Effects of neferine on cytosolic free calcium concentration in corpus cavernosum smooth muscle cells of rabbits

Abstract

To study the relaxation mechanisms of neferine (Nef) on the corpus cavernosum smooth muscle (CCSM), the CCSM cells from New Zealand White rabbits were cultured in vitro. [Ca(2+)](i) was measured by fluorescence ion digital imaging system (FIDIS), using Fluo-2/AM as a Ca(2+)-sensitive fluorescent indicator. Nef (0.1, 1 and 10 micromol l(-1)) had no effect on the resting [Ca(2+)](i) (P > 0.05). In the presence of extracellular Ca(2+) (2.5 mmol l(-1)), Nef (0.1, 1 and 10 micromol l(-1)) inhibited [Ca(2+)](i) elevation induced by high K(+) and phenylephrine (PE) in a concentration-dependent manner (P < 0.05). In calcium free solution containing egtaic acid (EGTA), Nef (0.1 micromol l(-1)) had no inhibitory effects on [Ca(2+)](i) elevation induced by PE (P > 0.05). However, Nef (1 and 10 micromol l(-1)) inhibited [Ca(2+)](i) elevation induced by PE (P < 0.05). These data suggest that Nef inhibited [Ca(2+)](i) in CCSM cells via blocking voltage-dependent Ca(2+) channel, alpha(1)-adrenoceptor-operated Ca(2+) channel and Ca(2+) release from intracellular Ca(2+) pool. This inhibitory action on [Ca(2+)](i) might be one of the relaxation mechanisms of Nef on the CCSM.

Bisbenzylisoquinoline alkaloids of lotus ( Nelumbo nucifera Gaertn.) seed embryo inhibit lipopolysaccharide-induced macrophage activation via suppression of Ca 2+ -CaM/CaMKII pathway

“…Since changes in intracellular Ca 2+ levels can lead to the activation of NF-κB in LPS-stimulated macrophages (Zhou et al, 2006), and Neferine, Liensinine and Isoliensinine are inhibitors of Ca 2+ in ventricular myocytes and corpus cavernosum smooth muscle cells (Chen et al, 2007;Feng et al, 2002;Wang et al, 2000), the effects of Neferine, Liensinine and Isoliensinine on the level of cytosolic free Ca 2+ in LPS-activated RAW264.7 macrophages were investigated. Flow cytometry analysis showed that increased cytosolic free Ca 2+ level in LPS-stimulated macrophages was significantly inhibited by treatment with Figure 1.…”

“…However, the potential antiinflammatory targets of these three alkaloids in the upstream pathway of NF-κB are still unclear. Previous studies on various cell types have shown that bisbenzylisoquinoline alkaloids, such as Neferine, Liensinine, Isoliensinine, Dauricine, Tetrandrine, Cepharanthine and Berbamine, are inhibitors of Ca 2+ (Chen et al, 2007;Feng, Yu, & Xiao, 2002;Murakami, Okajima, & Uchiba, 2000;Qian, 2002;Ren et al, 2008;Wang, Nong, Yao, & Jiang, 2000). In addition, several bisbenzylisoquinoline alkaloids have been found to bind to CaM and inhibit the activity and expression of CaM (Hu, Chen, Hao, Huang, & Peng, 1989;Sun, Hu, & Xu, 1990;Xu & Zhang, 1986).…”

A Comprehensive Review on Chemical Profiling of Nelumbo Nucifera: Potential for Drug Development

“…Because AD is highly associated with ChEs and BACE1 activities, inflammation, and oxidative stress, N. nucifera can be considered as a preventive measure to inhibit the progression of AD. Using different in vivo animal models, Jung et al demonstrated the requirement of anti-oxidant, antiinflammatory, and the inhibitory effects of neferine (Tomita et al, 1962;Kunitomo et al, 1973;Liu et al, 2006) 8 Norarmepavine Leaves, Seeds (Kupchan et al, 1963;Duan et al, 2014) 9 Bromo methyl armepavine Leaves, Seeds (Rai et al, 2006) 10 Methoxymethy lisoquinoline Leaves, Seeds (Rai et al, 2006) 11 Lotusine Embryo, Seeds, Fruit Wang et al, 1991) 12 Higenamine Plumule (Zhou et al, 2013) (Chao et al, 1962;Furukawa et al, 1965;Guo and Chen, 1984;Chen et al, 2007b;Furukawa, 1966) 19 Isoliensinine (Isolie) Seeds, Fruit, Embryo (Tomita et al, 1965;Chen et al, 2007b;Furukawa, 1966) 20 Neferine Seeds, Fruit, Embryo (Chen et al, 2007a;Zhou et al, 2013;Sugimoto et al, 2015;Zhang et al, 2015b;Furukawa, 1966) Fruits and seeds (Toyoda, 1966;Mukherjee et al, 1997aMukherjee et al, , 1997b 189Asparagine…”

“…The authors reported that neferine increased cAMP concentration in a dose-dependent manner (Chen et al, 2008). Neferine relaxes corpus caverosum smooth muscle cells by inhibiting calcium elevation in the cells via blocking voltage-dependent calcium channel, alpha 1 adrenoceceptor operated calcium channel, and calcium release from intracellular calcium pool (Chen et al, 2007a). Furthermore, Mutreja et al (Mutreja et al, 2008) found that hydroalcoholic extract of N. nucifera seeds has an anti-estrogenic effect without altering the general physiology of reproductive system.…”

Effects of plant extract neferine on cyclic adenosine monophosphate and cyclic guanosine monophosphate levels in rabbit corpus cavernosum in vitro

“…Because of the complex chemical ingredients, it remains unclear which ingredient exactly, and by which mechanisms, has a positive effect in treating ED. Recent studies have found that some extracts from traditional Chinese medicines, containing alkaloids, coumarin and saponin compounds, can relax the smooth muscle of corpus cavernosum [3][4][5][6][7][8][9][10][11][12][13][14][15], which provides an open window for developing new drugs for the treatment of ED.…”