of JMJD3, myeloperoxidase (MPO), F4/80 and CD3 positive cells, expression of cleaved caspase-3 and Bax, and expression of IL-1β, IL-6, TNF-α and monocyte chemotactic protein 1 (MCP-1) mRNA (by reverse transcription polymerase chain reaction). The damage to the renal tubules was scored. Results:Compared with C group, the serum BUN and Cr concentrations and renal tubular damage score were significantly increased, the number of JMJD3, myeloperoxidase (MPO), F4/80 and CD3 + positive cells was increased, the number of apoptotic cells was increased, and the expression of Bax, cleaved caspase-3, JMJD3 and IL-1β, TNF-α, IL-6 and MCP-1 mRNA was up-regulated in AKI group ( P<0.05), and no significant change was found in the parameters mentioned above in GSKJ4 group ( P>0.05). Compared with AKI group, the serum BUN and Cr concentrations and renal tubular damage score were significantly decreased, the number of JMJD3, myeloperoxidase (MPO), F4/80 and CD3 positive cells was decreased, the number of apoptotic cells was decreased, and the expression of Bax, cleaved caspase-3, JMJD3, and IL-1β, TNF-α, IL-6 and MCP-1 mRNA was down-regulated in GSKJ4-AKI group ( P<0.05). Conclusions:The mechanism of drug-associated AKI may be related to up-regulation of JMJD3 expression and thus induces cell apoptosis and inflammatory responses in mice.
4.Effects of modified posterior nasal nerve neurectomy combined with accessory posterior nasal nerve neurectomy on controlling intractable allergic rhinitis
Yunfangzi GU ; Benquan YU ; Xin WAN ; Yu GU ; Ganghua ZHU ; Zi'an XIAO
Journal of Central South University(Medical Sciences) 2023;48(3):404-413
Objective: A subset of intractable allergic rhinitis (iAR) patients experience severe symptoms which cannot be effectively controlled by standard drug therapy and/or antigen specific immunotherapy. In recent decades, endoscopy vidian neurectomy and posterior nasal nerve neurectomy (PNNN) were introduced as treatments of iAR that have shown to be highly successful at symptom management in a number of patients. But some patients experience relapse or suboptimal symptom control postoperation. To improve the effectiveness of PNNN to control iAR, a modified PNNN surgical approach (mPNNN) combined with accessory posterior nasal nerve neurectomy (aPNNN), which called as mPNNN-aPNNN was used. This study aims to compare the effects between mPNNN-aPNNN and PNNN on controlling the symptoms of iAR and evaluate the surgical effectiveness and safety of mPNNN-aPNNN. Methods: The patients with iAR experienced mPNNN-aPNNN or PNNN surgery at the department of Otolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were analyzed retrospectively. The approach of PNNN, a selective resection of the posterior nasal nerve branches, was modified to the neurectomy of total branches of posterior nasal nerve at the sphenopalatine foramen, and combined the operation of aPNNN in which the accessory posterior nasal nerve at the palatine bone perpendicular plate was resect in our study. Daily Nasal Symptom Scores (DNSS), Total Rhinitis Medication Score (TRMS), and the Rhinoconjunctivitis Qualities of Life Questionnaires Scores (RQLQS) were used to evaluate the complications during the operation and after the operation at the 3rd, 6th, 12th, and 24th month postoperatively. Total Nasal Symptom Scores (TNSS) was used to assess the total effective rate and markedly effective rate of the operations. Results: A total of 140 iAR patients experienced mPNNN-aPNNN or PNNN. Those with concomitant septoplasty and/or inferior turbinate reduction, and were absent during the postoperative follow-up were excluded. The final 62 patients with mPNNN-aPNNN and 34 with PNNN were enrolled. DNSS, TNSS, TRMS, and RQLQS at the postoperation were significantly improved compared with the preoperation in all patients (all P<0.001). Compared with PNNN, the postoperative DNSS, TNSS, and TRMS of mPNNN-aPNNN were obviously improved (all P<0.001). There was a persisted relief of symptoms at the postoperation in all patients with mPNNN-aPNNN. The total effective rate and markedly effective rate at the postoperative 24th month were 100% and 83.3%, respectively. Furthermore, the postoperative RQLQS decreased significantly (P<0.001). Only 5 sides of all patients (5/192, 2.6%) reported upper palate numbness during the first week after operation, with all recovered spontaneously in 1 month without treatment. No other postoperative complications occurred in mPNNN-aPNNN and PNNN.Conclusion: The surgery of mPNNN-aPNNN improve TNSS more significantly than PNNN. The operation of mPNNN-aPNNN is safe and effective to control iAR symptoms.