The role of PCSK9 on lipid accumulation and injury in the kidney of C57BL/6 mice / 中华肾脏病杂志

Objectives To evaluate the role of PCSK9 (proprotein convertase subtilisin kexin type 9) on the lipid accumulation and kidney injury of C57BL/6 mice. Methods The 24 h urine of 12 weeks old wide type C57BL/6 mice and PCSK9 knockout (KO) mice were collected through a metabolic cage, followed by perfusion and sacrifice. Urinary microalbumin?to?creatinine ratio (UACr), total cholesterol and triglyceride in kidney tissues were measured by ELISA. BODIPY 493/503 staining and standard transmission electron microscopy (TEM) of kidney tissues was performed for evaluating lipid accumulation and podocyte foot effacement in the kidney. Kidney tissues were also evaluated by PAS stain and TUNNEL stain. PCSK9, podocin and nephrin were quantified through real?time PCR, and the Bcl?2, Bax and cleaved caspase 3 were evaluated by Western blotting. Results Total cholesterol and triglyceride contents were higher in the kidneys of PCSK9 KO mice than controls (P<0.05). The level of lipid accumulation in glomeruli and tubules through BODIPY 493/503 stain, and the amount of lipid drop in TEM were more serious in PCSK9 KO mice. UACr and podocyte foot process effacement were increased, and the transcription of podocin and nephrin were decreased in the kidneys of PCSK9 KO mice (all P<0.05). The expression of Bcl?2 was decreased, and Bax and cleavedcaspase 3 were increased in the kidney samples of PCSK9 KO mice. Conclusion PCSK9 might be reversely involved in lipid homeostasis and accumulation, resulting in injury and apoptosis in the kidneys of C57BL/6 mice.

Effects of 12-lipoxygenase and angiotensin Ⅱ on p21, p27 and p57 in rat diabetic glomeruli / 中华肾脏病杂志

Objective To investigate the effects of 12-lipoxygenase (12-LO) and angiotensin Ⅱ (Ang Ⅱ) on the CIP/KIP family of cyclin-dependent kinase inhibitors (CKIs) p21,p27 and p57 related to cell hypertrophy.Methods Mesangial cells were treated with high glucose for 24 hours and 48 hours respectively.12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] and Ang Ⅱ were infused to rats by osmotic mini-pump for 1 week and 2 weeks respectively.Rats fed high fat diet were received low dose streptozotocin (STZ) to make type 2 diabetes (DN).The rats were divided into normal control group,DN group,DN+Ang Ⅱ type 1 receptor blocker (ARB) group or 12-LO inhibitor (CDC) group.DN+ARB rats were treated by losartan for 6 weeks,and DN+CDC rats were treated for 8 weeks.Urine albumin and protein expressions of p21,p27 and p57 were detected by ELISA and Western blotting respectively.Glomeruli injury and expressions of p21 and p27 were detected by PAS staining and immunohistochemistry respectively.Results High glucose increased p21 and p27 protein expression in mesangial cells significantly compared with the relative control (all P ＜ 0.05),but had no effect on p57.Ang Ⅱ increased p27 protein expression in gloneruli significantly (P ＜ 0.05),but had no effect on p21 and p57 protein expression.12(S)-HETE increased both p21 and p27 protein expression in glomeruli significantly (all P ＜ 0.05),but had no effect on p57 protein expression.Blood glucose,kidney/body weight,urinary protein,and glomerular p21 and p27 protein expressions were increased in DN group (all P ＜ 0.05) compared with those in control group,with little change of p57 protein expression (P ＜ 0.05).Moreover,glomerular hypertrophy and extra cellular matrix accumulation were observed in DN group.However,urine protein,kidney/body weight,renal injury,but not blood glucose,were decreased in DN+ARB group and DN+CDC group compared with DN group respectively (P＜ 0.05).Further DN+CDC rats had decreased both p21 and p27 protein expressions in glomeruli,but DN+ ARB rats only had decreased p27 protein expression (all P ＜ 0.05).Conclusions 12-LO may induce both p21 and p27 protein expression in DN glomeruli,but Ang Ⅱ may induce only p27 expression.

Protective effects of antioxidants on chronic intermittent hypoxia-induced cardiac remodeling in mice / 中华心血管病杂志

<p><b>OBJECTIVE</b>Chronic intermittent hypoxia (CIH) animal model was used to mimic the status of obstructive sleep apnea (OSA) in order to investigate the pathological mechanism of CIH-induced cardiac remodeling and observe the protective effect of antioxidants.</p><p><b>METHODS</b>FVB mice (8-10 weeks-old) were randomly divided into control (saline, i.p.) group and CIH group, reduced form of nicotinamide adenine dinucleotide phosphate oxidase inhibitor, apocynin (APO, 3 mg×kg(-1)×d(-1), i.p.) alone or CIH+APO, SOD mimic MnTMPyP (SODM, 5 mg×kg(-1)×d(-1), i.p.) alone or CIH+SODM (n = 5 each). After 4 weeks, cardiac function and structure were determined by echocardiography, cardiac inflammation, apoptosis, cardiac fibrosis and cardiac MDA contents were examined by Western blot and chemical-biological methods, respectively.</p><p><b>RESULTS</b>(1) Heart weight, LVIDd and LVIDs were increased while LVEF and FS were reduced in CIH group compared to control group (all P < 0.05). (2) Myocardial protein expression of ANP and VCAM-1 was significantly upregulated, myocardial MDA content and apoptosis as well as myocardial fibrosis marker CTGF and PAI-1 were increased in CIH group compared to control group (all P < 0.05). (3) Above parameters were similar between APO and CIH+APO as well as SODM and CIH+SODM (all P > 0.05).</p><p><b>CONCLUSION</b>CIH could induce cardiac remodeling and CIH-induced cardiac inflammation, cardiac oxidative injury, cardiac apoptosis and cardiac fibrosis serve as the pathological mechanisms of CIH-induced cardiac remodeling. The protective effects of the two antioxidants suggest that the main mechanism of CIH-induced cardiac injury is oxidative stress.</p>

Effect of metallothionein on the renal injury induced by chronic intermittent hypoxia in mice / 中华肾脏病杂志

Objective To investigate the mechanism of chronic intermittent hypoxia (CIH)-induced renal injury and the protection of metallothionein (MT).Methods 8-10 weeks old male MT-1 transgenic (MT-TG) mice (n=12) and the wide type (WT) mice (n=12) were randomly divided into two groups respectively,Air mimic control(Ctrl) group (n=6) and CIH group (n=6).The period of chronic intermittent hypoxia was continued for 8 weeks.The CIH paradigm consisted of 20.9％ O2 and 8％ O2 fraction of inspiration O2 (FiO2) alternation cycles (30 episodes per hour) with 20 seconds at the nadir FiO2 for 12 hours/day during daylight.The nadir hemoglobin oxygen saturations mainly ranged from 60％ to 70％.Urine,blood,kidney were collected at the end of study respectively.Histopathology,Western blotting and colorimetric method for related target were performed respectively.Results In WT mice,renal fibrosis,the expression of connective tissue growth factor (CTGF),type-1 plasminogen activator inhibitor (PAI-1),hypoxia-inducible factor 1α (HIF-1α),transforming growth factor β1 (TGF-β1),phosphorylated Smad2 and the MDA content were significantly increased by CIH (P ＜ 0.01).In WT mice,the expression of MT detected by using Western blotting was significantly decreased by CIH (P ＜ 0.01).However,in MT-TG mice,above-mentioned indicators showed no significant difference between CIH and Ctrl group.Conclusions Oxidative stresses is the main mechanism of CIH-induced renal injury.The possible molecular mechanism of CIH-induced renal injury is that CIH increases the expression of HIF-1α in kidney tissue,then activate the TGF-β1-Smad2 signaling pathway and lead to the renal fibrosis.The protection of MT on CIH-induced renal injury may be via its antioxidant effect.

Clinical Observation on Insomnia Treated by Wei-Defensive Qi-Regulating and Brain-Strengthening Needling Technique / 针灸推拿医学（英文版）

In order to make a report on 90 cases of insomnia treated by the Wei-defensive qi-regulating and brain-strengthening needling technique,as a clinical subject of the multiple center,for assessing the preliminary standardized indications and operating methods and making an initial summary for writing the draft of the technique operation.Methods:By ratio of 1:1,the patients in conformity with the criteria were randomly divided into the experimental group and control group.In the experimental group,45 cases were treated with the Wei-defensive qi-regulating and brain-strengthening needling technique on Baihui (GV 20),Dazhui(GV 14),Shenmai(BL 62),Zhaohai(KI 6),plus ear points.In the control group,45 cases were treated with routine acupuncture on Sishencong(Ex-HN 1),Shenmen (HT 7),Sanyinjiao(SP 6),etc.The therapeutic effects in the two groups were observed and assessed in comparison.Results:The results showed cure in 15 cases.remarkable effect in 27 cases,effect in 42 cases,failure in 6 cases,and the total eriective rate in 94.4% in 90 cases of insomnia.In the three centers,the statistic management was performed based upon PSQI integral and showed significant difference(P＜0.01),and no significant difference among various groups(P＞0.01).Conclusion:The therapeutic effect in the treatment of insomnia by Wei-defensive qi-regulating and brain-strengthening needling technique is better than the control group and the therapeutic effect and methods in various centers are stable.without any deviation.

Effect of cisplatin on ototoxicity and expression of caspase-3 in mouse cochlea / 西安交通大学学报(医学版)

Objective To establish a mice model of cisplatin-induced ototoxicity and to investigate the effect of cisplatin on the expression of caspase-3 in mouse cochlea.Methods Totally 69 Kunming mice were randomly divided into control group,cisplatin 2.5mg/(kg?d) group,cisplatin 3.5mg/(kg?d) group and cisplatin 4.5mg/(kg?d)group.Mice were injected intraperitoneally for 5 days.Auditory brainstem response(ABR) was measured to observe the change of hearing.Envision method of immunohistochemistry was applied to detect the expression of caspase-3 in cochlea.Results The weight and hearing of mice in different dose cisplatin groups were declined significantly as compared with those in control group(P