1.Cosmc and T-synthase mRNA expression in peripheral B lymphocytes and serum levels of galactose-deficient IgA1 in patients with Henoch-Sch?nlein purpura
Shaomin SHI ; Yun YANG ; Ding LUO ; Jianli HUO ; Dahu WANG ; Yaohui MA ; Haixia MA ; Cuimin REN ; Bingshen GUO ; Xin LI ; Xingnan WANG ; Qiang LIU
Chinese Journal of Dermatology 2020;53(4):285-289
Objective:To investigate changes in the expression of Cosmc and T-synthase in peripheral B lymphocytes and in serum levels of galactose-deficient IgA1 (Gd-IgA1) in patients with Henoch-Sch?nlein purpura (HSP) .Methods:From January to August 2014, 56 patients with HSP were collected from outpatient or inpatient department of dermatology and venereology in the Second Hospital of Hebei Medical University, and were divided into 4 groups, including skin type group (22 cases) , joint type group (9 cases) , abdominal type group (12 cases) and renal type group (13 cases) . Twenty healthy volunteers served as healthy controls. Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of Cosmc and T-synthase in peripheral B lymphocytes, and a lectin-based enzyme-linked immunosorbent assay (ELISA) to detect the serum level of Gd-IgA1. Comparisons among multiple groups were performed using one-way analysis of variance or Kruskal-Wallis H test, multiple comparisons were performed using least significant difference (LSD) - t test or Nemenyi test, and correlation analysis was performed using Spearman rank correlation analysis. Results:There was a significant difference in the duration from disease onset to the clinic visit ( χ2= 26.19, P < 0.05) among the skin type group (6.27 ± 3.09 d) , joint type group (5.56 ± 3.05 d) , abdominal type group (6.75 ± 3.75 d) , and renal type group (26.23 ± 14.12 d) , and the duration from disease onset to the clinic visit was significantly longer in the renal type group than in the other 3 groups (all P < 0.05) . The Cosmc mRNA expression significantly differed among the skin type group, joint type group, abdominal type group, renal type group and healthy control group (0.849 ± 0.239, 0.767 ± 0.181, 0.719 ± 0.183, 0.459 ± 0.121, 1.146 ± 0.232, F= 23.37, P < 0.05) , was significantly lower in the 4 patient groups than in the healthy control group ( P < 0.01) , and lower in the renal type group than in the other 3 patient groups (all P < 0.01) . There was no significant difference in the T-synthase mRNA expression in peripheral B lymphocytes among the patient groups and healthy control group ( F= 1.05, P > 0.05) . The serum level of Gd-IgA1 significantly differed among the skin type group, joint type group, abdominal type group, renal type group and healthy control group ( F= 7.06, P < 0.05) . Moreover, the Gd-IgA1 level was significantly higher in the patient groups than in the healthy control group (all P < 0.05) , and higher in the renal type group than in the other 3 patient groups (all P < 0.05) . The serum level of Gd-IgA1 in the HSP patients was significantly and negatively correlated with the mRNA expression of Cosmc ( rs=-0.50, P < 0.01) . Conclusion:Decreased mRNA expression of Cosmc and increased serum levels of Gd-IgA1 were observed in patients with HSP, and there was a negative correlation between the two indices.
2.Earthworm Protein Protects Vascular Endothelial Function in Spontaneously Hypertensive Rats via PI3K/Akt/Nrf2 Signaling Pathway
Huilin ZHAO ; Xiaojing LI ; Lirong WANG ; Yaohui HUO ; Yun LI ; Tianlong DING ; Kai LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):118-126
ObjectiveTo observe the effect of earthworm protein on the expression of phosphatidylinositol 3-kinase/protein kinase B/nuclear factor E2-related factor 2 (PI3K/Akt/Nrf2) pathway in the aorta of spontaneously hypertensive rats (SHR) and explore mechanism of earthworm protein in treating hypertensive vascular endothelial dysfunction (VED). MethodTen 10-week-old Wistar Kyoto (WKY) rats and fifty SHR rats were selected for a week of adaptive feeding. WKY rats were selected as the normal group, and fifty SHR rats were randomized according to body weight into model, valsartan (8×10-3 g·kg-1·d-1), and high-, medium-, and low-dose (0.2, 0.1, 0.05 g·kg-1·d-1, respectively) earthworm protein groups. The normal and model groups were administrated with equal volume of double distilled water by gavage. During the drug intervention period, the general situations of rats in each group were observed and their blood pressure was monitored at specific time points every other week before and after administration. After 8 weeks of drug intervention, enzyme-linked immunosorbent assay was employed to measure the levels of angiotensin-Ⅱ (Ang-Ⅱ) and endothelin-1 (ET-1) in the serum of rats in each group. The corresponding kits were used to determine the levels of nitric oxide (NO), malondialdehyde (MDA), glutathione peroxidase (GPX), superoxide dismutase (SOD), and ferrous ion (Fe2+). Hematoxylin-eosin (HE) staining was employed to observe the changes in the intima of the aorta. Fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to measure the mRNA levels of PI3K, Akt, Nrf2, heme oxygenase-1 (HO-1), and glutathione peroxidase 4 (GPX4) in the aortic tissue. Western blotting was used to determine the protein levels of p-PI3K (Tyr467/199), PI3K, p-Akt (Ser473), Akt, Nrf2, HO-1, and GPX4 in the thoracic aorta. ResultCompared with the normal group, the model group had decreased body mass, increased irritability, severe endothelial damage, elevated blood pressure and serum levels of Ang-Ⅱ, ET1, MDA, and Fe2+ (P<0.01), lowered NO level (P<0.01), and down-regulated mRNA and protein levels of p-PI3K (Tyr467/199), PI3K, p-Akt (Ser473), Akt, Nrf2, HO-1, and GPX4 in the aortic tissue (P<0.01). Compared with the model group, drug intervention caused no significant change in the body mass, calmed the rats, alleviated the endothelial damage, lowered blood pressure and serum levels of Ang-Ⅱ, ET1, MDA, and Fe2+ (P<0.01), elevated the NO level (P<0.05), and up-regulated the mRNA and protein levels of p-PI3K (Tyr467/199), PI3K, p-Akt (Ser473), Akt, Nrf2, HO-1, and GPX4 (P<0.05). ConclusionThe earthworm protein can exert antihypertensive effects by ameliorating VED in SHR. Specifically, it may regulate the PI3K/Akt/Nrf2 signaling pathway to inhibit oxidative stress and ferroptosis.