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Research article Special Issues

Screening revealed the strong cytotoxic activity of Alchemilla smirnovii and Hypericum alpestre ethanol extracts on different cancer cell lines

  • Received: 29 October 2022 Revised: 15 December 2022 Accepted: 16 December 2022 Published: 22 December 2022
  • Compounds of plant origin are considered promising alternative approaches in the development of medicines for the prevention and treatment of cancer. The large diversity of herbal species still requires careful exploration as a source for new anticancer compounds. The goal of the study was to screen different herbal extracts traditionally used in Armenian folk medicine for their cytotoxic effect against some cancer cell lines, and to find the prospective plant species among them. The cytotoxicity of the plant ethanol extracts was evaluated with MTT test against HeLa (human cervical carcinoma) and A549 (human lung adenocarcinoma) cells. Antioxidant properties were assessed with DPPH free radical scavenging assay. Five of the tested ten herbal extracts exhibited significant growth-inhibiting activity on HeLa cells. Moreover, Alchemilla smirnovii and Hypericum alpestre extracts also showed potent cytotoxicity on human lung adenocarcinoma cells. These two plants possessed high antiradical activity as well. Their DPPH stoichiometric values were 0.4234 and 0.14437 respectively, meaning that 1 µg of plant extract brought the reduction of DPPH equal to the respective stoichiometric values in µg. Thus, A. smirnovii and H. alpestre extracts expressed themselves as potent cytotoxic and antioxidant agents and could have promising anticancer potential. Further evaluation of their in vivo anticancer properties has much interest.

    Citation: Mikayel Ginovyan, Svetlana Hovhannisyan, Hayarpi Javrushyan, Gohar Sevoyan, Zaruhi Karabekian, Narine Zakaryan, Naira Sahakyan, Nikolay Avtandilyan. Screening revealed the strong cytotoxic activity of Alchemilla smirnovii and Hypericum alpestre ethanol extracts on different cancer cell lines[J]. AIMS Biophysics, 2023, 10(1): 12-22. doi: 10.3934/biophy.2023002

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  • Compounds of plant origin are considered promising alternative approaches in the development of medicines for the prevention and treatment of cancer. The large diversity of herbal species still requires careful exploration as a source for new anticancer compounds. The goal of the study was to screen different herbal extracts traditionally used in Armenian folk medicine for their cytotoxic effect against some cancer cell lines, and to find the prospective plant species among them. The cytotoxicity of the plant ethanol extracts was evaluated with MTT test against HeLa (human cervical carcinoma) and A549 (human lung adenocarcinoma) cells. Antioxidant properties were assessed with DPPH free radical scavenging assay. Five of the tested ten herbal extracts exhibited significant growth-inhibiting activity on HeLa cells. Moreover, Alchemilla smirnovii and Hypericum alpestre extracts also showed potent cytotoxicity on human lung adenocarcinoma cells. These two plants possessed high antiradical activity as well. Their DPPH stoichiometric values were 0.4234 and 0.14437 respectively, meaning that 1 µg of plant extract brought the reduction of DPPH equal to the respective stoichiometric values in µg. Thus, A. smirnovii and H. alpestre extracts expressed themselves as potent cytotoxic and antioxidant agents and could have promising anticancer potential. Further evaluation of their in vivo anticancer properties has much interest.


    Abbreviations

    DMEM:

    Dulbecco's modified Eagle medium; 

    DPPH:

    1,1-diphenyl-2-picrylhydrazyl; 

    DW:

    Dry weight; 

    FBS:

    Fetal bovine serum; 

    GAE:

    Gallic acid equivalent; 

    MTT:

    3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; 

    PBS:

    Phosphate-buffered saline; 

    ASI:

    Accumulated survival index; 

    ROS:

    Reactive oxygen species; 

    NF-κB:

    Nuclear factor kappa B; 

    MAPK:

    Mitogen-activated protein kinase

    In the general framework, Caputo and Fabrizio [1] proposed a new fractional derivative now called Caputo-Fabrizio (CF) fractional derivative in 2015. Compared with previous Riemann-Liouville (RL) and Riemann-Caputo (RC) fractional derivatives, this derivative has exponential kernel and non-singularity. The following comparison will reflect their differences. As we all know, when 0<γ<1, (tτ)γ and eγ1γ(tτ) are the kernels of RC- and CF-fractional derivative with γ-order, respectively. Decidedly, (tτ)γ (singular) and eγ1γ(tτ)1 (non-singular), as τt. In other words, CF-fractional derivative has unique advantages in eliminating singularity. Therefore, many scholars have carried out detailed and in-depth research on the CF-fractional differential equation. For example, some of them have applied CF-fractional differential equations to describe closed groundwater flows [2], population dynamics [3,4], electrical circuit [5,6], epidemics [7,8,9] and others [10,11,12]. There have been some papers dealing with some theoretical problems of CF-fractional calculus. Tarasov [13] explored whether CF-fractional derivative operators represent memory or distributed-delay from the definition of CF-fractional derivative. Pan [14] studied the chaotic behavior of a four dimensional CF-fractional differential system. Zhang [15] investigated the exponential Euler schemes for numerical solutions of CF-fractional differential equation. Tariq et al. [16] obtained the new fractional integral inequalities for CF-fractional integral operators. Abbas et al. [17] studied a fractional differential equations with non instantaneous impulses. They applied measures of noncompactness and two fixed point theorems to obtain the existence of solutions. In addition, the study of Hilfer fractional differential equations as a generalization of fractional derivatives is one of the recent focuses. Alsaedi et al. [18] considered a ψ-Hilfer fractional integral boundary value problem with the p-Laplacian operator. The authors studied the existence and uniqueness of solutions by using Banach's contraction mapping principle. Zhou and He [19,20] studied the mild solutions to two fractional evolution equations by analytic semigroup theory.

    In 1940s, Hyers and Ulam [21,22] put forward a new stability named Ulam and Hyers (UH) stability. After in-depth analysis of the UH-stability structure, some researchers have extended the concept of UH-stability, such as generalized UH-stability, Ulam-Hyers-Rassias stability, generalized Ulam-Hyers-Rassias, etc. The study on the UH-type stability of various dynamic systems has received great attention. Of course, the UH-type stability of fractional differential systems is also favored. There have been many papers dealing with UH-type stability of fractional differential system (see some of them [23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39]). However, there are rare works on the UH-type stability of CF-fractional differential system. It is worth to inquire into the stability of system with CF-fractional derivatives. In addition, when describing complex systems affected by many factors, fractional differential equations are more detailed and accurate than a single fractional differential equation. However, the study of the former is much more difficult than the latter. As far as I am concerned, there are no papers combining CF-fractional derivative with coupling Laplacian system, which is an interesting and challenging problem. Therefore, we emphasize on the below nonlinear CF-fractional coupled Laplacian equations

    {CFDν10+[Φp1(CFDμ10+U1(t))]=F1(t,U1(t),U2(t)),t(0,l],CFDν20+[Φp2(CFDμ20+U2(t))]=F2(t,U1(t),U2(t)),t(0,l],U1(0)=a1,U2(0)=a2,CFDμ10+U1(0)=b1,CFDμ20+U2(0)=b2, (1.1)

    where a1,a2,b1,b2R, l>0, 0<μ1,μ2,ν1,ν21 and p1,p2>1 are some constants, CFD0+ stands for the –order Caputo-Fabrizio (CF) fractional derivative. Φpj(z)=|z|pj2z(j=1,2) is pj–Laplacian. It is well known that the inverse of Φpj is Φqj, and 1pj+1qj=1, j=1,2. The nonlinear function FjC([0,l]×R2,R), j=1,2.

    This manuscript focuses on the solvability and stability of (1.1). In Section 2, we need to review some necessary knowledge of CF-fractional calculus. In Section 3, we apply the contraction mapping principle to prove that system (1.1) has a unique solution. We further established the GUH-stability of the system (1.1) in Section 4. Section 5 provides an example to illustrate the correctness of our major outcomes. We make a brief conclusion in Section 6.

    In this section, we first need to introduce the definitions of CF-fractional derivative and integral and some basic properties of p-Laplacian operator.

    Definition 2.1. [40] For 0α1, l>0 and UH1(0,l), the left–sided α–order Caputo–Fabrizio fractional integral of function U is defined by

    CFIα0U(t)=1αN(α)U(t)+αN(α)t0U(s)ds,

    where N(α) is a normalisation constant with N(0)=N(1)=1.

    Definition 2.2. [1] For 0α1, l>0 and UH1(0,l), the left–sided α–order Caputo–Fabrizio fractional derivative of U is defined by

    CFDα0+U(t)=N(α)1αt0eα1α(ts)U(s)ds.

    Lemma 2.1. [40] Let 0α1 and HC[0,). Then the unique solution of the following IVP

    {CFDα0+U(t)=H(t),t0,U(0)=U0.

    is expressed as

    U(t)=U0+1αN(α)[H(t)H(0)]+αN(α)t0H(s)ds.

    Lemma 2.2. Let p>1. The p–Laplacian operator Φp(z)=|z|p2z admits the properties as follows:

    (i) If z0, then Φp(z)=zp1, and Φp(z) is increasing with respect to z;

    (ii) For all z,wR, Φp(zw)=Φp(z)Φp(w);

    (iii) If 1p+1q=1, then Φq[Φp(z)]=Φp[Φq(z)]=z, for all zR;

    (iv) For all z,w0, zwΦq(z)Φq(w);

    (v)0zΦ1q(w)0Φq(z)w;

    (vi)|Φq(z)Φq(w)|{(q1)¯Mq2|zw|, q2,0z,w¯M; (q1)M_q2|zw|, 1<q<2,z,wM_0.

    The following lemma is crucial to establishing our main results later.

    Lemma 2.3. Let a1,a2,b1,b2R, l>0, 0<μ1,μ2,ν1,ν21 and p1,p2>1 are some constants, FjC([0,l]×R2,R), j=1,2. Then the nonlinear CF–fractional coupled Laplacian Eq (1.1) is equivalent to the following integral equations

    {U1(t)=a1+1μ1N(μ1)[Φq1(G1(t,U1(t),U2(t)))b1]+μ1N(μ1)t0Φq1(G1(s,U1(s),U2(s)))ds,t[0,l],U2(t)=a2+1μ2N(μ2)[Φq2(G2(t,U1(t),U2(t)))b2]+μ2N(μ2)t0Φq2(G2(s,U1(s),U2(s)))ds,t[0,l], (2.1)

    where 1pj+1qj=1(j=1,2), and

    G1(t,U1(t),U2(t))=Φp1(b1)+1ν1N(ν1)[F1(t,U1(t),U2(t))F1(0,a1,a2)]+ν1N(ν1)t0F1(τ,U1(τ),U2(τ))dτ,
    G2(t,U1(t),U2(t))=Φp2(b2)+1ν2N(ν2)[F2(t,U1(t),U2(t))F2(0,a1,a2)]+ν2N(ν2)t0F2(τ,U1(τ),U2(τ))dτ.

    Proof. Assume that (U1(t),U2(t))C([0,l],R)×C([0,l],R) satisfies the Eq (1.1). Then, from Lemma 2.1 and the first equation of (1.1), we have

    Φp1(CFDμ10+U1(t))=Φp1(CFDμ10+U1(0))+1ν1N(ν1)[F1(t,U1(t),U2(t))F1(0,U1(0),U2(0))]+ν1N(ν1)t0F1(τ,U1(τ),U2(τ))dτ,t[0,l]. (2.2)

    It follows from (2.2) and (iii) in Lemma 2.2 that

    CFDμ10+U1(t)=Φq1(Φp1(CFDμ10+U1(0))+1ν1N(ν1)[F1(t,U1(t),U2(t))F1(0,U1(0),U2(0))]+ν1N(ν1)t0F1(τ,U1(τ),U2(τ))dτ),t[0,l], (2.3)

    where 1p1+1q1=1, p1>1. Denote G1(t,U1(t),U2(t)) by

    G1(t,U1(t),U2(t))=Φp1(CFDμ10+U1(0))+1ν1N(ν1)[F1(t,U1(t),U2(t))F1(0,U1(0),U2(0))]+ν1N(ν1)t0F1(τ,U1(τ),U2(τ))dτ. (2.4)

    Combined (2.3), (2.4) and Lemma 2.1, we obtain

    U1(t)=U1(0)+1μ1N(μ1)[Φq1(G1(t,U1(t),U2(t)))Φq1(G1(0,U1(0),U2(0)))]+μ1N(μ1)t0Φq1(G1(s,U1(s),U2(s)))ds,t[0,l]. (2.5)

    Similar to (2.2)–(2.5), one derives from the second equation of (1.1) that

    U2(t)=U2(0)+1μ2N(μ2)[Φq2(G2(t,U1(t),U2(t)))Φq2(G2(0,U1(0),U2(0)))]+μ2N(μ2)t0Φq2(G2(s,U1(s),U2(s)))ds,t[0,l], (2.6)

    where 1p2+1q2=1,p2>1, and

    G2(t,U1(t),U2(t))=Φp2(CFDμ20+U2(0))+1ν2N(ν2)[F2(t,U1(t),U2(t))F2(0,U1(0),U2(0))]+ν2N(ν2)t0F2(τ,U1(τ),U2(τ))dτ. (2.7)

    Substituting the initial value conditions U1(0)=a1, U2(0)=a2, CFDμ10+U1(0)=b1 and CFDμ20+U2(0)=b2 into (2.4)–(2.7), one easily gets (2.1), that is, (U1(t),U2(t))C([0,l],R)×C([0,l],R) is a solution of the integral equations (2.1). Noticing that zΦp(z) is reversible, one knows that the above derivation is completely reversible. Vice versa, if (U1(t),U2(t))C([0,l],R)×C([0,l],R) is the solution of the integral Eq (2.1), then it is also a solution of (1.1). The proof is completed.

    This section mainly applies the contraction mapping principle to discuss the existence and uniqueness of solution to (1.1).

    Lemma 3.1. (contraction mapping principle [41]) Let X be a Banach space and ϕEX be closed. If T:EE is contract, then T admits a unique fixed point uE.

    According to Lemma 2.3, we take X=C([0,l],R)×C([0,l],R). For all w=(u,v)X, define the norm w=(u,v)=max{sup0tl|u(t)|,sup0tl|v(t)|}, then (X,) is a Banach space. Subsequently, we will inquire into the solvability and stability of (1.1) on (X,). For convenience, we introduce the following conditions and symbols.

    (H1)a10 or a20, l,b1,b2>0, 0<μ1,μ2,ν1,ν21 and p1,p2>1 are some constants, FjC([0,l]×R2,R), j=1,2.

    (H2) For all t[0,l], u,vR, there exist some constants mj,Mj>0 such that

    mjFj(t,u,v)Mj,j=1,2.

    (H3) For all t[0,l], u,¯u,v,¯vR, there exist some continuous functions Lj1(t), Lj2(t)0 such that

    |Fj(t,u,v)Fj(t,¯u,¯v)|Lj1(t)|u¯u|+Lj2(t)|v¯v|.

    Denote

    Mj_=bpj1j1νjN(νj)(Mjmj)+νjN(νj)mjl,
    ¯Mj=bpj1j+1νjN(νj)(Mjmj)+νjN(νj)Mjl,
    Θj=(1μj)(1νj)N(μj)N(νj)+(1μj)νjlN(μj)N(νj)+(1νj)μjlN(μj)N(νj)+μjνjl2N(μj)N(νj),
    ¯κj=Θj(qj1)¯Mjqj2(Lj1l+Lj2l),
    κj_=Θj(qj1)Mj_qj2(Lj1l+Lj2l),
    Ljil=max{Lji(t):0tl},j,i=1,2.

    In this position, we present one of our main results as follows.

    Theorem 3.1. Assume that (H1)(H3) and Mj_>0(j=1,2) are true. Further assume that one of the conditions holds as follows: when q1,q22, ¯κ1,¯κ2<1; or q12,1<q2<2, ¯κ1,κ2_<1; or 1<q1<2,q22, κ1_,¯κ2<1; or 1<q1,q2<2, κ1_,κ2_<1. Then system (1.1) has a unique nonzero solution (U1(t),U2(t))X.

    Proof. (U1(0),U2(0))=(a1,a2)(0,0) indicates (U1(t),U2(t))(0,0),t[0,l], that is, the solution of (1.1) is nonzero. For all (U1,U2)X, based on Lemma 2.3, we define the vector operator T:XX as

    T(U1,U2)(t)=(T1(U1,U2)(t),T2(U1,U2)(t)), (3.1)

    where

    T1(U1,U2)(t)=a1+1μ1N(μ1)[Φq1(G1(t,U1(t),U2(t)))b1]+μ1N(μ1)t0Φq1(G1(s,U1(s),U2(s)))ds,t[0,l], (3.2)
    T2(U1,U2)(t)=a2+1μ2N(μ2)[Φq2(G2(t,U1(t),U2(t)))b2]+μ2N(μ2)t0Φq2(G2(s,U1(s),U2(s)))ds,t[0,l], (3.3)

    G1(t,U1(t),U2(t)) and G2(t,U1(t),U2(t)) are the same as (2.1).

    For all U=(U1,U2) and t[0,l], we derive from (2.1), (H1) and (H2) that

    G1(t,U1(t),U2(t))bp111+1ν1N(ν1)(M1m1)+ν1N(ν1)M1l=¯M1, (3.4)
    G1(t,U1(t),U2(t))bp1111ν1N(ν1)(M1m1)+ν1N(ν1)m1l=M1_, (3.5)
    G2(t,U1(t),U2(t))bp212+1ν2N(ν2)(M2m2)+ν2N(ν2)M2l=¯M2, (3.6)

    and

    G2(t,U1(t),U2(t))bp2121ν2N(ν2)(M2m2)+ν2N(ν2)m2l=M2_. (3.7)

    Obviously, M1_¯M1, M2_¯M2. Thus, for all U=(U1,U2), ¯U=(¯U1,¯U2)X, and t[0,l], it follows from (3.2), (3.4), (3.5), (H3) and (vi) of Lemma 2.2 that

    |T1(U1,U2)(t)T1(¯U1,¯U2)(t)|=|1μ1N(μ1)[Φq1(G1(t,U1(t),U2(t)))Φq1(G1(t,¯U1(t),¯U2(t)))]+μ1N(μ1)t0[Φq1(G1(s,U1(s),U2(s)))Φq1(G1(s,¯U1(s),¯U2(s)))]ds|1μ1N(μ1)|Φq1(G1(t,U1(t),U2(t)))Φq1(G1(t,¯U1(t),¯U2(t)))|+μ1N(μ1)t0|Φq1(G1(s,U1(s),U2(s)))Φq1(G1(s,¯U1(s),¯U2(s)))|ds. (3.8)

    When q12, (3.8) gives

    |T1(U1,U2)(t)T1(¯U1,¯U2)(t)|1μ1N(μ1)(q11)¯M1q12|G1(t,U1(t),U2(t))G1(t,¯U1(t),¯U2(t))|+μ1N(μ1)(q11)¯M1q12t0|G1(s,U1(s),U2(s))G1(s,¯U1(s),¯U2(s))|ds1μ1N(μ1)(q11)¯M1q12[1ν1N(ν1)|F1(t,U1(t),U2(t))F1(t,¯U1(t),¯U2(t))|+ν1N(ν1)t0|F1(τ,U1(τ),U2(τ))F1(τ,¯U1(τ),¯U2(τ))|dτ]+μ1N(μ1)(q11)¯M1q12t0[1ν1N(ν1)|F1(s,U1(s),U2(s))F1(s,¯U1(s),¯U2(s))|+ν1N(ν1)s0|F1(τ,U1(τ),U2(τ))F1(τ,¯U1(τ),¯U2(τ))|dτ]ds.1μ1N(μ1)(q11)¯M1q12[1ν1N(ν1)[L11(t)|U1(t)¯U1(t)|+L12(t)|U2(t)¯U2(t)|]+ν1N(ν1)t0[L11(τ)|U1(τ)¯U1(τ)|+L12(τ)|U2(τ)¯U2(τ)|]dτ]+μ1N(μ1)(q11)¯M1q12t0[1ν1N(ν1)[L11(s)|U1(s)¯U1(s)|+L12(s)|U2(s)¯U2(s)|]+ν1N(ν1)s0[L11(τ)|U1(τ)¯U1(τ)|+L12(τ)|U2(τ)¯U2(τ)|]dτ]ds.1μ1N(μ1)(q11)¯M1q12[1ν1N(ν1)[L11lU¯U+L12lU¯U]+ν1N(ν1)l0[L11lU¯U+L12lU¯U]dτ]+μ1N(μ1)(q11)¯M1q12l0[1ν1N(ν1)[L11lU¯U+L12lU¯U]+ν1N(ν1)l0[L11lU¯U+L12lU¯U]dτ]ds=[(1μ1)(1ν1)N(μ1)N(ν1)+(1μ1)ν1lN(μ1)N(ν1)+(1ν1)μ1lN(μ1)N(ν1)+μ1ν1l2N(μ1)N(ν1)]×(q11)¯M1q12(L11l+L12l)U¯U=¯κ1U¯U. (3.9)

    When 1<q1<2, similar to (3.9), (3.8) leads

    |T1(U1,U2)(t)T1(¯U1,¯U2)(t)|[(1μ1)(1ν1)N(μ1)N(ν1)+(1μ1)ν1lN(μ1)N(ν1)+(1ν1)μ1lN(μ1)N(ν1)+μ1ν1l2N(μ1)N(ν1)]×(q11)M1_q12(L11l+L12l)U¯U=κ1_U¯U. (3.10)

    It is similar to (3.8)–(3.10) that

    |T2(U1,U2)(t)T2(¯U1,¯U2)(t)|[(1μ2)(1ν2)N(μ2)N(ν2)+(1μ2)ν2lN(μ2)N(ν2)+(1ν2)μ2lN(μ2)N(ν2)+μ2ν2l2N(μ2)N(ν2)]×(q21)¯M2q22(L21l+L22l)U¯U=¯κ2U¯U,q22, (3.11)

    and

    |T2(U1,U2)(t)T2(¯U1,¯U2)(t)|[(1μ2)(1ν2)N(μ2)N(ν2)+(1μ2)ν2lN(μ2)N(ν2)+(1ν2)μ2lN(μ2)N(ν2)+μ2ν2l2N(μ2)N(ν2)]×(q21)M2_q22(L21l+L22l)U¯U=κ2_U¯U,1<q2<2. (3.12)

    From (3.9)–(3.12), we obtain

    T(U1,U2)(t)T(¯U1,¯U2)(t){max{¯κ1,¯κ2}U¯U,q1,q22,max{¯κ1,κ2_}U¯U,q12,1<q2<2,max{κ1_,¯κ2}U¯U,1<q1<2,q22,max{κ1_,κ2_}U¯U,1<q1,q2<2. (3.13)

    Let κj{¯κj,κj_},j=1,2, then 0<max{κ1,κ2}<1. So (3.13) means that T:XX is contractive. Hence, we conclude from Lemma 3.1 and Lemma 2.2 that T has a unique fixed point U(t)=(U1(t),U2(t))X, which is the solution of (1.1). The proof is completed.

    In the portion, we mainly discuss the GUH-stability of (1.1) by direct analysis methods. We first give the definitions of UH- and GUH-stability corresponding to problem (1.1) as follows.

    Let U=(U1,U2)X and ϵ>0. Consider the following inequality

    {CFDν10+[Φp1(CFDμ10+U1(t))]F1(t,U1(t),U2(t))ϵ,t(0,l],CFDν20+[Φp2(CFDμ20+U2(t))]F2(t,U1(t),U2(t))ϵ,t(0,l],U1(0)=a1,U2(0)=a2,CFDμ10+U1(0)=b1,CFDμ20+U2(0)=b2. (4.1)

    Definition 4.1. Assume that, ϵ>0 and U=(U1,U2)X satisfying (4.1), there exist a unique U=(U1,U2)X satisfying (1.1) and a constant ω1>0 such that

    U(t)U(t)ω1ϵ,

    then problem (1.1) is called Ulam-Hyers (UH) stable.

    Definition 4.2. Assume that, ϵ>0 and U=(U1,U2)X satisfying (4.1), there exist a unique U=(U1,U2)X satifying (1.1) and ϑC(R,R+) with ϑ(0)=0 such that

    U(t)U(t)ϑ(ϵ),

    then problem (1.1) is called generalized Ulam-Hyers (GUH) stable.

    Remark 4.1. U=(U1,U2)X is a solution of inequality (4.1) iff there exists ϕ=(ϕ1,ϕ2)X such that

    (1)|ϕ1(t)|ϵ, and |ϕ2(t)|ϵ, 0<tl;

    (2)CFDν10+[Φp1(CFDμ10+U1(t))]=F1(t,U1(t),U2(t))+ϕ1(t),0<tl;

    (3)CFDν20+[Φp2(CFDμ20+U2(t))]=F2(t,U1(t),U2(t))+ϕ2(t),0<tl;

    (4)U1(0)=a1,U2(0)=a2,CFDμ10+U1(0)=b1,CFDμ20+U2(0)=b2.

    Theorem 4.1. If all conditions of Theorem 3.1 hold, then problem (1.1) is GUH-stable.

    Proof. Based on Lemma 2.3 and Remark 4.1, the inequality (4.1) is solved by

    {U1(t)=a1+1μ1N(μ1)[Φq1(Gϕ1(t,U1(t),U2(t)))b1]+μ1N(μ1)t0Φq1(Gϕ1(s,U1(s),U2(s)))ds,t[0,l],U2(t)=a2+1μ2N(μ2)[Φq2(Gϕ2(t,U1(t),U2(t)))b2]+μ2N(μ2)t0Φq2(Gϕ2(s,U1(s),U2(s)))ds,t[0,l], (4.2)
    Gϕ1(t,U1(t),U2(t))=Φp1(b1)+1ν1N(ν1)[F1(t,U1(t),U2(t))+ϕ1(t)F1(0,a1,a2)ϕ1(0)]+ν1N(ν1)t0[F1(τ,U1(τ),U2(τ))+ϕ1(τ)]dτ, (4.3)
    Gϕ2(t,U1(t),U2(t))=Φp2(b2)+1ν2N(ν2)[F2(t,U1(t),U2(t))+ϕ2(t)F2(0,a1,a2)ϕ2(0)]+ν2N(ν2)t0[F2(τ,U1(τ),U2(τ))+ϕ(τ)]dτ. (4.4)

    According to Theorem 3.1 and Lemma 2.3, the unique solution U(t)=(U1(t),U2(t))X of (1.1) satisfies (2.1). For all ϵ>0(ϵ small enough), from (H1), (H2) and (1) of Remark 4.1, it similar to (3.4)–(3.7) that

    Gϕ1(t,U1(t),U2(t))bp111+1ν1N(ν1)(M1m1+2ϵ)+ν1N(ν1)(M1+ϵ)l=¯M1(ϵ), (4.5)
    Gϕ1(t,U1(t),U2(t))bp1111ν1N(ν1)(M1m12ϵ)+ν1N(ν1)(m1ϵ)l=M1_(ϵ)>0, (4.6)
    Gϕ2(t,U1(t),U2(t))bp212+1ν2N(ν2)(M2m22ϵ)+ν2N(ν2)(M2+ϵ)l=¯M2(ϵ), (4.7)

    and

    Gϕ2(t,U1(t),U2(t))bp2121ν2N(ν2)(M2m22ϵ)+ν2N(ν2)(m2ϵ)l=M2_(ϵ)>0. (4.8)

    Clearly, 0<M1_(ϵ)<M1_<¯M1<¯M1(ϵ), 0<M2_(ϵ)<M2_<¯M2<¯M2(ϵ).

    Similar to (3.8) and (3.9), when q12, we derive from (2.1), (4.2), (4.3) and (4.5) that

    |U1(t)U1(t)|=|1μ1N(μ1)[Φq1(Gϕ1(t,U1(t),U2(t)))Φq1(G1(t,U1(t),U2(t)))]+μ1N(μ1)t0[Φq1(Gϕ1(s,U1(s),U2(s)))G1(s,U1(s),U2(s)))]ds|1μ1N(μ1)|Φq1(Gϕ1(t,U1(t),U2(t)))Φq1(G1(t,U1(t),U2(t)))|+μ1N(μ1)t0|Φq1(Gϕ1(s,U1(s),U2(s)))G1(s,U1(s),U2(s)))|ds1μ1N(μ1)(q11)¯M1(ϵ)q12|Gϕ1(t,U1(t),U2(t))G1(t,U1(t),U2(t))|+μ1N(μ1)(q11)¯M1(ϵ)q12t0|Gϕ1(s,U1(s),U2(s))G1(s,U1(s),U2(s))|ds1μ1N(μ1)(q11)¯M1(ϵ)q12[1ν1N(ν1)[|F1(t,U1(t),U2(t))F1(t,U1(t),U2(t))|+2ϵ]+ν1N(ν1)t0[|F1(τ,U1(τ),U2(τ))F1(τ,U1(τ),U2(τ))|+2ϵ]dτ]+μ1N(μ1)(q11)¯M1(ϵ)q12t0[1ν1N(ν1)[|F1(s,U1(s),U2(s))F1(s,U1(s),U2(s))|+2ϵ]+ν1N(ν1)s0[|F1(τ,U1(τ),U2(τ))F1(τ,U1(τ),U2(τ))|+2ϵ]dτ]ds.1μ1N(μ1)(q11)¯M1(ϵ)q12[1ν1N(ν1)[L11(t)|U1(t)U1(t)|+L12(t)|U2(t)U2(t)|+2ϵ]+ν1N(ν1)t0[L11(τ)|U1(τ)U1(τ)|+L12(τ)|U2(τ)U2(τ)|+2ϵ]dτ]+μ1N(μ1)(q11)¯M1(ϵ)q12t0[1ν1N(ν1)[L11(s)|U1(s)U1(s)|+L12(s)|U2(s)U2(s)|+2ϵ]+ν1N(ν1)s0[L11(τ)|U1(τ)U1(τ)|+L12(τ)|U2(τ)U2(τ)|+2ϵ]dτ]ds.1μ1N(μ1)(q11)¯M1(ϵ)q12[1ν1N(ν1)[L11lUU+L12lUU+2ϵ]+ν1N(ν1)l0[L11lUU+L12lUU+2ϵ]dτ]+μ1N(μ1)(q11)¯M1(ϵ)q12l0[1ν1N(ν1)[L11lUU+L12lUU+2ϵ]+ν1N(ν1)l0[L11lUU+L12lUU+2ϵ]dτ]ds=[(1μ1)(1ν1)N(μ1)N(ν1)+(1μ1)ν1lN(μ1)N(ν1)+(1ν1)μ1lN(μ1)N(ν1)+μ1ν1l2N(μ1)N(ν1)](q11)¯M1(ϵ)q12×[(L11l+L12l)U¯U+2ϵ]=¯Υ1(ϵ)U¯U+2¯Δ1(ϵ)ϵ, (4.9)

    where ¯Υ1(ϵ)=Θ1(q11)¯M1(ϵ)q12(L11l+L12l), ¯Δ1(ϵ)=Θ1(q11)¯M1(ϵ)q12.

    Analogy to (4.9), we apply (4.6)–(4.8) to obtain

    |U2(t)U2(t)|¯Υ2(ϵ)UU+2¯Δ2(ϵ)ϵ,q22, (4.10)
    |U1(t)U1(t)|Υ1_(ϵ)UU+2Δ1_(ϵ)ϵ,1<q1<2, (4.11)

    and

    |U2(t)U2(t)|Υ2_(ϵ)UU+2Δ2_(ϵ)ϵ,1<q2<2, (4.12)

    where ¯Υ2(ϵ)=Θ2(q21)¯M2(ϵ)q22(L21l+L22l), Υ1_(ϵ)=Θ1(q11)M1_(ϵ)q12(L11l+L12l), Υ2_(ϵ)=Θ2(q21)M2_(ϵ)q22(L21l+L22l). ¯Δ2(ϵ)=Θ2(q21)¯M2(ϵ)q22, Δ1_(ϵ)=Θ1(q11)M1_(ϵ)q12, and Δ2_(ϵ)=Θ2(q21)M2_(ϵ)q22.

    For all ϵ>0 (ϵ small enough), we have 0<¯Υ1(ϵ),Υ1_(ϵ),¯Υ2(ϵ),Υ2_(ϵ)<1. Take Υj(ϵ){¯Υj(ϵ),Υj_(ϵ)}, and Δj(ϵ){¯Δj(ϵ),Δj_(ϵ)}, j=1,2, then it follows from (4.9)–(4.12) that

    UU2max{Δ1(ϵ),Δ2(ϵ)}1max{Υ1(ϵ),Υ2(ϵ)}ϵ. (4.13)

    Therefore, we know from (4.13) and Definition 4.2 that problem (1.1) is GUH-stable. The proof is completed.

    The purpose of this section is to verify the correctness and applicability of our main results through an illustrative example.

    To do so, consider the following specific nonlinear CF-fractional coupled Laplacian system

    {CFD0.40+[Φp1(CFD0.70+U1(t))]=2+cos(U1(t))100+150|sin(t)|U2(t)1+U2(t)2,t(0,1],CFD0.80+[Φp2(CFD0.30+U2(t))]=2+sin(3t)100[3π4+arctan(U1(t)+U2(t))],t(0,1],U1(0)=1,U2(0)=1,CFD0.70+U1(0)=1,CFD0.30+U2(0)=1. (5.1)

    Obviously, l=1, μ1=0.7, ν1=0.4, μ2=0.3, ν2=0.8, a1=1, a2=b1=b2=1, F1(t,u,v)=2+cos(u)100+150|sin(t)|v1+v2, F2(t,u,v)=2+sin(3t)100[3π4+arctan(u+v)]. Take N(x)=1x+xΓ(x),0<x1, then N(0)=N(1)=1. By a simple calculation, we have

    1100F1(t,u,v)4100,π400F2(t,u,v)15π400,
    |F1(t,u,v)F1(t,¯u,¯v)|1100|u¯u|+|sin(t)|100|v¯v|,
    |F2(t,u,v)F2(t,¯u,¯v)|2+sin(3t)100[|u¯u|+|v¯v|].

    Thus, the conditions (H1)(H3) are true. Consequently, m1=1100, M1=4100, m2=π400, M2=15π400, L11(t)=1100, L12(t)=|sin(t)|100, L21(t)=L22(t)=2+sin(3t)100, L11l=1100, L12l=sin(1)100, L21l=L22l=3100, and

    Θ1=(1μ1)(1ν1)N(μ1)N(ν1)+(1μ1)ν1lN(μ1)N(ν1)+(1ν1)μ1lN(μ1)N(ν1)+μ1ν1l2N(μ1)N(ν1)1.5269,
    Θ2=(1μ2)(1ν2)N(μ2)N(ν2)+(1μ2)ν2lN(μ2)N(ν2)+(1ν2)μ2lN(μ2)N(ν2)+μ2ν2l2N(μ2)N(ν2)1.4085.

    Case 1: When p1=32, p2=54, then q1=3>2, q2=5>2, and

    M1_=bp1111ν1N(ν1)(M1m1)+ν1N(ν1)m1l0.9821>0,
    M2_=bp2121ν2N(ν2)(M2m2)+ν2N(ν2)m2l0.9823>0,
    ¯M1=bp111+1ν1N(ν1)(M1m1)+ν1N(ν1)M1l1.0436,
    ¯M2=bp212+1ν2N(ν2)(M2m2)+ν2N(ν2)M2l1.1239,
    ¯κ1=Θ1(q11)¯M1q12(L11l+L12l)0.0587<1,
    ¯κ2=Θ2(q21)¯M2q22(L21l+L22l)0.3202<1.

    Thus, all conditions of Theorem 3.1 are fulfilled. From Theorem 3.1 and Theorem 4.1, we claim that system (5.1) has a unique solution and is GUH-stable.

    Case 2: When p1=32, p2=5, then q1=3>2, 1<q2=54<2, and the values of M1_, M2_, ¯M1, ¯M2 and ¯κ1 are same as Case 1, as well as

    κ2_=Θ2(q21)M2_q22(L21l+L22l)0.0214<1.

    Thus, all conditions of Theorem 3.1 are fulfilled. From Theorem 3.1 and Theorem 4.1, we claim that system (5.1) has a unique solution and is GUH-stable.

    Case 3: When p1=3, p2=54, then 1<q1=32<2, q2=5>2, and the values of M1_, M2_, ¯M1, ¯M2 and ¯κ2 are same as Case 1, as well as

    κ1_=Θ1(q11)M1_q12(L11l+L12l)0.0142<1,

    Thus, all conditions of Theorem 3.1 are fulfilled. From Theorem 3.1 and Theorem 4.1, we claim that system (5.1) has a unique solution and is GUH-stable.

    Case 4: When p1=3, p2=5, then 1<q1=32<2, 1<q2=54<2, and the values of M1_, M2_, ¯M1, ¯M2, κ1_ and κ2_ are same as Cases 1–3. Thus, all conditions of Theorem 3.1 are fulfilled. From Theorem 3.1 and Theorem 4.1, we claim that system (5.1) has a unique solution and is GUH-stable.

    The integer order differential equation with p-Laplacian is a class of special second-order ordinary differential equations that have been extensively and deeply studied. Some scholars have also conducted some research on Riemann-Liouville or Caputo fractional differential equations with p-Laplacian. However, the study on CF-fractional differential equations p-Laplacian has not been seen so far. Therefore, it is novel and interesting for us to choose the system (1.1) as the research object. We establish the existence, uniqueness, and GUH-stability of the solution for problem (1.1) by using the Banach's contraction mapping principle and the direct analysis method. From the proof of Lemma 2.3 and Theorem 3.1, it can be seen that our difficulty lies in establishing the integral equation corresponding to system (1.1) and verifying the contractility of vector operator T defined by (3.1)–(3.3). The methods and steps used in this manuscript can be summarized as follows: (i) Convert differential system (1.1) to integral system (2.1); (ii) Define an operator T according to integral system (2.1); (iii) Prove that the operator T is contractive. The above methods and steps can be used for reference in the study of other types of fractional differential equations. In addition, illuminated by some of the latest achievements [42,43,44,45,46,47,48], we intend to apply fractional calculus theory and diffusion partial differential equation theory to the study of some ecosystems in the future.

    The author would like to express his heartfelt gratitude to the editors and reviewers for their constructive comments. The APC was funded by research start-up funds for high-level talents of Taizhou University.

    All authors declare that they have no competing interests.


    Acknowledgments



    This work was supported by the Science Committee of RA, in the frames of the research project № 20TTSG-1F004 as well as Basic support from Science Committee of RA, Ministry of Education, Science, Culture and Sports of RA.

    Conflict of interest



    The authors declare no conflict of interest.

    Author contributions



    All authors contributed to the study's conception and design. GS, MG, NA, HJ, SH, NZ and NS carried out the investigations and analyzed the outcomes. NZ identified plant samples. MG and NS wrote the manuscript. MG, NA, ZK and NS directed the experiments, corrected, and edited the manuscript. All authors revised and accepted the final version of the manuscript.

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