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Objective:To investigate the significance and level of serum GRP78 and GRP94 in the early stage ofacute pulmonary embolism.Methods:Ninety APE patients (APE group) and forty healthy controls (control group) in Affiliated Zhongshan Hospital of Dalian University from January 2018 to March 2020 were recruited. The level of serum D-Dimer, GRP78 and GRP94 were compared between the two groups. The pulmonary embolism group was divided into three groups according to guidelines for the diagnosis and treatment of pulmonary embolism in ESC in 2019: low risk, medium risk, high risk, The level of serum D-Dimer, GRP78, GRP94, blood gas, pulmonary arterial pressure, PESI mark of three groups were compared.Results:Two groups had no significant difference in sex, age, body mass index ( P>0.05), but the level of serum D-Dimer, GRP78 and GRP94 in control group and APE group were higher than those in the control group: (3.86 ± 1.82) mg/L vs. (0.31 ± 0.15) mg/L, (2.68 ± 0.71) μg/L vs. (1.64 ± 0.38) μg/L, (1.31 ± 0.29) μg/L vs. (0.98 ± 0.13) μg/L ( P<0.05). The levels of serum D-Dimer, GRP78, GRP94, pulmonary arterial pressure, PO2, PESI score of different degree group (low risk, medium risk, high risk) were higher than those of low risk group and medium risk group: (5.63 ± 1.75) mg/L vs. (2.29 ± 0.51) and (3.64 ± 1.02) mg/L, (3.24 ± 0.76) μg/L vs. (2.17 ± 0.41) μg/L and (2.64 ± 0.47) μg/L, (1.57 ± 0.33) μg/L vs. (1.12 ± 0.13) and (1.26 ± 0.14) μg/L, (47.75 ± 6.98) mmHg (1 mmHg = 0.133 kPa) vs. (26.15 ± 4.63) and (35.21 ± 5.85) mmHg, (123.5 ± 20.59) scores vs. (85.5 ± 14.31) and (102.5 ± 13.32) scores ( P<0.05), and that of medium risk group were higher than those of low risk group ( P<0.05). But PO 2 of high risk group was lower than that of low risk group and medium risk group ( P<0.05), and PO 2 of medium risk group was lower than that of low risk group ( P< 0.05). pH of three group had no significant difference ( P>0.05). PCO 2 of high risk group was lower than that of low risk group and medium risk group ( P<0.05), and PCO 2 of medium risk group and low risk group had no significant difference ( P>0.05). The level of serum D-Dimer, GRP78, GRP94 were positively correlated withPESI score ( r = 0.610, 0.622, 0.627, P<0.01). After treatment, the levels of D-Dimer, GRP78 and GRP94 were significantly decreased ( P<0.05). Conclusions:The level of serum GRP78 and GRP94 are connected with acute pulmonary embolism, and it can reflect the severity of the disease.
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@#Glucose-regulated protein 94(Grp94), an endoplasmic reticulum resident Hsp90 paralog, has a limited set of client proteins. Selective inhibition of Grp94 has emerged as a new direction for the development of drugs targeting the Hsp90 chaperone system. Now Grp94-Probe, an affinity-based probe of Grp94, was designed and synthesized based on DDO-5813, a most potent Grp94-selective inhibitor we found previously. Using fluorescence polarization(FP)assay and double staining assay with ER-Red in cells, we confirmed the binding of Grp94-Probe with ER Grp94. The FR results showed that the probe exhibited high affinity for Grp94N(EC50=117. 9 nmol/L)without exhibiting obvious Hsp90α inhibition, Moreover, as a fluorescence probe molecule, Grp94-Probe could better distinguish the inhibitory activity of compounds for Grp94N. The results of fluorescence analysis in cells showed that Grp94-Probe could co-stain with ER-Red in the endoplasmic reticulum, and the fluorescence did not decay rapidly with time after 4 h of staining, which further indicated the binding of Grp94-Probe with Grp94 in cells. This Grp94 selective probe can be further used for biology evaluation of Grp94 inhibitor and exploration of Grp94 biological functions.
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Objective To study the expression of protein 94 (GRP94) and C/EBP-homologous protein (CHOP) in myocardial tissue of hypertensive rats and to investigate the effects of amlodipine on endoplasm retieulum stress ( ERS) and ventricular hypertrophy in abdominal aortic banded rats .Methods One hundred and twenty adult male SD rats with criteria were divided randomly into three groups:sham-operated group , abdominal aortic banding ( AAB) and AAB treated with amlodipine (AAB+Aml)groups(n=40).In sham-operated rats, the abdominal aorta was isola-ted but not constricted , while the abdominal aorta were constricted in AAB rats .The AAB+Aml group was treated by abdominal aortic constriction and treated with amlodipine [10 mg/(kg· d)].According to the time of surgery, each group was further divided into 2, 4 and 8-week postoperative subgroups ( n=6 ) .Mean arterial pressure (MAP), the 1eft ventricular mass (LVM) and body mass (BM) were measured and (LVM/BM) was calculated. The morphology of cardiomyocytes was observed by HE staining .The protein level of GRP 94 and CHOP was ana-lyzedbyimmunohistochemistryandwesternblot.Results 1)Withtimeaftersurgery,MAPandLVM/BWof AAB group increased gradually , and they were obviously higher than the sham operation group [ MAP: 2 weeks (144 ±10)vs(118 ±9), 4 weeks (163±8)vs(120±7), 8 weeks(177±10)vs(120±6)mmHg;LVW/BW:2 weeks (2.21±0.17) vs (1.91±0.12), 4 weeks (2.45±0.16) vs (2.01±0.14), 8 weeks (2.68±0.15) vs ( 2.05 ±0.09 ) mmHg;( P<0.05 ) ] .The cardiomyocytes in AAB group were hypertrophic as compared to the sham group.The expression of GRP94 in AAB group increased significantly at 2 weeks post-operation, and reached peak level at 4 weeks after the surgery and was on the decline thereafter .The expression of CHOP and GRP94 in AAB rats were significantly higher than sham group, and reached the peak at the 8 weeks after surger-y.2)Treatment with amlodipine significantly reduced MAP and LVM/BW in AAB rats[(MAP:2 weeks (126± 6) vs (144±10), 4 weeks (125±8) vs (163±8), 8 weeks (128±5) vs (177±10)mmHg;LVM/BW:2 weeks ( 1.94 ±0.15 ) vs ( 2.21 ±0.17 ) , 4 weeks ( 2.13 ±0.08 ) vs ( 2.45 ±0.16 ) , 8 weeks ( 2.18 ±0.10 ) vs ( 2.68 ± 0.15)mg/g;(P<0.05)].The myocardial hypertrophic was alleviated in AAB+Aml group.The level of GRP94 and CHOP in AAB+Aml group was lower than those in AAB rats ( all P<0.05 ) .Conclusions Endoplasmic reticulum stress is involved in the ventricular remodeling caused by hypertension , and use of amlodipine can reduce ventricu-lar hypertrophy in hypertension models possibly by inhibiting endoplasmic reticulum stress via down -regulation of GRP94 and CHOP .
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Objective:To study the expression levels of tumor-associated glycoprotein (TAG72) and glucose-regulated protein 94 (GRP94) in the breast benign and malignant lesions and detect their clinicopathological significance. Methods:EnVisionTM immunohistochemistry was used for assaying expressive levels of TAG72 and GRP94 in conventional paraffin-embedded sections from specimens of breast cancer (n=60) and benign lesions (n=30). The survival analysis was performed by using Kaplan-Meier method.Results:The positive rates and scores of TAG72 and GRP94 were significantly higher in breast cancer than those in benign lesions (TAG72: 68.3% vs 10.0%, 2.53±1.86 vs 0.37±1.13; GRP94: 63.3% vs 13.3%, 2.73±2.23 vs 0.47±1.22; P=0.000). The positive rates and scores of TAG72 and GRP94 were significantly lower in the intra-ductal,histological grade Ⅰ or Ⅱ , with maximal diameter ≤3 cm, without lymph node metastasis, estrogen receptor (ER)-positive, p53 and CA15-3-negative breast cancer than those in the ductal invasive (P3 cm (P<0.05), with lymph node metastasis (P<0.01), ER-negative (P<0.05) , p53-positive (P<0.01), and CA15-3-positive (P<0.01) breast cancer tissues. Kaplan-Meier survival analysis showed that the median survival time were significantly shorter in the TAG72-positive, GRP94-positive or TAG72 and GRP94-positive breast cancer patients than those with negative TAG72 and/or GRP94 expression (P<0.01). Multivariate COX regression analysis showed that TAG72 and/or GRP94-positive expression were negatively associated with postoperative survival rate and positively related with death rate. Both of them were independent prognostic factors for breast cancer. Conclusion:The expressive levels of TAG72 and/or GRP94 might have important effects on the carcinogenesis, progression, biologic behaviors and prognosis of breast cancer. The breast cancer patients with over-expression of TAG72 and/or GRP94 might have poor prognosis.
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Objective To explore the expression of sterol regulatory element binding protein 1C (SREBP-1C) and glucose-regulated protein 94(GRP-94)in hyperhomocysteinmia and to evaluate the effects of endoplasmic reticulum stress proteins on hepatocytes lipids metabolism. Methods After hyperhomocysteinmia C57BL/6 mice model being induced by high methionine diet, TGE and CHO of Hepatocytes were determined, and the expression of SREBP-1C and GRP-94 was assessed by RT-PCR and Western blot. All data were compared to those in control group′s. Results The level of plasmic homocysteine(Hcy) and hepatocytes TGE or CHO of high methionine diet mice at different time point significantly ascended(P