ABSTRACT
OBJECTIVE Chronic kidney disease (CKD) has become a global public health problem with 10％-15％incidence rate, and inhibiting the renal interstitial fibrosis is considered to be a potential strategy to delay the progression of CKD. Z-Guggulsterone (Z-GS), an active compound from derived from Commiphora mukul, has been proved to be effective in various diseases. The present study aimes to determine the protective effect and the molecular mechanism of Z-GS on renal fibrosis. METHODS Unilateral ureteral obstruction (UUO) mice and hypoxia-induced HK-2 cells were used to simulate renal fibrosis in vitro and in vivo, respectively. The mice and cells were treated with different doses of Z-GS to observe the pharmacological action. Renal function, including Scr, BUN, and UA, were detected by commercial kits. H&E and Masson staining were performed to observe histopathological changes of kidney. Cell viability and LDH release of HK-2 cells were detected by commercial kits. Cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Fibrosis markers were detected by immunohistochemistry and immunofluorescence analysis. Cell cycle related proteins and Klotho/p53 signaling were analyzed by Western blotting. RESULTS The results showed that Z-GS decreased the rise of Scr, BUN, and UA and lightened renal histopathological injury, which were induced by UUO. Besides, Z-GS administration alleviated renal fibrosis in mice by inhibiting the expressions of α-SMA, TGF-β and colla?genⅣ, and delayed G2/M cell cycle arrest by promoting the expressions of CDK1 and cyclinD1/B1 rate. Experiments in vitro indicated that Z-GS treatment significantly increased the cell viability while decreased the LDH release in hypoxia-induced HK-2 cells. In addition, hypoxia induced fibrosis and G2/M cycle arrest in HK-2 cells were retarded by Z-GS. The study of its possible mechanism exhibited that Z-GS treatment increased the level of Klotho and inhibited P53 level. Nev?ertheless, the effect of Z-GS on Klotho/P53 signaling was reversed by siRNA-Klotho. Moreover, siRNA-Klotho treatment eliminated the effects of Z-GS on G2/M cell cycle arrest and fibrosis. CONCLUSION This study clarified that Z-GS allevi?ated renal fibrosis and G2/M cycle arrest through Klotho/P53 signaling pathway. People who have suffered CKD may potentially benefit from treatment with Z-GS.
ABSTRACT
OBJECTIVE： To study the improvement effects of Z-guggulsterone （Z-GL） combined with acetyl-11-keto-β- boswellic acid （AKBA） on cerebral ischemia-reperfusion injury model rats. METHODS： Male SD rats were randomly divided into sham operation group， model group， Z-GL+AKBA low-dose and high-dose groups （25， 50 mg/kg）， with 10 rats in each group. Except for sham operation group， middle cerebral artery occlusion/reperfusion injury model was induced by suture method in other groups. Administration groups were given relevant medicine intragastrically after reperfusion； sham operation group and model groups were given constant volume of DMSO intragastrically， every 12 h， for consecutive 7 d. The neurological deficits were evaluated with modified Longa score； HE staining was performed to observe the pathological changes of cerebral tissue in rats； the area of cerebral infarction was measured by TTC， and the percentage of cerebral infarction area； TUNEL staining was performed to detect apoptotic neurons. The expression of CD34， VEGF and DLL4 were detected by immunofluoresence and immunoblotting assay， respectively. RESULTS： Compared with sham operation group， the number of cortical cells in the model group decreased and arranged irregularly， with obvious infarct area and obvious decrease of neovascularization； the neurological deficit score， the percentage of cerebral infarction area and TUNEL positive cells increased significantly， while the expression of CD34， VEGF and DLL4 decreased significantly （P＜0.05 or P＜0.01）. Compared with model group， the above symptoms of the rats in each administration group were significantly improved， the neurological deficit score， the percentage of cerebral infarction area and the number of TUNEL positive cells were significantly reduced； the expression levels of CD34， VEGF and DLL4 were significantly increased； the neurological deficit score， the percentage of cerebral infarction area and the number of TUNEL positive cells in Z-GL+AKBA high-dose group were significantly lower or less than low dose group； the expression of CD34 and DLL4 in high-dose group was significantly higher than low-dose group （P＜0.05 or P＜0.01）. CONCLUSIONS： Z-GL combined with AKBA can relieve neurological deficit and cerebral injury of cerebral ischemia-reperfusion injury model rats， which may be related to promoting angiogenesis and up-regulating the expression of VEGF and DLL4 protein， with a certain dose-dependent effect.
ABSTRACT
OBJECTIVE:To study the improvement effect of Z-Guggulsterone(Z-GL)on blood coagulation and vascular endo-thelial functions of acute blood-stasis model rats and its mechanism. METHODS:40 rats were randomly divided into normal group,model group,positive group(Aspirin tablet,50 mg/kg)and Z-GL low-dose and high-dose groups(25,50 mg/kg),with 8 rats in each group. They were given relevant medicine intragastrically every 12 h,and normal group and model group were given constant volume of normal saline intragastrically for consecutive 7 times. After the fifth administration,except for normal group, those groups were given adrenalin hydrochloride subcutaneously+ice-cold water to induce acute blood-stasis model. Within 30 min after the last administration,aorta abdominalis sample were selected to detect the coagulation parameters [prothrombin time(PT), thrombin time (TT),activated partialthromboplastin time (APTT),fibrinogen (FIB)],and pathological changes of carotid artery were observed. Human umbilical vein endothelial cells (HUEVCs) were divided into blank group (normal saline),model group (normal saline) and Z-GL low-concentration and high-concentration groups (25,50 μmol/L). After culturing for 24 h,the cells were exposed to glucose and oxygen deprivation for 6 h in model group and Z-GL groups. The expression of p-eNOS protein was detected. Other cells were selected,grouped,administrated and treated as above cells,and the NO level of these cells were detect-ed. RESULTS:Compared with normal group,PT,TT and APTT were all shortened in model group,while FIB content was in-creased (P<0.01);vascular endothelium was injured,and endothelial cells fell off from the wall. Compared with model group, PT,TT and APTT were prolonged in administration groups,while FIB content was decreased(P<0.05 or P<0.01);vascular en-dothelium injury was relieved. Results of p-eNOS protein and NO levels determination showed that compared with model group, p-eNOS protein and NO levels were increased in Z-GL groups(P<0.05 or P<0.01). CONCLUSIONS:Z-GL can significantly im-prove coagulation and vascular endothelium functions of blood-stasis model rats,and its mechanism may be associated with the acti-vation of eNOS and the increase of NO level.