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ObjectiveTo investigate the effects of Biling Qutong prescription （BLQT） on serum levels of NOD-like receptor thermal protein domain associated protein 3 （NLRP3）， purinergic ligand-gated ion channel 7 receptor （P2X7R）， fibronectin （FN）， and hepatic steatosis in patients with type 2 diabetes mellitus （T2DM） complicated with gouty arthritis （GA）. MethodSixty-four patients diagnosed with T2DM comorbid with GA and treated at the First Affiliated Hospital of Anhui University of Chinese Medicine from January 2019 to December 2022 were enrolled and randomly divided into a BLQT group （Chinese medicine group， 32 cases） and the ibuprofen group （western medicine group， 32 cases）. Thirty healthy individuals who underwent routine health examinations during the same period were assigned to the control group. The BLQT group and the western medicine group received basic treatment along with BLQT and ibuprofen， respectively. After 8 weeks of continuous treatment， the traditional Chinese medicine syndrome score （TCMSS） of the patients was evaluated before and after treatment. The differences in fasting plasma glucose （FPG）， 2-hour postprandial plasma glucose （2 h PG）， glycated hemoglobin （HbA1c）， fasting insulin （FINS）， homeostasis model assessment of insulin resistance （HOMA-IR）， serum uric acid （SUA）， serum creatinine （SCr）， total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， controlled attenuation parameter （CAP）， liver stiffness measurement （LSM）， NLRP3， P2X7R， and FN levels before and after treatment were compared. Adverse drug reactions that occurred during treatment were recorded. ResultThe TCMSS for joint redness， swelling， pain， joint burning， yellow urine， and red tongue with yellow and greasy coating， as well as total score were significantly reduced in both the BLQT group and the western medicine group as compared with those before treatment （P<0.05， P<0.01）. The BLQT group also showed a significant reduction in symptom scores such as dry mouth， polyuria， polydipsia， and slippery and rapid pulse （P<0.01）. Compared with the western medicine group after treatment， the BLQT group exhibited a more significant reduction in all symptom scores and total score （P<0.05， P<0.01）. The BLQT group and the western medicine group showed a decrease in FPG， 2 h PG， HbA1c， SCr， SUA， TG， TC， and LDL-C levels （P<0.05， P<0.01） after treatment， and the BLQT group showed decreased HOMA-IR， ALT， AST， and HDL-C levels （P<0.05， P<0.01） compared with those before treatment. When compared with the western medicine group after treatment， the BLQT group showed a more significant reduction in all laboratory parameters except for HDL-C （P<0.05， P<0.01）. Before treatment， NLRP3， P2X7R， and FN levels in both the BLQT group and the western medicine group were higher than those in the control group （P<0.01）. After treatment， NLRP3 and P2X7R levels in both groups significantly decreased （P<0.01）， and FN levels in the BLQT group also decreased significantly （P<0.01）. When compared with the western medicine group after treatment， the BLQT group showed a more significant reduction in NLRP3， P2X7R， and FN levels （P<0.01）. Before treatment， CAP and LSM levels in both the BLQT group and the western medicine group were higher than those in the control group （P<0.01）. After treatment， CAP and LSM levels in both groups decreased （P<0.05， P<0.01）. Compared with the western medicine group after treatment， the BLQT group showed a more significant reduction in CAP and LSM （P<0.01）. The incidence of adverse reactions was 3.13% （1/32） in the BLQT group and 15.63% （5/32） in the western medicine group， with no significant difference. ConclusionBLQT has good efficacy in patients with T2DM complicated with GA， which can significantly alleviate joint redness， swelling， heat， pain， limited mobility， dry mouth， and polydipsia， reduce blood glucose， uric acid， and lipid levels， suppress the high expression of NLRP3， P2X7R， and FN， and improve hepatic steatosis.
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Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.
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Objective To investigate the effect of mixed-skin grafting with autologous microskin and allogenetic acellular dermal matrix microskin on wound healing in rats,and to make a further study on the related mechanism.Methods Wistar rats were served as a allogenetic acellular dermal matrix donor rats,and SD rats as acceptors with mould of full thickness skin defects on their back.The ninety SD rats were divided into 5 groups with 18 rats in each group.Group 1 was transplanted with autologous microskin,and group 2 with allogenetic acellular dermal matrix microskin.Groups 3,4 and 5 were grafted with mixed-skin ratio between autologous microskin and allogenetic acellular dermal matrix microskin 1 ∶ 1,1 ∶ 0.5 and 1 ∶ 0.25,repectively.The rate of wound healing was measured,wound samples collected,hematoxylin and eosin stain carried out,fibronectin (FN) and laminin (LN)detected,and intergroup comparison made,respectively,2,3 and 4 weeks after skin grafting.Results The wound healing rates and FN and LN expression of mixed-skin grafting groups were higher than those of the group with autologous microskin grafting.The group of 1 ∶ 0.25 obviously increased (P＜0.05 or P＜0.01).Conclusions The wound healing rate with mixed-skin grafting is higher than that with autologous microskin grafting.The best effect is achieved when the skin ratio between autologous microskin and allogenetic acellular dermal matrix microskin is 1 ∶ 0.25.It is possibly due to the increase of FN and LN on wound skin surface.
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Objective To investigate the changes of actin (HHF35), serum albumin (A1), fibronectin (Fn) in cardiac cells and cardiac conducting sysytem (CCS) in order to probe into postmortem diagnosis of sudden manhood death syndrome (SMDS). Methods Immunohistochemical detections of actin, serum albumin and fibronectin (Fn) in the cardiac cells and CCS were performed by S-P method in cases with SMDS and cases died from coronary heart disease and non-cardiovascular diseases which were used as the control. Results Of the 22 cases with SMDS, 17 were found of actin-negative staining, 18 serum albumin-positive and 15 Fn-positive staining with evident pathological lesions in CCS in 8 cases. Conclusion It is suggested that SMDS is associated with early myocardial ischemia or infarction, which results in abnormality in actin, serum albumin and fibronectin (Fn) in the cardiac cells and CCS.
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Objective To explore the molecular and cellular mechanism of Zaohuang Capsule No.3 (ZHC3) in treating chronic renal failure through observing the effect of ZHC3 on the human glomerular mesangial cells (GMC). Methods The proliferation of human GMC fostered in ZHC3 was detected by MTT, and the fibronectin (FN), interleukin 6 (IL-6) and transforming growth factor ?1 (TGF-?1) secretion was detected by ELISA. Results ZHC3 significantly inhibited GMC proliferation (P
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Objective: To investigate effects of Wuling Pills and Capsules on immunological liver injury in mice. Methods: The rat liver fibrosis model was induced by carbon tetrachloride. The hydroxyproline (HYP) level in serum was measured by chemical method. The precollagen Ⅰ(PCⅠ) and Ⅲ(PCⅢ) levels in serum and hepatic homogenate were measured by radioimmunoassay. The expressions of tumor necrosis factor (TNF) and fibronectin (FN) in liver tissue were detected by histochemistry method. Moreover, the hepatic histomrphology was observed by HE staining. Results: Both of Wuling Pills and Wuling Capsules could greatly decrease PCⅠ, PCⅢ and HYP levels. The FN expression of liver tissue in Wuling Capsule group was obviously lower than that of the model group. The TNF and FN in liver tissues of rats in the Wuling Pill group and Capsule group showed strong positive. Proliferative kupffer cells and activated endothelial cells were also observed in those two groups. Conclusion: Wuling Capsules has the effect of anti liver fibrosis. Both of Wuling Pills and Capsules can stabilize hepatocyte membrane, and prevent liver cells from the effect of TNF and FN.
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Objective To study the relationship between the expression of FN in heart, lung and kidney and the mechanism of multiple organ dysfunction syndrome (MODS). Method The Fn in heart, lung and kidney in 20 autopsy cases of MODS were studied immunohistochemically by using anti-FN as the first antibodies. Results The expression of FN in the MODS group were significantly decreased as compared with the control group (P
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In order to explore the value of fibronectin (Fn) in the postmortem diagnosis of myocardial infarction.The changes of Fn staining in normal, infarcted and other non infarcted myocardial injuries resulted from myocarditis, mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion and organophosphate poisoning were studied with immunohistochemistry and image analysis system. The results showed that positive Fn staining could only be observed in groups of myocardial infarction and myocarditis, but could not be found in groups of mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion, organophosphate poisoning. It is indicated that positive reaction of Fn could be affected only by myocarditis, so it is quite specific for the diagnosis of myocardial infarction.
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The experiment was designed to compare the changes of cytoskeleton (including tubulin and actin) and cell surface fibronectin (FN) between NIH 3T3 cell line and transformed NIH 3T3 cell line by genome DNA of human lung adenocarcinoma cell line AGZY. We stained and observed these cells using immunohistochemical methods with antibody against bovine brain tubulin, phalloidin and self-made affinity column purified antibody against porcine plasma FN.Our results showed that the bundles of actin and tubulin are damaged seriously, demonofrating an unclear cytoskeleton structure and diffused fluorescence over the cells when they were transformed. The amount of membrane FN on transformed cell surface decreases significantly which is only 1/9 of NIH 3T3. The FN distribution altered markedly from thin threadlike network to spots and speckles.These results suggested that cell transformation was a complex event including the changes of cytoskeleton system and cell surface glycoproteins. In addition, it might also indicate that cause and effect relationship existed between these changes and the alteration of cell phenotype and loss of growth control.