Comparative study on pathological characteristics of four different antigen-induced rheumatoid arthritis mouse models / 药学学报

Rheumatoid arthritis (RA) is an autoimmune disease driven by antigens and mediated by T cells. Collagen II (CII) and fibrinogen (Fib) are the two main antigens in the pathogenesis of RA. The antigen produced after citrulline modification (Cit) is also one of the inducements to induce the body to produce a pathogenic anti-citrulline protein antibody (ACPA). To provide a reference for RA-related research, this study intends to establish an RA animal model by using CII, Cit-CII, Fib, and Cit-Fib antigens, emulsification with complete Freund's adjuvant and immunization with DBA/1 mice, respectively, to compare the pathological characteristics of RA models induced by different antigens from the aspects of pathology, imaging and serum biochemistry. Animal welfare and experimental process are in accordance with the regulations of the Experimental Animal Ethics Committee of the China Academy of Chinese Medical Sciences. The results showed that the CII, Cit-CII, and Cit-Fib induced mice all had symptoms such as joint redness and swelling, and toe deformation and the clinical score and incidence rate were higher than those of the normal group. The CII group had the most serious lesions, with a incidence rate of 100%, and the Cit-CII and Cit-Fib groups had mild symptoms, with a incidence rate of 25% and 37.5%, respectively; pathological and imaging examination results showed that the joints of mice in CII-induced group showed severe synovial inflammation, cartilage and bone destruction, while those in Cit-CII and Cit-Fib group showed only slight inflammatory infiltration, joint cavity stenosis and bone destruction; the results of serum antibody detection showed that CII, Cit-CII and Cit-Fib groups all produced high levels of anti-cyclic citrullinated peptide (CCP) antibodies, among which, Cit-Fib group > Cit-CII group > CII group > Fib group, and both Cit-CII and Cit-Fib groups produced high levels of citrullinated epitope-specific antibodies, while the total IgG level was the highest in CII group; serum ELISA and RT-PCR analysis of joint tissue showed that the expression of pro-inflammatory factors and bone destruction-related molecules increased most significantly in the CII-induced group, followed by Cit-Fib and Cit-CII. The above results showed that among the four different antigens, the symptoms and conditions of arthritis in RA mice induced by CII were the most serious, and IgG instead of anti-CCP antibody was its typical immunological feature, and CII could be the first choice for the model of RA mice; Cit-Fib has certain immunogenicity, can partially induce the symptoms and conditions of RA arthritis in mice, and produce high-level anti-CCP antibody and anti-Cit-Fib antibody, which is more suitable for the study of citrulline-related RA; although Cit-CII has certain immunogenicity, the incidence, and severity of RA arthritis induced by Cit-CII in mice are low.

Effect of Pudilan Xiaoyan Oral Liquid on Acute Lung Injury Rat Induced by Lipopolysaccharide / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of Pudilan Xiaoyan oral liquid(PDL) on the acute lung injury rat induced by lipopolysaccharide (LPS). Method:The 72 Wistar rats were randomly divided into control group, model group, dexamethasone group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group according to body weight.The acute lung injury model was made through inhalation with lipopolysaccharide in the model group, hexadecadrol group, PDL 7, 3.5, 1.75 g·kg-1·d-1 group.To examining each rat alveolar lavage fluid (BALF) of the total number of white blood cells, enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of nuclear transcription factors-kappa B (NF-κB) and interleukin-10 (IL-10).Hematoxylin-eosin(HE) staining was used to observe morphological changes of lung tissue and explore different doses of PDL effect on acute lung injury in rats. Result:Compared with model group, the account of leukocyte in BALF decreased significantly in PDL 7 g·kg-1·d-1 group and PDL 3.5 g·kg-1·d-1 group (PκB significantly decreased in PDL 7, 3.5, 1.75 g·kg-1·d-1 group (P-1·d-1 group (P-1·d-1 group, the inflammation, edema and congestion in lung tissue reduced (PConclusion:PDL has a significant protective effect on the inflammation of acute lung injury model, and its mechanism is related to the expressions of NF-κB and IL-10. PDL could also repair the injury of lung in acute lung injury model.

Pharmacodynamic Evaluation of Tanreqing Inhalation Solution / 中国实验方剂学杂志

Objective: To clarify the antitussive, expectorant, antipyretic and anti-inflammatory effects of Tanreqing inhalation solution, and provide basis and data support for further research and development of this preparation. Method: The methods of cough induced by ammonia and tracheal phenol red excretion were used to observe the antitussive and expectorant effects of Tanreqing inhalation solution in mice. The fever model of rats was established by intraperitoneal injection of bacterial lipopolysaccharide(LPS) to observe the antipyretic effect of the Tanreqing inhalation solution, the acute pneumonia model of rats was established by atomizing LPS inhalation, and the anti-inflammatory effect of Tanreqing inhalation solution was observed. Result: Tanreqing inhalation solution could reduce the number of coughs in mice induced by ammonia water, increase the amount of phenol red excretion in mouse trachea, decrease the levels of body temperature and its related regulatory factors of prostaglandin E2(PGE2) and cyclic adenosine monophosphate(cAMP) of rats induced by LPS, decrease the white blood cell(WBC) count and the neutrophil ratio(NEUT) in bronchoalveolar lavage fluid(BALF) of rats with LPS-induced acute pneumonia, and reduce the levels of nuclear transcription factor-κB(NF-κB) and interleukin-1β(IL-1β) in lung tissue. Conclusion: Tanreqing inhalation solution has obvious antitussive, expectorant, antipyretic and anti-inflammatory effects, which is worthy of further development and promotion.

Effect of Different Administration Methods with Bleomycin on Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:Compare the effects of 3 administration methods (tracheal perfusion, tail vein injection and aerosol inhalation) with bleomycin (BLM) in inducing pulmonary fibrosis in rats, in order to find out the optimal administration methods. Method:Eighty sprague-dawley (SD) male rats with SPF were randomly divided into aerosol inhalation blank group, single tracheal perfusion group(10 mg·kg-1), multiple tracheal perfusion group(5 mg·kg-1), single intravenous injection group(150 mg·kg-1), multiple intravenous injection group(50 mg·kg-1), single aerosol inhalation group (30 min)and multiple aerosol inhalation group(30 min). The mortality and body weight of rats in each group were observed at 7 d, 14 d and 28 d after the administration. And 28 days later after the administration, the lung coefficients of rats in each group were observed, paraffin sections were prepared, hematoxylin-eosin staining (HE) and Masson staining were performed, and the contents of hydroxyproline (HYP) and plasminogen activator inhibitor-1 (PAI-1) in lung tissues were detected by enzyme-linked immunosorbent assay (ELISA), so as to evaluate the alveoli inflammation and pulmonary fibrosis of rats in each group. Result:Compared with the aerosol inhalation blank group, the rats in the trachea perfusion group had the highest mortality among the drug treatment groups. The pulmonary coefficients of rats in the multiple intravenous injection group and the multiple inhalation group were significantly higher than those in the blank group(PPPConclusion:Bleomycin was inhaled repeatedly to establish pulmonary fibrosis model. The pathological injury and physiological indexes of the model rats were relatively stable, which conforms with the evolution process of pulmonary fibrosis.

Therapeutic effect of Qingkailing and methylprednisolone injection alone or combined on the acute lung injury induced by oleic acid in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of Qingkailing and Methylprednisolone (MP) injection alone or combined on the acute lung injury (ALI) induced by oleic acid in rabbits.</p><p><b>METHOD</b>The rabbits were randomly divided into 11 groups: oleic acid group; control group; treatment groups including low, middle and high dosage groups of Qingkailing and MP alone and combined, respectively. ALI model was established by i.v. oleic acid (0.05 mL x kg(-1)) in these groups, and then i.v. above drugs respectively, while in control group, the same volume of normal saline was given. The respiratory amplitude and rate were observed, and blood samples were taken from cervical artery for blood-gas analysis before and at 30, 60, 120 min after oleic acid or normal saline administration. At the end of experiment, the concentration of LDH, CAT and MDA in the lung tissue were measured and pathologic changes of lung tissue were observed microscopically.</p><p><b>RESULT</b>Compared with oleic acid group, the respiratory amplitude markedly enhanced (P < 0.05) and respiratory rate lowered (P < 0.05) in the low, middle and high dose groups of Qingkailing and MP injection. On the 30 min of treatment, PaO2 increased significantly (P < 0.05) in the low and middle dose groups of combined Qingkailing and MP injection; PaCO2 decreased markedly (P < 0.05) on the 120 min of treatment in each treatment group. The level of LDH significantly increased (P < 0.05), CAT and MDA decreased (P < 0.05) in the middle and high groups of Qingkailing and MP injection. The low and middle dose groups of combined Qingkailing and MP injection can alleviate the pathological changes induced by oleic acid.</p><p><b>CONCLUSION</b>The curative effect of the low dose group of combined Qingkailing and MP for the ALI induced by oleic acid was better than Qingkailing and MP alone, while the big dose groups of Qingkailing and MP alone better than the combination at the same dosage.</p>

Therapeutic effect of qingkailing and shengmai injection alone or combined on the acute lung injury induced by oleic acid in rabbits / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of Qingikailing and Shengmai injection alone or combined on the acute lung injury (AL) induced by oleic acid in rabbits.</p><p><b>METHOD</b>The rabbits were randomly divided into 11 groups: oleic acid group; control group; treatment groups including low, middle and high dosage groups of Qingkailing and Shengmai injection alone and combined, respectively. ALI model was established by iv oleic acid (0.05 mL x kg(-1)) in these groups, and then iv above drugs respectively,while in control group, the same volume of normal saline was given. The respiratory amplitude and rate were observed, and blood samples were taken from cervical artery for blood-gas analysis before and at 30, 60, 120 min after oleic acid or normal saline administration. At the end of experiment, the concentration of LDH, CAT and MDA in the lung tissue were measured and pathologic changes of lung tissue were observed microscopically.</p><p><b>RESULT</b>Compared with oleic acid group, the respiratory amplitude markedly enhanced (P < 0.05) in the low and high dose groups of Qingkailing and Shengmai injection. PaO2 increased significantly (P < 0.05) in the low dose group of combined Qingkailing and Shengmai injection, PaCO2 decreased markedly (P < 0.05) in the low dose groups of Qingkailing and Shengmai injection alone and combined. The level of MDA significantly decreased (P < 0.05) in the each group of Qingkailing and Shengmai injection alone, the level of MDA significantly decreased (P < 0.05) and CAT increased (P < 0.05) in the low dose group of combined Qingkailing with Shengmai injection. The low dose group of combined Qingkailing and Shengmai injection can alleviate the pathological changes induced by oleic acid.</p><p><b>CONCLUSION</b>The curative effect of the low dose group of combined Qingkailing with Shengmai injection for the ALI induced by oleic acid was better than Qingkailing and Shengmai injection alone at the same dosage.</p>

Experimental study of chronic renal tubular-interstitial injury induced by radix aristolochiae fangchi extract in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the acute and chronic renal toxicity induced by Radix Aristolochiae Fangchi Extract (RAFE) in different doses in rats.</p><p><b>METHOD</b>The conventional method of acute toxicity was used. RAFE at the dose of 25.0 mg x kg(-1) x d(-1), 120.0 mg x kg(-1) x d(-1) and 200.0 mg x kg(-1) x d(-1) and aristolochic acid (AA, 10.0 mg x kg(-1) x d(-1)) were interruptedly administrated to rats for 13 week by gastric tube, and the sample of blood, urine and kidney were collected at 4 week, 8 week and 13 week respectively. The indexes of renal function were measured and the morphology of kidney was observed.</p><p><b>RESULT</b>LD50 of RAFE was 36.8 g x kg(-1) (the crude drug) and the 95% confidence limit was 38.8 - 28.9 g x kg(-1). The changes of renal functions were azotemia, massive proteinuria and the increase of urinary NAGase (beta-N-acetylglucosaminidase) in the earlier period of administration with RAFE in rats. Pathological changes of renal tissue were as follows: acute renal tubular necrosis mainly in the boundary of cortex and medulla was observed in the earlier period, and with the elongation of administration, the pathological process of renal interstitial fibrosis observed in the middle and high groups of RAFE and AA group.</p><p><b>CONCLUSION</b>RAFE at middle and high doses administrated by interrupted gavage above 13 week can cause the injury of renal tubular functions in rats. NAGase can be used as one of observation targets in the earlier period of renal injury.</p>

Experimental study of chronic renal tubular-interstitial injury induced by Radix Aristolochiae Fangchi Extract in rats / 中国中药杂志

<p><b>OBJECTIVE</b>Following the former report, we continue to observe the chronic renal tubular-interstitial injury induced by Radix Aristolochiae Fangchi Extract(RAFE) in rats in order to understand whether RAFE in different doses causes the renal tubular-interstitial injury or not.</p><p><b>METHOD</b>RAFE at the dose of 25.0 mg x kg(-1) x d(-1), 120.0 mg kg(-1) x d(-1) and 200.0 mg x kg(-1) x d(-1) and aristolochic acid (AA, 10.0 mg x kg(-1) d(-1)) was interruptedly administrated by gastric tube for 22 w and 4 w durg withdrawal. Blood, urine and kidney were taken out respectively in 17 w, 22 w and 26 w to measure the indexes of renal function. The morphology of kidney was observed, and Masson staining of kidney were made respectively to compare RAFE groups with AA group.</p><p><b>RESULT</b>Pathological changes of renal tissue forms were as follows: All RAFE groups and AA group could develop the pathological process of renal tubular injury-chronic renal interstitial fibrosis. The pathologic changes of RAFE were similar with AA.</p><p><b>CONCLUSION</b>RAFE at all doses administrated interruptedly by gastric tube above 13 w caused chronic renal tubulo-interstitium fibrosis. The renal injury in functions and tissue forms in rats were similar with AA closely. The results showed that AA was the main toxic composition of RAFE.</p>