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Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K+ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β2-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.
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Objective:To observe the effect of Xueniaoan tablets on the expression of complement regulatory protein CR1 in IgA nephropathy rats. Methods:Totally 36 SD rats were randomized into the normal group,the model group and Xueniaoan group after 4-day adaptive feeding. Complex method was used to establish IgA nephropathy model,and Xueniaoan group was given Xueniaoan at the dose of 7.0 g·kg-1during the 10thweek and the 12thweek,while the normal group and the model group were fed with normal saline, and the administration course was 3 weeks. The 24-h urine protein was detected,CR1 in red blood cells was detected by ELISA,the renal pathological changes were observed under a light microscope after HE staining,and the expression of CR1 was detected by immu-nohistochemistry. Results:Compared with that in the model group,the 24-h proteinuria in Xueniaoan group was significantly reduced, the number of red blood cell CR1 increased significantly (P<0.05), CR1 in renal tissue increased and the renal tissue injury was light. Conclusion:Xueniaoan tablets can delay the renal damages in IgA nephropathy rats through promoting the expression of CR1.
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Objective To discuss the effects of Liuwei Dihuang Decoction on expressions of TGF-β1 and complement regulatory protein CR1 of the renal on IgA nephropathy rats.Methods Forty SD rats were randomized into normal group, model group, tripterygium glycosides group and Liuwei Dihuang Decoction group after adaptive feeding for one week. Complex method was used to establish IgA nephropathy model. All administration groups were given relevant medicine for gavage for 4 weeks;urine routine, the number of urine erythrocyte and 24 h urine protein were detected;creatinine, urea nitrogen, total serum protein and serum alburin were detected from blood;the renal pathological changes were observed under light microscope by HE and Masson staining;the expressions of TGF-β1 and CR1 were detected by immunohistochemistry.Results Compared with model group,Liuwei Dihuang Decoction could significantly decrease the proteinuria and hematuria levels and renal fibrosis changes of IgA nephropathy model rats, reduce the expression of TGF-β1, and raise the expression of CR1 (P<0.05).Conclusion Liuwei Dihuang Decoction can delay the renal fibrosis in IgA nephropathy rats through reducing the degree of TGF-β1 and promoting the expression of CR1.
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Objective To evaluate the role of membrane-bound complement regulatory protein expression in spinal cord dorsal horn in the development of neuropathic pain (NP) in rats.Methods Forty-eight male Sprague-Dawley rats,weighing 200-250 g,in which intrathecal catheters were successfully implanted without complications,were randomly divided into 4 groups (n =12 each):sham operation group (group S),group NP,normal saline group (group NS) and minocycline group (group M).NP was induced by chronic constrictive injury of the sciatic nerve in NP,NS and M groups.Minocycline 50 μg was injected intrathecally once a day for 7 consecutive days starting from 1 day before ligation of the sciatic nerve in M group,while the equal volume of normal saline was given in stead of minocycline in NS group.Mechanical pain threshold and thermal pain threshold were measured at 1 day before ligation of the sciatic nerve (baseline,T0) and 1,3,7 days after ligation of the sciatic nerve (T1-3).Then the rats were sacrificed at T3 and the lumbar segment (L4,5) of spinal cord was removed for determination of the expression of CD46,CD55,CD59 protein and mRNA in the dorsal horn of spinal cord by Western-blot and RT-PCR,respectively.Results Compared with S group,mechanical pain threshold and thermal pain threshold were significantly decreased at T1-3 and the expression of CD46,CD55 and CD59 protein and RNA was down-regulated at T3 in NP,NS and M groups (P ＜ 0.05).Compared with groups NS and NP,mechanical pain threshold and thermal pain threshold were significantly increased at T2.3 and the expression of CD46,CD55 and CD59 protein and RNA was up-regulated at T3 in M group (P ＜ 0.05).Conclusion The down-regulated expression of membrane-bound complement regulatory proteins in spinal cord dorsal horn and abnormal activation of the complement are involved in the development of NP in rats.
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Objective To evaluate the role of MCP/DAF expression in the spinal cord in the development of neuropathic pain (NP) induced by chronic constrictive injury (CCI) of the sciatic nerve in rats.Methods Twenty-four Sprague-Dawley rats transfected with MCP/DAF,weighing 200-250 g,were randomly divided into 2 groups (n =12 each) using a random number table:sham operation of transfected rat group (Rsham group) and CCI of transfected rat group (RCCI group).Twenty-four healthy male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 2 groups (n =12 each) using a random number table:sham operation of normal rat group (Nsham group) and CCI of normal rat group (NCCI group).The right sciatic nerve was exposed and 4 loose ligatures wen placed on the sciatic nerve at 1 mm intervals with 4-0 catgut in RCCI and NCCI groups.The right sciatic nerve was only exposed in Rsham and Nsham groups.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured at 1 day before operation (baseline) and 1,3 and 7 days after operation.The animals were sacrificed after measurement of pain threshold on 7 days after operation and the L4,5 segment of the spinal cord was removed for determination of the expression of OX-42 (by immuno-histochemistry) and MCP mRNA and DAF mRNA (by RT-PCR).Results Compared with Nsham group,the PWT and PWL were significantly decreased on 1,3 and 7 days after operation,the expression of OX-42 was up-regulated,and the expression of MCP mRNA and DAF mRNA was down-regulated in NCCI group (P ＜ 0.05),and no significant changes were found in the PWT and PWL on 1,3 and 7 days after operation and expression of OX-42(P ＞ 0.05),and the expression of MCP mRNA and DAF mRNA was up-regulated in Rsham and RCCI groups (P ＞ 0.05).Compared with NCCI group,the PWT and PWL were significantly increased on 1,3 and 7 days after operation,the expression of OX-42 was down-regulated,and the expression of MCP mRNA and DAF mRNA was up-regulated in RCCI group (P ＜ 0.05).Conclusion Up-regulation of MCP/ DAF expression in the spinal cord can inhibit the development of NP in rats and regulation of activation of microglias in the spinal cord is involved in the mechanism.
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Sertoli cells (SC) are known to contain immunoprotective properties, which allow them to survive as allografts without the use of immunosuppressive drugs. Experiments were designed to determine which factors are related to prolonged survival of allogeneic SC. Balb/c derived Sertoli (TM4) and colon cancer (CT-26) cell lines were implanted beneath the kidney capsule of non-immunosuppressed C57BL/6 mice and compared their survival as allografts. Compared to TM4 graft, which survived more than 7 days after transplantation, CT-26 showed massive infiltration of polymorphonuclear cells, necrosis and enlargement of draining lymph nodes. Cultured cell lines showed no differences in their expression patterns of FasL, TGF beta1, clusterin and two complement regulatory proteins (CRP, i.e., membrane cofactor protein, MCP; decay accelerating factor, DAF), but protectin (CD59), another member of CRP was expressed only on TM4. These results suggest that CD59 and unknown factors may contribute to the prolonged survival of SC in non-immunoprivileged sites.
Subject(s)
Mice , Male , Female , Animals , Transplantation, Homologous/immunology , Transforming Growth Factor beta1/immunology , Sertoli Cells/immunology , Mice, Inbred C57BL , Graft Survival/immunology , Fas Ligand Protein/immunology , Complement System Proteins/immunology , Clusterin/immunology , Cells, Cultured , Cell SurvivalABSTRACT
Objective:To determine the expression of complement regulatory proteins(CRPs) C_1-INH,MCP(CD46) in the nasal mucosa of allergic rhinitis rats and to study the role of CRPs in the pathogenesis of allergic rhinitis.Methods: Nine healthy SD rats were sensitized and intranasally challenged with ovalbumin and Al(OH)_3(as supplement) to establish allergic rhinitis models and another 9 SD rats treated with saline were taken as control.The nasal mucosa in respiratory area of both groups was obtained.Then Western blotting was performed to investigate the expression levels of C_1-INH and CD46.Results: Western blotting showed that both C_1-INH and CD46 had been detected in rat nasal mucosa.Expression level of CD46 in the nasal mucosa of allergic rhinitis was significantly higher than that in control rats(P
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Objective:To investigate the expression of membrane cofactor protein (MCP)?decay accelerating factor (DAF) and homologous restriction factor 20 (HRF 20) on the surface of CD16 +monocyte/macrophage (Mo/M?) in the urine of patients with glomerulonephritis(GN) Methods:The expression of MCP?DAF and HRF 20 on the surface of urinary CD16 +Mo/M? were tested by Flow Cytometry and the levels of urinary sC5b 9 by ELISA in 134 patients with GN that were divided into mini change (MC)?glomerulosclerosis (GS)?membranous nephropathy (MN) and proliferative glomerulonephritis (PGN) Results:The expressions of MCP?DAF and HRF 20 on the surface of urinary CD16 +Mo/M? and the levels of urinary sC5b 9 in the patients with GS ?MN or PGN were significantly higher than those in controls (P
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Objective:To investigate the expression of membrane cofacto protein(MCP)?decay accelerating factor(DAF) and homologous restricition factor 20(HRF 20) on the surface of CD16 + monocyte/macrophage(Mo/M?) in patients with glomerulonephritis(GN).Methods:Tested the expression levels of MCP?DAF and HRF 20 on the surface of CD16 +Mo/M? by Flow Cytometry and the levels of serum C3d and C3d immunologic complex(C3d IC) by ELISA in 136 patients with GN,divided into mini change(MC)?glomerulosclerosis(GS)?membranous nephropathy(MN) and proliferative glomerulonephritis(PGN).Results:The expressions of MCP?DAF and HRF 20 on the surface of CD16 +Mo/M? and the serum levels of C3d in the patients with GS?MN or PGN were significantly higher than those in controls(P