The principal results in this paper are concerned with the description of domains of infinitesimal generators of strongly continuous groups of isometries in non-commutative operator spaces $E(\mathcal{M},\tau)$, which are induced by $\mathbb{R}$-flows on $\mathcal{M}$. In particular, we are concerned with the description of operator functions which leave the domain of such generators invariant in all symmetric operator spaces, associated with a semi-finite von Neumann algebra $\mathcal{M}$ and a separable function space $E$ on $(0,\infty)$. Furthermore, we apply our results to the study of operator functions for which $[D,x]\in E(\mathcal{M},\tau)$ implies that $[D,f(x)]\in E(\mathcal{M},\tau)$, where $D$ is an unbounded self-adjoint operator. Our methods are partly based on the recently developed theory of double operator integrals in symmetric operator spaces and the theory of adjoint $C_{0}$-semigroups.