1.Role of Beclin-1 in the autophagic regulation of podocyte
Chinese Journal of Applied Clinical Pediatrics 2014;29(17):1281-1284
Autophagy is the cytoplasm and organelles of eukaryotic cells through degradation of itself,aself digest,a series of biochemical process is a kind of programmed cell death,its main function is to remove and degrade its damaged organelles and redundant biological macromolecules,and use the degradation products provide energy,reconstruction and cellular structure,in the steady state of cell and cell life activities play an important role.In recent years,autophagy in glomerular cells play a role of regulation and control have become the focus in the confrontation podocyte damage,including Beclin-1 autophagy related genes in the role of autophagy regulation has caused wide public concern.This article chooses of Beclin-1 in sertoli cell autophagy related research in the role of regulation are reviewed.
2.Influence of dexamethasone on the expression and distribution of transient receptor potential cation channel 6 in glomerular podocytes
Huiyang WANG ; Li YU ; Shengyou YU ; Zhihong HAO ; Yao ZHANG
Chinese Journal of Nephrology 2014;30(5):377-383
Objective To observe the changes of foot processes,expression and distribution of transient receptor potential cation channel 6 (TRPC6) in podocytes by puromycin aminonucleoside (PAN) and dexamethasone (DEX) intervention,then to investigate the function of TRPC6 in podocytes and its relation to proteinuria in kidney diseases.Methods Podocytes cultured in vitro were divided into three group:control group,PAN stimulation group and DEX intervention group.Mouse podocyte cell line (MPC5) were cultured in 0.02% dimethyl sulfoxide (DMSO) in control group,subjected to PAN (50 μg/ml) treatment alone or with DEX (1 μmol/L) in other two groups for 8 h,24 h,48 h.The podocyte morphology was observed and took pictures by phase-contrast microscope,then the differences of morphology and areas were analyzed.The distribution,mRNA expression and protein expression of TRPC6 were detected by indirect immunocyto-fluorescence,real-time quantitative PCR and Western blotting,respectively.Results The well-developed podocyte arborization and interconnection was formed in control group,but PAN led to significant shrinkage of podocytes (P < 0.05),together with podocyte foot process retraction,effacement and loss of cell contact.DEX significantly prevented the shrinkage and apoptosis of podocytes.The apoptosis rate was significantly increased after PAN stimulated 48 h (P < 0.05).Real-time quantitative PCR and Western blotting found TRPC6 mRNA and protein expression were prone to increase in PAN group compared with control group (P < 0.05).The distribution of TRPC6 becamed abnormal in PAN group.DEX decreased TRPC6 mRNA and protein expression at 48 h compared with PAN group (P < 0.05).The abnormal distribution of TRPC6 was also alleviated by the protection of DEX.Conclusion DEX exerts a direct action to podocyte which retains the integrity of slit diaphragm against podocyte injury,and alleviates proteinuria via stabilizing mRNA,protein expression and distribution of TRPC6.
3.Distribution and expression of α-actin-4 in puromycin aminonueleoside injured mouse podocyte cell line
Huiyang WANG ; Li YU ; Xiaoyan LI ; Shengyou YU ; Yao ZHANG ; Zhihong HAO
Chinese Journal of Applied Clinical Pediatrics 2014;29(17):1296-1300
Objective To observe the expression and distribution of α-actin-4 mRNA through puromycin aminonueleoside(PAN) injury podocyte,and discuss the relation between α-actin-4 and podocyte damage.Methods Podocytes were cultured in vitro,and 2 groups were set up:control group and PAN stimulation group.The control group was cultured with concentration of 100 mL/L FBS RPMI 1640 nutrient solution culture,while the PAN group was cultivated to PAN(50 mg/L) treatment,and cell morphology,extraction of total RNA of α-actin-4 were observed in 8 h,24 h and 48 h.The podocyte morphology was observed and pictures were taken through phase-contrast microscope,then the differences of morphology and areas between the 2 groups were analyzed.The distribution and mRNA expression of α-actin-4 were detected by indirect immunocyto-fluorescence and real-time quantitative PCR,respectively.Results The well-developed podocyte arborization was formed after the in vitro induction,and the PAN treatment led to the podocyte foot process retraction and effacement together with the mouse podocyte cell line shrinkage and the loss of cell contact.The above time point α-actin-4 mRNA expressions between the 2 groups were compared,and there was no significant difference in 8 h (P > 0.05),but significant difference was found in 24 h,48 h,α-actin-4 higher mRNA expression,with statistical significance(all P <0.01).α-actin-4 in the control group had thin filaments evenly distributed in the cytoplasm,but a radioactive distribution in foot process.In the experimental group,α-actin-4 pressure silk fiber was shorter,with disordered arrangement,and PAN stimulus after 24 h,α-actin-4 distribution in cytoplasm was decreased significantly,while cytoplasmic distribution was missing after 48 h.Conclusions The abnormal of distribution and mRNA expression of α-actin-4 is time-related to the PAN injury podocyte,and α-actin-4 is an important part of podocyte damage mechanism.
4.Expression and significance of podocyte molecules mRNA in Doxorubicin-induced nephrotic rats
Yihui HUANG ; Li YU ; Huiyang WANG ; Shengyou YU ; Luo JIANG ; Yao ZHANG
Chinese Journal of Applied Clinical Pediatrics 2016;31(5):345-348
Objective To observe the expression of mRNA of podocin,nephrin,CD2AP and α - actin - 4 in Doxorubicin - induced nephrotic(ADN)rats,and explore the possible mechanisms of podocyte molecule during the de-velopment of proteinuria. Methods Forty - eight Sprague - Dawley(SD)rats were divided into ADN model group(in-jected with 6. 5 mg/ kg Doxorubicin in tail vein,n = 24)and control group(injected with saline solution in tail vein,n =24). After the nephropathy model was established,6 rats were killed at the end of 1st ,2nd ,4th ,6th week in each group. The changes of the following indicators were observed:(1)24 - hour urinary protein,serum albumin and cholesterol were detected;(2)mRNA expression of nephrin,podocin,CD2AP and α - actin - 4 in cortex of kidney were examined by real time fluorescence quantification PCR. Results The model group came out massive proteinuria(15. 66 ± 1. 50) mg/ 24 h,(45. 98 ± 1. 45)mg/ 24 h,(65. 58 ± 4. 68)mg/ 24 h,(82. 83 ± 8. 43)mg/ 24 h in 1,2,4,6 weeks respec-tively,hypoalbuminemia(27. 4 ±2. 5)g/ L,(23. 6 ±2. 9)g/ L,(20. 6 ±1. 5)g/ L,(6. 9 ± 2. 3)g/ L in 1,2,4,6 weeks respectively and hypercholesterolaemia(2. 00 ± 0. 25)mmol/ L,(2. 16 ± 0. 44)mmol/ L,(4. 02 ± 0. 81)mmol/ L, (7. 54 ± 1. 12)mmol/ L in 1,2,4,6 weeks respectively,and the differences of proteinuria,plasma albumin and total cholesterol compared with control group at each time point had statistical significance(all P ﹤ 0. 01). Compared with the control group,podocin mRNA expression in the model group decreased at the end of 1st week(10. 56 ± 3. 62),de-creased significantly at the end of 2nd week(20. 44 ± 9. 03),and decreased at the end of 4th week(2. 19 ± 0. 18)com-pared with the control group;nephrin mRNA expression decreased at the end of 1st week(2. 41 ± 1. 10)and reached to the peak value,decreased at the end of 4th week(0. 52 ± 0. 18);CD2AP mRNA expression did not change significantly in the 1st week(4. 17 ± 0. 79),increased at the end of 2nd week(6. 74 ± 1. 53),reached to the peak value at the end of 4th week(6. 91 ± 1. 13),but did not change significantly at the end of 6th week(4. 04 ± 0. 82);α - actin - 4 mRNA ex-pression did not change significantly at the end of 1st week(1. 75 ± 0. 48),decreased at the end of 2nd week(2. 01 ± 0. 55),reached to the peak value at the end of 4th week(2. 24 ± 0. 81),but did not change significantly at the end of 6th week(1. 39 ± 0. 18). Compared with the control group,the difference had statistical significance( all P ﹤ 0. 05). Conclusion The abnormal expression of podocyte molecules mRNA in ADN rats may be an important molecular mechanism in the development of proteinuria.
5.Modified transanal repair of vesicorectal fistula after radical prostatectomy
Shengsong HUANG ; Chao LI ; Ying LIU ; Gang WU ; Weidong ZHOU ; Wei LE ; Chengdang XU ; Huiyang JIANG ; Yicong YAO ; Denglong WU
Chinese Journal of Urology 2021;42(1):1-5
Objective:To investigate the effect of modified transanal approach in the repair of vesicorectal fistula after radical prostatectomy.Methods:From September 2011 to December 2019, 32 cases of vesicorectal fistula after radical prostatectomy were retrospectively analyzed. All patients underwent cystostomy before repair operation. The average diameter of the fistulas was 19 (3-40) mm. There was only one fistula in 24 cases and 8 cases with more than 2 fistulas. The operation was performed in the jack knife position, and the fistula was prepared by resection of the fistula through the anus with bipolar resectoscope. Then bladder wall and rectum wall were separated by the loop and sutured respectively. After operation, the patients were treated with antispasmodic and anti-infective treatment, and the catheter was retained. Cystography and cystoscopy were reexamined 3 months after operation. Catheter was removed in the successful cases, and the failure was repaired again.Results:All operations were completed successfully. The mean operation time was 67(55-125) min, and the median follow-up was 22 (6-30) months. Thirty-one cases (96.8%) were successfully repaired, of which 25 cases were successfully repaired at the first operation, and 6 cases were successfully repaired again (all by transanal route). One case failed to be repaired. He had received external pelvic radiotherapy before operation. After the failure of repair, cystoscopy showed large fistula and stiff surrounding tissue. Then bilateral ureteral skin stoma and cystectomy were performed.Conclusions:Modified transanal approach in the repair of vesicorectal fistula after radical prostatectomy is an effective method. This kind of operation has less trauma, fewer complications and can be operated repeatedly. It is suitable for patients with low position, small fistula and without radiotherapy.
6.Clinical analysis of 6 cases of systemic lupus erythematosus complicated by transverse myelitis.
Li-fu TAN ; Hao LI ; Liu-qin LIANG ; Zhong-ping ZHAN ; Fan LIAN
Journal of Southern Medical University 2011;31(2):313-316
OBJECTIVETo summarize the clinical features and therapeutic approach of systemic lupus erythematosus (SLE) complicated by transverse myelitis (TM).
METHODSThe clinical characteristics, laboratory examinations, treatment and prognosis of 6 SLE cases with TM were retrospectively analyzed with review of the literatures.
RESULTSThe 6 patients consisted of 5 females and 1 male aged 14 to 36 years (mean 23 years). The mean duration from symptom onset of SLE to TM was 8 months (1 to 13 months). All the patients had lower limb hypodynamia, and 3 of them developed upper limb hypodynamia. MRI scanning of the spine identified lesions in the cervical spinal cord in 2 cases, thoracic lesions in 3 cases, and multiple involvement of the cervical, thoracic and lumbar cord in 1 case. Examination of the cerebrospinal fluid yielded no specific findings except for leukocytosis in 1 case and hypoglycemia in another. Five cases were treated with high-dose MP+CTX, and the other case was treated with MP (80 mg/day)+CTX. Five patients responded favorably to the treatment, while the other showed no obvious improvement.
CONCLUSIONTM is a rare complication of SLE affecting mostly young patients and occurring in the early stage of the disease. Early diagnosis and aggressive treatment might improve the prognosis.
Adolescent ; Adult ; Female ; Humans ; Lupus Erythematosus, Systemic ; complications ; Magnetic Resonance Imaging ; Male ; Myelitis, Transverse ; complications ; Spinal Cord ; pathology ; Young Adult
7.TCM Treatment of Podocyte Injury in IgA Nephropathy Based on Multiple TCM Theories: A Review
Yongfang LIU ; Huiyang LIU ; Bangming CHEN ; Li ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):198-208
IgA nephropathy is the most common primary glomerular disease in China. Its clinical manifestations are mainly proteinuria, hematuria, hypertension, edema, hyperuricemia, etc. Most patients have hidden onset. 30%-40% of patients develop into end stage renal disease 10-20 years after diagnosis and rely on dialysis or kidney transplantation to maintain their lives, which is extremely harmful. Proteinuria is a common clinical manifestation of this disease, and most patients have small-to-moderate amounts of proteinuria, while 10%-24% of patients have large amounts of proteinuria. Proteinuria is the main risk factor affecting the progression of renal function in IgA nephropathy. Podocytes are the terminal part of the glomerular filtration barrier, and various factors can affect the fusion and detachment of podocyte processes that occur after podocyte injury. They are common histological lesions in IgA nephropathy and are key factors leading to proteinuria and the continuous progression of the disease. At present, Western medicine lacks targeted treatment for podocyte injury, with limited intervention methods. Drugs such as glucocorticoids are often used for treatment, and there are many adverse reactions. Based on the physiological function of podocytes, pathological and physiological changes after injury, and histological morphology of this disease, it is believed that it is closely related to traditional Chinese medicine's "Xuanfu Theory" "Kidney Collateral Syndrome" "Collateral Disease Theory", and "Dry Blood Theory". More and more studies have shown that traditional Chinese medicine, which has the characteristics of multiple links, pathways, and targets, has a significant therapeutic effect on podocyte injury in IgA nephropathy. It can protect podocytes and reduce proteinuria and has good application and research prospects. This article systematically summarizes the mechanism and morphological changes of podocyte injury in IgA nephropathy, the understanding of podocyte injury in traditional Chinese medicine theory, and the research progress in traditional Chinese medicine treatment of podocyte injury in IgA nephropathy, so as to provide a reference for further research and application of traditional Chinese medicine intervention in podocyte injury in IgA nephropathy.
8.Role of NLRP3 Inflammasome in IgA Nephropathy and Chinese Medicine Intervention: A Review
Yongfang LIU ; Li ZHOU ; Huiyang LIU ; Rui HUANG ; Zhiying FENG ; Tiantian YIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):269-279
IgA nephropathy is recognized as the most common primary glomerular disease, with up to 20%-40% of patients developing end-stage kidney disease within 20 years of onset. The deposition of IgA1-containing immune complexes targeting glycosylation defects in the mesangial region and the subsequent inflammation caused by T lymphocyte activation are considered as the main causes of IgA nephropathy, and innate immunity is also involved in the pathogenesis. Nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) is a newly discovered pattern recognition receptor expressed in renal intrinsic cells such as renal tubular epithelial cells, mesangial cells, and podocytes. Activated by external stimuli, NLRP3 can form NLRP3 inflammasomes with apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC). The NLRP3 inflammasome can activate cysteine aspartate-specific protease-1 (Caspase-1), causing the maturation and release of interleukin-18 (IL-18) and interleukin-1β (IL-1β) involved in inflammation. Increasing evidence has suggested that NLRP3 inflammasomes are involved in the pathogenesis and progression of IgA nephropathy and associated with the damage of renal intrinsic cells such as podocytes, mesangial cells, endothelial cells, and renal tubular epithelial cells. Chinese medicine can regulate inflammatory cytokines and their signaling pathways by acting on NLRP3 inflammasomes and related molecules, exerting therapeutic effects on IgA nephropathy. This article introduces the role of NLRP3 inflammasomes in IgA nephropathy and reviews the clinical and experimental research progress of Chinese medicine intervention in IgA nephropathy via NLRP3 inflammasomes, aiming to provide a reference for further research and application of Chinese medicine intervention in the NLRP3 inflammasome as a new therapeutic target.
9.Highly sensitive H2O2-scavenging nano-bionic system for precise treatment of atherosclerosis.
Xiaoyu LIANG ; Huiyang LI ; Xuanling LI ; Xinxin TIAN ; Aiai ZHANG ; Qingzhi LUO ; Jianwei DUAN ; Youlu CHEN ; Liyun PANG ; Chen LI ; Xing-Jie LIANG ; Yong ZENG ; Jing YANG
Acta Pharmaceutica Sinica B 2023;13(1):372-389
In atherosclerosis, chronic inflammatory processes in local diseased areas may lead to the accumulation of reactive oxygen species (ROS). In this study, we devised a highly sensitive H2O2-scavenging nano-bionic system loaded with probucol (RPP-PU), to treat atherosclerosis more effectively. The RPP material had high sensitivity to H2O2, and the response sensitivity could be reduced from 40 to 10 μmol/L which was close to the lowest concentration of H2O2 levels of the pathological environment. RPP-PU delayed the release and prolonged the duration of PU in vivo. In Apolipoprotein E deficient (ApoE‒/‒) mice, RPP-PU effectively eliminated pathological ROS, reduced the level of lipids and related metabolic enzymes, and significantly decreased the area of vascular plaques and fibers. Our study demonstrated that the H2O2-scavenging nano-bionic system could scavenge the abundant ROS in the atherosclerosis lesion, thereby reducing the oxidative stress for treating atherosclerosis and thus achieve the therapeutic goals with atherosclerosis more desirably.
10.Establishment of the fingerprints of Yinhuang solution for inhalation and content determination of phenolic acids
Jin GAO ; Cui LI ; Ruizhuo YIN ; Xincheng MA ; Huiyang WANG ; Chunhui GONG ; Chengyu CHEN ; Hui CAO
China Pharmacy 2022;33(2):160-164
OBJE CTIVE To establish the finger prints for Yinhuang solution for inhalation and determine the contents of neochlorogenic acid ,chlorogenic acid and cryptochlorogenic acid simultaneously. METHODS Using baicalin as reference ,the fingerprints of Yinhuang solution for inhalation were established by high performance liquid chromatography (HPLC). Relative correction factors of neochlorogenic acid and cryptochlorogenic acid were calculated by slope correction method ,using chlorogenic acid as reference ;the contents of them were calculated according to relative correction factor. The results of quantitative analysis of multi-components by single marker (QAMS)were compared with those of external standard method (ESM). RESULTS There were 18 common peaks in the fingerprints of 10 batches of Yinhuang solution for inhalation ,and their similarities with reference fingerprint were higher than 0.90. A total of 7 common peaks were identified as baicalin ,neochlorogenic acid ,chlorogenic acid , cryptochlorogenic acid ,isochlorogenic acid B ,3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid. The linear range of neochlorogenic acid ,chlorogenic acid and cryptochlorogenic acid were 0.025 0-1.247 4 μg(r=0.999 7),0.039 3-1.178 7 μg(r= 0.999 9),0.031 6-1.184 1 μg(r=0.999 9),respectively. RSDs of precision ,reproducibility and stability tests (48 h)were all lower than 1.0%. The average recoveries were 93.92%(RSD=1.32% ,n=6),94.46%(RSD=1.45%,n=6),93.93%(RSD= 1.57%,n=6). Relative correction factors of neochlorogenic acid and cryptochlorogenic acid were 1.068 and 1.233. The contents of neochlorogenic acid and cryptochlorogenic acid determined by QAMS method were 0.301 8-0.386 3 and 0.262 5-0.362 5 mg/mL, respectively. The contents of neochlorogenic acid ,chlorogenic acid and cryptochlorogenic acid by ESM were 0.302 6-0.387 2, 0.231 0- 0.334 0,0.261 6-0.361 3 mg/mL,respectively. The deviations of the content determination results of the two methods(except for chlorogenic acid )were both not higher than 0.20%. CONCLUSIONS Established HPLC fingerprints are stable and feasible. Established QAMS method is accurate and rapid. HPLC fingerprint combined with QAMS can be used for the quality control for Yinhuang solution for inhalation .