Background/Aims It is known that chronic low-grade inflammation contributes to the initiation and development of both diabetes and diabetic nephropathy (DN), so we designed this study to investigate the role of P2X7R and NLRP3 inflammasome in DN pathogenesis and the antagonistic effects of artificially cultivated Ophiocordyceps sinensis (ACOS). Methods A rat model of DN caused by high-fat-diet feeding and low-dose streptozotocin injection and a mouse podocyte injury model induced by high-glucose (HG) stimulation were established, and the intervention effects of ACOS on them were observed. The biological parameters of serum and urine and the pathological manifestations of kidney tissue were examined. The expression of mRNA and protein of P2X7R and NLRP3 inflammasome (NLRP3, ASC, and caspase-1) and downstream effectors (IL-1β and IL-18), as well as podocyte-associated molecules, was determined by real-time quantitative PCR and Western blot assay, respectively. Results The DN rats showed to have developed insulin resistance, elevated fasting blood glucose, increased urinary protein excretion, and serum creatinine level as well as corresponding glomerular pathological alterations including podocyte damages. ACOS significantly antagonized the above changes. The experiments in vivo and in vitro both displayed that the mRNA and protein expression of P2X7R, NLRP3, ASC, caspase1 (procaspase-1 mRNA in the gene level and active caspase-1 subunit P10 in the protein level), IL-1β, and IL-18 was significantly upregulated and the mRNA and protein expression of podocyte-associated molecules was significantly changed (downregulation of nephrin, podocin, and WT-1 expression and upregulation of desmin expression) indicating podocyte injury in the kidney tissue of DN rats and in the HG-stressed mouse podocytes, respectively. ACOS also significantly antagonized all the above changes. Conclusion Our research work suggests that P2X7R and NLRP3 inflammasome are involved in the pathogenesis of DN, and ACOS can effectively inhibit the high expression of P2X7R and the activation of NLRP3 inflammasome, which may contribute to the therapeutic effects of Ophiocordyceps sinensis.

Background:The nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome composed of NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), and caspase-1 is engaged in the inflammatory response of many kidney diseases and can be activated by purinergic 2X7 receptor (P2X7R). This study was conducted to explore whether P2X7R plays a pathogenic role in the podocyte damage of obesity-related glomerulopathy (ORG) and whether this role is mediated by the activation of NLRP3 inflammasome.Methods:A mouse model of ORG was established by high-fat diet feeding. The conditionally immortalized mouse podocytes were cultured with leptin or with leptin and P2X7R antagonist (KN-62 or A438079). The mRNA and protein expression of the P2X7R and NLRP3 inflammasome components including NLRP3, ASC, and caspase-1, as well as the podocyte-associated molecules including nephrin, podocin, and desmin in mouse renal cortex or cultured mouse podocytes were tested by real-time-polymerase chain reaction and Western blot analysis, respectively.Results:The significantly upregulated expression of P2X7R and NLRP3 inflammasome components and the NLRP3 inflammasome activation were observed in the renal cortex (in fact their location in podocytes was proved by confocal microscopy) of ORG mice in vivo, which were accompanied with the morphological changes of podocyte damage and the expression changes of podocyte-associated molecules. Similar changes in the expression of P2X7R and NLRP3 inflammasome components as well as in the expression of podocyte-associated molecules were also observed in the cultured podocyte studies treated by leptin in vitro, and all of the above changes were significantly attenuated by the P2X7R antagonist KN-62 or A438079.Conclusions:P2X7R could trigger the activation of NLRP3 inflammasome, and the activated P2X7R/NLRP3 inflammasome in podocytes might be involved in the podocyte damage of ORG.

Background Intermediate coronary lesion (ICL) is a critical stage affecting coronary artery disease’s progression and prognosis and may quickly progress to acute coronary syndrome. However, percutaneous coronary intervention therapy and conventional drug therapy have certain limitations. Qing-Xin-Jie-Yu Granules (QXJYG), a Chinese herbal prescription with a pre-research basis, has the potential to be a complementary treatment for ICL. Previous studies have shown that QXJYG combined with conventional drug therapy could alleviate angina symptoms and reduce the incidence of composite ‘hard’ endpoint in treating stable coronary artery disease. However, for ICL, the effect of this prescription on the degree of coronary stenosis, plaque stability, and long-term efficacy remains unclear. Therefore, we designed this study to evaluate the efficacy and safety of QXJYG in patients with ICL. Methods This is a multi-center, block-randomized, double-blinded, placebo-controlled trial. One hundred and twenty participants with ICL will be randomly assigned to two groups in a 1:1 ratio. Based on conventional intervention, the participants of the treatment group will receive QXJYG orally, and the participants of the control group will receive placebo for six successive months. The primary outcomes involve the degree of coronary stenosis, including the percentage of diameter stenosis and the percentage of area stenosis measured by coronary CT angiography. The secondary outcomes involve coronary artery calcification score, Gensini score, CT-fractional flow reserve, angina symptom score, traditional Chinese medicine syndrome score, blood lipids, inflammatory factors, carotid artery ultrasound parameters, and major adverse cardiovascular events. Safety will be assessed by adverse events and laboratory examinations. Measurements will be performed at baseline (visit 1), months 2 (visit 2), months 4 (visit 3), months 6 (visit 4), and months 12 (visit 5). Discussion The results of this trial will verify the efficacy and safety of QXJYG in treating ICL, thus adding to the existing knowledge for resolving the problem in ICL. Trial registration: This trial is registered at the Chinese Clinical Trial Registry (www.chictr.org.cn). Registration number: ChiCTR2200059262. Register date: April 27, 2022.